Inducing Activity Research Articles

Tryptophan metabolism reprogramming contributes to the prothrombotic milieu in mice and humans infected with SARS-CoV-2.

SARS-CoV-2 infection disturbs the coagulation balance in the blood, triggering thrombosis and contributing to organ failure. The role of prothrombotic metabolites in COVID-19-associated coagulopathy remains elusive. Leveraging K18-hACE2 mice infected with SARS-CoV-2, we observed higher levels of the tryptophan metabolite, kynurenine, compared to controls. SARS CoV-2 infected mice showed a significant upregulation of enzymes controlling Kynurenine biogenesis, such as indoleamine 2,3-dioxygenase (IDO-1) and tryptophan 2,3-dioxygenase levels in kidneys and liver, respectively, as well as changes in the enzymes involved in kynurenine catabolism, including kynurenine monooxygenase and kynurinase. Consistent with the agonistic role of these metabolites in Aryl Hydrocarbon Receptor (AHR) signaling, AHR activation and its downstream mediator, tissue factor (TF), a highly potent procoagulant factor, was observed in endothelial cells (ECs) of lungs and kidneys of infected mice. These findings were validated in humans, where compared to controls, sera of COVID-19 patients showed increased levels of Kynurenine, kynurenic acid, anthranilic acid, and quinolinic acid. Activation of the AHR-TF axis was noted in the kidneys and lungs of COVID-19 patients, and COVID-19 sera showed higher IDO-1 activity than controls. Levels of Kyn in COVID-19 patients correlated strongly with the TF inducing activity of COVID-19 sera on ECs. A specific IDO-1 inhibitor or AHR inhibitor separately or in combination suppressed COVID-19 sera-induced TF activity in ECs. Together, we identified IDO-1 as upregulated by SARS-CoV-2 infection, resulting in augmented Kyn and its prothrombotic catabolites, thereby suggesting the Kyn AHR-TF axis as possibly a new diagnostic and/or therapeutic target.

Investigation of antiproliferative and apoptotic effects of Rosmarinus officinalis essential oil obtained by hydrodistillation on neuroblastoma cells

This study was designed to determine the potential antiproliferative and apoptosis inducing activities of Rosmarinus officinalis essential oil (RE) on neuroblastoma cancer cells. For this purpose, different concentrations of RE were applied to SH-SY5Y cells for 24 hours and cell viability was determined by MTT. In addition, the percentage of early, late and non-apoptotic cells was determined by AnnexinV/propodium iodide staining to determine the induction of apoptosis. In addition, the composition of RE was determined by GC-MS. In MTT assay, it was determined that the viability of SH-SY5Y cells decreased dose-dependently as a result of the application of different concentrations of RE. Moreover, 200 mg/ml RE treatment increased the percentage of cells in the late apoptotic phase. The main compounds of RE were determined as (1R)-2,6,6-Trimethylbicyclo[3.1.1]hept-2-ene, Bicyclo[3.1.1]hept-3-en-2-one, 4,6,6-trimethyl, 1,8-Cineole, Camphor by GC-MS. In conclusion, RE is thought to be an important source of anti-proliferative and apoptosis inducing activity on neuroblastoma cells.

The impacts of flagellin on larval metamorphosis of mussel Mytilus coruscus varied with protein structure

Flagellin produced by Pseudoalteromonas marina has been shown to induce larval metamorphosis in mussels. The flagellin protein FliC was composed of four subunits encoded by the genes fliC-02330, fliC-02345, fliC-02346, and fliC-02347. In this study, we investigated the effects of these four FliC proteins on biofilm formation and larval metamorphosis. The inducing activity of the recombinant proteins FliC-02345, FliC-02346, and FliC-02347 was similar and significantly higher (p < 0.05) than that of FliC-02330. When compared to the wild-type strain, biofilm formation was significantly enhanced in the four fliC mutants, whereas the inducing activity of four ΔfliC biofilms decreased. Among these mutants, ΔfliC-02330 exhibited the highest thickness, density, and c-di-GMP levels among its respective biofilms. The inducing activity of the ΔfliC-02330 biofilm increased by 30–40% compared to the inducing activities of other mutant strains. Furthermore, structural analysis revealed differences in protein structure and phosphorylation sites between FliC-02330 and its counterparts (FliC-02345, FliC-02346, and FliC-02347), suggesting that variations in protein structure contribute to differing impacts on both biofilm formation and larval metamorphosis. These findings provide new insights into the interaction between bacterial flagellin protein and the larval metamorphosis of marine invertebrates.

