Nuclear fusion is essential for the sexual reproduction of various organisms, including plants, animals, and fungi. During the life cycle of flowering plants, nuclear fusion occurs three times: once during female gametogenesis and twice during double fertilization, when two sperm cells fertilize the egg and the central cell. Haploid nuclei migrate in an actin filament-dependent manner to become in close contact, then two nuclei fuse. The nuclear fusion process in plant reproduction is achieved by the sequential nuclear membrane fusion events. Recent molecular genetic analyses using Arabidopsis thaliana showed the conservation of nuclear membrane fusion machinery between plants and the budding yeast Saccharomyces cerevisiae. These include the heat shock protein 70 in the endoplasmic reticulum and conserved nuclear membrane proteins. Analyses of A. thaliana mutants of these components show that completion of the sperm nuclear fusion at fertilization is essential for proper embryo and endosperm development.

Maxillary sinus augmentation with a lateral approach (MSA) is a well-established treatment. In this prospective study, we evaluated risk factors for postoperative bone-graft displacement and report the clinical application of long-term resorbable GMEM-B-2 (L-lactic acid/-caprolactone [P(LA/CL)]) as a barrier membrane to cover the open window in the lateral wall in MSA. Twenty-four patients underwent MSA according to the relevant criteria; CT data obtained before and 1 week (1W) and 5–6 months (5M) post-MSA, bone height changes, bone height reduction rates at 1W and 5M post-MSA, bone-graft displacement measurements, and risk factors were examined. All patients showed bone height increments (p < 0.005). However, no difference was observed between 1W and 5M post-MSA. Bone-graft displacement was observed in eight patients; the reduction rate from 1W to 5M post-MSA was 8.38% ± 4.88%. Sex, septa, maxillary sinus floor–palatal bone distance, and maxillary sinus floor–natural mouth distance were associated with bone-graft displacement (p < 0.05). Height from the maxillary sinus floor to the palatal bone and the sinus angle influenced the augmentation amount (p < 0.05). The L-lactic acid/ε-caprolactone [P(LA/CL)] membrane compared favourably with other membranes and may be useful as a barrier membrane for the MSA open window.

Insoluble contaminant or a varnish is a result of oil degradation by products and sometimes depleted additive molecules. This process is in most cases initiated by thermal stress placed on the oil. In tribology varnish becomes a significant problem for modern complex machinery lubrication systems as it has severe debilitating effects such as loss of operating clearances and heat transfer. Problems arising from varnish build-up in lubricants can be inhibited by timely oil analysis which gives us important information about oil-degradation level and current insoluble contaminants potential of oil. There are several laboratory testing methods to describe varnish potential. The most applied one is a colorimetric analysis, also known as MPC (Membrane Patch Colorimetry) as it tends to be reasonably quick and cost-effective. This study applies principles generally used for MPC test of turbine oils to measure lubricant generated insoluble contaminants of hydraulic fluids. For this purpose, in-service oil samples were taken from hydraulic circuits of rubber vulcanizing presses and analyzed using testing method based on method defined by standard ASTM D7843 – 21. Accuracy of this testing method is dependent on development time of tested sample. Therefore, we not only try to proof applicability of this method for hydraulic oils but also want to determine incubation time needed for efficient and accurate determination of concentration of insoluble contaminants in the sample.

Mitochondria are highly dynamic organelles that constantly undergo fusion and fission events to maintain their shape, distribution, and cellular function. Mitofusin 1 and 2 proteins are two dynamin-like GTPases involved in the fusion of outer mitochondrial membranes (OMM). Mitofusins are anchored to the OMM through their transmembrane domain and possess two heptad repeat domains (HR1 and HR2) in addition to their N-terminal GTPase domain. The HR1 domain was found to induce fusion via its amphipathic helix, which interacts with the lipid bilayer structure. The lipid composition of mitochondrial membranes can also impact mitochondrial fusion. However, the precise mode of action of lipids in mitochondrial fusion is not fully understood. In this study, we have examined the role of the mitochondrial lipids phosphatidylethanolamine (PE), cardiolipin (CL) and phosphatidic acid (PA) in membrane fusion induced by the HR1 domain, both in the presence and absence of divalent cations (Ca2+ or Mg2+). Our results show that PE, as well as PA in the presence of Ca2+, effectively stimulate HR1-mediated fusion, while CL has a slight inhibitory effect. By considering the biophysical properties of these lipids in the absence or presence of divalent cations, we infer that the interplay between divalent cations and specific cone-shaped lipids creates regions with packing defects in the membrane, which provides a favorable environment for the amphipathic helix of HR1 to bind to the membrane and initiate fusion.

Hoechst 33342 (H33342) is a fluorescent probe that is commonly used to stain DNA of living cells. To do so, it needs to interact with and permeate through cell membranes. The pH of biological media may vary quite significantly and this influences H33342 ionization state, and may change its rate of permeation through biomembranes. In this work, we address the effect of pH in the association of H33342 with lipid bilayers using a combined experimental and computational approach. The partition of H33342 to 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) lipid membranes was experimentally quantified using fluorescence spectroscopy and isothermal titration calorimetry (ITC) measurements. Quantum chemical calculations were performed to select the most stable isomer of H33342 for the overall charges 0, +1, and +2, expected to predominate across the 5 < pH < 10 range. The interaction of these isomers with POPC bilayers was then studied by both unrestrained and umbrella sampling molecular dynamics simulations. While probe insertion occurs for all ionization states, with similar overall equilibrium location, orientation was found to depend on the probe charge, with the neutral form lying mostly parallel and the doubly charged displaying a significant tilt relative to the membrane plane. Both experimental results and computational free energy profiles indicate that the partition coefficient of H33342 displays a small variation over a wide pH range, not exceeding one order of magnitude. The enthalpy of interaction is negative and is larger at lower pH values. This suggests efficient hydrogen bonding between the probe and the lipid, namely for the protonated +2 form. The relatively high lipophilicity obtained for the charged species contrasts with the decrease in their general hydrophobicity as estimated from water and octanol partition. This highlights the distinction between lipophilicity and hydrophobicity, and the importance of considering association with lipid bilayers when predicting the affinity for biomembranes.

The present work is focused on functionalizing Montmorillonite (Mt) with poly dimethyl siloxane (PDMS) to develop a hybrid membrane reactor for the removal of Co (II) and Ni (II) ions from model wastewater samples. The specific surface area of the novel material was found to be 444 m2 /g. X-Ray Diffraction (XRD) and Fourier Transform Infra-Red (FTIR) spectra reveal the formation of exfoliated composite material. The efficiency of this membrane material was determined by subjecting it to continuous filtration experiments. It was able to remove 50% of Co (II) and 75% of Ni (II) ions, respectively, in one cycle of filtration in less than three minutes of operation with a permeate flux rate of 675 L/m2/H and 975 L/m2/H respectively, for both the ions.

Introduction: Diclofenac is the most prescribed non-steroidal anti-inflammatory drug worldwide and used to reliev pain and inflammation for inflammatory arthritis. Diclofenac do not slows disease progression and cartialge damage of Rheuamtoid Arthritis individuals. Moreover, it associated with seriuos adverse effects even using regular dose regimens. Drug delivery systems can overcome this issues reducing adverse effects and optmizing efficacy. Objectives: to evaluate the activity of a lipid-core nanocapsule loaded of Diclofenac (DIC-LNC) in an experimental model of adjuvant-induced arthritis and its anti-arthritic properties at the joint components. Methods: The diclofenac nanoformulation was obtained by self-assembling methodology. The stereology analysis aproach was applied for morphological quantification of the volume, density and cellular profile count of the metatarsophalangeal joints of rats induced to adjuvant arthritis. Proinflamatory cytokines and biochemical profile was also obtained. Results: DIC-LNC is able to reduce arthitis compared to control group (p<0.0001) and DIC group (p=0.009). The TNF and IL1 cytokine as well as C-reative protein and Xanthine-oxidade were efficiently reduced by DIC-LNC. Additionally, DIC-LNC reduces synovites and condrocytes lossing compared to DIC (p<0.05)and control group (p<0.05). The synovial space volume was higher for DIC-LNC compared to DIC (p<0.05) and Control (p<0.05). These data are suggesting that DIC-LNC is showing anti-arthritic actvity preserving deep joint components. Conclusion: DIC-LNC is a promissing nanoformulation for clinical use, since is able to reduce joint inflamation and synovits, avoiding damage of cartilage and synovial space at advjuvant athrits. Further studies and developments are necessary to achieve future clinical use.

A non-steady state mathematical model of amino acid (phenylalanine (Phe)) and mineral salt (NaCl) solution separation by neutralization dialysis (ND) carried out in a batch mode is proposed. The model takes into account the characteristics of membranes (thickness, ion-exchange capacity and conductivity) and solutions (concentration, composition), as well as the flow rate of the solution in the dialyzer compartments. As compared to previously developed models, the new one considers the local equilibrium of Phe protolysis reactions in solutions and membranes and, also, the transport of all the phenylalanine forms (zwitterionic, positively and negatively charged) through membranes. A series of experiments on ND demineralization of NaCl and Phe mixed solution was carried out. In order to minimize Phe losses, the solution pH in the desalination compartment was controlled by changing the concentrations of the solutions in the acid and alkali compartments of the ND cell. The validity of the model was verified by comparison of simulated and experimental time dependencies of solution electric conductivity and pH, as well as the concentration of Na+, Cl– ions and Phe species in the desalination compartment. Based on the simulation results, the role of Phe transport mechanisms in the losses of this amino acid during ND was discussed.

Background: Visceral leishmaniasis affects approximately 12 million people worldwide. Current existing drugs cause several side effects and resistance, the identification of new therapeutic targets for the development of new drugs is necessary. The vesicular trafficking in the parasite flagellar pocket is essential for parasite survival and proliferation, but little is known about the proteins involved in the endocytic/secretory pathways. In this work, we are describing the characterization of a transporter cargo protein SFT-2/Got-1 like in Leishmania infantum. Material and Methods: Using bioinformatics tools, the putative Got-1 protein A4HSK3 was characterized structurally and phylogenetically. Molecular biology was used to clone and express the protein and confocal fluorescence to determine its subcellular localization. Results: The Got-1 like protein presents four alpha helix TM conserved domains (aa38-aa147) characteristics of the Got-1/SFT-2 family proteins and an important Pen-2 conserved motif LGDYXXF located in the extracellular loop. Confocal microscopy suggests that Got-1/SFT-2 like is localized in the flagellar pocket, and is partially co-localized with the L. infantum presenilin-like protein. Phylogenetically, the Leishmania sp Got-1 like is contained in a subgroup of proteins clearly detached from the other Trypanosomatidae, such as T. cruzi and T. brucei. Conclusions: Our data show the presence of an important transported proteins, and its colocalization with the presenilin aspartyl protease in the flagellar pocket of the L. infantum open new perspectives to identify novel targets, which can be an alternative therapeutic strategy against leishmaniasis.

Background: Transarterial chemoembolization (TACE) is the standard treatment for BCLC-B hepatocellular carcinoma (HCC). A novel glass membrane emulsification device (GMD) produces a high percentage of water/oil emulsions with homogeneous and stable droplets. There are few reports on the efficacy of GMD-conventional-TACE (GMD-c-TACE)；therefore, we aimed to evaluate the effectiveness of GMD-c-TACE. Methods: Seventy-one patients with HCC with tumor diameter <5 cm who underwent c-TACE with and without GMD were included in this study to investigate local recurrence and hepatic functional reserve. Results: The local recurrence rates of TACE without GMD were 3.0% at 6 months, 16.7% at 12 months, and 35.0% at 18 months, around where it plateaued. Hence, the local recurrence rates in the GMD-c-TACE group were 7.7% at 14 months and 23.1% at 20 months, respectively. Thus, GMD-c-TACE had a significantly lower local recurrence. Multivariate analysis showed that GMD-c-TACE could suppress local recurrence and maintain hepatic reserve. Conclusions: GMD-c-TACE allows dense lipiodol accumulation in the tumor and attainment of good local control. Additionally, the inhibition of the release of anticancer drugs may maintain hepatic reserve. GMD-c-TACE is useful in preventing local recurrence and is expected to become the standard treatment form of c-TACE in the future.

This study compares the efficacy of the two most commonly used surgical methods for pterygium in the Polish population, conjunctival autograft versus amniotic membrane transplantation, and to evaluate the postopera-tive recurrence rate. We retrospectively analysed the medical records of 65 patients who underwent surgery for primary or recurrent pterygium at an ophthalmology clinic in Bialystok, Poland between 2016 and 2020. Surgical success (no regrowth) occurred in almost half of the amniotic membrane patients (44%) and in most of the conjunctival autograft patients (79%); this was a significant relationship. The odds of successful surgery were 79% lower for subjects with amniotic membranes than for those with conjunc-tival autografts (OR with 95% CI = 0.21 (0.05; 0.94]; p = 0.045). Our study confirms that in Polish Caucasian population the success rate of the pro-cedure using conjunctival autograft versus the use of amniotic membrane, is in favoured for the procedure with conjunctival graft.

Spider toxin SNX-482 is a cysteine-rich peptide that interferes with calcium channel activity by binding to voltage-sensing domains of CaV2.3 subtype. Two general binding mechanisms are present in nature: direct binding from the aqueous phase or through lateral diffusion from the membrane, the so-called reduction in dimensionality mechanism. In this work, via coarse-grained and atomistic molecular dynamics simulations, we have systematically studied the spontaneous partitioning of SNX-482 with membranes of different anionic compositions and explored via diffusional analysis both binding mechanisms. Our simulations revealed a conserved protein patch that inserts within the membrane, a preference for binding towards partially negatively charged membranes, and that electrostatics drives membrane binding. Finally, diffusivity calculations showed that the toxin diffusion along the membrane plane is an order of magnitude slower than the aqueous phase suggesting that the critical factor in determin-ing the SNX-482-CaV2.3 binding mechanism is the affinity between the membrane and SNX-482

PG9, PG16, PGT121, and PGT145 antibodies were identified from culture media of activated memory B-cells of an infected donor and shown to neutralize many HIV-1 strains. Since HIV-1 spreads via both free virions and cell-cell fusion, we examined the effect of the antibodies on HIV-1 Env-mediated cell-cell fusion. Clone69TRevEnv cells that express Env in the absence of tetracycline were labeled with Calcein-AM Green, and incubated with CD4+ SupT1 cells labeled with CellTrace™ Calcein Red-Orange, with or without antibodies. Monoclonal antibodies PG9, PG16, 2G12, PGT121, and PGT145 (at up to 50 µg/mL) had little or no inhibitory effect on fusion between HIV-Env and SupT1 cells. By contrast, Hippeastrum hybrid agglutinin completely inhibited fusion. Our results indicate that transmission of the virus or viral genetic material would not be inhibited by these broadly neutralizing antibodies. Thus, antibodies generated by HIV-1 vaccines should be screened for their inhibitory effect on Env-mediated cell-cell fusion.

