Acidic Region Research Articles

Utilizing the pH-Shift Method for Isolation and Nutritional Characterization of Mantis Shrimp (Oratosquilla nepa) Protein: A Strategy for Developing Value-Added Ingredients.

This study focused on the production of protein isolates from mantis shrimp (MS). The pH-shift method was investigated to understand its impact on the protein yield, quality, and properties of the produced isolates. The first step was determining how the pH affected the protein solubility profile, zeta potential, and brown discoloration. The pH-shift process was then established based on the maximum and minimum protein solubilization. The solubilization pH had a significant impact on the mass yield and color of the produced protein, with a pH of 1.0 producing the maximum mass in the acidic region, whereas a maximum was found at a pH of 12.0 in the alkaline region (p < 0.05). Both approaches yielded mantis shrimp protein isolates (MPIs) with precipitation at a pH of 4.0 and a mass yield of around 25% (dw). The TCA-soluble peptide and TBARS levels were significantly lower in the MPI samples compared to MS raw material (p < 0.05). The MPIs maintained essential amino acid index (EAAI) values greater than 90%, indicating a high protein quality, and the pH-shift procedure had no negative impact on the protein quality, as indicated by comparable EAAI values between the mantis shrimp protein isolate extract acid (MPI-Ac), mantis shrimp protein isolate extract alkaline (MPI-Al), and MS raw material. Overall, the pH-shift approach effectively produced protein isolates with favorable quality and nutritional attributes.

THE ROLE OF LIMING IN COPPER ADSORPTION OF ACIDIC SOILS

Investigate the impact of varying rates of liming on the availability and sorption properties of copper in a range of acid soils with differing physico-chemical futures. Soil samples with varying acidity levels were collected from different locations in the acidic Black Sea Region of Türkiye. Out of the nine soil samples collected, four were obtained from regions where hazelnuts are grown, while the other five were obtained from areas dedicated to tea growth. The sorption properties of soils limed at 0%, 50%, and 100% of lime requirement and incubated for 12 months, then the Langmuir and Freundlich adsorption isotherms were determined by batch sorption technique. The results showed that increasing the soil pH resulted in an increase in Cu adsorption. Experimental results of Cu adsorption were explained with high coefficients of determination (93.44-98.98%) of the Langmuir model. In the three experimental groups with full, half, and no lime application, pH increased significantly to 3.95, 4.46, and 5.89, respectively, while the maximum adsorption obtained from Langmuir adsorption curves, Qmax1 1553, Qmax2 2205, and Qmax3 3902 mg kg-1, respectively. While the maximum adsorption in the tea plantation areas varied between 1491 and 1514 mg kg-1, the maximum adsorption in hazelnut cultivation areas varied between 777 and 3333 mg kg-1. These findings suggest that liming or other methods to increase soil pH could be effective soil management in reducing Cu mobility in acidic soils.

The first structure of human Golm1 coiled coil domain reveals an unexpected tetramer and highlights its structural diversity

Golgi membrane protein 1 (Golm1), a transmembrane protein with diverse subcellular localizations, has garnered significant attention in recent years due to its strong association with the development and progression of liver diseases and numerous cancers. Interestingly, although Golm1 is a membrane protein, the C-terminal of Golm1, which contains a coiled coil domain and a flexible acid region, can also be detected in the plasma of patients with various liver diseases. Notably, the coiled coil domain of serum Golm1 is postulated to play a pivotal role in physiological and pathological functions. However, little is currently known about the structure of this coiled coil domain and the full-length protein, which may limit our understanding of Golm1. Therefore, this study aims to address this gap in knowledge and reports the first crystal structure of the coiled coil domain of Golm1 at a resolution of 2.28 Å. Meanwhile, we have also confirmed that the Golm1 coiled coil domain in solution can form tetramer. Our results reveal that Golm1 can form a novel tetrameric structure that differs from the previous reported dimeric structure Golm1 could assemble, which may provide novel insights into the diversity of physiological functions and pathological roles.