Chemotaxis gene of a bacterium impacts larval settlement and metamorphosis in the marine mussel Mytilus coruscus via c-di-GMP controlling extracellular protein production

Bacterial chemotaxis enhances bacterial adaptation to the environment and is important for biofilm formation. Biofilms play a key role in inducing larval settlement and metamorphosis in many marine invertebrates. However, the specific mechanisms by which bacterial chemotaxis influences larval settlement and metamorphosis in mussels remain unknown. The findings indicate that the absence of the chemotaxis gene cheW resulted in reduced motility of Pseudoalteromonas marina, accompanied by an increase in c-di-GMP content. The ΔcheW strain exhibited a higher capacity for biofilm formation compared to the wild-type strain. The extracellular protein content of the ΔcheW strain exhibited a significant 77% reduction, specifically in the flagellin content. The inducing activity of ΔcheW was reduced by 56% compared to the wild-type strain. This study highlights that the deficiency of the chemotaxis gene cheW inhibited larval settlement and metamorphosis in mussels through c-di-GMP regulation of extracellular protein production. It provides a novel ecological function of bacterial chemotaxis in regulating the larval settlement and metamorphosis of marine invertebrates.

Double-filtration plasmapheresis reduces type I interferon bioavailability and inducing activity in systemic lupus erythematosus

Type I interferons (IFN-Is) play a significant role in systemic lupus erythematosus (SLE) pathogenesis. Double-filtration plasmapheresis (DFPP) is a treatment option for SLE; however, its effect on IFN-Is remains unclear. Therefore, we investigated the effects of DFPP on IFN-Is. Plasma from patients with SLE (n = 11) who regularly underwent DFPP was analysed using a cell-based reporter system to detect the bioavailability and inducing activity of IFN-I. The concentration of plasma dsDNA was measured, and western blotting analysis was used to assess the phosphorylation of the STING pathway. A higher IFN-I bioavailability and inducing activity were observed in patients compared to healthy controls, and both parameters decreased after DFPP. The reduction in IFN-I-inducing activity was particularly prominent in patients with high disease activity. Notably, this reduction was not observed in STING-knockout reporter cells. Additionally, plasma dsDNA levels decreased after DFPP treatment, suggesting that inhibition of the STING pathway was responsible for the observed decrease in activity. Western blotting analysis revealed suppression of STING pathway phosphorylation after DFPP. DFPP reduced IFN-I bioavailability and the inducing activity of plasma. This reduction is likely attributable to the inhibition of the STING pathway through the elimination of dsDNA.

Camptothecin bioprocessing from Aspergillus terreus, an endophyte of Catharanthus roseus: antiproliferative activity, topoisomerase inhibition and cell cycle analysis