Micellar Casein Concentrate (MCC) is manufactured from microfiltration (MF) of skim milk utilizing ceramic or polymeric membrane filtration. While ceramic filtration has higher efficiency, use of polymeric is cost effective and the process is familiar to several US dairy processors. The aim of the present study was to develop an optimized membrane filtration process to produce MCC using spiral wound polymeric membrane filtration (SW MF) system by systematic selection of transmembrane pressure (TMP) and level of diafiltration (DF). Using skim milk as feed material, preliminary lab-scale MF experiments were conducted using 0.5 µm polyvinylidene fluoride (PVDF) membrane. Three TMP (34.5, 62.1, and 103.4 kPa) and three levels of DF (70, 100, and 150%) along with a process without DF as control were used in the study. Effect of TMP and effectiveness of DF on flux rates, SP removal, casein to total protein (CN/TKN) ratio, casein to true protein (CN/TP) ratio, rejection of casein (rej CN) and SP (rej SP) were evaluated. At all TMP values used in the study, the overall flux (O Flux) increased with the level of DF. Highest O Flux of 30.77 liter per meter square per hour (LMH) was obtained with 34.5 kPa pressure and 150% DF. The impact of DF was more pronounced at lower pressures than at the higher pressures used in the study. With controlled DF, instantaneous flux was maintained within 80% of initial flux for the entire process run. For all the experiments, casein has a rejection of 0.97 to 1.0, while serum protein has the lowest rejection of 0.10 at 34.5 kPa pressure and 150% DF level. Use of 34.5 kPa and DF level of 150 % contributed to 81.45% SP removal, and casein to true protein ratio of 0.96. SP removal data from the lab-scale experiments were fitted into a mathematical model using DF and square of TMP as factors. The model predicts SP removal within 90-95% of actual SP removal got from the pilot plant experiments.

The cellular model we teach and have theorized assumes that the cell is the basic unit of multicellular living beings. This fundamental element has been the subject of many theories concerning its properties and the exchanges that exist with its environment. In this article, we demonstrate that certain functional aspects, in particular the electrical aspects related to diffusion, have not been correctly assumed or that certain initial conditions have been purely ignored and are in contradiction with physics, chemistry and thermodynamics.

The neonatal Hub and Spoke (HandS) ECMO approach provides emergent ECMO implantation to support neonates in severe cardio-respiratory life-threatening conditions at secondary (Spoke) centers, and a safe transfer to the ECMO center (Hub). We report a retrospective review of 11 neonates fom January 2014 to January 2020, presenting with life-threatening conditions and treated by HandS ECMO team in Spoke hospitals. Protocols and checklists were arranged by a local group of ECMO experts and shared with all the hospitals located in North-Eastern Italy. At the emergent call, all patients were cared for at the Spoke hospitals, receiving maximal respiratory and cardiovascular support (including high frequency oscillatory ventilation, inhaled nitric oxide, and major inotropic intravenous infusion). All but 3 patients were affected by life-threatening meconium aspiration syndrome. Median duration of ECMO support and hospitalization were 4 days (range 2-32) and 30 days ( range 8-50), respectively. All but 2 patients with congenital diaphragmatic hernia, were weaned off ECMO and discharged home without complications. At a median follow up of 14.4 months (1.3-74.8), all survivors were alive and in good clinical conditions, without medications, and normal somatic growth. All but one had normal neuropsychological development. We conclude that HandS ECMO model for treating neonates in life-threatening conditions is effective and successful. A specialized multidisciplinary team and common organizational protocols shared between Hub and Spoke centers are the key-points for success. While excellent results occur with MAS (100% survival), treatment was not successful in CDH, in which other that HandS ECMO issues are implied. Lessons learned from our experience are presented and discussed in this article.

To investigate the effect of Polyvinylpyrrolidone (PVP) addition and consequently porosity, two different sets of membranes are manufactured, since PVP is a widely used poring agent which has an impact on the mechanical properties of the membrane material. The first set (PAN 1) includes PAN and the necessary solvent while the second set (PAN 2) is made of PAN and PVP. These membranes are put through several characterisation processes including tensile testing. The obtained data are used to model the static behaviour of the membranes with different geometries, but similar loading and boundary conditions that represent their operating conditions. This modelling process is undertaken by using finite element method. The main idea is to investigate how geometry affects the load-carrying capacity of the membranes. Alongside membrane modelling, their materials are modelled with representative elements with hexagonal and rectangular pore arrays (RE) to understand the impact of porosity on the mechanical properties. Exploring the results, the best geometry is found as the elliptic membrane with the aspect ratio 4 and the better RE as the hexagonal array which can predict the elastic properties with an approximate error of 12%.

(1) Introduction: Karstic springs are used worldwide by rural communities as sources of fresh water for human use and livestock survival. In Romania, 1/3 of the population has no direct access to the public water supply. The present study is part of a country-wide project on developing simple, quick and cheap methods for seasonal environmental and microbiological monitoring of karstic springs used as drinking water supply by rural populations. Critical steps for the monitoring workflow consist in the evaluation of water quality and choosing of suitable membrane filters to efficiently capture environmental DNA for further microbial diversity estimation by 16S rRNA gene-based metabarcoding; (2) Methods: Several commercial membrane filters of different composition and pore sizes were tested on the water sampled from three karstic springs in Romania, followed by water chemistry and whole community 16S rRNA gene-based metabarcoding analysis; (3) Results: We found that the different types of applied membrane filters provide a varying recovery of diversities and abundances of both overall and pathogenic bacteria; and (4) Conclusions: The result of the experiment with different filters shows which are the best for amplicon-based metabarcoding monitoring of karst springs.

A major parameter controlling the extent and rate of oral drug absorption is permeability through the lipid bilayer of intestinal epithelial cells. Here, a biomimetic artificial membrane permeability assay (Franz-Bampa) was validated using Franz cells apparatus. Both high and low permeability drugs (metoprolol and mannitol, respectively) were used as external standards. Biomimetic properties of Franz-Bampa were also characterized by electron paramagnetic resonance spectroscopy (EPR). Moreover, the permeation profile for the 14 BCS class I-IV drugs cited in the FDA guidance (including other drugs as acyclovir, cimetidine, diclofenac, ibuprofen, piroxicam, and trimethoprim) were measured across Franz-Bampa. Apparent permeability (Papp) was compared to literature fraction dose absorbed in humans (Fa%). Papp in Caco-2 cells and Corti artificial membrane were likewise compared to Fa% to assess Franz-Bampa performance. Mannitol and metoprolol Papp values across Franz-Bampa were lower (3.20 x 10-7 and 1.61 x 10-5 cm/s, respectively) than those obtained across non-impregnated membrane (2.27 x 10-5 and 2.55 x 10-5 cm/s, respectively), confirming lipidic barrier resistivity. Performance of the Franz cell permeation apparatus using an artificial membrane showed similar log linear correlation (R2 = 0.664) with Fa%, as seen for Papp in Caco-2 cells (R2 = 0.805). Data support the validation of the Franz-Bampa method for use during drug discovery process.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the disease COVID-19 that has decimated the health and economy of our planet. The virus causes the disease not only in people but also in companion and wild animals. People with diabetes are at risk of the disease. As yet we do not know why the virus is highly successful in causing the pandemic within 3 months of its first report. The structural proteins of SARS include, membrane glycoprotein (M), envelope protein (E), nucleocapsid protein (N) and the spike protein (S). The structure and function of the most abundant structural protein of SARS-CoV-2, the membrane (M) glycoprotein is not fully understood. Using in silico analyses we determined the structure and potential function of the M protein. The M protein of SARS-CoV-2 is 98.6% similar to the M protein of bat SARS-CoV, maintains 98.2% homology with pangolin SARS-CoV, and has 90% homology with M protein of SARS-CoV; whereas, the similarity was only 38% with the M protein of MERS-CoV. In silico analyses showed that the M protein of SARS-CoV-2 has a triple helix bundle, form a single 3-transmembrane domain (TM), and are homologous to the prokaryotic sugar transport protein semiSWEET. SemiSWEETs are related to the PQ-loop family that function as cargo receptors in vesicle transport, mediates movement of basic amino acids across lysosomal membranes, and is also involved in phospholipase flippase function. The advantage and role of the M protein having a sugar transport-like structure is not clearly understood. The M protein of SARS-CoV-2 interacts with S, E and N protein. The S protein of the virus is glycosylated. It could be hypothesized that the sugar transporter-like structure of the M protein influences glycosylation of the S protein. Endocytosis is critical for the internalization and maturation of RNA viruses, including SARS-CoV-2. Sucrose is involved in endosome and lysosome maturation and may also induce autophagy, pathways that help in the entry of the virus. Overall, it could be hypothesized that the semiSWEET sugar transporter-like structure of the M protein may be involved in multiple functions that may aid in the rapid proliferation, replication and immune evasion of the SARS-CoV-2 virus. Biological experiments would validate the presence and function of the semiSWEET sugar transporter.

The major stress-inducible Hsp70 (HSPA1A) is overexpressed in the cytosol of many highly aggressive tumor cells including glioblastoma multiforme and presented on their plasma membrane. Depending on its intracellular or membrane localization, Hsp70 either promotes tumor growth or serves as a target for NK cells. The kinetics of the membrane Hsp70 (mHsp70) density on human glioma cells (U87) was studied after different irradiation doses to define the optimal therapeutic window for Hsp70-targeting NK cells. To maintain the cells in the exponential growth phase during a cultivation period of 7 days different initial cell counts were seeded. Although cytosolic Hsp70 levels remained unchanged on days 4 and 7 after a sublethal irradiation with 2 Gy, 4 Gy and 6 Gy, a dose of 2 Gy resulted in an upregulated mHsp70 density in U87 cells which peaks on day 4 and starts to decline on day 7. Higher radiation doses (4 Gy, 6 Gy) resulted in an earlier and more rapid onset of the mHsp70 expression on days 2 and 1, respectively, followed by a decline on day 5. Membrane Hsp70 levels are higher in G2/M than in G1, however, an irradiation-induced cell cycle arrest on days 4 and 7, was not associated with an increase in the mHsp70 density. Extracellular Hsp70 concentrations increased significantly compared to sham (0 Gy) irradiated cells on days 4 and 7 but not on day 1. Functionally, elevated mHsp70 densities were associated with a significantly better lysis by Hsp70-targeting NK cells. In summary, the kinetics of changes in the mHsp70 density upon irradiation on tumor cells is time- and dose-dependent.

A pattern of NK cell heterogeneity in each individual determines proliferative and functional responses of NK cells to activating stimuli. Obtaining the progeny of a single cell by cloning original population is one of the ways to study the NK cell heterogeneity. In this work, we used single cell sorting into a plate and stimulation by IL-2 and gene-modified K562 feeder cells expressing membrane-bound IL-21 (K562-mbIL21) that led to generation of phenotypically confirmed and functionally active NK cell clones. We applied two models of clone cultivation, which differently affected their phenotype, lifespan and functional activity. The first model, which included weekly restimulation of clones with K562-mbIL21 and IL-2, resulted in the generation of relatively short-lived (5-7 weeks) clones of highly activated NK cells. HLA-DR expression in the expanded NK cells correlated strongly with IFN-γ production. The second model, in which NK cells were restimulated mainly with IL-2 alone, produced long-lived clones (8-14 weeks) that expanded up to 10⁷ cells with lower ability to produce IFN-γ. Our method is applicable for studying variability in phenotype, proliferative and functional activity of the certain NK cell progeny in response to the stimulation, which may help in selecting NK cells best suited for clinical use.

Scrub typhus threatens one billion people in the Asia-Pacific area and cases have emerged outside this region. It is caused by infection with any of the multitude of strains of the bacterium, Orientia tsutsugamushi. A vaccine that affords heterologous protection and a commercially available molecular diagnostic assay are lacking. Herein, we determined that the nucleotide and translated amino acid sequences of outer membrane protein A (OmpA) are highly conserved among 51 O. tsutsugamushi isolates. Molecular modeling revealed the predicted tertiary structure of O. tsutsugamushi OmpA to be very similar to that of the phylogenetically related pathogen, Anaplasma phagocytophilum, including the location of a helix that contains residues functionally essential for A. phagocytophilum infection. PCR primers were developed that amplified ompA DNA from all O. tsutsugamushi strains, but not from negative control bacteria. Using these primers in quantitative real-time PCR enabled sensitive detection and quantitation of O. tsutsugamushi ompA DNA from organs of mice that had been experimentally infected with the Karp or Gilliam strains. The high degree of OmpA conservation among O. tsutsugamushi strains evidences its potential to serve as a molecular diagnostic target and justifies its consideration as a candidate for developing a broadly protective scrub typhus vaccine.

Drug-target binding is an essential parameter in drug discovery and design to ensure drug efficacy and specificity. In 2022, the concept of a general intermolecular binding affinity calculator (GIBAC) was for the first time coined: $Kd = f(molecules, envPara)$. Technically, GIBAC represents a set of in silico approaches for structural and biophysical data generation towards a paradigm shift in precise drug discovery and design, providing a comprehensive framework for intermolecular binding affinity calculation with adequate accuracy, precision and efficiency. For the first time, this study reports a prototype of GIBAC (semaGIBAC), i.e., a one-dimensional semaglutide-GLP-1R-based mini static GIBAC, based on an experimental complex structure of semaglutide and GLP-1R. Semaglutide is a potent GLP-1 receptor agonist used to treat type 2 diabetes mellitus by regulating blood glucose levels and promoting weight loss. In 2021, a structural modification involving a Val27-Arg28 exchange was manually introduced to enhance semaglutide-GLP-1R binding affinity. This study employs a comprehensive structural and biophysical analysis aimed at thoroughly exploring the sequence space of semaglutide-GLP-1R to design analogues with improved binding affinity, leading to the identification of a promising semaglutide analogue, which binds to GLP-1R with an affinity that is more than two orders of magnitude (113.3 times) higher than native semaglutide. To sum up, this article puts forward a promising structural biophysical approach for developing GLP-1 receptor agonists with enhanced efficacy, and with a GIBAC prototype (semaGIBAC), this article argues again that the time is now ripe for the construction of a real GIBAC to be listed on the agenda of the drug discovery and design community.