Albumin incorporation into recognising layer of HER2-specific magnetic nanoparticles as a tool for optimal targeting of the acidic tumor microenvironment

Cancer is unquestionably a global healthcare challenge, spurring the exporation of novel treatment approaches. In recent years, nanomaterials have garnered significant interest with the greatest hopes for targeted nanoformulations due to their cell-specific delivery, improved therapeutic efficacy, and reduced systemic toxicity for the organism. The problem of successful clinical translation of nanoparticles may be related to the fact that most in vitro tests are performed at pH values of normal cells and tissues, ranging from 7.2 to 7.4. The extracellular pH values of tumors are characterized by a shift to a more acidic region in the range of 5.6–7.0 and represent a crucial target for enhancing nanoparticle delivery to cancer cells. Here we show the method of non-active protein incorporation into the surface of HER2-targeted nanoparticles to achieve optimal cellular uptake within the pH range of the tumor microenvironment. The method efficacy was confirmed in vitro and in vivo showing the maximum binding of nanoparticles to cells at a pH value 6.4. Namely, fluorescent magnetic nanoparticles, modified with HER2-recognising affibody ZHER2:342, with proven specificity in terms of HER2 recognition (with 62-fold higher cellular uptake compared to control nanoparticles) were designed for targeting cancer cells at slightly acidic pH values. The stabilizing protein, namely, bovine serum albumin, one of the major blood components with widespread availability and biocompatibility, was used for the decoration of the nanoparticle surface to alter the pH response of the targeting magnetic conjugates. The optimally designed nanoparticles showed a bell-shaped dependency of interaction with cancer cells in the pH range of 5.6–8.0 with maximum cellular uptake at pH value 6.4 close to that of the tumor microenvironment. In vivo experiments revealed that after i.v. administration, BSA-decorated nanoparticles exhibited 2 times higher accumulation in tumors compared to magnetic nanoparticles modified with affibody only. Thus, we demonstrated a valid method for enhancing the specificity of targeted nanoparticle delivery to cancer cells without changing the functional components of nanoparticles.

Molecular and functional characterization of ILYS-5, a major invertebrate lysozyme of Caenorhabditis elegans

To overcome bacterial invasion and infection, animals have evolved various antimicrobial effectors such as antimicrobial peptides and lysozymes. Although C. elegans is exposed to a variety of microbes due to its bacterivorous lifestyle, previous work on the components of its immune system mainly based on the description of transcriptional changes during bacterial challenges. Very few effector components of its immune system have been characterized so far. To investigate the role of lysozymes in terms of antibacterial defense and digestion, we studied a member of the widely neglected family of C. elegans invertebrate lysozymes (ILYS). We focused on the so far virtually undescribed ILYS-5, which we purified from protein extracts of C. elegans tracing its peptidoglycan-degrading activity and localized the tissue expression of the gene in vivo using a translational reporter construct. We recombinantly synthesized ILYS-5 and determined the physicochemical activity optimum and the antibacterial spectrum of a lysozyme from C. elegans for the first time. With an activity optimum at low ionic strength (≤100 mM) and at acidic pH (≤ pH 4.0), ILYS-5 is likely to be involved in killing and digestion of bacteria within acidified phagolysosomes and acidic regions of the gut, presumably secreted by lysosome-like vesicles. This notion is supported by potent activity against various live Gram-positive and Gram-negative bacteria. Notably, members of the natural associated microbiome of C. elegans are substantially less susceptible to ILYS-5. Ablation of the ilys-5 gene resulted in reduction of lifespan and fertility when cultured on the standard food bacterium Escherichia coli OP50, whereas exposure of the ilys-5 knock-out mutant to the host-associated bacterium Pseudomonas lurida MYb11 did not have a clear effect. These findings indicate a role of ILYS-5 in immunity and nutrition and a co-evolved adaptation of host and bacteria to the mutualistic nature of their interaction.

Molecular insights into TP53 mutation (p. Arg267Trp) and its connection to Choroid Plexus Carcinomas and Li-Fraumeni Syndrome.

