Increased Binding Capacity Research Articles

The First Report of The Growth Performance and Environmental Sustainability Effects of Dietary Insect Meal Application on The Jardine River Turtle (Emydura Subglobosa)

Abstract This study provides empirical data on the balancing, production, and quality of freshwater turtle diets containing partially defatted black soldier fly larvae Hermetia illucens meal (BSFL) as a fish meal (FM) replacement. A 70-day experiment was performed using 27 Jardine river turtle (Emydura subglobosa) juveniles (6 months old). Three dietary treatments were applied, namely, CON with 30% fish meal and no BSFL as the control, H75 with 22.5% fish meal and 7.5% BSFL, and H150 with 15% fish meal and 15% BSFL, with 3 replicates per treatment, 3 animals per replicate, and 9 specimens in total per treatment. Post-extrusion tests showed that feed technological parameters are dependent on the BSFL meal proportion in terms of the pellet length expansion rate, volume increase, and water binding capacity. The obtained experimental feeds were well accepted by the animals. During the entire experimental period, no turtle mortality, diet-related issues, or differences in shell development or growth performance were recorded among the treatments. However, the feed intake increased in comparison to CON when 7.5% BSFL meal was used (42.30 g vs. 50.40 g), and a lower feed conversion ratio was observed in the 15% BSFL treatment (1.51 vs. 1.38). For the first time, it was empirically proven that E. subglobosa can efficiently utilize BSFL meal for up to 15% of their diet. Moreover, the possibility of an increase in environmental sustainability during turtle husbandry due to a decrease in total marine resource use of 55.8% and a 57.4% decrease in the use of fish meal per kg of body weight gain were recorded.

Integrative chemical and multiomics analyses of tetracycline removal mechanisms in Pseudomonas sp. DX-21

Tetracycline (TC), widely found in various environments, poses significant risks to ecosystems and human health. While efficient biodegradation removes TC, the mechanisms underlying this process have not been elucidated. This study investigated the molecular mechanisms underlying TC biosorption and transfer within the extracellular polymeric substances (EPS) of strain DX-21 and its biodegradation process using fourier transform infrared spectroscopy, molecular docking, and multiomics. Under TC stress, DX-21 increased TC biosorption by secreting more extracellular polysaccharides and proteins, particularly the latter, mitigating toxicity. Moreover, specialized transporter proteins with increased binding capacity facilitated TC movement from the EPS to the cell membrane and within the cell. Transcriptomic and untargeted metabolomic analyses revealed that the presence of TC led to the differential expression of 306 genes and significant alterations in 37 metabolites. Notably, genes related to key enzymes, such as electron transport, peroxidase, and oxidoreductase, exhibited significant differential expression. DX-21 combated and degraded TC by regulating metabolism, altering cell membrane permeability, enhancing oxidative defense, and enhancing energy availability. Furthermore, integrative omics analyses indicated that DX-21 degrades TC via various enzymes, reallocating resources from other biosynthetic pathways. These results advance the understanding of the metabolic responses and regulatory mechanisms of DX-21 in response to TC.

Abstract 2732: NB203: A Novel therapeutic bispecific antibody targeting CEACAM1/VEGF for tumor immunity and angiogenesis modulation in the tumor microenvironment

Abstract The carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1) is detected on leukocytes, epithelia and endothelia, and involved in cell adhesion through homophilic or heterophilic binding. In various gastrointestinal cancers, NSCLC (non-small cell lung cancer), and melanoma, CEACAM1 is highly expressed on tumor cells and tumor stroma, with its levels increasing as the disease progresses. Notably, CEACAM1 has been found to dampen T and NK cell functions while promoting tumor angiogenesis and metastasis. In this study, we present NB203, a novel CEACAM1/VEGF bispecific antibody developed using Neologics CEACAM-plus, a bifunctional molecule engineering platform designed to target and modulate the tumor microenvironment (TME). NB203 effectively accumulated in the TME and hindered the homophilic or heterophilic interactions of CEACAM1. It demonstrated a significant enhancement in T cell activities and NK cell-mediated killing of CEACAM1-expressing tumor cells, and the inhibition of tumor cell migration and polyploidy through macrophages. More importantly, NB203 was specifically designed to impede tumor angiogenesis. It exhibited an increased binding capacity to both CEACAM1 and VEGF in pro-angiogenic TME, and undermined intratumoral vessel maturation by blocking VEGF and negatively regulating myeloid cell-dependent tumor angiogenesis. In summary, our preclinical data indicate that NB203 exhibits effective anti-tumor activity by mediating NK and cytotoxic T cell activities, improving the immune-suppressive TME, and suppressing tumor angiogenesis. The use of NB203 holds promise in overcoming resistance to anti-VEGF therapy and substantially improving the safety of therapeutic targeting of angiogenesis in cancer treatment. Citation Format: Jinyu Dong, Binbin Wang, Dong Wang, Jie Ni, Tingting Bu, Haojie Wang, Bishnu Nayak, Baiyang Wang, Xin Dong, Yu Zhang. NB203: A Novel therapeutic bispecific antibody targeting CEACAM1/VEGF for tumor immunity and angiogenesis modulation in the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2732.