In the intricate dance between viruses and host cells, RNA-binding proteins (RBPs) serve as crucial orchestrators of gene expression and cellular processes. We will delve into the riveting realm of viral hijacking, where viruses deftly exploit host RBPs to manipulate cellular machinery for their replication and pathogenesis. Through a masterstroke of molecular subterfuge, viruses co-opt RBPs involved in various facets of RNA metabolism - from transcription to degradation - to promote viral gene expression and evade host immune defenses. This manipulation leads to global alterations in cellular RNA metabolism, impacting essential host genes vital for immune responses and homeostasis.Unveiling this clandestine alliance between viruses and RBPs is not just a scientific pursuit, but a critical imperative for devising innovative antiviral strategies to disrupt these interactions. By unraveling the intricate interplay between viral proteins and host RBPs throughout the viral life cycle - from entry to assembly - researchers aim to identify vulnerabilities that can be targeted for therapeutic intervention. Strategies such as disrupting viral protein-RBP interactions hold promise for inhibiting viral replication and curbing pathogenesis, offering a beacon of hope in the battle against viral infections.Our manuscript elucidates the indispensable roles played by RBPs in viral replication, translation, and pathogenesis, shedding light on the molecular mechanisms driving viral success. Delving deeper, it explores how viruses intricately entwine with host RBPs, manipulating cellular signaling pathways to create a hospitable environment for viral spread. By dissecting these manipulative tactics, researchers uncover new targets for antiviral therapy, envisioning a future where tailored interventions disrupt viral-host RBP interactions with precision and efficacy.As the narrative unfolds, the therapeutic implications of targeting RBPs or their interactions with viral proteins emerge as a promising frontier in the fight against viral infections. From small molecule inhibitors to RNA-based therapeutics, innovative approaches are on the horizon, offering new avenues for combatting viral diseases. We set the stage for future research, beckoning researchers to delve deeper into the molecular intricacies of viral hijacking of RBPs and charting a course towards novel therapeutic interventions that promise to reshape the landscape of antiviral therapy.\Ultimately, it beckons the scientific community to embark on a voyage of discovery, unraveling the secrets of viral hijacking of RBPs and paving the way for transformative advances in antiviral therapeutics. The stage is set for a new chapter in the battle against viral infections, where knowledge becomes the sword and innovation the shield against the pernicious machinations of viral pathogens.

Silencing of the fragile X mental retardation 1 (FMR1) gene and consequently lack of synthesis of FMR protein (FMRP) are associated with fragile X syndrome, which is one of the most prevalent inherited intellectual disabilities. FMRP is a multifunctional protein involved in many cellular functions in nearly all subcellular compartments under normal conditions and under conditions of cellular stress in both neuronal and non-neuronal cell types. This is achieved through its trafficking signals, nuclear localization signal (NLS), nuclear export signal (NES), and nucleolar localization signal (NoLS), as well as its RNA and protein binding domains, and is modulated by various post-translational modifications such as phosphorylation, ubiquitination, sumoylation, and methylation. This review summarizes recent advances in understanding the interaction networks of FMRP with a special focus on FMRP stress-related functions, including stress granule formation, mitochondrion and endoplasmic reticulum plasticity, ribosome biogenesis, cell cycle control, and DNA damage response.

In archaea, the interaction of HerA helicase and NurA nuclease is required for 3' end resection during DNA double-stranded break repair, and its disruption results in unfaithful DNA repair. Interactions transpire by the four conserved domains, including the N-terminal HAS domain (HerA-ATP synthase), which serves as a binding site for the exonuclease NurA. As a HerA hexamer between its two subunits, the conserved C-terminal brace extension functions as an adjacent brace. DBD serves as a location for DNA binding, which causes the DNA strands to be propelled into the HerA pocket to begin unwinding. The putative pore entrance is made up of four helix coils, which provide a suitable charged surface for guiding DNA into the pore. The crystal structure of the complex HerA-NurA allows for the identification of key residues for protein-protein interaction. This allows us to predict that most archaeal HerA orthologs will be able to interact with one another. Archaeal HerA is a homo-hexameric assembly featuring a DNA-binding domain that is conserved. It also features distinctive ATP and NurA binding domains. Following the structure and sequence conservation criteria, we find one HerA ortholog in Pyrobaculum calidifontis and multiple HerA paralogs, but the role of none is established yet.Interestingly, orthologous HerA is part of an Mre11-Rad50 gene neighborhood while paralogous HerA is not, which could be misleading. As a result, it sought to describe the criteria that may be used as a possible fingerprint for identifying orthologous genes, as well as the cloning procedures and tactics that would aid in the confirmation of the HerA helicase in P. calidifontis. Their confirmation is based on PCR cloning and characterization following HerA construct creation.

The ability to simulate realistically the electronic structure of superconducting materials is important to understand and predict various properties emerging in both the superconducting topological and spintronics realms. We introduce a tight-binding implementation of the Bogoliubov-de Gennes method, parameterized from density functional theory, which we utilize to explore the bulk and thin films of Nb, known to host a significant superconducting gap. The latter is useful for various applications such as the exploration of trivial and topological in-gap states. Here, we focus on the simulations aspects of superconductivity and study the impact of temperature, Cooper-pair coupling and dimensionality on the size of the superconducting order parameter.

The role of calcium ions (Ca2+) is extensively documented and comprehensively understood in eukaryotic organisms. Nevertheless, emerging insights, primarily derived from studies on human pathogenic bacteria, suggest that this ion also holds a pivotal role in prokaryotes. In this review, our primary focus will be on unraveling the intricate Ca2+ toolkit within prokaryotic organisms, with particular emphasis on its implications in plant growth promoting rhizobacteria (PGPR). To pinpoint and identify homologues of Ca2+ channels, pumps, exchangers, and Calcium-Binding Proteins (CaBPs) we undertook an in silico exploration in the genomes of four distinct PGPR strains using as query prokaryotic sequences that were experimentally confirmed to play a role in regulating intracellular calcium concentration ([Ca2+]i) or sensing and transducing this essential cation. These investigations were conducted in Pseudomonas chlororaphis subsp. aurantiaca SMMP3, P. donghuensis SVBP6, Pseudomonas sp. BP01, and Methylobacterium sp. 2A. which have been isolated and characterized within our research laboratories. We also present experimental data to evaluate the influence of exogenous Ca2+ concentrations ([Ca2+]ex) on the growth dynamics of these isolated strains

Understanding the binding behavior and conformational dynamics of intrinsically disordered proteins (IDPs) is crucial for unraveling their regulatory roles in biological processes. However, their lack of stable 3D structures poses challenges for analysis. To address this, we propose an algorithm that explores IDP binding behavior with protein complexes by extracting topological and geometric features from the protein surface model. Our algorithm identifies a geometrically favorable binding pose for the IDP and plans a feasible trajectory to evaluate its transition to the docking position. We focus on IDPs from Homo sapiens and Mus-musculus, investigating their interaction with the Plasmodium Falciparum (PF) pathogen associated with malaria-related deaths. We compare our algorithm with HawkDock and HDOCK docking tools for quantitative (computation time) and qualitative (binding affinity) measures. Our results indicate that our method outperforms the compared methods in computation performance and binding affinity of experimental conformations.

We calculate energies of shallow donors confined in a rolled-up quantum well in the presence of the electric field by solving numerically the Schrödinger equation in natural curvilinear coordinates. It is found that the curves of density of states (DOS) are very sensitive to the variation of the donor position, the geometry of the spiral and the applied electric field value. Novel results for dependencies of donor’s dipole moment and its polarizability on the electric field strength and its orientation, for different donor positions are presented. Also, we found that anisotropic Stark effect of the first order provides in this structure a dependency of the polarizability on the external electric field in a spike-like shape, giving rise to a sharp variation of the dipole moment.

Zika virus (ZIKV) has been characterized as one of the potential pathogens and placed under future epidemic outbreaks by the WHO. However, lack of potential therapeutics can result in uncontrolled pandemic like other human pandemic viruses; therefore, prioritized effective therapeutics development has been recommended against ZIKV. In this context, the present study adopted the strategy to explore the lead compounds from Azadirachta indica against ZIKV via concurrent inhibition of the ZIKVpro and ZIKVRdRp proteins using molecular simulations. Initially, structure-based virtual screening of 44 bioflavonoids reported in Azadirachta indica against the crystal structures of targeted ZIKV proteins resulted in the identification of top four common bioflavonoids, viz. rutin, nicotiflorin, isoquercitrin, and hyperoside. These compounds showed substantial docking energy (-7.9 to -11.01 kcal/mol) and intermolecular interactions with essential residues of ZIKVpro (His51, Asp74, and Ser135) and ZIKVRdRp (Asp540, Ile799, and Asp665) by comparison to the reference compounds, O7N inhibitor (ZIKVpro) and Sofosbuvir inhibitor (ZIKVRdRp). Long interval molecular dynamics simulation (500 ns) on the selected docked poses reveals the stability of respective docked complexes contributed by intermolecular hydrogen bonds and hydrophobic interactions. The predicted complex stability was further supported by calculated end-point binding free energy using molecular mechanics generalized born surface area (MM/GBSA) method. Consequently, the identified common bioflavonoids are recommended as promising therapeutic inhibitors of ZIKVpro and ZIKVRdRp for further experimental assessment.

Multidrug resistance protein 4 (MRP4) is an energy-dependent membrane transporter that is responsible for cellular efflux of a broad range of xenobiotics and physiological substrates. In this trial, we aimed to investigate the co-effects of aging and MRP4 deficiency using gene expression microarray and morphological and electrophysiological analyses of the mouse retina. Mrp4-knockout (null) mice and wild-type (WT) mice were reared in the same condition to 8–12 wk (young) or 45–55 wk (aged). Microarray analysis identified 186 differently expressed genes from the retinas of aged Mrp4-null mice as compared to that from aged WT mice, and subsequence gene ontology and KEGG pathway analyses showed that differently expressed genes were related to lens, eye development, vision, and transcellular barrier function that are involved in metabolic pathways or viral infection pathways. No significant change in thickness was observed for each retinal layer among young/aged WT mice and young/aged Mrp4-null mice. Moreover, immunohistochemical analyses of retinal cell type did not exhibit an overt change in the cellular morphology or distribution among the 4 age/genotype groups, and the electroretinogram responses showed no significant differences in the amplitude or the latency between aged WT mice and aged Mrp4-null mice. Aging would be an insufficient stress to cause some damage to the retina in the presence of MRP4 deficiency.

In this paper we propose the generation of synthetic small and more sophisticated molecule structures that optimize the binding affinity to a target (ASYNT-GAN). To achieve this we leverage on three important achievements in A.I.: Attention, Deep Learning on Graphs and Generative Adversarial Networks. Similar to text generation based on parts of text we are able to generate a molecule architecture based on an existing target. By adopting this approach, we propose a novel way of searching for existing compounds that are suitable candidates. Similar to question and answer Natural Language solutions we are able to find drugs with highest relevance to a target. We are able to identify substructures of the molecular structure that are the most suitable for binding. In addition, we are proposing a novel way of generating the molecule in 3D space in such a way that the binding is optimized. We show that we are able to generate compound structures and protein structures that are optimised for binding to a target.

RNA interference (RNAi) has become a widely utilised method to study gene function, yet despite this, many of the mechanisms surrounding RNAi remain elusive. The core RNAi machinery is relatively well understood, however many of the systemic mechanisms, particularly double stranded RNA (dsRNA) transport, are not. Here, we demonstrate that dsRNA binding proteins in the serum contribute to systemic RNAi, and may be the limiting factor in RNAi capacity for species such as spiny lobsters where gene silencing is not functional. Incubating serum from a variety of species across phyla with dsRNA led to a gel mobility shift in species where systemic RNAi has been observed, with this response being absent in species where systemic RNAi has never been observed. Proteomic analysis suggested lipoproteins may be responsible for this phenomenon, and may transport dsRNA to spread the RNAi signal systemically. Following this, we identified the same gel shift in the slipper lobster Thenus australiensis and subsequently silenced the insulin androgenic gland hormone, marking the first time RNAi has been performed in any lobster species. These results pave the way for inducing RNAi in spiny lobsters, and better understanding the mechanisms of systemic RNAi in Crustacea, as well as across phyla.

At the molecular level, response to an external factor or an internal condition causes reprogramming of temporal and spatial transcription. When an organism undergoes physiological and/or morphological changes, several signaling pathways are activated simultaneously. Examples of such complex reactions are the response to temperature changes, dehydration, various biologically active substances, and others. Synergistic action of multiple pathways greatly complicates the experimental study of the molecular genetic mechanisms of the organism's reactions. As a result, a significant part of the regulatory ensemble in such complex reactions remains unidentified. We developed metaRE, an R package for the systematic search for cis-regulatory elements enriched in the promoters of the genes significantly changed their transcription in a complex reaction. metaRE mines multiple expression profiling datasets generated to test the same organism's response and identifies simple and composite cis-regulatory elements systematically associated with differential expression of genes. Here we showed metaRE performance for identification of cold stress-responsive cis-regulatory code in Arabidopsis thaliana. MetaRE identified potential binding sites for known as well as unknown cold response regulators. Software with source files, documentation, and example data files are freely available online at the repository (https://github.com/cheburechko/MetaRE).

Two X chromosomes of female mammals randomly inactivate one of paternal or maternal X chromosome in early embryonic development and all the daughter cells produced from these cells retain the same feature of X chromosome inactivation, which is called X chromosome inactivation (XCI). Studying the mechanisms of XCI is important for understanding epigenetic that plays an important role in age-associated diseases. The previous studies have demonstrated that binding of RNAs and DNAs may play a role in activating gene expression. In this paper, our study aims to explore whether the mechanisms of XCI involve the RNA binding strength to X chromosome DNAs. The bioinformatics analyses based on big data were used to analyze the simulated binding strength of RNAs (RNA binding strength) to 23 chromosomes (including X chromosome and 22 human autosomes) and the characteristics of repetitive sequences in the X-inactivation centre. The results revealed that RNA binding strength of the long arm of the X chromosome that is almost entirely inactivated in XCI was significantly lower than that of all autosomes and the short arm of X chromosome, meanwhile the RNA binding strengths of inactivation regions in X chromosome were significantly lower than that of regions escaping from XCI. Different repetitive sequence clusters within the center of XCI presented a cross distribution characteristic. To further prove whether the repetitive sequences in human X chromosome involve in XCI, we cloned long interspersed element (LINE-1, L1) and short interspersed element (Alu) from human Xq13, the center of XCI, and constructed expression vectors carrying sense-antisense combination repetitive sequences (L1s or Alus). Effects of combined L1 or combined Alu sequences on expression of EGFP reporter gene were examined in stably transfected HeLa cells, which simulates the effects of repetitive sequences located on chromosomes. The results of experiments revealed transcribed L1 repetitive sequences activated EGFP reporter gene expression, so did the Alus. The experiment results suggested repetitive sequences activated genes by interaction of transcribed RNAs and DNAs. Since the binding of RNAs and DNAs can activate gene, so the low RNA binding strength of human X chromosome may be one of reasons of XCI. The cross distribution characteristics of different repetitive sequence clusters leading to a cascade of gene activation or gene inactivation may be the reason of transcriptional silencing one of the X chromosomes in female mammals.