Choroid plexus carcinomas (CPCs) are rare malignant tumors primarily affecting pediatric patients and often co-occur with Li-Fraumeni Syndrome (LFS), an inherited predisposition to early-onset malignancies in multiple organ systems. LFS is closely linked to TP53 mutations, with germline TP53 gene mutations present in approximately 75% of Li-Fraumeni syndrome families and 25% of Li-Fraumeni-like syndrome families. Individuals with TP53 mutations also have an elevated probability of carrying mutations in BRCA1 and BRCA2 genes. To investigate the structural and functional implications of the TP53: 799C > T, p. (Arg267Trp) missense mutation, initially identified in a Saudi family, and understand its impact on TP53 functionality and related intermolecular interactions. Computational analyses were conducted to examine the structural modifications resulting from the TP53: 799C > T, p. (Arg267Trp) mutation. These analyses focused on the mutation's impact on hydrogen bonding, ionic interactions, and the specific interaction with Cell Cycle and Apoptosis Regulator 2 (CCAR2), as annotated in UniProt. The study revealed that the native Arg267 residue is critical for a salt bridge interaction with glutamic acid at position 258. The mutation-induced charge alteration has the potential to disrupt this ionic bonding. Additionally, the mutation is located within an amino acid region crucial for interaction with CCAR2. The altered properties of the amino acid within this domain may affect its functionality and disrupt this interaction, thereby impacting the regulation of catalytic enzyme activity. Our findings highlight the intricate intermolecular interactions governing TP53 functionality. The TP53: 799C > T, p. (Arg267Trp) mutation causes structural modifications that potentially disrupt critical ionic bonds and protein interactions, offering valuable insights for the development of targeted mutants with distinct functional attributes. These insights could inform therapeutic strategies for conditions associated with TP53 mutations.

HcN57, A Novel Unusual Acidic Silk-Like Matrix Protein from Hyriopsis cumingii, Participates in Framework Construction and Nacre Nucleation During Nacreous Layer Formation.

In the classic molecular model of nacreous layer formation, unusual acidic matrix proteins rich in aspartic acid (Asp) residues are essential for nacre nucleation due to their great affinity for binding calcium. However, the acidic matrix proteins discovered in the nacreous layer so far have been weakly acidic with a high proportion of glutamate. In the present study, several silk-like matrix proteins, including the novel matrix protein HcN57, were identified in the ethylenediaminetetraacetic acid-soluble extracts of the nacreous layer of Hyriopsis cumingii. HcN57 is a highly repetitive protein that consists of a high proportion of alanine (Ala, 34.4%), glycine (Gly, 22.5%), and serine (Ser, 11.4%). It forms poly Ala blocks, GlynX repeats, an Ala-Gly repeat, and a Ser-Ala-rich region, exhibiting significant similarity to silk proteins found in spider species. The expression of HcN57 was specifically located in the dorsal epithelial cells of the mantle pallium and mantle center. Notably, expression of HcN57 was relatively high during nacreous layer regeneration and pearl nacre deposition, suggesting HcN57 is a silk matrix protein in the nacreous layer. Importantly, HcN57 also contains a certain content of Asp residues, making it an unusual acidic matrix protein present in the nacreous layer. These Asp residues are mainly distributed in three large hydrophilic acidic regions, which showed inhibitory activity against aragonite deposition and morphological regulation of calcite in vitro. Moreover, HcN57-dsRNA injection resulted in failure of nacre nucleation in vivo. Taken together, our results show that HcN57 is a bifunctional silk protein with poly Ala blocks and Gly-rich regions that serve as space fillers within the chitinous framework to prevent crystallization at unnecessary nucleation sites and Asp-rich regions that create a calcium ion supersaturated microenvironment for nucleation in the center of nacre tablets. These observations contribute to a better understanding of the mechanism by which silk proteins regulate framework construction and nacre nucleation during nacreous layer formation.

