RESUMO
Cell fate specification is essential for every major event of embryogenesis, and subsequent cell maturation ensures individual cell types acquire specialized functions. The mechanisms that regulate cell fate specification have been studied exhaustively, and each technological advance in developmental biology ushers in a new era of studies aimed at uncovering the most fundamental processes by which cells acquire unique identities. What is less appreciated is that mechanisms are in place to ensure cell identity is maintained throughout the life of the organism. The body wall musculature in the Drosophila embryo is a well-established model to study cell fate specification, as each hemisegment in the embryo generates and maintains thirty muscles with distinct identities. Once specified, the thirty body wall muscles fuse with mononucleate muscle precursors that lack a specific identity to form multinucleate striated muscles. Multinucleate body wall muscles do not fuse with each other, which maintains a diversification of muscle cell identities. Here we show the serine/threonine kinase Back seat driver (Bsd) prevents inappropriate muscle fusion to maintain cell identity. Thus, the regulation of cell fusion is one mechanism that maintains cell identity.
Assuntos
Proteínas de Drosophila , Animais , Proteínas de Drosophila/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Fusão Celular , Drosophila/metabolismo , Músculo Esquelético/metabolismo , Serina/metabolismo , Regulação da Expressão Gênica no DesenvolvimentoRESUMO
Root parasitic plants of the Orobanchaceae, broomrapes and witchweeds, pose a severe problem to agriculture in Europe, Asia and especially Africa. These parasites are totally dependent on their host for survival, and therefore, their germination is tightly regulated by host presence. Indeed, their seeds remain dormant in the soil until a host root is detected through compounds called germination stimulants. Strigolactones (SLs) are the most important class of germination stimulants. They play an important role in planta as a phytohormone and, upon exudation from the root, function in the recruitment of symbiotic arbuscular mycorrhizal fungi. Plants exude mixtures of various different SLs, possibly to evade detection by these parasites and still recruit symbionts. Vice versa, parasitic plants must only respond to the SL composition that is exuded by their host, or else risk germination in the presence of non-hosts. Therefore, parasitic plants have evolved an entire clade of SL receptors, called HTL/KAI2s, to perceive the SL cues. It has been demonstrated that these receptors each have a distinct sensitivity and specificity to the different known SLs, which possibly allows them to recognize the SL-blend characteristic of their host. In this review, we will discuss the molecular basis of SL sensitivity and specificity in these parasitic plants through HTL/KAI2s and review the evidence that these receptors contribute to host specificity of parasitic plants.
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Micorrizas , Orobanche , Striga , Raízes de Plantas/microbiologia , Especificidade de Hospedeiro , Lactonas , Plantas , GerminaçãoRESUMO
Helicobacter pylori is a leading cause of stomach cancer and peptic ulcers. Thus, identifying epitopes in H. pylori antigens is important for disease etiology, immunological surveillance, enhancing early detection tests, and developing optimal epitope-based vaccines. We used immunoinformatic and computational methods to create a potential CagW epitope candidate for H. pylori protection. The cagW gene of H. pylori was amplified and cloned into pcDNA3.1 (+) for injection into the muscles of healthy BALB/c mice to assess the impact of the DNA vaccine on interleukin levels. The results will be compared to a control group of mice that received PBS or cagW-pcDNA3.1 (+) vaccinations. An analysis of CagW protein antigens revealed 8 CTL and 7 HTL epitopes linked with AYY and GPGPG, which were enhanced by adding B-defensins to the N-terminus. The vaccine's immunogenicity, allergenicity, and physiochemistry were validated, and its strong activation of TLRs (1, 2, 3, 4, and 10) suggests it is antigenic. An in-silico cloning and immune response model confirmed the vaccine's expression efficiency and predicted its impact on the immune system. An immunofluorescence experiment showed stable and bioactive cagW gene expression in HDF cells after cloning the whole genome into pcDNA3.1 (+). In vivo vaccination showed that pcDNA3.1 (+)-cagW-immunized mice had stronger immune responses and a longer plasmid DNA release window than control-plasmid-immunized mice. After that, bioinformatics methods predicted, developed, and validated the three-dimensional structure. Many online services docked it with Toll-like receptors. The vaccine was refined using allergenicity, antigenicity, solubility, physicochemical properties, and molecular docking scores. Virtual-reality immune system simulations showed an impressive reaction. Codon optimization and in-silico cloning produced E. coli-expressed vaccines. This study suggests a CagW epitopes-protected H. pylori infection. These studies show that the proposed immunization may elicit particular immune responses against H. pylori, but laboratory confirmation is needed to verify its safety and immunogenicity.