PHA synthases (PhaC) are grouped into four classes based on the kinetics and mechanisms of reaction. The grouping of PhaC enzymes into four classes is dependent on substrate specificity, according to the preference in forming short chain length (scl) or medium chain length (mcl) polymers: class I, class III, and class IV produce scl-PHAs depending on propionate, butyrate, valerate and hexanoate precursors, while class II phaC synthesize mcl-PHAs based on the alkane (C6 to C14) precursors. PHA synthases of class I, in particular PhaC_Cs from Chromobacterium USM2 and PhaC_Cn/RePhaC1 from Cupriavidus necator/R. eutropha, have been analysed and the crystal structures of the C-domains have been determined. PhaC_Cn/RePhaC1 was also studied by small angle X-ray scattering (SAXS) analysis. Models have been proposed for dimerization, catalysis mechanism, substrate recognition and affinity, product formation and product egress route. The assays based on amino acid substitution by mutagenesis have been useful to validate the hypothesis on the role of amino acids in catalysis and in accommodation of bulky substrates, for the synthesis of PHB co-polymers and medium chain length-PHA polymers with optimized chemical properties.

In this work α-synuclein amyloid fibrils, formation of which is a biomarker of the Parkinson’s disease, were investigated with the use of fluorescent probe thioflavin T (ThT). Experimental conditions of the protein fibrillogenesis were chosen so that a sufficient number of continuous measurements can be performed to characterize and analyze all stages of this process. The reproducibility of fibrillogenesis and the structure of the obtained aggregates (that is a critical point for their further investigation) were proved using a wide range of physical-chemical methods. For determination of ThT—α-synuclein amyloid fibrils binding parameters sample and reference solutions were prepared with the use of equilibrium microdialysis. By absorption spectroscopy of these solutions ThT—fibrils binding mode with the binding constant about 10⁴ M⁻¹ and stoichiometry of ThT per protein molecule about 1:8 was observed. Fluorescence spectroscopy of the same solutions with the subsequent correction of the recorded fluorescence intensity on the primary inner filter effect allowed to determine another mode of ThT binding to fibrils with the binding constant about 10⁶ M⁻¹ and stoichiometry about 1:2500. Analysis of photophysical characteristics of the dye molecules bound to the sites of different binding modes allowed to assume the possible localization of these sites. Obtained differences in the ThT binding parameters to amyloid fibrils formed from α-synuclein and other amyloidogenic proteins, as well as in the photophysical characteristics of the bound dye, confirmed the hypothesis of amyloid fibrils polymorphism.

The antioxidant activity of molecules constitutes an important factor for the regulation of redox homeostasis and reduction of oxidative stress. Cells affected by oxidative stress can undergo genetic alteration, causing structural changes and promoting the onset of chronic diseases, such as cancer. The in silico study performed here was developed to evaluate the antioxidant potential of two molecules, ZINC08706191 (Z91) and ZINC08992920 (Z20), with recognized epithelial anticancer potential. Molecular docking, chemical-quantum calculations and Pearson's correlation were performed. The Z91 and Z20 molecules showed lower binding free energy (ΔG) values for the receptor-ligand interaction than the reference molecules (caffeine – CAF and ascorbic acid – AA), and better results for values of molecular descriptors correlated with ΔG, resulting in a decrease of ΔG. Strong correlations were observed between ΔG values for the five receptors evaluated and ΔG values of the potential epithelial anticancer activity available in literature. These results attest to the significant antioxidant potential of the Z91 and Z20 molecules and their strong relation with the potential epithelial anticancer activity and may be indicated for further analysis in relation to the control of oxidative stress and epithelial anticancer activity.

Nuclear export of viral mRNAs, is an essential step in the HIV replication cycle. This role is played by a small regulatory protein of HIV-1 called Rev.The N-terminal region of Rev contains an arginine-rich sequence. The arginine-rich motif (ARM) is located between amino acids 38-50 and forms an alpha-helical secondary structure. Expression of the structural proteins of human immunodeficiency virus type 1 requires the direct interaction of multiple copies of the viral Rev protein with its highly structured RNA target sequence, the Rev Response element (RRE). The major viral proteins are not produced if this transport of RNA is stopped. Therefore, knowledge of Rev structure is essential for understanding of its cooperative binding to the RRE, for understanding the mechanism of HIV infection and for the development of antiviral drugs that interfere with Rev’s essential functions and for acknowledgment of good candidate drugs for treatment of AIDS. To understand how REV interact with RRE element of HIV-RNA and its formation of oligomeric complex it is better to characterize the domain wise structure of REV with regard in function of each domain. Due to lack of structural data on Rev no single compound is reported as inhibitor of REV expect antiviral drugs. Identification of a high-affinity RNA-binding site for the human immunodeficiency virus type 1 Rev Protein is much more important. The ARM is a highly specific sequence which allows for the multimerization of Rev and also binding of REV with RNA. Here we are first time exploring the structural characteristics of REV protein both in free form and in complex with RNA at domain function level especially explore the role of ARM motif in REV HIV-1 protein as RNA binding sites by molecular dynamics (MD) simulation and homology modeling studies. Results indicate that the arginine-rich motif (ARM) is crucial in stability of this complex. The residues ARG38, 39, 41, 43, 44, 48, 50, and ASN40 are most interacting with nucleobases of RRE in Crystal structure of Rev and Rev-response-element RNA complex. Our study plays a major role in elaboration of binding of RNA with REV and pave the way for further investigation for therapeutically agent for HIV.

We have previously reported that 60% sucrose diet-fed ChREBP knockout mice (KO) showed body weight loss resulting in lethality. We aimed to elucidate whether sucrose and fructose metabolism are impaired in KO. Wild type mice (WT) and KO were fed a diet containing 30% sucrose with/without 0.08% miglitol, an α-glucosidase inhibitor, and these effects on phenotypes were tested. Furthermore, we compared metabolic changes of oral and peritoneal fructose injection. Thirty percent sucrose diet feeding did not affect phenotypes in KO. However, miglitol induced lethality in 30% sucrose-fed KO. Thirty percent sucrose plus miglitol diet-fed KO showed increased cecal contents, increased fecal lactate contents, increased growth of lactobacillales and Bifidobacterium and decreased growth of clostridium cluster XIVa. ChREBP gene deletion suppressed the mRNA levels of sucrose and fructose related genes. Next, oral fructose injection did not affect plasma glucose levels and liver fructose contents; however, intestinal sucrose and fructose related mRNA levels were increased only in WT. In contrast, peritoneal fructose injection increased plasma glucose levels in both mice; however, the hepatic fructose content in KO was much higher owing to decreased hepatic Khk mRNA expression. Taken together, KO showed sucrose intolerance and fructose malabsorption owing to decreased gene expression.

This study describes the synthesis and characterization of mixed hydrotalcite–organosilica membrane for gas separation. Hydrotalcite exhibits the capability to adsorb carbon dioxide at elevated temperatures. High surface area and favorable coating properties are essential to harness its potential for practical applications. Stable alcohol-based dispersions are needed for thin film applications of mixed membranes containing hydrotalcite. Currently, producing such dispersions without the need for delamination and dispersion agents is a challenging task. This work introduces for the first time a manufacturing approach to overcoming the drawbacks mentioned above. It includes a synthesis of hydrotalcite nanoparticles, followed by agent-free delamination of their layers and final dispersion into alcohol without dispersion agents. Further, the hydrotalcite-derived sorption agent is dispersed in a matrix based on organo-silica gels derived from 1,2-bis(triethoxysilyl)ethane (BTESE). The analytical results indicate that the interconnection between hydrotalcite and BTESE-derived gel occurs via forming a strong hydrogen bonding system between the interlayer species (OH groups, CO32-) of hydrotalcite and oxygen and silanol active gel centers. These findings lay the foundation for applications involving incorporating hydrotalcite-like compounds into silica matrices, ultimately enabling the development of materials with exceptional mass transfer properties. In part 2 of this study, gas separation performance of the organo-silica and the hydrotalcite-like materials and their combined form will be investigated

The pyrolysis process has emerged as an attractive option for waste treatment. In this technology, inorganic compounds play catalytic effects in the pyrolytic reaction of organic materials, increa-sing the char's yield, porosity, and physicochemical properties. The present research investigates and compares the slow pyrolysis of spent coffee grounds (SCG) using potassium hydroxide (KOH) (weight ratio of 1:1) and landfill leachate membrane concentrate (LLMC) residue (weight ratio of 1:1) as pyrolitic additives. We hypothesized that the high content of alkali and alkaline earth metals in LLMC could enhance the yield or improve the quality of the carbon-based material produced in pyrolysis. This strategy could be a sustainable alternative for valorizing SCG and landfill wastes, replacing catalyzing agents such as steam, CO2, and chemical additives used on an industrial scale. The char yields in slow pyrolysis ranged from 18 to 24%. The SCG had a low specific surface area (4.5 m2 g-1), contrasting with the notable surface areas observed in both ac-tivated chars. In particular, the KOH-activated char displayed a higher surface area than the LLMC-activated char (1960 vs. 1130 m2 g-1). On the other hand, the combustion enthalpy of the LLMC-activated material was estimated at 22 MJ kg-1. This value was 22% higher than the energy enthalpy of SCG and KOH-activated char used for comparison. Our findings confirm that pyrolysis of SCG with KOH produces a microporous material with a high specific surface area. In contrast, the resulting LLMC-activated char demonstrates a higher calorific value. Our findings could contribute to establishing potential industrial applications for SCG and LLMC residues to produce energy and high-value materials.

The removal of chromium(VI) from acidic (HCl) medium through non-dispersive extraction with strip dispersion (NDXSD) was investigated using a microporous PVDF membrane support in a permeation cell. The ionic liquid Cyphos IL102 (phosphonium salt) in Solvesso 100 was used as organic phase. In NDXSD the stripping phase (NaOH) is dispersed in the organic phase in the cell side with an impeller stirrer adequate to form a strip dispersion. This pseudo-emulsion phase (organic + strip solutions) provides a constant supply of the Cyphos IL102/Solvesso 100 to the membrane phase. Various hydrodynamic and chemical parameters, such as variation in the feed and pseudo-emulsion stirring speeds, HCl and Cr(VI) concentrations in the feed phase, and carrier concentration, were investigated. From the experimental data, several mass transfer coefficients were estimated, as well as it was investigated the performance of the present system against other ionic liquids and the presence of base metals in the feed phase.

Chronic kidney disease (CKD) is characterized by a continuous decline in kidney function, often persisting for years or even a lifetime. This study presents a new exploration into the extraction and application of insect chitosan (Tenebrio molitor) targeting hemodialysis (HD). Initially, the potential of insect-derived chitosan as a sustainable and effective alternative to traditional sources (i.e. crustaceans) is demonstrated. Efficient extraction methods were developed (totaling between 6 h and 12 h), producing chitosan. Firstly, chitin was efficiently isolated from T. molitor through deproteinization and demineralization, with yields of approximately 5% (w/w). Subsequently, chitosan was obtained from the extracted chitin resulting in yields of between 65.0 ± 0.8% (w/w) and 79.3 ± 0.8% (w/w). Characterization using Fourier Transform Infrared Spectroscopy (FTIR) confirmed structural similarities with commercial chitosan and degrees of deacetylation in the 73 - 75% range. The bioactive properties of chitosan obtained from T. molitor, including antimicrobial and antioxidant activities, were evaluated. All the microorganisms tested were inhibited, exhibiting minimum lethal concentrations between 2 and 8 mg/mL, confirming the antimicrobial activity of the extracted chitosan. In addition, chitosan showed antioxidant activity in the range of 60 to 65 µmol Trolox equivalent/g, suggesting its viability for various medical applications.This study also developed a chitosan-based membrane (CH - M) culminating in a proof-of-concept phase for hemodialysis. The permeation characteristics of CH-M for urea and albumin were studied in vitro to assess their suitability as HD membranes. Urea was permeable to values of over 70% and albumin was retained.

This research aimed to synthesize Polyvinyl fluoride (PVDF) membranes and coat them with Tannic acid (TA) nanoparticles and Polyethylene glycol (PEG) additives so that the membrane's removal efficacy for humic acid (HA) pollutant from agricultural wastewater was investigated. Thus, six membranes with PEG:TA ratios of 0:0, 1:0, 0:1, 1:1, 4:1 and 1:4 were synthesized. Then the membranes' characteristics were identified by FT-IR ATR, FESEM, and AFM analysis, and HA's particle size and zeta potential were also investigated. Based on optimizing effective parameters, the operating pressure of 1.5 bar and HA concentration of 80 ppm were selected as optimal values. The membrane with PEG:TA=4:1, as the optimally modified membrane, had a pure water flux of 446.03 L/m2.h, effluent flux of 72.43 L/m2.h and pollutant removal rate of 86.62% at pH=7 After 60 min had passed. These values for the pristine membrane (PEG:TA=0:0) were 265.64 L/m2.h, 89.39 L/m2.h, and 75.59%, respectively. The results showed that although the effluent flux was lower in the optimized modified membrane than in the pristine membrane, HA removal percentage was increased.

Hepatocellular carcinoma (HCC) is a heterogeneous malignancy with complex carcinogenesis. Although there has been significant progress in the treatment of HCC over the past decades, drug resistance to chemotherapy remains a major obstacle in its successful management. In this study we were able to reduce chemoresistance in cisplatin-resistant HepG2 cells by either silencing the expression of transglutaminase type 2 (TG2) using siRNA or by pre-treatment of cells with the TG2 enzyme inhibitor cystamine. Further analysis revealed that, whereas the full-length TG2 isoform (TG2-L) was almost completely cytoplasmic in its distribution, the majority of the short TG2 isoform (TG2-S) was membrane-associated in both parental and chemoresistant HepG2 cells. Following induction of cisplatin toxicity in non-chemoresistant parental cells, TG2-S together with cisplatin quickly relocated to the cytosolic fraction. Conversely, no cytosolic relocalisation of TG2-S or nuclear accumulation cisplatin was observed following identical treatment of chemoresistant cells, where TG2-S remained predominantly membrane-associated. This suggests that deficient subcellular relocalisation of TG2-S from membranous structures into the cytoplasm may limit the apoptic response to cisplatin toxicity in chemoresistant cells. Structural analysis of TG2 revealed the presence of binding motifs for interaction of TG-S with the membrane scaffold protein LC3/LC3 homologue that could contribute to a novel mechanism of chemotherapeutic resistance in HepG2 cells.

Background: We aimed to explore microvascular changes evaluated with optical coherence tomography angiography (OCTA) in patients undergoing epiretinal membrane (ERM) pars-plana vitrectomy (PPV) combined with intravitreal Ozurdex implantation, compared with standard PPV. (2) Methods: Prospective interventional analysis on 25 eyes undergoing PPV+Ozurdex (Group A) and 25 eyes undergoing PPV alone. Best corrected visual acuity (BCVA) and OCTA parameters, such as vessel density (VD) of the superficial and deep capillary plexi (SCP and DCP) in the whole 6.4x6.4 mm and fovea area, were evaluated preoperatively and 3 months after surgery. (3) Results: Postoperative BCVA significantly improved in both groups. No cases of post-operative cystoid macular edema (CME) were reported in Group A vs. 2 eyes in Group B. In Group A we found a statistically significant increase of SCP’s VD in either the whole image (from 42.1±4.1 to 45.6±4.3 %, p=0.01) and the fovea image (from 38.5±7.5 to 41.7±4.2%, p=0.03). In Group B, we reported no significant variations in the SCP’s VDs. In the DCP, VD significantly increased only in the whole image in Group A. Stage 4 ERMs showed the greatest improvement in VD, especially in Group A. (4) Conclusions: In conclusion, intraoperative Ozurdex prompted a significant BCVA recover and limited the occurrence of postoperative CME compared to the standard procedure. Moreover, Ozurdex implant is associated with a better restoration of microvascular structure in either SCP and DCP.