In-situ synthesis of bimetallic Co/Zn-ZIF on the graphite-felt for electrocatalytic degradation of pharmaceutical pollutants

In the electro-Fenton process, one of the important challenges is to adjust the pH of the environment in the acidic region so that the system's performance is not affected by side reactions. In this case, it will be necessary to neutralize the solution after the process. One of the solutions is to use a heterogeneous catalyst to widen the pH range. However, recovering the used catalyst itself can be another challenge. Modifying the electrode using electroactive materials can answer both of these challenges. This work modified the bare graphite-felt (GF) electrode using zeolitic imidazolate frameworks (ZIF)-derived carbon material doped with Co and Zn through a rapid, room-temperature, and water-based synthesis method. Various analyses, including XRD, SEM, XPS, and LSV, confirmed the proper modification of bare GF carbon fibers. The modified Co/Zn-N-C@GF electrode showed good performance in removing rifampicin (RFP) during the electro-Fenton process, so about 90 % of the pollutant was removed within 30 min without adding a homo-/heterogeneous catalyst. Optimum conditions for this process were 1.0 V for applied voltage, 40 mg/L for initial concentration of RFP, and 5.6 for pH. Also, LC-MS analysis was performed to confirm the degradation of RFP and investigate its degradation pathway.

JEV infection leads to dysfunction of lysosome by downregulating the expression of LAMP1 and LAMP2

Japanese Encephalitis Virus (JEV), the predominant cause of viral encephalitis in many Asian countries, affects approximately 68,000 people annually. Lysosomes are dynamic structures that regulate cellular metabolism by mediating lysosomal biogenesis and autophagy. Here, we showed that lysosome-associated membrane protein 1 (LAMP1) and LAMP2 were downregulated in cells after JEV infection, resulting in a decrease in the quantity of acidified lysosomes and impaired lysosomal catabolism. What’s more, JEV nonstructural protein 4B plays key roles in the reduction of LAMP1/2 via the autophagy-lysosome pathway. JEV NS4B also promoted abnormal aggregation of SLA-DR, an important component of the swine MHC-II molecule family involved in antigen presentation and CD4+ cell activation initiation. Mechanistically, NS4B localized to the ER during JEV infection and interacted with GRP78, leading to the activation of ER stress-mediated autophagy. The 131–204 amino acid (aa) region of NS4B is essential for autophagy induction and LAMP1/2 reduction. In summary, our findings reveal a novel pathway by which JEV induces autophagy and disrupts lysosomal function.

Spectroscopic detection of chromatin uncoiling and chromosome alignment in neuronal-like cells under exposure to low intensity magnetic fields

In this manuscript we reported the results of our studies concerning the response of vibration bands in deoxyribonucleic acid region in neuronal-like cells under exposure to static and 50 Hz electromagnetic fields using Fourier Transform Infrared spectroscopy. This technique was chosen because it is able to detect alteration of vibration bands of chemical compounds even induced by small stress agents. The aim of this study was to investigate the response of chromatin and chromosome to low intensity electromagnetic fields at values similar to manmade electromagnetic fields. The phosphate bands of asymmetric and symmetric stretching mode (representative of deoxyribonucleic acid spectral region) were observed to decrease after 3 h exposure at 1 mT (both static and 50 Hz magnetic field). Prolonging time exposure or increasing the intensity of applied magnetic field induced a low increasing in intensity of these vibration bands. This finding can be explaining assuming that chromatin uncoiling occurred at low intensity of the field and that increasing time exposure or magnetic field intensity caused an increasing of the torque on chromosome inducing an alignment along the direction of the field.

STUDYING THE COMPLEXATION OF V(V) WITH 3-[2-HYDROXY-3- SULFO-5-NITROPHENYLAZO]-PENTADIONE-2,4 IN THE PRESENCE OF A THIRD COMPONENT