Assuntos
Infecções por Helicobacter , Helicobacter pylori , Vacinas , Animais , Camundongos , Helicobacter pylori/genética , Epitopos Imunodominantes , Infecções por Helicobacter/prevenção & controle , Simulação de Acoplamento Molecular , Escherichia coli , Epitopos/genéticaRESUMO
DWARF14 (D14) and HTL/KAI2 (KAI2) are paralogous receptors in the α/ß-hydrolase superfamily. D14 is the receptor for a class of plant hormones, strigolactones (SLs), and KAI2 is the receptor for the smoke-derived seed germination inducer, Karrikin (KAR), in Arabidopsis. Germinone (Ger) was previously reported as a KAI2 agonist with germination-inducing activity for thermo-inhibited Arabidopsis seed. However, Ger was not specific to KAI2, and could also bind to D14. It was reported that SL analogs with a desmethyl-type D-ring structure are specifically recognized by KAI2. On the basis of this observation, we synthesized a desmethyl-type germinone (dMGer). We found that dMGer is highly specific to KAI2. Moreover, dMGer induced Arabidopsis seed germination more effectively than did Ger. In addition, dMGer induced the seed germination of Arabidopsis in a manner independently of GA, a well-known germination inducer in plants.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Germinação , Proteínas de Arabidopsis/metabolismo , Giberelinas/farmacologia , Giberelinas/metabolismo , Sementes/metabolismo , Hidrolases/metabolismo , Lactonas/farmacologiaRESUMO
Sewage sludge management poses challenges due to its environmental impact, varying composition, and stringent regulatory requirements. In this scenario, hydrothermal liquefaction (HTL) is a promising technology for producing biofuel and extracting phosphorus from sewage sludge. However, the toxic nature of the resulting process water (HTL-PW) raises concerns about integrating HTL into conventional wastewater treatment processes. This study investigated the inhibitory effects of HTL-PW on the activity of the main microbial functions in conventional activated sludge. Upon recirculation of the HTL-PW from the excess sludge into the wastewater treatment plant, the level of COD in the influent is expected to increase by 157 mgO2â L-1, resulting in 44% nitrification inhibition (IC50 of 197 mgâ L-1). However, sorption of inhibitory compounds on particles can reduce nitrification inhibition to 27% (IC50 of 253 mgâ L-1). HTL-PW is a viable carbon source for denitrification, showing nearly as high denitrification rates as acetate and only 17% inhibition at 157 mgO2â L-1 COD. Under aerobic conditions, heterotrophic organic nitrogen and organic matter conversion remains unaffected up to 223 mgO2â L-1 COD, with COD removal higher than 94%. This study is the first to explore the full integration of HTL in wastewater treatment plants for biofuel production from the excess activated sludge. Potential nitrification inhibition is concerning, and further long-term studies are needed to fully investigate the impacts.
Assuntos
Nitrificação , Purificação da Água , Esgotos , Biocombustíveis , Óxido de Magnésio , Purificação da Água/métodos , Nitrogênio , Reatores Biológicos , Desnitrificação , Eliminação de Resíduos Líquidos/métodosRESUMO
The gating of the Voltage-Dependent Anion Channel (VDAC) is linked to oxidative stress through increased generation of mitochondrial ROS with increasing mitochondrial membrane potential (ΔΨm). It has been already reported that H2O2 increases the single-channel conductance of VDAC on a bilayer lipid membrane. On the other hand, homocysteine (Hcy) has been reported to induce mitochondria-mediated cell death. It is argued that the thiol-form of homocysteine, HTL could be the plausible molecule responsible for the alteration in the function of proteins, such as VDAC. It is hypothesized that HTL interacts with VDAC that causes functional abnormalities. An investigation was undertaken to study the interaction of HTL with VDAC under H2O2 induced oxidative stress through biophysical and electrophysiological methods. Fluorescence spectroscopic studies indicate that HTL interacts with VDAC, but under induced oxidative stress the effect is prevented partially. Similarly, bilayer electrophysiology studies suggest that HTL shows a reduction in VDAC single-channel conductance, but the effects are partially prevented under an oxidative environment. Gly172 and His181 are predicted through bioinformatics tools to be the most plausible binding residues of HTL in Rat VDAC. The binding of HTL and H2O2 with VDAC appears to be cooperative as per our analysis of experimental data in the light of the Hill-Langmuir equation. The binding energies are estimated to be - 4.7 kcal mol-1 and - 2.8 kcal mol-1, respectively. The present in vitro studies suggest that when mitochondrial VDAC is under oxidative stress, the effects of amino acid metabolites like HTL are suppressed.