The present study focuses on the development and optimization of a highly efficient unique hybrid technique — membrane-assisted gas absorption (MAGA) in designing the separation unit, which provides the improvement in mass-transfer of a target component during the natural gas processing in terms of sweetening stage. A novel design of membrane-assisted gas absorption cell is used to enhance the ratio between absorbent amount and membrane surface available for permeation. The performance tests were implemented on example of two gas mixtures separation: one is model ternary gas mixture and the second is quasi real natural gas, which includes number of hydrocarbons, nitrogen, acid impurities and xenon. For both cases, the MAGA unit demonstrated high acid gases removal efficiency, and high recovery of hydrocarbons. The ultimate efficiency of studied system was up to 96 and 61 % of removed acid gases with hydrocarbons losses up to 1 % at highest productivity. In this way, the residual sum of acid gases content may be lowered to 1 vol.% using the heat-free hybrid membrane-assisted gas absorption unit maintaining suitable recovery rate of methane, ethane, propane and n-butane. Nevertheless, the overall efficiency of the process may be enhanced using the specific absorption agents with aqueous amino alcohol solutions.

The Spiking Neural Network (SNN) has attracted more and more attention recently. It adopts binary spike signals to transmit information. Benefitting from the information passing paradigm of SNNs, the multiplications of activations and weights can be replaced by additions, which are more energy-efficient. However, its ``Hard Reset" mechanism for the firing activity would ignore the difference among membrane potentials when the membrane potential is above the firing threshold, causing information loss. Meanwhile, quantifying the membrane potential to 0/1 spikes at the firing instants will inevitably introduce the quantization error thus bringing about information loss too. To address these problems, we propose to use the ``Soft Reset" mechanism for the supervised training-based SNNs, which will drive the membrane potential to a dynamic reset potential according to its magnitude, and Membrane Potential Rectifier (MPR) to reduce the quantization error via redistributing the membrane potential to a range close to the spikes. Results show that the SNNs with the ``Soft Reset" mechanism and MPR outperform their vanilla counterparts on both static and dynamic datasets.

Needle abscission in balsam fir has been linked to both cold acclimation and changes in lipid composition. The overall objective of this research is to uncover lipid changes in balsam fir during cold acclimation and link those changes with postharvest abscission. Branches were collected monthly from September to December and assessed for cold tolerance via membrane leakage and chlorophyll fluorescence changes at -5, -15, -25, -35, and -45°C. Lipids were extracted and analyzed using mass spectrometry while postharvest needle abscission was determined gravimetrically. Cold tolerance and needle retention each significantly (P < 0.001) improved throughout autumn in balsam fir. There were concurrent increases in DGDG, PC, PG, PE, and PA throughout autumn and a decrease in MGDG. Those same lipids were strongly related to cold tolerance, though MGDG had the strongest relationship (R2 = 55.0% and 42.7% from membrane injury and chlorophyll fluorescence, respectively). There was a similar, albeit weaker, relationship between MGDG:DGDG and needle retention (R2 = 24.3%). Generally, a decrease in MGDG:DGDG ratio resulted in better cold tolerance and higher needle retention in balsam fir possibly due to increased membrane stability.

Abstract Aims: In recent years, electro kinetic (EK) remediation has become more popular as a novel method for removing chromium contamination from soil. This approach, however, is ineffective since it uses both cationic and anionic forms of chromium. In this work, a membrane –based technique was employed to increase the efficiency of the electro kinetic removal of chromium. Methods: Chromium removal from polluted sludge was studied in four bench-scale experiments, two of which used distilled water (EK-1, EK2 & Membrane) and two of which used acetic acid as a catholyte (EK-3, EK4 &Membrane). Results: The pH, total chromium, and fractionation of chromium in the sludge were measured after remediation. In the EK-1, EK-2 & Membrane and EK-3, EK-4 & Membrane trials, the average removal efficiency of total chromium was 47.6%, 58.6%, and 74.4%, 79.6%, respectively. Conclusion: In contrast to the electro kinetic remediation strategy, which left approximately 80% of the sludge neutral or alkaline after treatment, the use of the membrane created acidic soil conditions throughout the sludge. For example, the high field intensity used in the membrane tests may have helped to facilitate chromium desorption, dissolution and separation from the sludge, as well as enhance chromium mobility. The findings show that the membrane can improve the effectiveness of chromium removal from sludge when utilized in the EK remediation process.

The established classical method of treating oil refinery effluent is flotation followed by biological treatment. Membrane bioreactors (MBRs) offer more advanced treatment, producing a clarified and potentially reusable treated effluent, but demand robust pretreatment to remove oil and grease (O&amp;G) down to consistent, reliably low levels. An analysis of a full-scale conventional oil refinery ETP based on flotation alone, coupled with projected performance, energy consumption and costs associated with a downstream MBR, have demonstrated satisfactory performance of flotation-based pre-treatment. The flotation processes, comprising an API separator followed by dissolved air flotation (DAF), provided ~90% removal of both total suspended solids (TSS) and O&amp;G coupled with 75% COD removal. The relative energy consumption and cost of the pretreatment, normalised against both the volume treated and COD removed, was considerably less for the API-DAF sequence compared to the MBR. The combined flotation specific energy consumption in kWh was found to be almost an order of magnitude lower than for the MBR (0.091 vs 0.86 kWh per m3 effluent treated), and the total cost (in terms of the net present value) around one sixth that of the MBR. However, the nature of the respective waste streams generated and end disposal of waste solids differs significantly between the pretreatment and MBR stages.

Extracorporeal membrane oxygenation (ECMO), in conjunction with its life-saving benefits, carries a significant risk of acute brain injury (ABI). Hypoxic-ischemic brain injury (HIBI) is one of the most common types of ABI in ECMO patients. Various risk factors such as history of hypertension, high day 1 lactate level, low pH, cannulation technique, large peri-cannulation PaCO2 drop (∆PaCO2), and early low pulse pressure, have been associated with the development of HIBI in ECMO patients. The pathogenic mechanisms of HIBI in ECMO are complex and multifactorial, attributing to the underlying pathology requiring initiation of ECMO and the risk of HIBI associated with ECMO itself. HIBI is likely to occur in the peri-cannulation or peri-decannulation time secondary to underlying refractory cardiopulmonary failure before or after ECMO. Current therapeutics target pathological mechanisms, cerebral hypoxia and is-chemia, by employing targeted temperature management in the case of extracorporeal cardiopulmonary resuscitation (eCPR), and optimizing cerebral O2 saturations and cerebral perfusion. This review describes the pathophysiology, neuromonitoring, and therapeutic techniques to improve neurological outcomes in ECMO patients so as to prevent and minimize the morbidity of HIBI. Further studies aimed at stand-ardizing the most relevant neuromonitoring techniques, optimizing cerebral perfusion, and minimizing the severity of HIBI once it occurs will improve long-term neurological outcomes in ECMO patients.

Ion current rectification (ICR) is the ratio of ion current by forward bias to by backward bias and is a critical indicator of diode performance. In previous studies, many attempts have been continued to improve the performance of this ICR, but there is the intrinsic problem for geometric changes that induce ionic rectification due to their fabrication problem. Additionally, the high ICR could be achieved in the narrow salt concentration range only. Here, we propose a multi-layered bipolar ionic diode based on an asymmetric nanochannel network membrane (NCNM), which is realized by soft lithography and self-assembly of homogenous-sized nanoparticles. Owing to the freely changeable geometry based on soft lithography, the ICR performance can be explored according to the variation of microchannel shape. Interestingly, very stable ICR performance can be achieved using the multi-layered 3D configuration for the bipolar diode in a broad range of salt concentrations (0.1mM ~ 100 mM). This demonstrates the promising potential of multi-layered NCNM for applications in highly concentrated electrolytes, such as biosensors, desalination, or energy harvesting.

Transmembrane transport of small organic and inorganic molecules is one of the cornerstones of cellular metabolism. Among transmembrane transporters, solute carrier (SLC) proteins form the largest, albeit very diverse, superfamily with over 400 members. It has been early on recognized that xenobiotics can directly interact with SLCs and that this interaction can fundamentally determine their efficacy, including bioavailability and intertissue distribution. Apart from the well-established prodrug strategy, the chemical ligation of transporter substrates to nanoparticles of various chemical composition has recently been used as a means to enhance their targeting or absorption. In this review, we summarize efforts in drug design exploiting interactions with specific SLC transporters to optimize their therapeutic effects. Furthermore, we describe current and future challenges as well as new directions for the advanced development of therapeutics that target SLC transporters.

Fouling represents a bottleneck problem for promoting the use of membranes in filtration and separation applications. It becomes even more persistent when it comes to the filtration of fluid emulsions. In this case, a gel-like layer that combines droplets, impurities, salts, and other materials form at the membrane's surface, blocking its pores. It is, therefore, a privilege to combat fouling by minimizing the accumulation of these droplets that work as seeds for other incoming droplets to cluster and coalesce with. In this work, we explore the use of the newly developed and novel periodic feed pressure technique (PFPT) in combating the fouling of ceramic membranes upon the filtration of oily water systems. The PFPT is based on alternating the applied transmembrane pressure (TMP) between the operating one and zero. A PFPT cycle is composed of a filtration half-cycle and a cleaning half-cycle. Permeation occurs when the TMP is set at its working value, while the cleaning occurs when it is zero. Three PFPT patterns were examined over two feeds of oily water systems with oil contents of 100 and 200ppm, respectively. The results show that the PFPT is very effective in minimizing the problem of fouling compared to a non-PFPT normal filtration. Furthermore, the overall drops in permeate flux during the cleaning half cycles are compensated by appreciable enhancement due to the significant elimination of fouling development such that the overall production of filtered water is even increased. Inspection of the internal surface of the membrane post rinsing at the end of the experiment proves that all PFPT cycles maintained the ceramic membranes as clean after a 2-hours operation. This can ensure a prolonged lifespan of the ceramic membrane use and a continuous greater permeate volume production. The advantage of the PFPT is that it can be implemented on existing units with minimal modification, ease of operation, and saving energy.

Understanding the ionic channel network of proton exchange membranes, which dictate fuel cell performance, is crucial when developing proton exchange membrane fuel cells. However, itis difficult to characterize due to complicated nano structure and differing changes to their structure with different amounts of water uptake. Electrostatic force microscopy (EFM) can map surface charge distribution as nano special resolution by measuring the electrostatic force between a vibrating conductive tip and a charged surface under an applied voltage, . In this study, the ionic channel network of a proton exchange membrane is analyzed using EFM. A mathematical approximation model of the ionic channel network is first derived, to explain changes in force gradient on the surface using EFM. The phase lag of dry and wet Nafion under stepwise changes to bias voltage is then measured. Based on the model, variations in the ionic channel network of Nafion with different amounts of water uptake are analyzed numerically. The mean surface charge density of both membranes, which is connected with the ionic channel network, is calculated using the model. The results show that the difference between the mean surface charge of the dry and wet membranes is consistent with the variation in their proton conductivity.

The Ras family of proteins is tethered to the inner leaflet of the cell membranes which play an essential role in signal transduction pathways that promote cellular proliferation, survival, growth, and differentiation. KRas-4B, the most mutated Ras isoform in different cancers, has been under extensive study for more than two decades. Here we have focused our interest on the influence of cholesterol on the orientations that KRas-4B adopts with respect to the plane of the anionic model membranes. How cholesterol in the bilayer might modulate preferences for specific orientation states is far from clear. Herein, after analyzing data from in total 4000 ns-long MD simulations for four KRas-4B systems, properties such as the area per lipid and thickness of the membrane as well as selected radial distribution functions, penetration of different moieties of KRas-4B, and internal conformational fluctuations of flexible moieties in KRas-4B have been calculated. It has been shown that high cholesterol content in the PM favors OS1, exposing the effector-binding loop for signal transduction in the cell from the atomic level. We confirm that high cholesterol in the PM helps KRas-4B mutant stay in its constitutively active state, which suggests that high cholesterol intake can increase mortality and may promote cancer progression for cancer patients. We propose that during the treatment of KRas-4B-related cancers, reducing the cholesterol level in the PM and sustaining cancer progression by controlling the plasma cholesterol intake might be taken into account in anti-cancer therapies.

The Coronavirus Disease 2019 (COVID-19) is a new viral infection caused by the severe acute respiratory coronavirus 2 (SARS-CoV-2). Genomic analyses have revealed that SARS-CoV-2 is related to Pangolin and Bat coronaviruses. In this report, a structural comparison between the Sars-CoV-2 Envelope and Membrane proteins from different human isolates with homologous proteins from closely related viruses is described. The analyses here reported show the high structural similarity of Envelope and Membrane proteins to the counterparts from Pangolin and Bat coronavirus isolates. However, the comparisons have also highlighted structural differences specific of Sars-CoV-2 proteins which may be correlated to the cross-species transmission and/or to the properties of the virus. Structural modelling has been applied to map the variant sites onto the predicted three-dimensional structure of the Envelope and Membrane proteins.

As of today, there is not any direct report yet of the degree to which missing residues exist for experimentally determined membrane protein (MP) structures, which constitute more than half of current drug targets. With a chain- and position-specific visualisation and a statistical analysis of all MP structures inside PDB (as of September 25, 2019), this article argues that the experimentally uncharted territories (EUTs, i.e., consisting of missing residues) within PDB are pluggable and should be plugged with an experimental data-driven hybrid approach, and calls for continued development of MP structural determination with less and less EUTs, in light of MPs' crucial role in biological and biomedical research, both fundamental and pharmaceutical.