In the present study, we investigated the formation of the complex vanadium c 3-[2-hydroxy-3- sulfo-5-nitrophenylazo]-pentadione-2,4 (R) in the presence of cationic surfactants (CPAS) including cetylpyridinium chloride (CPCl), cetylpyridinium bromide (CPBr), and cetyltrimethylammonium bromide (CTMABr). The study on the pH dependence of complexation revealed that the release of the VR complex occurred at pH=4 with a maximum absorption at λ=425 nm, while the reagent exhibited a maximum absorption at 383 nm. In the presence of the third component, three-component compounds VR-СPBr, VR-CPBr, and VR-CTMABr were formed. The absorption maxima of the mixed-ligand complexes V(V) were bathochromic relative to the absorption maxima of the binary complex, with λmax values of 437 nm, 449 nm, and 454 nm, respectively. The optimal pH for complexation was found to be in the acidic region at pH 4.0, 3.0, and 3.0, respectively. The impact of the third component and reagent concentrations on complex formation was also investigated, with the maximum yield of VR complex was achieved at a concentration of 8·10-5 M R, VR-CPCl at 8·10-5 M R and 8•10-4 M CPCl, VR-CPBr at 8•10-5 M R and 8•10-4 M CPBr, and VR-CTMABr at 8•10-5 M R and 8•10-4 M CTMABr. The optimal ratio of reacting components was determined to be 1:2 and 1:2:2, respectively. The compliance with Beer's law was confirmed, and molar absorption coefficients were calculated from saturation curves. The developed method was applied to the determination of vanadium (V) in the waters of the Akstafa rivers in the Kazakh region of the Azerbaijan Republic. Keywords: complexation of vanadium(V), cationic surfactants, cetylpyridinium chloride, cetylpyridinium bromide, cetyltrimethylammonium bromide.

An N-terminal acidic β-sheet domain is responsible for the metal-accumulation properties of amyloid-β protofibrils: a molecular dynamics study

The influence of metal ions on the structure of amyloid-β\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\beta $$\\end{document} (Aβ) protofibril models was studied through molecular dynamics to explore the molecular mechanisms underlying metal-induced Aβ aggregation relevant in Alzheimer’s disease (AD). The models included 36-, 48-, and 188-mers of the Aβ42 sequence and two disease-modifying variants. Primary structural effects were observed at the N-terminal domain, as it became susceptible to the presence of cations. Specially when β-sheets predominate, this motif orients N-terminal acidic residues toward one single face of the β-sheet, resulting in the formation of an acidic region that attracts cations from the media and promotes the folding of the N-terminal region, with implications in amyloid aggregation. The molecular phenotype of the protofibril models based on Aβ variants shows that the AD-causative D7N mutation promotes the formation of N-terminal β-sheets and accumulates more Zn2+, in contrast to the non-amyloidogenic rodent sequence that hinders the β-sheets and is more selective for Na+ over Zn2+ cations. It is proposed that forming an acidic β-sheet domain and accumulating cations is a plausible molecular mechanism connecting the elevated affinity and concentration of metals in Aβ fibrils to their high content of β-sheet structure at the N-terminal sequence.Graphic abstract

Pathological investigation and morpho-molecular characterisation of Lasiodiplodia theobromae causing fruit rot diseases on Hydnocarpus longipedunculatus – A novel host record from India

Hydnocarpus longipedunculatus, a medically significant tree species, is threatened and has a narrow distribution, being endemic to the Southern Western Ghats of India. A case of fruit rot disease was observed in a solitary tree of H. longipedunculatus in the Kulamavu forest areas of Kerala, India. The causative fungus, Lasiodiplodia theobromae, was isolated and identified through a combination of morpho-cultural characteristics and molecular sequence analysis involving the internal transcribed spacer (ITS), partial translation elongation factor-1-α (TEF1-α), partial β-tubulin (TUB2), and large subunit ribosomal ribonucleic acid (LSU) regions. Additionally, concatenated multigene (ITS-TEF1-α-TUB2) phylogenetic analysis was done. Pathogenicity tests were conducted in vitro using mycelial disc methods and confirmed the pathogenic nature of the fungus through re-isolation and morphocultural analysis, satisfying Koch's postulates. Previous studies neither have reported H. longipedunculatus as a host for L. theobromae, nor has any genus within Hydnocarpus. Therefore, this represents the first documented instance of H. longipedunculatus as a novel host record for L. theobromae from India.