Assuntos
Peróxido de Hidrogênio , Canais de Ânion Dependentes de Voltagem , Animais , Homocisteína/análogos & derivados , Homocisteína/metabolismo , Homocisteína/farmacologia , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/farmacologia , Mitocôndrias/metabolismo , Estresse Oxidativo , Ratos , Canais de Ânion Dependentes de Voltagem/químicaRESUMO
BACKGROUND: The risk of graft-induced dyskinesias (GIDs) presents a major challenge in progressing cell transplantation as a therapy for Parkinson's disease. Current theories implicate the presence of grafted serotonin neurons, hotspots of dopamine release, neuroinflammation and established levodopa-induced dyskinesia. OBJECTIVE: To elucidate the mechanisms of GIDs. METHODS: Neonatally desensitized, dopamine denervated rats received intrastriatal grafts of human embryonic stem cells (hESCs) differentiated into either ventral midbrain dopaminergic progenitor (vmDA) (n = 15) or ventral forebrain cells (n = 14). RESULTS: Of the eight rats with surviving grafts, two vmDA rats developed chronic spontaneous GIDs, which were observed at 30 weeks post-transplantation. GIDs were inhibited by D2 -like receptor antagonists and not affected by 5-HT1A/1B/5-HT6 agonists/antagonists. Grafts in GID rats showed more microglial activation and lacked serotonin neurons. CONCLUSIONS: These findings argue against current thinking that rats do not develop spontaneous GID and that serotonin neurons are causative, rather indicating that GID can be induced in rats by hESC-derived dopamine grafts and, critically, can occur independently of both previous levodopa exposure and grafted serotonin neurons. © 2021 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.
Assuntos
Discinesia Induzida por Medicamentos , Discinesias , Doença de Parkinson , Animais , Antiparkinsonianos/efeitos adversos , Dopamina , Discinesia Induzida por Medicamentos/etiologia , Discinesias/complicações , Humanos , Levodopa/efeitos adversos , Neurônios , Doença de Parkinson/complicações , Ratos , Ratos Sprague-Dawley , SerotoninaRESUMO
PEDOT: PSS is a commonly used hole-transport layer (HTL) in inverted perovskite solar cells (PSCs) due to its compatibility with low-temperature solution processing. However, it possesses lower conductivity than other conductive polymers and metal oxides, along with surface defects, limiting its photovoltaic performance. In this study, we introduced two-dimensional Ti3C2Tx (MXene) as an additive in the PEDOT:PSS HTL with varying doping concentrations (i.e., 0, 0.03, 0.05, and 0.1 wt.%) to tune the electrical conductivity of PEDOT:PSS and to modify the properties of the perovskite film atop it. We noted that the grain size of the CH3NH3PbI3 (MAPI3) perovskite layer grown over an optimal concentration of MXene (0.03 wt.%)-doped PEDOT:PSS increased from 250 nm to 400 nm, reducing charge recombination due to fewer grain boundaries. Ultraviolet photoelectron spectroscopy (UPS) revealed increased work function (WF) from 4.43 eV to 4.99 eV with 0.03 wt.% MXene doping, making the extraction of holes easier due to a more favorable energy level alignment with the perovskite. Quantum chemical investigations based on density functional theory (DFT) were conducted at the ωB97XD/6-311++G(d,p) level of theory to provide more insight into the stability, bonding nature, and optoelectronic properties of the PEDOT:PSS-MXene system. The theoretical investigations revealed that the doping of PEDOT:PSS with Ti3C2Tx could cause a significant effect on the electronic properties of the HTL, as experimentally demonstrated by an increase in the electrical conductivity. Finally, the inverted PSCs employing 0.03 wt.% MXene-doped PEDOT:PSS showed an average power conversion efficiency (PCE) of 15.1%, up from 12.5% for a reference PSC employing a pristine PEDOT:PSS HTL. The champion device with a 0.03 wt.% MXene-PEDOT:PSS HTL achieved 15.5% PCE.