Introduction: In rural communities in regions with limited resources the provision of clean water remains challenging. Fecal contamination of water is very common and results in a high incidence of diarrhea, subsequent acute kidney injury and mortality particularly in the very young and old. Membrane filtration is a practical solution to this problem and recent innovation allows membrane filtration using recycled hemodialyzers. We, Easy Water for Everyone, have quantified the systematic effect on health outcomes. Material and Methods: Between 02/2018 and 12/2018, 4 communities in rural Ghana (in the Greater-Accra region) were each provided with a high-volume membrane filtration devices (NUF 500; NuFiltration using recycled hemodialyzers). Health data from montly household surveys and chart review in local healthcare facilities were collected with approval from Ghana Health Services. Specifically, data was collected on gastrointestinal disease, acute kidney injury and therapeutic interventions. Incidence rates for a five-months period before and after implementation of the device were calculated and compared to rates during the same months from 4 neighboring communities that were not yet provided with the device. Results: Acceptance of the devices and the purified water in the studied villages was good and self-reported data of 1130 villagers over 10 months from 9 studied communities in rural Ghana (11% younger than 5 years and 14 % older than 65 years) were included in this analysis. The overall monthly incidence rate of diarrhea showed a decline following the implementation of the device in the 4 study villages from a mean of 0.18 to 0.05 cases per person-month for a reduction in rates by 72% (rate ratio = 0.27). By contrast, the control group of 4 villages in the same region showed no decline in mean rates during the same months as the study period with mean rates changing not significantly from 0.11 to 0.08 cases per person-month. Discussion: Provision of a hemodialyzer membrane filtration device markedly improves health outcomes as measured by diarrhea incidence within rural communities. While our data awaits confirmation in a larger population and further statistical analyses accounting for village characteristics, seasonality and subject demographics, the obvious decline in incidence rates supports widespread use of hemodialyzer membrane filtration devices, particularly in rural regions. Rollout of the device in further sites will likely increase our understanding in terms of risk and other preventive factors modifying the incidence of diarrhea and subsequent acute kidney injury.

Porous asphalt (PA) pavement systems with and without a geotextile layer were investigated in laboratory experiments to determine the impacts of the geotextile layer on processes leading to lead ion (Pb2+) removal from stormwater runoff. Two types of geotextile membranes placed separately at upper and lower levels within the PA systems were tested in an artificial rainfall experiment using synthetic rainwater. The effect of storage capacity within the system on Pb2+ removal was also investigated. Results indicated that the use of a geotextile layer resulted in a longer delay to the onset of effluent. The non-woven geotextile membrane placed below the reservoir course improved the Pb2+ removal rate by 20% over removal efficiency of the system using a woven geotextile placed just below the surface but before the choker course. Pb2+ ions were reduced by over 98% in the effluent after being held for 24 hours in reservoir storage. Results suggest that temporary storage of stormwater in the reservoir course of a PA system is essential to improving Pb2+ ion removal capability.

Gonorrhea is a major global public health problem with emergence of multiple drug-resistant strains with no effective vaccine. This retrospective cohort study aimed to estimate the effectiveness of the New Zealand meningococcal B vaccine against gonorrhea associated hospitalization. The cohort consisted of individuals born 1984-1999 residing in New Zealand, therefore eligible for meningococcal B vaccination during 2004-2008. Administrative datasets of demographics, customs, hospitalization, education, income tax and immunization, were linked using the national Integrated Data Infrastructure. The primary outcome was hospitalization with a primary diagnosis of gonorrhea. Cox’s proportional hazards models were applied with a Firth correction for rare outcomes to generate estimates of hazard ratios. Vaccine effectiveness estimates were calculated as 1-Hazard Ratio expressed as percent. There were 1,143,897 eligible cohort members, with 135 missing information on gender, 16,245 missing ethnicity and/or 197,502 missing deprivation hence 935,496 were included in the analysis. After adjustment for gender, ethnicity and deprivation, vaccine effectiveness (MeNZB™) against hospitalization caused by gonorrhea was estimated to be 24% (95% CI 1-42%). In conclusion, vaccination with MeNZB™ significantly reduced the rate of hospitalization from gonorrhea. This supports prior research indicating possible cross protection of this vaccine against gonorrhea acquisition and disease in the outpatient setting.

Recent years have witnessed a breakthrough in identification of a trimer-of-hairpins motif within viral envelopes that triggers a broad range of virus-host fusion. Identifying a domain capable of controlling virus-host fusion remains a challenge due to sequence diversity, heavy glycan shielding and multiple conformations. Here, we report that HR2, a prevalent heptad repeat sequence comprising an alpha-helical coil anchored in viral membranes, is an accessible site to triterpenes, a class of widely distributed natural products. Triterpenes and their derivatives inhibit the entry of Ebola, HIV, and influenza A viruses with distinct structure-activity relationships. Specifically, triterpenoid probes, upon activation by ultraviolet light, capture the viral envelope via crosslinking the HR2 coil. Profiling the Ebola HR2 sequence using amino acid substitution, surface plasmon resonance (SPR) and nuclear magnetic resonance (NMR) spectroscopy disclosed six constitutive residues that are accessible to triterpenoids, leading to wrapping of the hydrophobic helix by triterpenoids and blocking of the HR1-HR2 interaction, which is critical in the trimer-of-hairpins formation. This finding was also observed in the envelopes of HIV and influenza A viruses and might potentially extend to a broader variety of viruses. Our findings might translate into a shared mechanism that host utilize natural product triterpenoids to antagonize membrane fusion of respective viruses, complementing the current repertoire of antiviral agents.

Oleogels are of high interest as promising substitutes for trans fats in foods. An emulsion-templated method was used to trap olive oil in the chitosan crosslinked with vanillin matrix. Oil in water emulsions (50:50 w/w) with different chitosan content (0.7 and 0.8% w/w) with constant vanillin/chitosan ratio (1.3) were air-dried at different temperatures (50, 60, 70 and 80ºC) and freeze-dried (-26ºC and 0.1 mbar) to produce oleogels. Only falling-rate periods were determined during air-drying kinetics and were successfully modelled with empirical and diffusional models. At drying temperature of 70ºC the drying kinetics were the fastest. Viscoelasticity of oleogels showed that elastic modulus significantly increased after drying at 60 and 70ºC, and those dried at 50ºC and freeze-dried were weaker. All oleogels showed high oil binding capacity (&gt; 91%), but the highest values (&gt; 97%) were obtained in oleogels with a threshold elastic modulus (50,000 Pa). Oleogels color depended on drying temperature and chitosan content (independent of the drying method). Significant differences were observed between air-dried and freeze-dried oleogels with respect to oxidative stability. Oxidation increased with air drying time regardless of chitosan content. Found results indicated that drying conditions must be carefully selected to produce oleogels with specific features.

Frontotemporal lobar degeneration (FTLD) belongs to a heterogeneous group of highly complex neurodegenerative diseases and represents the second cause of presenile dementia in individuals under 65. Frontotemporal-TDP is a subgroup of frontotemporal dementia characterized by the aggregation of abnormal protein deposits, predominantly transactive response DNA-binding protein 43 (TDP-43), in the frontal and temporal brain regions. These deposits lead to progressive degeneration of neurons resulting in cognitive and behavioral impairments. Limbic age-related encephalopathy (LATE) pertains to age-related cognitive decline primarily affecting the limbic system which is crucial for memory, emotions, and learning. Though distinct, emerging research suggests a potential overlap in pathogenic processes, with some cases of limbic encephalopathy displaying TDP-43 pathology. Genetic factors play a pivotal role in both disorders. Mutations in various genes, such as progranulin (GRN) and chromosome 9 open reading frame 72 (C9orf72), have been identified as causative in frontotemporal-TDP. Similarly, specific genetic variants have been associated with an increased risk of developing LATE. Understanding these genetic links provides crucial insights into disease mechanisms and the potential for targeted therapies.

Interactions between proteins and ions are essential for various biological functions like structural stability, metabolism, and signal transport. Given that more than half of all proteins bind to ions, it becomes crucial to identify ion-binding sites. Accurate identification of protein-ion binding sites helps us to understand proteins’ biological functions and plays a significant role in drug discovery. While several computational approaches have been proposed, this remains a challenging problem due to the small size and high versatility of metals and acid radicals. In this study, we propose IonPred, a sequence-based approach that employs ELECTRA (Efficiently Learning an Encoder that Classifies Token Replacements Accurately) to predict ion binding sites using only raw protein sequences. We successfully fine-tuned our pretrained model to predict the binding sites for nine metal ions (Zn2+, Cu2+, Fe2+, Fe3+, Ca2+, Mg2+, Mn2+, Na+, and K+) and four acid radical ion ligands (CO32−, SO42−, PO43−, NO2−). IonPred surpassed six current state-of-the-art tools by over 44.65% and 28.46% respectively in F1 score and MCC when compared on an independent test dataset. Our method is more computationally efficient than existing tools producing prediction results for a hundred sequences for a specific ion in under ten minutes.

Plasminogen (Plg) is the inactive form of plasmin (Plm) that exists in two major glycoforms, referred to as glycoforms I and II (GI and GII). In the circulation, Plg assumes an activation-resistant 'closed' conformation via interdomain interactions and is mediated by the lysine binding site (LBS) on the kringle (KR) domains. These inter-domain interactions can be readily disrupted when Plg binds to lysine/arginine residues on protein targets or free L-lysine and analogues. This causes Plg to convert into an 'open' form which is crucial for activation by host activators. In this study, we investigated how various ligands affect the kinetics of Plg conformational change using small-angle X-ray scattering (SAXS). We began by examining the open and closed conformations of Plg using size-exclusion chromatography (SEC) coupled with SAXS. Next, we developed a high throughput (HTP) 96-well SAXS assay setup. This method enables us to determine the Kopen value, which is used to compare ligands’ effect on Plg conformation directly. Based on our analysis using Plg GII, we have found that the Kopen for ε-aminocaproic acid (EACA) is approximately three times greater than that of Tranexamic acid (TXA), which is widely recognized as a highly effective ligand. We demonstrated further that Plg undergoes a conformational change when it binds to the C-terminal peptides of the inhibitor α2-antiplasmin and receptor Plg‑RKT. Our findings suggest that, besides the C-terminal lysine, internal lysine(s) are also necessary for the formation of open Plg. Finally, we compared the conformational changes of Plg GI and GII directly and found that the closed form of GI, which has an N-linked glycosylation, is less stable. To summarize, we have successfully determined the response of Plg to various ligand/receptor peptides by directly measuring the kinetics of its conformational changes.

Researches are moving to iron and ammonia elimination from ground water. Here, we are using poly acrylic–poly acrylamide hydrogel which grafted with 3-chloroaniline. This copolymer was synthesized by addition polymerization. Effect of agitation time, dosage and adsorbents temperature on the removal process sensitivity has been investigated. The copolymer was described experientially and theoretically. Isothermal, kinetic adsorption models and were discussed. This hydrogel could be regenerated efficiently (98.3%removal of iron and 100% removal of ammonia). Density functional theory DFT method using B3LYP/6-311G(d,p),LANL2DZ level of the theory were managed to investigate stationary states of grafted co-polymer and the complexation energy of the hydrogel with the studied cations. NBO analysis is using DFT to investigate the negative centers on the hydrogel. The complexation energy showed selectively of hydrogel to studied cations.

The Duffy protein, a transmembrane molecule, functions as a receptor for various chemokines and facilitates attachment between the reticulocyte and the Plasmodium Duffy antigen-binding protein. Duffy expression correlates with the Duffy receptor gene for the chemokine, located on chromosome 1, and exhibits geographical variability worldwide. Traditionally, researchers have described the Duffy negative genotype as a protective factor against phenotypic malaria expression. However, recent studies suggest this microorganism's evolution potentially diminishes this protective effect. Nevertheless, there is currently insufficient global data to demonstrate this phenomenon. This study aimed to evaluate the relationship between the Duffy genotype/phenotype and the prevalence of Plasmodium vivax infection. The protocol for the systematic review was registered in PROSPERO as CRD42022353427 and involved reviewing published studies from 2012 to 2022. Medline, Web of Science, Scopus, and Scielo databases were consulted. Assessments of study quality were conducted using the STROBE and GRADE tools. A total of 34 studies were included, with Africa accounting for most recorded studies. The results varied significantly regarding the relationship between the Duffy genotype/phenotype and Plasmodium vivax invasion. Some studies predominantly featured the negative Duffy genotype yet reported no malaria cases. Other studies identified minor percentages of infections. Conversely, certain studies observed a higher prevalence (99%) of Duffy-negative individuals infected with Plasmodium vivax. In conclusion, no evidence of a gender-specific distribution of malaria between Duffy-negative men and women was found. However, evidence supports that the homozygous Duffy genotype positive for the A allele (FY*A/*A) is associated with a higher incidence of Plasmodium infection. Furthermore, the negative Duffy genotype does not confer protection against this disease.

The effects of hypothermia on neonatal encephalopathy may vary regionally, spatially, and cytopathologically in the gyrencephalic neocortex with manifestations potentially influenced by seizures that alter the severity and distribution of neuropathology. We developed a neonatal piglet survival model of hypoxic-ischemic (HI) encephalopathy and hypothermia with continuous encephalographic (cEEG) monitoring for seizures to study injury in neocortex. Neonatal piglets were randomized to naïve, HI-normothermia (NT), overnight hypothermia (HT) initiated 2 hours after HI, sham-NT, or sham-HT treatments. Some piglets within sham and HI groups received cEEG monitoring during recovery. Survival was 2-7 days (piglets were unmedicated and those with poor recovery and unresolving seizures were euthanized early); there was no differences in survival among groups (p&gt;0.078). Neuropathology was assessed by hematoxylin and eosin staining and immunohistochemistry for RNA Binding FOX-1 Homolog 3 (Rbfox3/NeuN). Normal and ischemic-necrotic neurons were counted (layers II-VI collectively) in somatosensory, motor, and prefrontal cortices, identified by cytoarchitecture and connectomics, and in inferior parietal cortex by layer. Seizure burden was determined. HI-NT piglets had reduced normal/total neuron ratio and increased ischemic-necrotic/total neuron ratio relative to naïve, sham-NT, and sham-HT piglets in anterior and posterior motor and somatosensory cortices. Frontal cortex was vulnerable in the prefrontal lateral bank after HI-NT with ischemic-necrosis. HI-HT piglets had higher normal/total neuron ratios and lower ischemic-necrotic/total neuron ratios than HI-NT piglets in anterior/posterior motor and somatosensory cortices. Total normal neuron density in layer III of inferior parietal cortex was reduced in HI-NT piglets compared to sham piglets and was protected by HT. Laminar analysis of Rbfox3 in somatosensory cortex revealed three types of neurons: Rbfox3-positive/normal, Rbfox3-positive/ischemic-necrotic, and Rbfox3-depleted. HI piglets had increased Rbfox3-depleted/total neuron ratio in layers II and III compared to sham-NT. Cortical neuron Rbfox3-depletion was partly rescued by HT. Seizure burden was more severe in HI piglets compared to sham piglets, but HI-NT and HI-HT piglets were similar. We conclude that 1) the neonatal piglet neocortex has a suprasylvian spatial vulnerability to HI and seizures; 2) HT protects against neuropathology in functionally different regions of the neonatal gyrencephalic neocortex; 3) neurons in neonatal neocortex have a limited cytopathology repertoire seen by H&amp;E staining; 4) higher seizure burden correlates with more ischemic-necrotic neurons in somatosensory cortex; 5) seizure presence associates with damage spread to inferior parietal cortex; 6) seizure presence is insensitive to HT, and 7) Rbfox3 immunophenotyping identifies a novel neuronal RNA splicing protein nuclear-depletion pathology that appears reversible by HT. This work demonstrates that HT protection of neocortex in neonatal HI is topographic and laminar, seizure unmitigating, and restores depleted neuronal RNA splicing factor.