Challenging of ECMO application in pediatric restrictive cardiomyopathy: case report of a novel TNNI3 variant

BackgroundRestrictive cardiomyopathy (RCM) represents a rare cardiovascular disorder stemming from filament-associated genes. Nonetheless, treating RCM presents considerable challenges, particularly concerning device implantation and mechanical support. Furthermore, elucidating the molecular function of specific variants holds promise in benefiting patients and enhancing prognosis, given the significant heterogeneity among RCM variants.Case presentationThe proband, an eight-year-old female, was admitted to our hospital post cardiopulmonary resuscitation due to sudden cardiac arrest. Echocardiography revealed bilateral atrial enlargement. Whole-exome sequencing uncovered a novel heterozygous mutation (c.509G&amp;gt;A, p.R170Q) in TNNI3. Evaluation using the MutationTaster application deemed c.509G&amp;gt;A pathogenic (probability = 0.99). Following clinical manifestations, imaging assessments, and genetic screening, the proband received an RCM diagnosis. ECMO was recommended along with continuous renal replacement therapy. However, persistent atrial flutter ensued post-ECMO withdrawal. Attempts to restore cardiac rhythm with cardioversion, metoprolol, and amiodarone proved futile. Subsequent heart failure led to the patient's demise due to cardiac shock. Based on crystal protein structural analysis, we observed that cTnI-R170Q and R170W exerted similar impacts on protein structural stability and formation. However, both differed significantly from cTnI-R170G, primarily influencing amino acid regions 32–79 and 129–149, involved in TnC and actin binding. Therefore, cTnI-R170Q was revealed to induce RCM via the same molecular mechanism as cTnI-R170W.ConclusionManaging RCM remains a critical challenge. This study underscores the discouragement of device implantations for cardiac pump functional support in RCM, particularly for non-short-term scheduled HTx. Additionally, considering catheter ablation for atrial fibrosis-induced AFs is recommended. Mechanistically, cTnI-R170Q primarily diminishes troponin-actin interactions and destabilizes thin filaments.

Receptor-mediated cargo hitchhiking on bulk autophagy

While the molecular mechanism of autophagy is well studied, the cargoes delivered by autophagy remain incompletely characterized. To examine the selectivity of autophagy cargo, we conducted proteomics on isolated yeast autophagic bodies, which are intermediate structures in the autophagy process. We identify a protein, Hab1, that is highly preferentially delivered to vacuoles. The N-terminal 42 amino acid region of Hab1 contains an amphipathic helix and an Atg8-family interacting motif, both of which are necessary and sufficient for the preferential delivery of Hab1 by autophagy. We find that fusion of this region with a cytosolic protein results in preferential delivery of this protein to the vacuole. Furthermore, attachment of this region to an organelle allows for autophagic delivery in a manner independent of canonical autophagy receptor or scaffold proteins. We propose a novel mode of selective autophagy in which a receptor, in this case Hab1, binds directly to forming isolation membranes during bulk autophagy.

Production and characterization of novel monoclonal antibodies against pathological human TDP-43 proteins.

The RNA/DNA-binding protein TDP-43 plays a pivotal role in the ubiquitinated inclusions characteristic of TDP-43 proteinopathies, including most cases of frontotemporal lobar degeneration (FTLD-TDP) and Alzheimer disease (AD). To understand the mechanisms of pathological TDP-43 processing and identify potential biomarkers, we generated novel phosphorylation-independent monoclonal antibodies (MAbs) using bacteria-expressed human full-length recombinant TDP-43. Remarkably, we identified a distinctive MAb, No. 9, targeting an epitope in amino acid (aa) region 311-360 of the C-terminus. This antibody showed preferential reactivity for pathological TDP-43 inclusions, with only mild reactivity for normal nuclear TDP-43. MAb No. 9 revealed more pathology in FTLD-TDP type A and type B brains and in AD brains compared to the commercial p409/410 MAb. Using synthetic phosphorylated peptides, we also obtained MAbs targeting the p409/410 epitope. Interestingly, MAb No. 14 was found to reveal additional pathology in AD compared to the commercial p409/410 MAb, specifically, TDP-43-immunopositive deposits with amyloid plaques in AD brains. These unique immunopositivities observed with MAbs No. 9 and No. 14 are likely attributed to their conformation-dependent binding to TDP-43 inclusions. We expect that this novel set of MAbs will prove valuable as tools for future patient-oriented investigations into TDP-43 proteinopathies.