Predicting Natural Evolution in the RBD Region of the Spike Glycoprotein of SARS-CoV-2 by Machine Learning.

Machine learning (ML) is a key focus in predicting protein mutations and aiding directed evolution. Research on potential virus variants is crucial for vaccine development. In this study, the machine learning software PyPEF was employed to conduct mutation analysis within the receptor-binding domain (RBD) of the Spike glycoprotein of SARS-CoV-2. Over 48,960,000 variants were predicted. Eight prospective variants that could surface in the future underwent modeling and molecular dynamics simulations. The study forecasts that the latest variant, ISOY2P5O1, may potentially emerge around 17 November 2023, with an approximate window of uncertainty of ±22 days. The ISOY8P5O2 variant displayed an increased binding capacity in the dry assay, with a total predicted binding energy of -110.306 kcal/mol. This represents an 8.25% enhancement in total binding energy compared to the original SARS-CoV-2 strain discovered in Wuhan (-101.892 kcal/mol). Reverse research confirmed the structural significance of mutation sites using ML models, particularly in the context of protein folding. The study validated regression methods (SVR, RF, and PLS) with different data structures. This study investigates the effectiveness of the "ML-Guided Design Correctly Predicts Combinatorial Effects Strategy" compared to the "ML-Guided Design Correctly Predicts Natural Evolution Prediction Strategy". To enhance machine learning, we created a timestamping algorithm and two auxiliary programs using advanced techniques to rapidly process extensive data, surpassing batch sequencing capabilities. This study not only advances machine learning in guiding protein evolution but also holds potential for forecasting future viruses and vaccine development.

Multifunctional graphitic carbon nitride/manganese dioxide/epoxy nanocomposite coating on steel for enhanced anticorrosion, flame retardant, mechanical, and hydrophobic properties

Benzidine (Bz) was used to modify the nanofiller, manganese dioxide (MnO2), and the resulting Bz/MnO2 was then encased in pure epoxy resin (EP) with graphitic carbon nitride (GCN). The effectiveness of epoxy-coated mild steel in protecting against varying concentrations of GCN/Bz-MnO2 was assessed through the use of electrochemical techniques in natural seawater. The flame retardancy capabilities of the epoxy coating were significantly improved by the incorporation of GCN/Bz-MnO2. The EP-GCN-Bz/MnO2 nanocomposites display peak heat release rate (PHRR) and total heat release (THR) values which are 79 % and 62 % lesser than that of pure EP, respectively. When compared to a pure epoxy coating, the results of salt spray tests indicated that the addition of GCN/Bz-MnO2 improved corrosion protection and decreased water absorption. Electrochemical impedance spectroscopy (EIS) studies showed that the epoxy-GCN/Bz-MnO2 coating still had an improved resistance of 6.40E10 Ω.cm2 even after 30 days of exposure to seawater. Scanning electrochemical microscopy (SECM) displayed lowest ferrous ion dissipation (1.0 I/nA) in the EP-GCN/Bz-MnO2 nanocomposite coated steel. Potentiodynamic polarization testing showed that EP-GCN-Bz/MnO2 had a lower current density (icorr: 0.03 µA/cm2) in comparison with other coatings. This is mainly because the Bz alteration of the MnO2 nanoparticles and their successful coordination with GCN sheets result in a noticeably more compact nanostructure that lowers aggregation and increases binding capacity. Surface-modified GCN was found to be improved in its breakdown products, leading to the production of a strong, inert nanolayered covering, according to Scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM/EDX) analysis. Water contact angle (WCA) of 160° demonstrates the exceptional water resistance of the recently developed EP-GCN/Bz-MnO2 coating. In terms of hardness and adhesion strength, Bz/MnO2 wrapped in GCN had good mechanical characteristics on the epoxy substrate. The improved adhesive strength (18.3 MPa) for mild steel coated with EP-GCN/Bz-MnO2 was attained prior to being immersed in seawater. The EP-GCN/Bz-MnO2 crystallizes form a robust, inert coating that prevents ions from accessing the specimen. The coating has increased adhesive strength as a result, and it can withstand deep immersions without losing its integrity. Hence, the EP-GCN/Bz-MnO2 nanocomposite might work as a useful coating component in the automotive industry.