The detection and quantification of protein–protein interactions (PPIs) is a crucial technique that often involves the use of recombinant proteins with fusion-protein tags, such as maltose-binding protein (MBP) and glutathione-S-transferase (GST). In this study, we improved the cohesive and sticky properties of gelatinized starch by supplementing it with agarose, resulting in a harder gel that could coat the bottom of a microtiter plate. The resulting gelatinized starch/agarose mixture allowed for the efficient immobilization of MBP-tagged proteins on the coated plates, enabling the use of indirect ELISA-like PPI assays. By using the enzymatic activity of GST as an indicator, we succeeded in determining the dissociation constants between MBP-tagged and GST-tagged proteins on 96-well microtiter plates and a microplate reader without any expensive specialized equipment.

Three finger toxins (3FTX) are a group of peptides that affect multiple receptor types. One group of proteins affected by 3FTX are nicotinic acetylcholine receptors (nAChR). Structural information on how neurotoxins interact with nAChR is limited and are confined to a small group of neurotoxins. Therefore, in silico methods are valuable in understanding the interactions between 3FTX and different nAChR subtypes, but there are no established protocols to model 3FTX – nAChR interactions. We developed a homology modeling and protein docking protocol to address this issue and tested its success on three different systems. First, neurotoxin peptides co-crystallized with acetylcholine binding protein (AChBP) were re-docked to assess whether Rosetta protein – protein docking can reproduce the native poses. Second, experimental data on peptide binding to AChBP was used to test whether the docking protocol can qualitatively distinguish AChBP-binders from non-binders. Finally, we docked eight peptides with known α7 and muscle-type nAChR binding properties to test whether the protocol can explain the differential activities of the peptides at the two receptor subtypes. Overall, our docking protocol succeeded in predicting both qualitative and specific aspects of 3FTX binding to nAChR and shed light on some unknown aspects of 3FTX binding to different receptor subtypes.

A SARS-CoV receptor-binding domain (RBD) recombinant protein was developed and manufactured under current good manufacturing practices in 2016. The protein known as RBD219-N1 when formulated on Alhydrogel®, induced high-level neutralizing antibodies and protective immunity with minimal immunopathology in mice after a homologous virus challenge with SARS-CoV (MA15 strain). In this report, we examined published evidence in support of whether the SARS-CoV RBD219-N1 could be repurposed as a heterologous vaccine against Coronavirus Infectious Disease (COVID)-19. Our findings include evidence that convalescent serum from SARS-CoV patients can neutralize SARS-CoV-2. Additionally, a review of published studies using monoclonal antibodies (mAbs) raised against SARS-CoV RBD and that neutralize the SARS-CoV virus in vitro, finds that some of these mAbs bind to the receptor-binding motif (RBM) within the RBD, while others bind to domains outside this region within RBD. This information is relevant and supports the possibility of developing a heterologous SARS-CoV RBD vaccine against COVID-19, especially due to the finding that the overall high amino acid similarity (82%) between SARS-CoV and SARS-CoV-2 spike and RBD domains is not reflected in RBM region (59%). However, the high similarity (94%) in the region outside of RBM offers the potential of conserved neutralizing epitopes between both viruses.

Various peptides or non-structural amino acids are recognized by their specific target proteins and perform biological role in various pathways in vivo. Understanding the interactions between target protein and peptides (or non-structural amino acids) provides key information on the molecular interactions, which can be potentially translated to the development of novel drugs. However, it is experimentally challenging to determine the crystal structure of protein-peptide complexes. To obtain structural information on substrate recognition of peptide-recognizing enzyme, X-ray crystallographic studies were performed using triglycine (Gly-Gly-Gly) as main-chain of peptide. The crystal structure of Parengyodontium album Proteinase K in complex with triglcyine was determined at 1.4 Å resolution. Two different bound conformations of triglycine were observed at the substrate recognition site. The triglycine backbone forms stable interactions with β5-α4 and α5-β6 loops of main-chain. One of the triglycine-binding conformations was identical with the binding mode of a peptide-based inhibitor from a previously reported crystal structure of Proteinase K. Triglycine has potential application X-ray crystallography to identify substrate recognition sites in peptide binding enzymes.

At nuclear scale, we present three heuristic relations pertaining to strong and electroweak coupling constants. With these relations, close to beta stability line, it is possible to study nuclear binding energy with a single energy coefficient of magnitude $\left(\frac{1}{{\alpha }_s}\right)\left[\frac{e^2}{4\pi {\epsilon }_0{R}_0}\right]\approx 10.0\text{MeV}\text{.}$ With reference to up and down quark masses, it is also possible to interpret that, nuclear binding energy is proportional to the mean mass of $\left[\left(2m_u+m_d\right)\text{and}\left(m_u+2m_d\right)\right]\approx 10.0\text{MeV}.$

Abstract: Background: In 2021, two randomized controlled trials (RCTs), TheraP and VISION, demonstrated that 177Lu-PSMA-617 as monotherapy was more effective for enhancing the decline of PSA than the comparator third-line treatments. Methods: Our review summarizes new RCTs that add to the use of radioligand therapy (RLT) for patients with high-risk prostate cancer (PCa). Results: Five past and present RCTs include 1139 patients. A RCT, ENZA-p, studied first-line treatment of patients with metastatic castration-resistant PCa (mCRPC). A combination of enzalutamide (ENZA) and 177Lu-PSMA-617 gave longer progression-free survival than ENZA as monotherapy. Other RCTs of patients with mCRPC, including the PSMAfore, and SPLASH trials, showed 177Lu-PSMA-617 as second-line treatment gave better progression-free survival than androgen receptor pathway inhibitors (combined p value 6 x 10-8). Conclusion: Patients with PCa may gain if they are given PSMA-RLT early in the treatment of PCa and as part of combination therapies.

Traumatic brain injury (TBI) affects millions of people each year. Previous studies using the central fluid percussion injury (CFPI) model in adult male rats indicated that elevated intracranial pressure (ICP) was associated with long-term effects, including neuronal cell loss and increased sensory sensitivity post-injury and secondary ICP elevation, that were not seen following injury alone. Investigations also indicated that Cathepsin B (Cath B), a lysosomal cysteine protease, may play a role in the pathological progression of neuronal membrane disruption; however, the specific impact of Cath B inhibition following CFPI remains unknown. Thus, the focus of this study was to evaluate the effects of Cath B inhibition via intracerebroventricular infusion of CA-074Me 2w following injury with or without secondary elevation of ICP. This was accomplished using adult male rats continuously infused with CA-074Me or 10% DMSO as a vehicle control for 2w following either sham injury, a CFPI only, or a CFPI with subsequent ICP elevation to 20mmHg. We assessed Cath B activity in the lateral neocortices and liver, evaluated protein levels of Cath B and Cath B binding partners AIF, Bcl-XL, and Bak. We also conducted histological analyses of total cell counts to assess for cell loss, membrane disruption, and Cath B localization. Finally, we investigated somatosensory changes with the whisker nuisance task. Overall, this study demonstrated that Cath B is not a direct driver of membrane disruption, however, administration of CA-074Me alters Cath B localization and reduced hypersensitivity, emphasizing Cath B being an important component in late secondary pathologies.

The present work reports the effective method for removal of inorganic and organic pollutants using membranes based on different carbonaceous materials. The membranes were prepared based on cellulose acetate (18 wt. %), polyvinylpyrrolidone as a pore generating agent (2 wt. %) and activated carbon (1 wt.%). Activated carbons were developed from residues after extraction of mushroom Inonotus obliguus using microwave radiation. It has been shown that addition of activated carbon the physical (porosity, equilibrium water content and permeability) and chemical (content of the surface oxygen group) properties of the membranes. The addition of carbon material has a positive effect on the removal of copper ions from their aqueous solutions by the cellulose-carbon composites obtained. Moreover, the membranes were proved to be more effective in removal of copper ions than iron ones and phenol. The membranes were found to show higher effectiveness in copper removal from a solution of the initial concentration 800 mg/L. The most efficient in copper ions removal was the membrane containing urea-enriched activated carbon.

Membrane gas absorption technology has been considered as a promising approach to mitigate CO2 emissions from power plants. The aim of this study is to evaluate the environmental impacts of CO2 absorption and desorption processes by hollow fiber membrane contactors using life cycle assessment methodology. On the basis of ReCipe 2016 Midpoint and ReCipe 2016 Endpoint methods, the research results show that, membrane gas absorption system exhibits the lowest environmental impacts across the majority assessed categories in comparison with chemical absorption and membrane gas separation systems. The CO2 capture process via membrane gas absorption has the most significant impact on METP category, with heat consumption as the primary contributing factor accounting for 55%, followed by electricity consumption accounting for 43.1%. According to the sensitivity analysis, heating by natural gas shows a better performance than other heat supply sources in improving overall environmental impacts. In addition, the increasing utilization of renewable energy in electricity supply reduces the global warming potential, fossil resource consumption and ozone formation.

As marine traffic is contributing to pollution and mostly have predictable route with repetitive load profiles, to reduce their impact on environment, hybrid system with proton exchange membrane fuel cells (PEMFC-s) and battery pack, is promising replacement. For this purpose, the numerical model of repowered existing vessel that have two diesel engines with 300 kWe is developed. The goal of this paper is to develop numerical model which analyzes and determine equivalent hybrid ship propulsion system for known traveling route. The developed numerical model consists of an interconnected system with PEMFC stack and battery pack as power sources. Numerical model is developed and optimized to meet minimal required power demand for successful route demand, which has variable loads and daily sails six-time same voyage- in total 54 nautical miles. The results have shown that equivalent hybrid power system consists of 300 kWe PEMFC stack and battery pack with 424 kWh battery and state of charge varying between 20 and 87%. To power this new hybrid power system, hydrogen tank of 7200 l holding 284.7 kg at pressure of 700 bar is needed, compared to previous system that spend 1524 kg of diesel and generate 4886 kg of CO2.

In this study, the hybrid biological ion exchange (BIEX) resin and gravity-driven membrane (GDM) process was employed for the treatment of coloured and turbid river water. The primary objective was to investigate the impact of both physical and chemical cleaning methods on ceramic and polymeric membranes in terms of stabilised flux, flux recovery after physical/chemical cleaning, and permeate quality. To address these objectives, two types of MF and UF membranes were utilised (M1 = polymeric MF, M2 = polymeric UF, M3 = ceramic UF, and M4 = lab-made ceramic MF).Throughout extended operation, the resin functioned initially in the primary ion exchange (IEX) region (NOM displacement with pre-charged chloride) and progressed to a secondary IEX stage (NOM displacement with bicarbonate and sulphate), while membrane flux remained stable. Subsequently, physical cleaning involved air/water backwash with two different flows and pressures, and chemical cleaning utilised NaOH at concentrations of 20 and 40 mM, as well as NaOCl at concentrations of 250 and 500 mg Cl2/L. These processes were carried out to assess flux recovery and identify fouling reversibility. The results indicated an endpoint of 1,728 bed volumes (BV) for the primary IEX region, while the secondary IEX continued up to 6,528 BV. At the end of the operation, DOC and UVA254 removal in the effluent of the BIEX columns were 68% and 81%, respectively, compared to influent water. This was followed by 30% and 57% DOC and UVA254 removal by M4 (ceramic MF). The stabilised flux remained approximately 3.8 – 5.2 LMH both before and after the cleaning process, suggesting that membrane materials do not play a pivotal role. The mean stabilised flux of polymeric membranes increased after cleaning, whereas that of ceramics decreased. Enhanced air–water backwash flow and pressure resulted in increased removal of hydraulic reversible fouling, which was identified as the dominant fouling type. Ceramic membranes exhibited higher removal of reversible hydraulic fouling than polymeric membranes. Chemical cleaning had a low impact on flux recovery; therefore, we recommend solely employing physical cleaning.

In this study, high-performance FAU (NaY type) zeolite membranes were successfully synthesized using small-sized seeds of 50 nm, and their gas separation performance was systematically evaluated. Employing nano-sized NaY seeds and an ultra-dilute reaction solution with a molar composition of 80 Na2O: 1Al2O3: 19 SiO2: 5000H2O, the effects of synthesis temperature, crystallization time, and porous support (α-Al2O3 or mullite) on the formation of FAU membranes were investigated. The results illustrated that further extending the crystallization time or increasing the synthesis temperature led to the formation of a NaP impurity phase on the FAU membrane layer. The most promising FAU membrane with a thickness of 2.7 µm was synthesized on an α-Al2O3 support at 368 K for 8 h, and had good reproducibility. The H2 permeance of the membrane was as high as 5.34×10-7 mol/(m2 s Pa), and the H2/C3H8 and H2/i-C4H10 selectivities were 183 and 315, respectively. The C3H6/C3H8 selectivity of the membrane was as high as 46 with a remarkably high C3H6 permeance of 1.35× 10-7 mol/(m2 s Pa). The excellent separation performance of the membrane is mainly attributed to the thin, defect-free membrane layer and relatively wide pore size (0.74 nm).

Composite polymolecular separation membranes were prepared by combining multi-branched ZIF-L with high porosity electrospinning nanofibers PI. Meanwhile, PDA and PEI were introduced into the membrane in order to improve its adhesion. The new membrane is so called “PI@PDA@PEI/ZIF-L-4” composite membrane; compared with PI@PDA@PEI/ZIF-8 composite membrane, the new membrane’s filtration rates for heavy metal ions such as Cd2+, Cr3+ and Pb2+ were increased by 7.0%, 6.6% and 9.3%, respectively. Furthermore, the new membrane has a permeability up to 1,140.0 L·m−2·h−1·bar−1, and were presenting a very stable performances after four repeated uses. The separation mechanism of PI@PDA@PEI/ZIF-L composite membranes was further analyzed in order to provide a base support for producing separation membranes with high barrier rate and high flux.

Energy demand has exponentially increased with the increase in the world population and ur-banization. Renewable energy is clean, renewable, and environmentally friendly. Hydrogen production through Proton Exchange Membrane (PEM) electrolyzer is an efficient way to produce renewable energy. The efficiency and energy consumption of the PEM electrolyzer depends on the feed water quality. In this report, we study the effect of pH, TDS, and conductivity on hydrogen production as well as energy consumption during the process. pH values (3, 7, 9), TDS values (300ppm, 600ppm, 900ppm), and conductivity values (30mS/cm, 70mS/cm, 100mS/cm) were studied to understand and optimize the hydrogen production using PEM electrolyzer. The in-vestigations showed that hydrogen production is significantly affected by pH, total dissolved solids, and conductivity, and the best levels of each variable were identified by extensive testing. The findings show both the scientific significance of proton exchange electrolysis in meeting the growing energy demands of society as well as the importance of knowing the influence that di-verse conditions have on the creation of hydrogen. Our research, in general, contributes to the expanding body of scientific knowledge on the topic of the effective production of hydrogen and offers useful insights for the continuation of investigation in this field.