Abstract 3081: Competitive Inhibition Of Tlt-1/fibrinogen Molecular Interaction Disrupts Platelet Function In Ex Vivo Platelet Assays

Background: Platelets facilitate thrombosis, hemostasis, and immunity through their interaction with fibrinogen. Fibrinogen is the ligand for the platelet receptors αIIbβ3 and T REM- L ike T ranscript-1 (TLT-1), TLT-1 is an Ig-domain containing receptor released from platelet α-granules upon platelet activation. Unlike αIIbβ3, little is known about the TLT-1/fibrinogen interaction. Aims: Delineate the TLT-1/fibrinogen molecular interaction and determine if, peptides representing this interaction can regulate platelet function and inflammatory hemostasis. Methods: Octet Qke Bio-layer Interferometry and atomic force microscopy (AFM) were used to evaluate soluble TLT-1 binding to fibrinogen. Immunoprecipitation and a peptide array were used to determine which fibrinogen peptides bound TLT-1. Flow cytometry using FITC-labelled fibrinogen binding to activated human platelets and human platelet aggregometry in the presence of these peptides were used in competitive inhibition assays. Additionally, competitive ELISAs using peptides representing regions the TLT-1 extracellular domain incubated with soluble TLT-1 in a fibrinogen coated wells were used to identify the region(s) on TLT-1 interaction with fibrinogen. Results: We obtained an equilibrium dissociation constant (KD) of 3.02 ± 0.20 nM for the TLT-1/Fibrinogen interaction. AFM suggests that sTLT-1 mediates fibrinogen polymerization. Competitive inhibition assays suggest complementary determining loop-1 and LP-17 (a 17 amino acid region of TLT-1) mediate TLT-1’s interaction to fibrinogen. The 3D modeling of our identified peptides show that they form a binding complex on the B module of the D-domain rather than a single binding site. These peptides competitively reduce platelet aggregation in aggregation assays and decrease fibrinogen binding to platelets in flow cytometry assays compared to controls. Cell binding and clotting assays are underway and the current status of projected will be reported here. Conclusions: TLT-1 uses LP17 and CDR-1 to interact with the newly identified TLT-1 binding complex on fibrinogen. This TLT-1 binding surface may represent a targetable intervention to regulate thrombosis and inflammation.

Effect of Various Nano PGR on Growth and Productivity of wheat (Triticum aestivum Var. Shriram Super 252)

Nano PGR is a multifaceted field and has vast application in all spheres of life. But in agriculture the application and utility islimited. Use of nano PGR can assist to tackle these issues to some extent. by bieng more efficient and have longer lasting nutrient release and a higher water holding capacity reducing soil erosion, It also release multiple types of nutrient simultaneously. These nano particles are having high reactivity, better catalytic surface, rapid chemical reaction, rapidly dispersible and adsorb abundant water. The present study was conducted in the agricultural experimental farm of Calcutta University, Baruipur, 24 Parganas (south), West Bengal during winter season of 2016 and 2017 to study the “effect of various nano PGR on growth and productivity of wheat (Shriram super 252)” being conducted under slightly acidic lower gangetic alluvial region of West Bengal. The experiment was laid out in a Randomized Complete Block Design with three replications. The treatments of the experiment was T1 was control ; T2 = NPK @ 100:50:50 and foliar spray of green Nano PGR @ 0.5ml L-1; T3 = NPK @ 75:30:30 and foliar spray of Yellow Nano PGR @ 0.5ml L-1; T4 = NPK @ 75:30:30 and foliar spray of Red Nano PGR @0.5ml L-1; T5 = NPK @ 75:30:30 and foliar spray of Green Nano PGR @ 1ml L-1; T6 = NPK @ 75:30:30 foliar spray of Yellow Nano PGR @ 1ml L-1; T7 = NPK @ 75:30:30 and foliar spray of red Nano PGR @ 1ml L-1; T8 = NPK @ 75:30:30 and foliar spray of Green Nano PGR @ 1.5ml L-1; T9 = NPK @ 75:30:30 and foliar spray of Yellow Nano PGR @ 1.5ml L-1 ; T10 = NPK @ 75:30:30 and foliar spray of Red Nano PGR @ 1.5ml L-1 .The recommended dose of inorganic fertilizer was 100:50:50 and 75:30:30 of N, P2O5 and K2O kg ha-1respectively.All the yield attributing characters showed that the usage of different doses of green nano PGR through foliar application has positive effects compared to yellow and red nano PGR and the lowest was from control.