Black soybean cooking water (aquasoya) powder as a novel clean‐label ingredient in plant‐based vegan patties

SummaryBlack soybean cooking water (BSCW), a by‐product of the soybean paste industry, is a novel foaming and emulsifying material. Its application, however, is limited to sauces and bakery goods. To expand its use, this research explored its binding and texturising abilities on a plant‐based patty. This vegan patty was developed including methylcellulose (MC) at 3% or BSCW powder at 3, 6 and 12%, and their functional and textural characteristics were analysed. Adding BSCW powder at 12% resulted in enhanced fluid retention parameters, namely, decreased cooking loss (9.98 ± 0.72%; control: 12.64 ± 0.36%) and increased binding capacity (86.18 ± 0.47%; control: 83.26 ± 1.94%). Texturally, BSCW powder enhanced the hardness and adhesiveness of the patties. Consequently, BSCW powder could improve the characteristics of a minced‐type vegan food similar to MC constituting an alternative plant‐based ingredient that is environmentally friendly due to being sourced from an upcycled material.

Starch oxidation and study of changing its properties for use as an adhesive component for the production of corrugated cardboard

It is known that for sizing papers and cardboards, starch is required, which has a lower viscosity compared to natural, increased stability and increased binding capacity. This is necessary so that the starch dispersions in the size press have a sufficiently high concentration, are process able and remain stable. The choice of an oxidizing agent and the conditions for the method of oxidizing corn starch are theoretically substantiated. The mechanism and regularities of the process of starch oxidation have been studied. Changes in the gel-forming and sorption properties of starch during its oxidation were revealed. The intervals of quantitative parameters of the components for the modification process are determined. The dependence of the surface tension of starch pastes on its concentration in solution has been obtained.

The effect of adjacent double-strand DNA on the G-triplex-ThT complex fluorescence intensity enhancement and its application in TNOS and Hg2+ detection

In this paper, we have found that adjacent double-strand DNA (dsDNA) can enhance the fluorescence intensity of the G-triplex (G31)-thioflavin T (ThT) complex. By connecting the newly formed dsDNA with the G31 sequence, terminator nopaline synthase (TNOS) gene and Hg2+ were detected. The intermolecular duplex hybridization (e.g., TNOS gene and its complementary DNA), or the intramolecular mismatched thymine (T)-Hg2+-T pairs induced the formation of dsDNA with planar morphology, which resulted in the increased binding capacity of adjacent G31 to ThT, as well as the enhanced fluorescence intensity of G31-ThT complex. A versatile ‘turn-on’ fluorescence scaffold was developed for discriminating transgenic and non-transgenic soybeans and detecting Hg2+ in lake water.

Epidemiological Impact of GII.17 Human Noroviruses Associated With Attachment to Enterocytes.

For the last 30 years, molecular surveys have shown that human norovirus (HuNoV), predominantly the GII.4 genotype, is one of the main causative agents of gastroenteritis. However, epidemiological surveys have revealed the worldwide emergence of GII.17 HuNoVs. Genetic analysis confirmed that GII.17 strains are distributed into three variants (i.e., Kawasaki 308, Kawasaki 323, and CS-E1). Here, virus-like particles (VLPs) were baculovirus-expressed from these variants to study putative interactions with HBGA. Qualitative analysis of the HBGA binding profile of each variant showed that the most recent and predominant GII.17 variant, Kawasaki 308, possesses a larger binding spectrum. The retrospective study of GII.17 strains documented before the emergence of the dominant Kawasaki 308 variant showed that the emergence of a new GII.17 variant could be related to an increased binding capacity toward HBGA. The use of duodenal histological sections confirmed that recognition of enterocytes involved HBGA for the three GII.17 variants. Finally, we observed that the relative affinity of recent GII.17 VLPs for HBGA remains lower than that of the GII.4-2012 variant. These observations suggest a model whereby a combination of virological factors, such as polymerase fidelity and increased affinity for HBGA, and immunological factors was responsible for the incomplete and non-persistent replacement of GII.4 by new GII.17 variants.