RESUMO
Helper T lymphocytes (HTLs) play a central role in cancer immunity because they can not only help the induction and proliferation of cytotoxic T lymphocytes (CTLs) but also their differentiation into cytotoxic CD4+ T cells and directly kill the target cells.This study describes the identification of three novel mouse Th epitope peptides, WT135-52, WT186-102 and WT1294-312, derived from WT1 protein, which is the most potent tumor-associated antigen. Compared to immunization with WT1 CTL peptide alone, immunization with the addition of these WT1-specific Th peptides strongly induced WT1-specific CTLs, continued to maintain them, and efficiently rejected the challenge of WT1-expressing tumor cells. Importantly, the majority of WT1-specific CTLs induced by the co-immunization with WT1 CTL and the WT1-specific Th peptides were CD44+CD62L- effector memory CD8+ T cells, which played a central role in tumor rejection. Establishment of mouse models suitable for the analysis of the detailed mechanism of these functions of HTLs is very important. These results clearly showed that WT1-specific HTLs perform an essential function in WT1-specific tumor immunity. Therefore, the WT1-specific Th peptides identified here should make a major contribution to elucidation of the mutual roles of WT1-specific CTLs and HTLs in cancer immunity in in vivo mouse models.
Assuntos
Antígenos de Neoplasias/imunologia , Epitopos de Linfócito T/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Proteínas WT1/imunologia , Animais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Linfócitos T Citotóxicos/imunologiaRESUMO
Salmonella enterica serovar Typhi (S. Typhi), a causative agent of typhoid fever, is a Gram-negative, human-restricted pathogen that causes significant morbidity and mortality, particularly in developing countries. The currently available typhoid vaccines are not recommended to children below six years of age and have poor long-term efficacy. Due to these limitations and the emerging threat of multidrug-resistance (MDR) strains, the development of a new vaccine is urgently needed. The present study aims to design a multiepitope-based subunit vaccine (MESV) against MDR S. Typhi str. CT18 using a computational-based approach comprising subtractive proteomics and immunoinformatics. Firstly, we investigated the proteome of S. Typhi str. CT18 using subtractive proteomics and identified twelve essential, virulent, host non-homologous, and antigenic outer membrane proteins (OMPs) as potential vaccine candidates with low transmembrane helices (≤1) and molecular weight (≤110 kDa). The OMPs were mapped for cytotoxic T lymphocyte(CTL) epitopes, helper T lymphocyte (HTL) epitopes, and linear B lymphocyte (LBL) epitopes using various immunoinformatics tools and servers. A total of 6, 12, and 11 CTL, HTL, and LBL epitopes were shortlisted, respectively, based on their immunogenicity, antigenicity, allergenicity, toxicity, and hydropathicity potential. Four MESV constructs (MESVCs), MESVC-1, MESVC-2, MESVC-3, and MESVC-4, were designed by linking the CTL, HTL, and LBL epitopes with immune-modulating adjuvants, linkers, and PADRE (Pan HLA DR-binding epitope) sequences. The MESVCs were evaluated for their physicochemical properties, allergenicity, antigenicity, toxicity, and solubility potential to ensure their safety and immunogenic behavior. Secondary and tertiary structures of shortlisted MESVCs (MESVC-1, MESVC-3, and MESVC-4) were predicted, modeled, refined, validated, and then docked with various MHC I, MHC II, and TLR4/MD2 complex. Molecular dynamics (MD) simulation of the final selected MESVC-4 with TLR4/MD2 complex confirms its binding affinity and stability. Codon optimization and in silico cloning verified the translation efficiency and successful expression of MESVC-4 in E. coli str. K12. Finally, the efficiency of MESVC-4 to trigger an effective immune response was assessed by an in silico immune simulation. In conclusion, our findings show that the designed MESVC-4 can elicit humoral and cellular immune responses, implying that it may be used for prophylactic or therapeutic purposes. Therefore, it should be subjected to further experimental validations.