LncRNA RGMB-AS1 inhibits HMOX1 ubiquitination and NAA10 activation to induce ferroptosis in non-small cell lung cancer

Ferroptosis, an iron-dependent regulated cell death caused by excessive lipid peroxide accumulation, has emerged as a promising therapeutic target in various cancers, including non-small cell lung cancer (NSCLC). In this study, we identified the long non-coding RNA RGMB-AS1 as a key regulator of ferroptosis in NSCLC. Mechanistically, RGMB-AS1 interacted with heme oxygenase 1 (HMOX1) and prevented its ubiquitination by the E3 ligase TRC8, leading to increased HMOX1 stability and enhanced ferroptosis. Additionally, RGMB-AS1 bound to the 82–87 amino acid region of N-alpha-acetyltransferase 10 (NAA10), stimulating its acetyltransferase activity and promoting the conversion of acetyl-CoA to HMG-CoA, further contributing to ferroptosis. The RGMB-AS1-HMOX1 and RGMB-AS1-NAA10 axes synergistically inhibited NSCLC growth both in vitro and in vivo. Clinically, low RGMB-AS1 expression was associated with advanced tumor stage and poor overall survival in NSCLC patients. Furthermore, adeno-associated virus-mediated RGMB-AS1 overexpression significantly suppressed tumor growth in mouse xenograft models. Our findings uncover a novel lncRNA-mediated regulatory mechanism of ferroptosis and highlight the potential of RGMB-AS1 as a prognostic biomarker and therapeutic target in NSCLC.

Dysfunction of Nrf2-regulated cellular defence system and JNK activation induced by high dose of fly Ash particles are associated with pulmonary injury in mouse lungs

The mechanisms of the exposure to fine particulate matter (PM) as a risk factor for pulmonary injury are not fully understood. The transcription factor, NF-E2-related factor 2 (Nrf2), plays a key role in protection lung against PM insult and cancer chemoprevention. In this study, F3-S fly ash particles from a municipal waste incinerator were evaluated as a PM model. We found that F3-S triggered hierarchical oxidative stress responses involving the prolonged activation of the cytoprotective Nrf2 transcriptional program via Keap1 Cys151 modification, and c-Jun NH2-terminal kinase (JNK) phosphorylation at higher doses. In mouse lungs exposed to fly ash particles at a low dose (10–20 mg/kg), Nrf2 signalling was upregulated, while in those exposed to a high fly ash particle dose (40 mg/kg), there was significant activation of JNK, and this correlated with Nrf2 phosphorylation and the downregulation of antioxidant response element (ARE)-driven genes. The JNK inhibitor, SP600125, reversed Nrf2 phosphorylation, and downregulation of detoxifying enzymes. Silencing JNK expression in mouse lungs using adenoviral shRNA inhibited JNK activation and Nrf2 phosphorylation, promoted ARE-driven gene expression, and reduced pulmonary injury. Furthermore, we found that the 452–515 amino acid region within the Neh1 domain of Nrf2 was required for its interaction with P-JNK. We demonstrated that Nrf2 was an important P-JNK target in fly ash-induced pulmonary toxicity. JNK phosphorylated Nrf2, leading to a dysfunction of the Nrf2-mediated defence system.