Bovine Serum Albumin as a Potential Carrier for the Protection of Bioactive Quercetin and Inhibition of Cu(II) Toxicity.

Considering the protective ability of proteins and the potential toxicity of free Cu(II), it was proposed herein that the co-presence of protein could play an important role in suppressing the toxicity of free Cu(II) to the stability of bioactive quercetin if a flavonoid-protein-Cu(II) complex could be formed. In this study, the interaction between quercetin (a major flavonoid in the human diet) and bovine serum albumin (BSA) was investigated in the absence and presence of free Cu(II). The results demonstrated that both quercetin and free Cu(II) had a strong ability to quench the intrinsic fluorescence of BSA through a static procedure (i.e., formation of a BSA-monoligand complex). Site marker competitive experiments illustrated that the binding of both quercetin and Cu(II) to BSA mainly took place in subdomain IIA. The quenching process of free Cu(II) with BSA was easily affected by quercetin, and the increased binding capacity possibly resulted from the generation of a ternary quercetin-BSA-Cu(II) complex. The stability and free radical scavenging activity of bioactive quercetin during incubation was promoted in the BSA-diligand complex relative to a quercetin-Cu(II) complex. A quercetin-Cu(II) system could generate reactive oxygen species such as hydrogen peroxide (H2O2) and hydroxyl radicals (•OH), which were significantly inhibited upon BSA binding. Consistently, the cytotoxicity of the quercetin-Cu(II) system to endothelial cells was decreased in the BSA-diligand complex, where the co-presence of BSA played an important role. These results suggest the possibility and advantage of developing albumin-based carriers for the protection of bioactive components and suppression of Cu(II) toxicity in their biomedical and nutritional applications.

Influence of metakaolin and limestone on chloride binding of slag activated by mixed magnesium oxide and sodium hydroxide

The layered double hydroxides (LDH) formed in alkali-activated slag (AAS), i.e. hydrotalcite and AFm phases, play a considerable role in its chloride binding behaviours. In addition to slag chemistry, the formation of LDH and chloride binding capacity of AAS are highly influenced by the activator composition and incorporation of other precursors. Considering their potential in enhancing LDH formation, this study investigates the influence of metakaolin and limestone incorporation on the phase assemblages, composition, and chloride binding capacity of AAS activated by mixed reactive magnesium oxide (MgO)-sodium hydroxide (NaOH). The results show that the chloride binding capacity of AAS is increased by up to 50% with 20% metakaolin or limestone substitution. However, different mechanisms for the binding enhancement are proposed for these two blended AAS systems. For metakaolin-blended AAS, the hydrotalcite formation is probably limited by the mobility of the dissolved Mg during MgO hydration and its interaction with dissolved Al from metakaolin; thus the increased chloride binding is primarily attributed to the sorption by C-A-S-H and formation of chloride-bearing chabazite. For limestone-blended AAS, carbonate-AFm phase is identified which enhances the formation of Friedel's salt and its polymorphs, thus showing increased binding capacity. These findings suggest that the enhanced chloride resistance of AAS materials could be achieved with the use of combinational raw precursors if appropriately designed.

Adsorption of enzymes with hydrolytic activity on polyethylene terephthalate

Polyethylene terephthalate (PET) degrading enzymes have recently obtained an increasing interest as a means to decompose plastic waste. Here, we have studied the binding of three PET hydrolases on a suspended PET powder under conditions of both enzyme- and substrate excess. A Langmuir isotherm described the binding process reasonably and revealed a prominent affinity for the PET substrate, with dissociation constants consistently below 150 nM. The saturated substrate coverage approximately corresponded to a monolayer on the PET surface for all three enzymes. No distinct contributions from specific ligand binding in the active site could be identified, which points towards adsorption predominantly driven by non-specific interactions in contrast to enzymes naturally evolved for the breakdown of insoluble polymers. However, we observed a correlation between the progression of enzymatic hydrolysis and increased binding capacity, probably due to surface modifications of the PET polymer over time. Our results provide functional insight, suggesting that rational design should target the specific ligand interaction in the active site rather than the already high, general adsorption capacity of these enzymes.