Extracorporeal membrane oxygenation (ECMO) can be a life-saving intervention in cases of potentially reversible refractory respiratory failure. One such indication can be bleomycin-induced lung injury. However, in some cases, the injury can be so severe that it becomes irreversible and creates complex medical decisions regarding life support and the continuation of care when no additional therapeutic options are feasible, particularly in cases of patients who were young and fully functional prior to an acute illness. In cases of full pulmonary replacement with mechanical support and the degree of functionality that can be attained utilizing modalities such as ECMO can obscure the true severity of illness and make end-of-life decisions significantly harder for families and caregivers.

Specialized classes of proteins, working together in a tightly orchestrated manner, induce and maintain highly curved cellular and organelles membrane morphology. Due to the various ex- perimental constraints, including the resolution limits of imaging techniques, it is non-trivial to accurately elucidate interactions among the various components involved in membrane deformation. The spatial and temporal scales of the systems also make it formidable to investigate them using simulations with molecular details. Interestingly, mechanics-based mesoscopic models have been used with great success in recapitulating the membrane defor- mations observed in experiments. In this review, we collate together and discuss the various mechanics based mesoscopic models for protein-mediated membrane deformation studies. In particular, we provide an elaborate description of a mesoscopic model where the membrane is modeled as a triangulated sheet and proteins are represented as either nematics or fila- ments. This representation allows us to explore the various aspects of protein-protein and protein-membrane interactions as well as examine the underlying mechanistic pathways for emergent behavior such as curvature-mediated protein localization and membrane deforma- tion. We also put forward current efforts in the field towards back-mapping these mesoscopic models to finer-grained particle based models - a framework that could be used to explore how molecular interactions propagate to physical scales and vice-versa. We end the review with an integrative-modeling based road map where experimental imaging micrograph and biochemical data are combined with mesoscopic and molecular simulations methods in a theoretically consistent manner to faithfully recapitulate the multiple length and time scales in the membrane remodeling processes.

All living cells maintain a charge distribution across their cell membrane (membrane potential) by carefully controlled ion fluxes. These bioelectric signals regulate cell behavior (such as migration, proliferation, differentiation) as well as higher-level tissue and organ patterning. Thus, voltage gradients represent an important parameter for diagnostics as well as a promising target for therapeutic interventions in birth defects, injury, and cancer. However, despite much progress in cell and molecular biology, little is known about bioelectric states in human stem cells. Here, we present simple methods to simultaneously track ion dynamics, membrane voltage, cell morphology, and cell activity (pH and ROS), using fluorescent reporter dyes in living human neurons derived from induced neural stem cells (hiNSC). We developed and tested functional protocols for manipulating ion fluxes, membrane potential, and cell activity, and tracking neural responses to wounding and re-innervation in vitro. Finally, using morphology sensor, we tested and quantified the ability of physiological actuators (neurotransmitters and pH) to manipulate nerve wound re-innervation. These methods are not specific to a particular cell type and should be broadly applicable to the study of bioelectrical controls across a wide range of combinations of models and endpoints.

Cell migration is essential for physiological, pathological and biomedical processes such as, in embryogenesis, wound healing, immune response, cancer metastasis, tumour invasion and inflammation. In light of this, quantifying mechanical properties during the process of cell migration is of great interest in experimental sciences, yet few theoretical approaches in this direction have been studied. In this work, we propose a theoretical and computational approach based on the optimal control of geometric partial differential equations to estimate cell membrane forces associated with cell polarisation during migration. Specifically, cell membrane forces are inferred or estimated by fitting a mathematical model to a sequence of images, allowing us to capture dynamics of the cell migration. Our approach offers a robust and potentially accurate framework to compute geometric mechanical membrane forces associated with cell polarisation during migration and also yields geometric information of independent interest, we illustrate one such example that involves quantifying cell proliferation levels which are associated with cell division, cell fusion or cell death.

To investigate the role of sulfhydryl groups and disulfide bonds in different protein-stabilized emulsions, N-ethylmaleimide (NEM) was used as sulfhydryl-blocking agent to be added in the emulsion. The addition of NEM to block the sulfhydryl groups resulted in a reduction of the content of disulfide bonds formation, which enabled destruction of the internal structure of the protein molecule, and then decreased the restriction of protein membrane on the oil droplets. Furthermore, with NEM content increasing in the emulsion, a reduction of protein emulsifying activity and emulsion stability also occurred. At the same time, the intermolecular interaction of the protein on the oil droplet interface membrane was destroyed, and the emulsion droplet size increased with the NEM content in the emulsion. Although NEM blocking sulfhydryl groups not to form disulfide bonds has similar effects on three types of protein emulsion, the degree of myofibrillar protein (MP), egg-white protein isolate (EPI), and soybean protein isolate (SPI) as emulsifier had a subtle difference.

BACKGROUND: The number of waitlisted patients requiring mechanical circulatory support (MCS) as a bridge to heart transplantation is increasing. The data concerning the results of double-bridge strategy are limited. We sought to investigate the post-transplant outcomes across the different bridge strategies. METHODS: We retrospectively reviewed a heart transplantation database from Jan 2009 to Jan 2019. Intra-aortic balloon pump (IABP), extracorporeal membrane oxygenation (ECMO), and ventricular assist devices (VAD) were the MCS that we investigated. The pre- and post-transplant characteristics and variables of patients bridged with the different types of MCS were collected. The post-transplant survival was compared using Kaplan-Meier survival analysis. RESULTS: A total of 251 heart transplants were reviewed; 115 without MCS and 136 with MCS. The patients were divided to five groups: Group 1 (no MCS): n=115; Group 2 (IABP): n=15; Group 3 (ECMO): n=33; Group 4 (ECMO-VAD): double-bridge (n=59); Group 5 (VAD): n=29. Survival analysis demonstrated that the 3-year post-transplant survival rates were significantly different among the groups (Log-rank p < 0.001). There was no difference in survival between group 4(ECMO-VAD) and group 1(no MCS)1 (p = 0.136), or between group 4(ECMO-VAD) and group 5(VAD) (p = 0.994). Group 3(ECMO) had significantly inferior 3-year survival than group 4(ECMO-VAD) and group 5(VAD). CONCLUSION: Double bridge may not lead to worse mid-term results in patients who could receive a transplantation. Initial stabilization with ECMO for critical patients before implantation of VAD might be considered as a strategy for obtaining an optimal post-transplant outcome.

An embryonic version of membrane theory can be date back to the Bernstein's work reported more than a hundred years ago. Such an originally old work has evolved conceptually and mathematically up until today, and it plays a central role in current membrane theory. Goldman-Hodgkin-Katz equation (GHK eq.) is one of the math-based monumental works, which constitutes the present membrane theory. Goldman theoretically derived GHK eq., but its physiological meaning was provided by the two renowned scientists, Hodgkin and Katz. These two employed an assumption that the electric field (EF) across the plasma membrane is constant to validate the GHK eq. physiologically. Proposal of Hodgkin-Huxley model (HH model) is another math-based monumental works developed from the membrane theory and now forms a fundamental part of the current membrane theory. GHK eq. and HH model are quite fundamental central concepts in the current physiology. Despite the broad acceptance of GHK eq. at present time, its prerequisite that the EF within the plasma membrane is constant is hardly believable. Especially when the action potential is generated, it sounds totally nonsense. Furthermore, the existence of constant EF within the plasma membrane is conceptually almost in conflict with the HH model. The authors will discuss those problematic issues the membrane theory inherits.

Severe COVID-19 is associated with viraemia and multiple organ disease. Similar clinicopathological features have been previously seen in SARS and MERS. Clinically, the severity of SARS, MERS and COVID-19 has been associated with the presence of SARS-CoV, MERS-CoV or SARS-CoV2 viraemia in affected patients. In vitro work has looked at the pattern of viral entry and release from polarised epithelial cells infected by coronaviruses. This work has demonstrated a correlation between the severity of a coronavirus infection and the ability of the virus to reach and infect the basal surface of host cells. It has been postulated that this ability helps the virus invade the bloodstream of the host, resulting in a systemic infection with multiple organ involvement. Here we propose that basal surface release and entry of COVID-19 into and out of cells at epithelial-endothelial interface plays a key pathogenic role in severe COVID-19 disease.

Molecular hydrogen (H2) has been shown to have antioxidant and anti-inflammatory activities that may reduce the development and progression of many diseases. In this study, Hydrogen-rich water (HRW) was obtained by reacting hybrid magnesium-carbon hydrogen storage materials with water. Then the effects of intake of HRW on the activities of xenobiotic-metabolizing enzymes, membrane transporters, and oxidative stress in rats were investigated. Rats were given HRW ad libitum for 4 weeks. Results showed that intake of HRW had no significant effect on the activities of various cytochrome P450 (CYP) enzymes (CYP1A1, 1A2, 2B, 2C, 2D, 2E1, 3A, 4A), glutathione-S-transferase and UDP-glucuronosyltransferase. Except for a slight lower plasma glucose concentration, intake of HRW had no effect on other plasma biochemical parameters in rats. P-Glycoprotein and multidrug resistance-associated protein (Mrp)2 protein expressions in liver were elevated after intake of HRW. However, HRW had no significant effects on glutathione, glutathione peroxidase, or lipid peroxidation in liver. Results from this study suggest that consumption of HRW may not affect xenobiotic metabolism or oxidative stress in liver. However, intake of HRW may increase the efflux of xenobiotics or toxic substances from the liver into bile by enhancing the expression of p-glycoprotein and Mrp2 protein.

Nowadays, the imidazolium-based ionic liquids containing acetate counter-ions are attracting much attention as both highly selective absorbents of the acidic gases and CO₂ carriers in the supported ionic liquid membranes. In this regard, the investigation of the gas transport properties of such membranes may be appropriate for better understanding of various factors affecting the separation performance and the selection of the optimal operating conditions. In this work, we have tested CH₄, CO₂ and H₂S permeability across the SILM impregnated by 1-butyl-3-methylimidazolium acetate (bmim[ace]) with the following determination of the ideal selectivity in order to compare the facilitated transport membrane performance with the SILM that dissolves acidic gases physically, namely, containing 1-butyl-3-methylimidazolium tetrafluoroborate (bmim[BF₄]). Both SILMs have showed modest individual gases permeability and ideal selectivity of CO₂/CH₄ and H₂S/CH₄ separation that achieves values up to 15 and 32, respectively. The effect of the feed gas mixture composition on the permeability of acidic gases and permeselectivity of the gas pair was investigated. It turned out that the permeation behavior for the bmim[ace]-based SILM toward the binary CO₂/CH₄, H₂S/CH₄ and ternary CO₂/H₂S/CH₄ mixtures was featured with high acidic gases selectivity due to the relatively low methane penetration through the liquid phase saturated by acidic gases.

We have developed IrO_x/M-SnO₂ (M = Nb, Ta, and Sb) anode catalysts, IrO_x nanoparticles uniformly dispersed on M-SnO₂ supports with fused-aggregate structures, which make it possible to evolve oxygen efficiently, even with a reduced amount of noble metal (Ir) in proton exchange membrane water electrolysis. Polarization properties of IrO_x/M-SnO₂ catalysts for the oxygen evolution reaction (OER) were examined at 80 °C in both 0.1 M HClO₄ solution (half cell) and a single cell with a Nafion^® membrane (thickness = 50 μm). While all catalysts exhibited similar OER activities in the half cell, the cell potential (E_cell) of the single cell was found to decrease with the increasing apparent conductivities (σ_{app, catalyst}) of these catalysts: an E_cell of 1.61 V (voltage efficiency of 92%) at 1 A cm^-2 was achieved in a single cell by the use of an IrO_x/Sb-SnO₂ anode (highest σ_{app, catalyst}) with a low Ir-metal loading of 0.11 mg_Ir cm^-2 and Pt supported on graphitized carbon black (Pt/GCB) as the cathode, with 0.35 mg_Pt cm⁻². In addition to the reduction of the ohmic loss in the anode catalyst layer, the increased electronic conductivity contributed to decreasing the OER overpotential due to the effective utilization of the IrO_x nanocatalysts on the M-SnO₂ supports, which is an essential factor in improving the performance with low noble metal loadings.

The function of the endoplasmic reticulum (ER) can be impaired by the alternation of the extra- and intracellular environment such as disruption of calcium homeostasis, expression of mutated proteins and oxidative stress. In response to disruptions to ER homeostasis, eukaryotic cells activate canonical branches of signal transduction cascades, collectively termed the unfolded protein response (UPR). The UPR attempts to recover the protein folding capacity and avoid irreversible cellular damage. Additionally, the UPR has been shown to play unique physiological roles in the regulation of diverse cellular events, including cell differentiation and development and lipid biosynthesis. Recent studies have indicated that these important cellular events are also regulated by contact and communication among organelles. These reports suggest strong involvement among the UPR, organelle communication and regulation of cellular homeostasis. However, the precise mechanisms for the formation of contact sites and the regulation of its dynamics by UPR remain unresolved. In this review, we summarized the current understanding of how the UPR regulates morphological changes to the ER and the formation of contact sites between the ER and other organelles. We also review how UPR-dependent connections between the ER and other organelles affect cellular and physiological functions.

Gas-liquid reactors pose transfer difficulties due to diffusion effects. It is necessary to master the aeration and hydrodynamics of the medium to conduct the reaction well and get a good performance. For this purpose, a study in a submerged membrane bioreactor with a useful volume of 30L, consisting of a microfiltration membrane with an average pore size of 0.14 mm having an effective surface area of 0.2 m2 and a PVC cylindrical air diffuser of radius 4 cm has been studied. The saline tracing method associated with a conductimetric follow-up made it possible to determine the residence times and the mixing time in the reactor at 4 different points both in recirculation and in the absence of recirculation. Gas retention was measured by the manometric method. The experiments were carried out at different temperatures of 25 ° C, at 45 ° C, with a variable air flow rate of 0.5 to 16 mL / s and different solutions (osmosis water, ammonium formate solution, solution ammonium formate + salt, synthetic rubber effluent). The results show that the mixing time varies from one point to another and the recirculation of the mixture reduces the mixing time. One of the positions is limiting, the transfer is done most by diffusion with a mixing time of 115 min without circulation and 65 min with circulation. Gas retention increases with aeration rate and temperature. On the other hand, the more the medium becomes rich in organic substances, the more the gaseous retention decreases. The homogeneous fine-bubble regime is obtained for an air flow rate of between 3 and 10 mL / s of aeration. Beyond this flow rate, the regime becomes heterogeneous without a transition phase for ammonium formate and formate ammonium + salt solutions.