Attenuation of camptothecin (CPT) productivity by fungi with preservation and subculturing is the challenge that halts fungi to be an industrial platform of CPT production. Thus, screening for novel endophytic fungal isolates with metabolic stability for CPT production was the objective. Catharanthus roseus is one of the medicinal plants with diverse bioactive metabolites that could have a plethora of novel endophytes with unique metabolites. Among the endophytes of C. roseus, Aspergillus terreus EFBL-NV OR131583.1 had the most CPT producing potency (90.2 μg/l), the chemical identity of the putative CPT was verified by HPLC, FT-IR, NMR and LC–MS/MS. The putative A. terreus CPT had the same molecular mass (349 m/z), and molecular fragmentation patterns of the authentic one, as revealed from the MS/MS analyses. The purified CPT had a strong activity against MCF7 (5.27 μM) and UO-31 (2.2 μM), with a potential inhibition to Topo II (IC50 value 0.52 nM) than Topo 1 (IC50 value 6.9 nM). The CPT displayed a high wound healing activity to UO-31 cells, stopping their metastasis, matrix formation and cell immigration. The purified CPT had a potential inducing activity to the cellular apoptosis of UO-31 by ~ 17 folds, as well as, arresting their cellular division at the S-phase, compared to the control cells. Upon Plackett–Burman design, the yield of CPT by A. terreus was increased by ~ 2.6 folds, compared to control. The yield of CPT by A. terreus was sequentially suppressed with the fungal storage and subculturing, losing ~ 50% of their CPT productivity by 3rd month and 5th generation. However, the productivity of the attenuated A. terreus culture was completely restored by adding 1% surface sterilized leaves of C. roseus, and the CPT yield was increased over-the-first culture by ~ 3.2 folds (315.2 μg/l). The restoring of CPT productivity of A. terreus in response to indigenous microbiome of C. roseus, ensures the A. terreus-microbiome interactions, releasing a chemical signal that triggers the CPT productivity of A. terreus. This is the first reports exploring the potency of A. terreus, endophyte of C. roseus” to be a platform for industrial production of CPT, with an affordable sustainability with addition of C. roseus microbiome.

Antiviral activity and interferon productivity of low-molecular inducers of interferon analogs of amixin and derivative of diphenyl

Antiviral activity and interferon productivity of low-molecular inducers of interferon analogs of amixin and derivative of diphenyl

Some Structural Elements of Bacterial Protein MF3 That Influence Its Ability to Induce Plant Resistance to Fungi, Viruses, and Other Plant Pathogens.

The ability of the MF3 protein from Pseudomonas fluorescens to protect plants by inducing their resistance to pathogenic fungi, bacteria, and viruses is well confirmed both in greenhouses and in the field; however, the molecular basis of this phenomenon remains unexplored. To find a relationship between the primary (and spatial) structure of the protein and its target activity, we analyzed the inducing activity of a set of mutants generated by alanine scanning and an alpha-helix deletion (ahD) in the part of the MF3 molecule previously identified by our group as a 29-amino-acid peptide working as the inducer on its own. Testing the mutants' inducing activity using the "tobacco-tobacco mosaic virus" pathosystem revealed that some of them showed an almost threefold (V60A and V62A) or twofold (G51A, L58A, ahD) reduction in inducing activity compared to the wild-type MF3 type. Interestingly, these mutations demonstrated close proximity in the homology model, probably contributing to MF3 reception in a host plant.

Ovulation Inducing Activity in PCOD Induced Female Wister Albino Rats and In vitro Studies of Free Radical Scavenging Activity of Kadugurogini chooranam (Picrorhiza scrophulariiflora)

Siddha system of medicine is oldest traditional system in south India. In these system herbals plays a vital role in prevention and curing the disease. Siddhar’s who told the amazing solution for aging process. Causes of aging are due to increased level of free radicals that damage the major constituents of the cell. Herbals naturally have free radical scavenging activity because Presence of phyto-constiuents like polyphenols generally indicates the rejuvenating properties. The present study also to validate the Estrodial induced PCOS female wistar albino rats to treat with extract of Kadugurogini Chooranam (Picrorhiza scrophulariiflora) and the level of hormones were detected for the ovulation inducing activity and analysis the anti-radical properties from the decoction of Kadugurogini Chooranam (Picrorhiza scrophulariiflora) through the four parameters superoxide, nitricoxide, lipid peroxidation, ABTS radical activity with standard antioxidant and positive control is used as a ascorbic acid. Therefore, aqueous root extracts of Kadugurogini Chooranam have considerable antioxidant properties. In preliminary study, the Kadugurogini Chooranam posse’s significant effects on hormonal level on PCOS induced female wister albino rats.