Assuntos
Proteômica , Salmonella typhi , Criança , Biologia Computacional , Epitopos de Linfócito B/genética , Epitopos de Linfócito T/genética , Escherichia coli , Humanos , Simulação de Acoplamento Molecular , Salmonella typhi/genética , Vacinas de Subunidades AntigênicasRESUMO
Two small molecular hole-transporting type materials, namely 4-(9,9-dimethylacridin-10(9H)-yl)-N-(4-(9,9-dimethylacridin-10(9H)-yl)phenyl)-N-phenylaniline (TPA-2ACR) and 10,10'-(9-phenyl-9H-carbazole-3,6-diyl)bis(9,9-dimethyl-9,10-dihydroacridine) (PhCAR-2ACR), were designed and synthesized using a single-step Buchwald-Hartwig amination between the dimethyl acridine and triphenylamine or carbazole moieties. Both materials showed high thermal decomposition temperatures of 402 and 422 °C at 5% weight reduction for PhCAR-2ACR and TPA-2ACR, respectively. TPA-2ACR as hole-transporting material exhibited excellent current, power, and external quantum efficiencies of 55.74 cd/A, 29.28 lm/W and 21.59%, respectively. The achieved device efficiencies are much better than that of the referenced similar, 1,1-Bis[(di-4-tolylamino)phenyl]cyclohexane (TAPC)-based device (32.53 cd/A, 18.58 lm/W and 10.6%). Moreover, phenyl carbazole-based PhCAR-2ACR showed good device characteristics when applied for host material in phosphorescent OLEDs.
RESUMO
A series of novel allosteric antagonists of the GLP-1 receptor (GLP-1R), exemplified by HTL26119, are described. SBDD approaches were employed to identify HTL26119, exploiting structural understanding of the allosteric binding site of the closely related Glucagon receptor (GCGR) (Jazayeri et al., 2016) and the homology relationships between GCGR and GLP-1R. The region around residue C3476.36b of the GLP-1R receptor represents a key difference from GCGR and was targeted for selectivity for GLP-1R.
Assuntos
Receptor do Peptídeo Semelhante ao Glucagon 1/antagonistas & inibidores , Compostos Heterocíclicos/química , Regulação Alostérica/efeitos dos fármacos , Sítio Alostérico , Sequência de Aminoácidos , Desenho de Fármacos , Simulação de Acoplamento Molecular , Estrutura Molecular , Ligação Proteica , Receptores de Glucagon/antagonistas & inibidores , Transdução de Sinais , Relação Estrutura-AtividadeRESUMO
HTL/KAI2, a member of the α/ß-fold hydrolase superfamily, is known to be a receptor-like protein of lactone compounds and that triggers seed germination of Arabidopsis. However, the endogenous ligand and physiological roles of HTL/KAI2 have remained unclear. To understand the mechanism underlying seed germination involved in HTL/KAI2 signaling, it is necessary to identify the endogenous ligand of HTL/KAI2. To date, even a biosynthetic mutant of the ligand has not yet been isolated. Because exogenous agonistic chemicals can only be purchased in small amounts at high prices, the limited supply of those chemicals has hampered any large-scale experiments, such as mutant screening. Therefore, easily synthesized and scalable artificial agonist would remove the limitation of the chemical supply and contribute to the identification of the endogenous ligand of HTL/KAI2 and/or the biosynthetic mutants. In this study, we demonstrated that designed chemicals with a phenoxyfuranone scaffold potently stimulated seed germination via HTL/KAI2 in Arabidopsis. As a result of screening of these chemicals, we selected a representative compound with convincing selectivity. Here in, we provide a new promising synthetic agonist of HTL/KAI2.