A review of concrete durability in marine environment

There are many concrete constructions in marine environments. Due to its salinity content, seawater is aggressive, both towards concrete and reinforcement. Concrete interacts with seawater which causes deterioration. The chemical attack occurs at least through 3 common mechanisms: sulfate-attack, carbonation, and chloride-induced corrosion of reinforcement. The chemical resistance of concrete is determined by the porosity and chemical composition of the concrete. Partly subsitution of cement with supplementary cementitious materials (SCM) known as an effective way to create the less permeable concrete. Concrete with fly ash as SCM has excellent potential to be applied to marine construction in Indonesia because the abundant availability of fly ash in Indonesia and the presence of aluminum species in fly ash provide an opportunity to increase binding capacity to chloride.

Von Willebrand factor with increased binding capacity is associated with reduced platelet aggregation but enhanced agglutination in COVID-19 patients: another COVID-19 paradox?

Patients with Coronavirus-associated disease-2019 (COVID-19) display alterations of the hemostatic system and the presence of a prothrombotic status frequently leading to vascular complications. However, the impact of COVID-19 on platelet activity, aggregation and agglutination still needs to be clarified. We measured total levels of von Willebrand factor (vWF) and vWF binding to the platelet glycoprotein (Gp) complex (GPIb-IX-V), in a cohort of COVID-19 patients admitted to the intensive care unit of our Institution. Moreover, we evaluated platelet aggregation in response to agonists (ADP, collagen, arachidonic acid) and platelet agglutination in response to ristocetin. We found that levels of vWF antigen and the active form of vWF binding to platelets (vWF:RCo), were markedly increased in these patients. These results were associated with higher agglutination rates induced by ristocetin, thereby indirectly indicating an increased capability of vWF to bind to platelets. Conversely, we found that platelet aggregation in response to both ADP and collagen was lower in COVID-19 patients compared to healthy volunteers. This study shows that COVID-19 is associated with increased vWF-induced platelet agglutination but reduced platelet responsivity to aggregation stimuli. Our findings have translational relevance since platelet adhesion to vWF may represent a marker to predict possible complications and better delineate therapeutic strategies in COVID-19 patients.Electronic Supplementary MaterialThe online version of this article (10.1007/s11239-020-02339-6) contains supplementary material, which is available to authorized users.

Abstract 6611: Functional characterization of a novel gene in the antigen presentation pathway allows specific immunotherapeutic targeting of glioblastoma cancer stem cells

Abstract Glioblastoma is the most prevalent and aggressive primary brain tumor in adults, and conventional treatment strategies have made minimal progress in prolonging overall survival. Cancer stem cells are critical for tumor initiation, growth, and maintenance, and are able to co-opt complex host mechanisms to evade immune recognition. Immune checkpoint inhibition has proven revolutionary for the treatment of several types of solid tumors, but has largely failed in attempts to improve outcomes in glioblastoma. Developing a deeper understanding of the functional immunomodulatory role of genes unique to glioma stem cells (GSCs) may help to elucidate this enigma, and inform more specific immunotherapeutic targeting. To better understand the ability of GSCs to promote immunosuppression and self-tolerance in the tumor niche, we interrogated RNA-seq data from GSCs and differentiated glioma cells (DGCs) and identified a novel gene in the antigen presentation pathway (APP). In vitro targeting of this gene increased sensitivity to CD8+ T cell killing and reduced expression of other T-cell inhibitory ligands in GSCs alone, decreasing tumor-mediated immunosuppression. Compared to DGCs, GSCs were found to be key promoters of immune self-tolerance and evasion, with unaltered GSCs exhibiting minimal peptide binding to MHC Class I and, therefore, decreased antigen presentation. Specific targeting of the identified APP gene led to a reversal of this interaction, with increased binding capacity and, correspondingly, tumor antigen recognition. In vivo administration of a specific APP gene targeting agent in human GSC xenograft mice displayed decreased tumor burden and a striking prolongation of overall survival compared to both controls and mice treated with known immune checkpoint inhibitors. High expression of the APP gene is associated with poor clinical prognosis, supporting the clinical relevance of this discovery. Thus, novel targeting of the antigen-presenting pathway may provide a therapeutic vulnerability unique to glioblastoma cancer stem cells. Citation Format: Reilly L. Kidwell, Xiqing Li, Jeremy N. Rich. Functional characterization of a novel gene in the antigen presentation pathway allows specific immunotherapeutic targeting of glioblastoma cancer stem cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6611.