Derivatives of Differentiation-Inducing Factor 1 Differentially Control Chemotaxis and Stalk Cell Differentiation in Dictyostelium discoideum.

Differentiation-inducing factors 1 and 2 (DIF-1 and DIF-2) are small lipophilic signal molecules that induce stalk cell differentiation but differentially modulate chemotaxis toward cAMP in the cellular slime mold Dictyostelium discoideum; DIF-1 suppresses chemotactic cell movement in shallow cAMP gradients, whereas DIF-2 promotes it. The receptor(s) for DIF-1 and DIF-2 have not yet been identified. We examined the effects of nine derivatives of DIF-1 on chemotactic cell movement toward cAMP and compared their chemotaxis-modulating activity and stalk cell differentiation-inducing activity in wild-type and mutant strains. The DIF derivatives differentially affected chemotaxis and stalk cell differentiation; for example, TM-DIF-1 suppressed chemotaxis and showed poor stalk-inducing activity, DIF-1(3M) suppressed chemotaxis and showed strong stalk-inducing activity, and TH-DIF-1 promoted chemotaxis. These results suggest that DIF-1 and DIF-2 have at least three receptors: one for stalk cell induction and two for chemotaxis modulation. In addition, our results show that the DIF derivatives can be used to analyze the DIF-signaling pathways in D. discoideum.

Novel topoisomerase II/EGFR dual inhibitors: design, synthesis and docking studies of naphtho[2′,3′:4,5]thiazolo[3,2-a]pyrimidine hybrids as potential anticancer agents with apoptosis inducing activity

Topoisomerases II are ubiquitous enzymes with significant genotoxic effects in many critical DNA processes. Additionally, epidermal growth factor receptor (EGFR) plays pivotal role in tumour growth and angiogenesis. A novel series of naphtho[2',3':4,5]thiazolo[3,2- a ]pyrimidine hybrids have been designed, synthesised and evaluated for their topo IIα/EGFR inhibitory and apoptotic inducer activities. Cytotoxicity of the synthesised hybrids was evaluated against MCF-7, A549 and HCT-116 cell lines. Of the synthesised hybrids, 6i, 6a and 6c experienced superior cytotoxic activity compared to doxorubicin and erlotinib against the tested cancer cells. The molecular mechanism of these hybrids revealed their ability to successfully inhibit topo IIα and EGFR activities in micromolar concentration and may serve as topo II catalytic inhibitor. Moreover, these hybrids significantly arrested cell cycle at G2/M phase together with increased p53, caspae-7, caspase-9 levels and Bax/Bcl-2 ratio. The synthesised hybrids showed efficient binding pattern in molecular docking study and have acceptable drug likeness characters.

Evaluation of inducing activity of CIP elicitors from diverse sources based on monosaccharide composition and physiological indicators

Application of elicitors can greatly enhance plant immune resistance against pathogens. However, it is still obscure whether elicitor activity is influenced by diverse sources. This study investigated the effect of foliar spraying of 19 batches of Chrysanthemum indicum polysaccharides (CIPs) on the disease resistance of Atractylodes macrocephala Koidz. (A. macrocephala) and explored the main reasons for the differences of inducing activity of CIP elicitors. PCA, OPLS-DA, grey relational analysis and entropy weight method had good predictability for the activity evaluation of CIP elicitors and other plant-derived elicitors. The results showed that 19 batches of CIPs had definite regional differences in inducing activity and monosaccharide content. CIP elicitors with high inducing activity could significantly increase the accumulation of Atractylenolide Ⅱ and Atractylenolide Ⅲ, the mRNA relative transcription level of CAT, POD, PAL genes, the amount of pH change in the medium and effectively reduce the disease index of A. macrocephala. Furthermore, CIP with high inducing activity exhibited the high contents of Rha, Ara and GalA, which might be the main contributor to their high activity. The evaluation procedure developed in this work can be applied for screening CIP elicitors with high inducing activity, and it lays a foundation for identifying more functional elicitors related to plant immune resistance.