Assuntos
Proteínas de Arabidopsis/agonistas , Arabidopsis/crescimento & desenvolvimento , Germinação , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Furanos/síntese química , Furanos/química , Furanos/farmacologia , Germinação/efeitos dos fármacos , Hidrolases/metabolismo , Ligantes , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Transdução de Sinais/efeitos dos fármacos , Relação Estrutura-Atividade , TemperaturaRESUMO
(ta-C) films coated through the filtered cathodic vacuum arc (FCVA) process as a hole transport layer (HTL) for perovskite solar cells (PSCs) and quantum dot light-emitting diodes (QDLEDs). The p-type ta-C film has several remarkable features, including ease of fabrication without the need for thermal annealing, reasonable electrical conductivity, optical transmittance, and a high work function. X-ray photoelectron spectroscopy and ultraviolet photoelectron spectroscopy examinations show that the electrical properties (sp3/sp2 hybridized bond) and work function of the ta-C HTL are appropriate for PSCs and QDLEDs. In addition, in order to correlate the performance of the devices, the optical, surface morphological, and structural properties of the FCVA-grown ta-C films with different thicknesses (5 ~ 20 nm) deposited on the ITO anode are investigated in detail. The optimized ta-C film with a thickness of 5 nm deposited on the ITO anode had a sheet resistance of 10.33 Ω-2, a resistivity of 1.34 × 10-4 Ω cm, and an optical transmittance of 88.97%. Compared to the reference PSC with p-NiO HTL, the PSC with 5 nm thick ta-C HTL yielded a higher power conversion efficiency (PCE, 10.53%) due to its improved fill factor. Further, the performance of QDLEDs with 5 nm thick ta-C hole injection layers (HIL) showed better than the performance of QDLEDs with different ta-C thicknesses. It is concluded that ta-C films have the potential to serve as HTL and HIL in next-generation PSCs and QDLEDs.
RESUMO
Tumor necrosis factor (TNF)-α is a pleiotropic cytokine with intense pro-inflammatory and immunomodulatory properties, and anti-TNF-α biologics are effective therapies for various inflammatory diseases such as inflammatory bowel disease (IBD) and sepsis. Snake venom, as a traditional Chinese medicine, has been used in the treatment of inflammatory diseases in China for centuries. In this research, we constructed a venom gland T7 phage display library of the sea snake Hydrophis cyanocinctus to screen bioactive compounds that antagonize TNF-α and identified a novel nine-amino-acid peptide, termed hydrostatin-TL1 (H-TL1). In enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) analyses, H-TL1 inhibited the interaction between TNF-α and TNF receptor 1 (TNFR1). Further, H-TL1 attenuated the cytotoxicity of TNF-α in L929 cells as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. H-TL1 also decreased the mRNA expression of TNF-α/TNFR1 downstream targets and suppressed the phosphorylation of well-characterized proteins of downstream signal transduction pathways in HEK-293 cells. In vivo data demonstrated that H-TL1 protects animals against dextran sodium sulfate (DSS)-induced acute colitis and lipopolysaccharide (LPS)-induced acute shock. Given its significant anti-inflammatory activity in vitro and in vivo, H-TL1 is a potential peptide for the development of new agents to treat TNF-α-associated inflammatory diseases.