The association of a single-nucleotide variant in the microRNA-146a with advanced colorectal cancer prognosis.

The aim of this study was to evaluate the association of single-nucleotide variant n.60G>C (rs2910164) of microRNA (miR)-146a, related to suppressing of BRCA1/2 DNA repair protein, with the risk and survival of colorectal cancer patients, as well as miR-146a and BRCA1/2 levels and miR binding efficiency. The genotypes were identified in 125 colorectal cancer patients and 276 controls using TaqMan polymerase chain reaction assay. The miR-146a and BRCA1/2 levels were assessed by quantitative-polymerase chain reaction protocols. Primary precursor of miR-146a containing G (wild-type) and C (variant) allele were cloned into pcDNA.3.3 vector and co-transfected in HT-29 colorectal cancer cell line. Luciferase reporter assay was performed to assess miR-146a binding to BRCA2 3'-untranslated region in HT-29. The differences between groups were calculated using chi-square or Fisher's exact test, logistic regression, and Mann-Whitney test. The prognostic impact of single-nucleotide variant genotypes on overall survival was evaluated by Kaplan-Meier estimate and Cox regression. The GC or CC genotypes prevalence was similar in patients and controls (50.4% vs 50.7%, p = 0.74). However, patients with tumors in advanced stage with miR-146a GG genotype had 2.41 more chance of dying than GC or CC genotypes. In addition, tumor tissues of patients with GG genotype presented higher miR-146a (p = 0.02) and lower BRCA1 (p = 0.01) and BRCA2 (p < 0.0001) levels when compared to those with GC or CC genotypes. In fact, pcDNA.3.3-miR-146a-G presented increased binding capacity to the 3'-untranslated region of BRCA2 (p = 0.001) compared to pcDNA.3.3-miR-146a-C. In addition, the G allele altered the binding affinity between miR-146a and its BRCA2 3'-untranslated region target (p < 0.001), thus enhancing suppression of BRCA2 expression. Our results suggest that single-nucleotide variant rs2910164 does not influence the colorectal cancer risk in Brazilian patients; however, the GG genotype could act as a factor of worse prognosis in patients with advanced disease due to suppression of BRCA1/2 modulated by miR-146a.

High-temperature tolerant enzyme based extraction of lignocellulose derived C5 and C6 sugars for the integrated production of a promising supplement for a diesel/jet-engine fuel

Development of a novel, integrated production of C8 ester butyl butyrate (BuB), a promising supplement for diesel/jet-engine fuels, from lignocellulose derived C5 and C6 sugars is attempted by means of smart and innovative process design. In the pathway, enzymatic hydrolysis of biomass with high solid loading is an important step which requires reduced amount of water supply and energy input. However, increased substrate loading lead to diffusion limitations of enzymes, on top of nonproductive absorption of enzymes. Addition of ß-glucosidase over cellulase is another reason to increase the cost of enzymes. Viscozyme can be considered as a troubleshooter in these cases where it can offset high substrate loading and increased monomeric sugar generation. In the present study, 17.61 g/L of glucose, along with 7.43 g/L of xylose, have been recovered with a viscozyme concentration of 54.4 g/L. Additionally, thermal deactivation of viscozyme has been investigated. The deactivation constant is found to be gradually increasing with escalated enzyme loading from 24.2 g/L to 54.4 g/L. In this study, it has been found that hydrolysis of biomass follows a first order pseudo-kinetics and the same is eventually used to determine the fractal kinetics of cellulose hydrolysis. Fractal kinetics showed that increment of enzyme concentration is associated with lower fractal dimensions and increased binding capacity of substrate and enzyme. Diffusion limitation of the present enzyme-substrate system has also been investigated. It is observed that faster diffusion, along with increased resistances, can be achieved with increased enzyme concentration.