Two-Component System Response Regulator ompR Regulates Mussel Settlement through Exopolysaccharides.

The outer membrane protein (OMP) is a kind of biofilm matrix component that widely exists in Gram-negative bacteria. However, the mechanism of OMP involved in the settlement of molluscs is still unclear. In this study, the mussel Mytilus coruscus was selected as a model to explore the function of ompR, a two-component system response regulator, on Pseudoalteromonas marina biofilm-forming capacity and the mussel settlement. The motility of the ΔompR strain was increased, the biofilm-forming capacity was decreased, and the inducing activity of the ΔompR biofilms in plantigrades decreased significantly (p < 0.05). The extracellular α-polysaccharide and β-polysaccharide of the ΔompR strain decreased by 57.27% and 62.63%, respectively. The inactivation of the ompR gene decreased the ompW gene expression and had no impact on envZ expression or c-di-GMP levels. Adding recombinant OmpW protein caused the recovery of biofilm-inducing activities, accompanied by the upregulation of exopolysaccharides. The findings deepen the understanding of the regulatory mechanism of bacterial two-component systems and the settlement of benthic animals.

A Pythium myriotylum Small Cysteine-Rich Protein Triggers Immune Responses in Diverse Plant Hosts

The oomycete Pythium myriotylum is a necrotrophic pathogen that infects many crop species worldwide, including ginger, soybean, tomato, and tobacco. Here, we identified a P. myriotylum small cysteine-rich protein, PmSCR1, that induces cell death in Nicotiana benthamiana by screening small, secreted proteins that were induced during infection of ginger and did not have a predicted function at the time of selection. Orthologs of PmSCR1 were found in other Pythium species, but these did not have cell death-inducing activity in N. benthamiana. PmSCR1 encodes a protein containing an auxiliary activity 17 family domain and triggers multiple immune responses in host plants. The elicitor function of PmSCR1 appears to be independent of enzymatic activity, because the heat inactivation of PmSCR1 protein did not affect PmSCR1-induced cell death or other defense responses. The elicitor function of PmSCR1 was also independent of BAK1 and SOBIR1. Furthermore, a small region of the protein, PmSCR186-211, is sufficient for inducing cell death. A pretreatment using the full-length PmSCR1 protein promoted the resistance of soybean and N. benthamiana to Phytophthora sojae and Phytophthora capsici infection, respectively. These results reveal that PmSCR1 is a novel elicitor from P. myriotylum, which exhibits plant immunity-inducing activity in multiple host plants. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Iruplinalkib (WX‑0593), a novel ALK/ROS1 inhibitor, overcomes crizotinib resistance in preclinical models for non-small cell lung cancer

Despite remarkable initial responses of anaplastic lymphoma kinase (ALK) inhibitors in ALK-positive non-small cell lung cancer (NSCLC) patients, cancers eventually develop resistance within one to two years. This study aimed to compare the properties of iruplinalkib (WX‑0593) with other ALK inhibitors and report the comprehensive characterization of iruplinalkib against the crizotinib resistance. The inhibitory effect of iruplinalkib on kinase activity was detected. A kinase screen was performed to evaluate the selectivity of iruplinalkib. The effect of iruplinalkib on related signal transduction pathways of ALK and c-ros oncogene 1 (ROS1) kinases was examined. The cellular and in vivo activities of ALK inhibitors were compared in engineered cancer-derived cell lines and in mice xenograft models, respectively. Human hepatocytes derived from three donors were used for evaluating hepatic enzyme inducing activity. HEK293 cell lines expressing transportors were used to invesigated the drug interaction potential mediated by several transporters. The results showed iruplinalkib potently inhibited the tyrosine autophosphorylation of wild-type ALK, ALKL1196M, ALKC1156Y and epidermal growth factor receptor (EGFR)L858R/T790M. The inhibitory effects of iruplinalkib in patient-derived xenograft and cell line-derived xenograft models were observed. Moreover, iruplinalkib showed robust antitumor effects in BALB/c nude mice xenograft models with ALK-/ROS1-positive tumors implanted subcutaneously, and the tumor suppressive effects in crizotinib-resistant model was significantly better than that of brigatinib. Iruplinalkib did not induce CYP1A2, CYP2B6 and CYP3A4 at therapeutic concentration, and was also a strong inhibitor of MATE1 and MATE2K transporters, as well as P-gp and BCRP. In conclusion, iruplinalkib, a highly active and selective ALK/ROS1 inhibitor, exhibited strong antitumor effects in vitro and in crizotinib-resistant models.