Assuntos
Anti-Inflamatórios/farmacologia , Colite/tratamento farmacológico , Venenos Elapídicos/farmacologia , Oligopeptídeos/farmacologia , Receptores Tipo I de Fatores de Necrose Tumoral/antagonistas & inibidores , Proteínas de Répteis/farmacologia , Choque Séptico/tratamento farmacológico , Venenos de Serpentes/química , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Doença Aguda , Animais , Anti-Inflamatórios/síntese química , Anti-Inflamatórios/isolamento & purificação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Colite/induzido quimicamente , Colite/genética , Colite/patologia , Colubridae/metabolismo , Sulfato de Dextrana , Venenos Elapídicos/síntese química , Venenos Elapídicos/isolamento & purificação , MAP Quinases Reguladas por Sinal Extracelular/química , MAP Quinases Reguladas por Sinal Extracelular/genética , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Regulação da Expressão Gênica , Células HEK293 , Humanos , Lipopolissacarídeos , Camundongos , Oligopeptídeos/síntese química , Oligopeptídeos/isolamento & purificação , Biblioteca de Peptídeos , Fosforilação , Receptores Tipo I de Fatores de Necrose Tumoral/química , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Proteínas de Répteis/síntese química , Proteínas de Répteis/isolamento & purificação , Choque Séptico/induzido quimicamente , Choque Séptico/genética , Choque Séptico/patologia , Transdução de Sinais , Venenos de Serpentes/isolamento & purificação , Transcrição Gênica , Fator de Necrose Tumoral alfa/química , Fator de Necrose Tumoral alfa/genética , Proteínas Quinases p38 Ativadas por Mitógeno/química , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
The human endothelin receptors are members of the rhodopsin class A of G-protein coupled receptors and key modulators of blood pressure regulation. Their functional in vitro characterization has widely been limited by the availability of high quality samples. We have optimized cell-free expression protocols for the human endothelin A and endothelin B receptors by implementing co-translational association approaches of the synthesized proteins with supplied liposomes or nanodiscs. Efficiency of membrane association and ligand binding properties of the receptors have systematically been studied in correlation to different membrane environments and lipid types. Ligand binding was analyzed by a number of complementary assays including radioassays, surface plasmon resonance and fluorescence measurements. High affinity binding of the peptide ligand ET-1 to both endothelin receptors could be obtained with several conditions and the highest Bmax values were measured in association with nanodiscs. We could further obtain the characteristic differential binding pattern of the two endothelin receptors with a panel of selected agonists and antagonists. Two intrinsic properties of the functionally folded endothelin B receptor, the proteolytic processing based on conformational recognition as well as the formation of SDS-resistant complexes with the peptide ligand ET-1, were observed with samples obtained from several cell-free expression conditions. High affinity and specific binding of ligands could furthermore be obtained with non-purified receptor samples in crude cell-free reaction mixtures, thus providing new perspectives for fast in vitro screening applications.
Assuntos
Endotelina-1/química , Lipossomos/química , Receptor de Endotelina A/química , Receptor de Endotelina B/química , Sistema Livre de Células/metabolismo , Detergentes/química , Endotelina-1/metabolismo , Expressão Gênica , Humanos , Cinética , Nanoestruturas/química , Ligação Proteica , Dobramento de Proteína , Receptor de Endotelina A/biossíntese , Receptor de Endotelina A/genética , Receptor de Endotelina B/biossíntese , Receptor de Endotelina B/genéticaRESUMO
Cancer immunotherapy has an outstanding position in cancer prevention and treatment. In this kind of therapy, the immune system is activated to eliminate cancerous cells. Multi-epitope peptide cancer vaccines are manifesting as the next generation of cancer immunotherapy. In the present study, we have implemented various strategies to design an efficient multi-epitope vaccine. CD8+ cytolytic T lymphocytes (CTLs) epitopes, which have a pivotal role in cellular immune responses, helper epitopes and adjuvant, are three crucial components of peptide vaccine. CTL epitopes were determined from two high immunogenic protein Wilms tumor-1 (WT1) and human papillomavirus (HPV) E7 by various servers, which apply different algorithms. CTL epitopes were linked together by AAY and HEYGAEALERAG motifs to enhance epitope presentation. Pan HLA DR-binding epitope (PADRE) peptide sequence and helper epitopes, which have defined from Tetanus toxin fragment C (TTFrC) by various servers, were used to induce CD4+ helper T lymphocytes (HTLs) responses. Additionally, helper epitopes were conjugated together via GPGPG motifs that stimulate HTL immunity. Heparin-Binding Hemagglutinin (HBHA), a novel TLR4 agonist was employed as an adjuvant to polarize CD4+ T cells toward T-helper 1 to induce strong CTL responses. Moreover, the EAAAK linker was introduced to N and C terminals of HBHA for efficient separation. 3D model of protein was generated and predicted B cell epitopes were determined from the surface of built structure. Our protein contains several linear and conformational B cell epitopes, which suggests the antibody triggering property of this novel vaccine. Hence, our final protein can be used for prophylactic or therapeutic usages, because it can potentially stimulate both cellular and humoral immune responses.