The identification and function of a Netrin-1 mutation in a pedigree with premature atherosclerosis

Background and aimsNeuroimmune guidance cues have been shown to play a role in atherosclerosis, but their exact role in human pathophysiology is largely unknown. In the current study, we investigated the role of a c.1769G > T variant in Netrin-1 in (premature) atherosclerosis. MethodsTo determine the effect of the genetic variation, purified Netrin-1, either wild type (wtNetrin-1) or the patient observed variation (mutNetrin-1), was used for migration, adhesion, endothelial barrier function and bindings assays. Expression of adhesion molecules and transcription proteins was analyzed by RT-PCR, Western blot or ELISA. To further delineate how mutNetrin-1 mediates its effect on cell migration, lenti-viral knockdown of UNC5B or DCC was used. ResultsBindings assays revealed a decreased binding capacity of mutNetrin-1 to the receptors UNC5B, DCC and β3-integrin and an increased binding capacity to neogenin, heparin and heparan sulfate compared to wtNetrin-1. Exposure of endothelial cells to mutNetrin-1 resulted in enhanced monocyte adhesion and expression of IL-6, CCL2 and ICAM-1 compared to wtNetrin-1. In addition, mutNetrin-1 lacks the inhibitory effect on the NF-κB pathway that is observed for wtNetrin-1. Moreover, the presence of mutNetrin-1 diminished migration of macrophages and smooth muscle cells. Importantly, UNC5B or DCC specific knockdown showed that mutNetrin-1 is unable to act through DCC resulting in enhanced inhibition of migration. ConclusionsOur data demonstrates that mutNetrin-1 fails to exert anti-inflammatory effects on endothelial cells and more strongly blocks macrophage migration compared to wtNetrin-1, suggesting that the carriers of this genetic molecular variant may well be at risk for premature atherosclerosis.

Stringent Selection of Knobby Plasmodium falciparum-Infected Erythrocytes during Cytoadhesion at Febrile Temperature.

Changes in the erythrocyte membrane induced by Plasmodium falciparum invasion allow cytoadhesion of infected erythrocytes (IEs) to the host endothelium, which can lead to severe complications. Binding to endothelial cell receptors (ECRs) is mainly mediated by members of the P. falciparum erythrocyte membrane protein 1 (PfEMP1) family, encoded by var genes. Malaria infection causes several common symptoms, with fever being the most apparent. In this study, the effects of febrile conditions on cytoadhesion of predominately knobless erythrocytes infected with the laboratory isolate IT4 to chondroitin-4-sulfate A (CSA), intercellular adhesion molecule 1 (ICAM-1), and CD36 were investigated. IEs enriched for binding to CSA at 40 °C exhibited significantly increased binding capacity relative to parasites enriched at 37 °C. This interaction was due to increased var2csa expression and trafficking of the corresponding PfEMP1 to the IE surface as well as to a selection of knobby IEs. Furthermore, the enrichment of IEs to ICAM-1 at 40 °C also led to selection of knobby IEs over knobless IEs, whereas enrichment on CD36 did not lead to a selection. In summary, these findings demonstrate that knobs are crucial for parasitic survival in the host, especially during fever episodes, and thus, that selection pressure on the formation of knobs could be controlled by the host.

A Review on D 614G Mutation with Bangladesh Scenario

With the COVID-19 pandemic, disparities between the infection rate and death rate in different countries become a major concern. In some countries, lower mortality rate compared to others can be explained by better testing capacity and intensive care facilities. Complete SARS-CoV-2 genome sequences from different countries of the world are continually submitted to Global Initiative for Sharing All Influenza Data using Next Generation Sequencing method. A SARS-CoV-2 variant with a D 614G Mutation in the spike (S) protein has become the most dominant form in the global pandemic. There are a number of ongoing studies trying to relate this mutation with the infectivity, mortality, transmissibility of the virus and its impact on vaccine development. This review aims to accumulate the major findings from some of these studies and focus its future implication. Some studies suggested D 614G strain has increased binding capacity, it affects more cells at a faster rate, so has a high transmissibility. Patients infected with this strain were found with high viral load. But still now there is no such evidence that this strain produces more severe disease as well as increased mortality. The structural change of spike protein produced by D 614G mutation was minor and did not hamper the vaccine efficacy. Some studies showed antibodies produced against D614 strain can neutralize G614 strain and vice versa. Whenever a mutation occurs in spike protein there are always chances of affecting the infectivity, transmissibility, vaccine efficacy. Therefore, more studies are required to find out the overall effect of D 614G mutation.