Mussel settlement mediated by bacterial VgrG proteins via extracellular outer membrane vesicles

T6SS plays an important role in biofilm formation, but how bacterial T6SS regulates biofilm formation and its effect on marine invertebrate settlement is still unclear. Here, the role of T6SS spike protein VgrG and VgrG protein synthesis gene on biofilm formation and Mytilus coruscus settlement was examined. Deletion of the core gene vgrG of T6SS resulted in a decrease in the growth capability of Leisingera aquaemixtae and inhibition of the ability of biofilm formation. The OMV content and inducing activity of ΔvgrG biofilms were decreased respectively by 20.72% and 59.70% compared with the wild-type biofilms. OMVs induced the settlement of M. coruscus, while VgrG proteins did not induce mussel settlement directly. Addition of VgrG proteins resulted in the upregulation of OMVs in the ΔvgrG biofilms. Adding OMVs caused no increasing inducing activity of ΔvgrG biofilms. Interestingly, the inducing activity of ΔvgrG biofilms both by adding VgrG proteins alone, and by adding VgrG proteins and OMVs in combination increased significantly, and restored to the inducing capability of wild-type biofilms. Thus, bacterial T6SS spike protein VgrG could mediate mussel settlement by increasing the OMV content of biofilms. The present findings shed new light on the interaction between T6SS and marine invertebrate settlement.

Evaluation of the properties of Bungarus caeruleus venom and checking the efficacy of antivenom used in Bangladesh for its bite treatment.

As a disaster-prone country with unique geographical features, snake biting is a major public health concern in Bangladesh. The primary reasons of mortality from snakebite include late presentation to the hospital, low efficacy of antivenom, and a lack of adequate management facilities. Because snake venom characteristics vary depending on geographical location, antivenom should be manufactured from snakes native to the region in which it would be administered. Bungarus caeruleus is a highly venomous snake contributing to the major snakebite issue in Bangladesh. Therefore, the neutralization efficacy of the antivenom against B. caeruleus venom was evaluated in the current study along with the characterization of venom. For biological characterization of venom, RP-HPLC and SDS-PAGE profiling, hemolytic activity, hemorrhagic activity, phospholipases A2 (PLA2) activity, edema inducing activity and histopathological observations were carried out following standard protocol. LD50 of the venom was calculated along with neutralization potency of Incepta antivenom through probit analysis. Results showed that venom possesses phospholipase A2 activity, hemolytic activity and edema inducing activity while hemorrhagic activity was absent in the skin of envenomed mice. Histopathological alterations including necrosis, congestion and infiltrations were observed in envenomed mice organs after hematoxylin and eosin staining. Neutralization study showed that Incepta polyvalent antivenom could neutralize (potency 0.53mg/ml) the lethal effect in in vitro study on mice. Further investigation on snakebite epidemiology and clinical observations of the envenomed patients will help in combating the snakebite problem more efficiently.