Assuntos
Vacinas Anticâncer/imunologia , Simulação por Computador , Epitopos/imunologia , Vacinas de Subunidades Antigênicas/imunologia , Sequência de Aminoácidos , Sítios de Ligação , Vacinas Anticâncer/química , Biologia Computacional , Epitopos/química , Epitopos de Linfócito B/química , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/imunologia , Antígenos HLA/química , Antígenos HLA/imunologia , Humanos , Ligantes , Modelos Moleculares , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/imunologia , Estrutura Secundária de Proteína , Reprodutibilidade dos Testes , Análise de Sequência de Proteína , Homologia de Sequência de Aminoácidos , Linfócitos T Citotóxicos/imunologia , Vacinas de Subunidades Antigênicas/químicaRESUMO
In perovskite solar cells (PSCs), the charge carrier recombination obstacles mainly occur at the ETL/perovskite and HTL/perovskite interfaces, which play a decisive role in the solar cell performance. Therefore, this study aims to enhance the flexible PSC (FPSC) efficiency by adding the newly designed CBz-PAI-interlayer (simply CBz-PAI-IL) at the perovskite/HTL interface. In addition, substantial work has been carried out on five different HTLs (Se/Te-Cu2O, CuGaO2, V2O5, and CuSCN, including conventional Spiro-OMeTAD as a reference HTL with and without CBz-PAI-IL), using drift-diffusion simulation to find suitable FPSC design to attain the maximum PCE. Interestingly, PET/ITO/AZO/ZnO NWs/FACsPbBrI3/CBz-PAI/Se/Te-Cu2O/Au device architecture demonstrates the highest achievable power conversion efficiency (PCE) of 27.9 %. The findings of this study confirmed that the reference device (without IL) displays a large valence band edge (VBE)/highest occupied molecular orbital (HOMO) energy level misalignment compared to the modified interface device (with CBz-PAI-IL that reduces VBE/HOMO level mismatch) that eases the hole transport, simultaneously, it reduces the charge carrier recombinations at the interface, resulting in diminished Voc losses in the device. Furthermore, the influence of perovskite absorber thickness and defect density, parasitic resistances, and working temperature are systematically examined to govern the superior FPSC efficiency and concurrently understand the device physics.
RESUMO
Researchers have recently shown a great deal of interest in molybdenum diselenide (MoSe2)-based solar cells due to their outstanding semiconducting characteristics. However, discrepancies in the band arrangement at the MoSe2/ETL (electron transport layer) and hole transport layer (HTL)/MoSe2 interfaces impede performances. In this research, a device combination with Ag/FTO/ETL/MoSe2/HTL/Ni is employed, where 7 HTLs and 3 different ETLs have been utilized to explore which device arrangement is superior. To achieve the most effective device arrangement, the effects of various device variables, such as thickness, donor density, acceptor density, defect density, temperature, series, and shunt resistance, are optimized. The computational evaluation under AM 1.5 light spectrums (100 mW/cm2) is performed using the SCAPS-1D simulator. When the several device parameters were optimized, the device that was correlated with Ag/FTO/SnS2/MoSe2/V2O5/Ni revealed the highest overall performances among the three different ETL (In2S3, SnS2, ZnSe)-based devices, with measuring a PCE of 34.07 %, a VOC of 0.918 V, a JSC of 42.565 mAcm-2, and an FF of 87.19 %. This recommended MoSe2-based solar cell exhibits outstanding efficiency in terms of maintenance and comparison to numerical thin film solar cells, highlighting MoSe2 as an attractive option for solar energy systems while eliminating toxicity challenges.
RESUMO
Perovskite solar cells (PSCs) have attracted increasing research interest, but their performance depends on both the choice of materials and the process used. The materials can typically be treated in solution, which makes them well suited for roll-to-roll processing methods, but their deposition under ambient conditions requires overcoming some challenges to improve stability and efficiency. In this review, we highlight the latest advancements in photonic curing (PC) for perovskite materials, as well as for hole transport layer (HTL) and electron transport layer (ETL) materials. We present how PC parameters can be used to control the optical, electrical, morphological, and structural properties of perovskite HTL and ETL layers. Emphasizing the significance of these advancements for perovskite solar cells could further highlight the importance of this research and underline its essential role in creating more efficient and sustainable solar technology.