Larval settlement and metamorphosis of Mytilus coruscus in response to varying bacterial capsular polysaccharide

Marine invertebrates are the backbone of marine biodiversity and play a pivotal role in the marine ecosystem. The life cycle of most marine invertebrates includes the settlement and metamorphosis stage, which is induced by marine biofilms, but the mechanism is still enigmatic. In the present study, we constructed the capsular polysaccharide (CPS) synthesis gene capC-deleted mutant of Pseudoalteromonas marina by gene knockout and then compared the phenotype, the biofilm-forming ability, the effect on settlement and metamorphosis of Mytilus coruscus, and the exopolysaccharide and CPS levels between the mutant and the wild-type strains to explicate the relationship between bacteria and mussels. The study presented that the phenotype and biofilm-forming ability between the wild-type and ΔcapC strains had no significant difference, but the inducing activity of ΔcapC biofilms on larval settlement and metamorphosis decreased significantly (p &amp;lt; 0.05). Compared with the wild-type, the CPS content of ΔcapC strain significantly decreased by 38.07%, accompanied by the increase of c-di-GMP. Meanwhile, the biomass of α-polysaccharides and β-polysaccharides on ΔcapC biofilms decreased significantly (p &amp;lt; 0.05). Thus, the CPS synthesis gene could modulate c-di-GMP, which regulates bacterial polysaccharide secretion, and then impact larval settlement and metamorphosis of mussels. This work brings an entry point to deeply understand the interaction between bacterial polysaccharide and larval recruitment.

A Relaxin-Like Gonad-Stimulating Peptide Appears in the Early Development of the Starfish Patiria pectinifera.

Relaxin-like gonad-stimulating peptide (RGP) is a hormone with gonadotropin-like activity in starfish. This study revealed that spawning inducing activity was detected in an extract of brachiolaria larvae of Patiria pectinifera. Spawning inducing activity in the extract was due to P. pectinifera RGP (PpeRGP), not 1-methyladenine. The expression of PpeRGP mRNA was also found in brachiolaria. Immunohistochemical observation with specific antibodies for PpeRGP showed that PpeRGP was distributed in the peripheral adhesive papilla of the brachiolaria arms. In contrast, PpeRGP was not detected in the adult rudiment or ciliary band regions, which are present in the neural system. These findings strongly suggest that RGP exists in the larvae before metamorphosis. Because gonads are not developed in starfish larvae, it seems likely that RGP plays another role other than gonadotropic action in the early development of starfish.

In silico design of a promiscuous chimeric multi-epitope vaccine against Mycobacterium tuberculosis

Tuberculosis (TB) is a global health threat, killing approximately 1.5 million people each year. The eradication of Mycobacterium tuberculosis, the main causative agent of TB, is increasingly challenging due to the emergence of extensive drug-resistant strains. Vaccination is considered an effective way to protect the host from pathogens, but the only clinically approved TB vaccine, Bacillus Calmette-Guérin (BCG), has limited protection in adults. Multi-epitope vaccines have been found to enhance immunity to diseases by selectively combining epitopes from several candidate proteins. This study aimed to design a multi-epitope vaccine against TB using an immuno-informatics approach. Through functional enrichment, we identified eight proteins secreted by M. tuberculosis that are either required for pathogenesis, secreted into extracellular space, or both. We then analyzed the epitopes of these proteins and selected 16 helper T lymphocyte epitopes with interferon-γ inducing activity, 15 cytotoxic T lymphocyte epitopes, and 10 linear B-cell epitopes, and conjugated them with adjuvant and Pan HLA DR-binding epitope (PADRE) using appropriate linkers. Moreover, we predicted the tertiary structure of this vaccine, its potential interaction with Toll-Like Receptor-4 (TLR4), and the immune response it might elicit. The results showed that this vaccine had a strong affinity for TLR4, which could significantly stimulate CD4+ and CD8+ cells to secrete immune factors and B lymphocytes to secrete immunoglobulins, so as to obtain good humoral and cellular immunity. Overall, this multi-epitope protein was predicted to be stable, safe, highly antigenic, and highly immunogenic, which has the potential to serve as a global vaccine against TB.