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1.
Food Funct ; 2021 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-33528481

RESUMO

Food processing can change the structure and immunoreactivity of purified allergens, but the effect of food processing on the immunoreactivity of the processed and purified allergen is still poorly understood. In this study, tropomyosin (TM) was obtained from Scylla paramamosain and purified after different treatments. A basophil activation test was employed to detect the allergenicity of allergens. The protein structure was detected by mass spectrometry, circular dichroism spectroscopy and surface hydrophobicity. Critical amino acids were identified by Dot blot. Heating obviously affects the biochemical characteristics of TM. The allergenicity of TM was decreased in high temperature-pressure-processed crabs, due to alteration in the protein structure (e.g. denaturation). Seven critical amino acids, namely, R21, E103, E104, E115, A116, E122 and E156, related to the maintenance of the IgE-binding activity of TM were identified. This research of thermal processing helps to accurately reduce or eliminate the immunoreactivity of crabs by food processing.

2.
Bioorg Chem ; 108: 104671, 2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33550072

RESUMO

Andrastones are unusual 6,6,6,5-tetracyclic meroterpenoids that are rarely found in nature. Previously, three andrastones were obtained from the rice static fermentation extract of the deep-sea-derived fungus Penicillium allii-sativi MCCC 3A00580. Inspired by one strain many compounds (OSMAC) approach, the oat static fermentation on P. allii-sativi was conducted. As a result, 14 andrastones were isolated by UV-guided isolation. The chemical structures of the nine new compounds (1-9) was established by comprehensive analysis of the NMR, MS, ECD, and X-ray crystallography and the five known ones (10-14) were assigned by comparing their NMR, MS, and OR data with those reported in literature. Compound 1 bears a novel hemiketal moiety while 2 is the first example to possess a novel tetrahydrofuran moiety via C-7 and C-15. All isolates were tested for anti-allergic bioactivity. Compound 10, 3-deacetylcitreohybridonol, significantly decreased degranulation with the IC50 value of 14.8 µM, compared to that of 92.5 µM for the positive control, loratadine. Mechanism study indicated 10 could decrease the generation of histamine and TNF-α by reducing the accumulation of Ca2+ in RBL-2H3 cells. These findings indicate andrastones could be potential to discover new anti-allergic candidate drugs.

3.
Langmuir ; 2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33499598

RESUMO

The study of ion specificities of charged-neutral random copolymers is of great importance for understanding specific ion effects on natural macromolecules. In the present work, we have investigated the specific anion effects on the thermoresponsive behavior of poly([2-(methacryloyloxy)ethyl trimethylammonium chloride]-co-N-isopropylacrylamide) [P(METAC-co-NIPAM)] random copolymers. Our study demonstrates that the anion specificities of the P(METAC-co-NIPAM) copolymers are dependent on their chemical compositions. The specific anion effects on the copolymers with high mole fractions of poly(N-isopropylacrylamide) (PNIPAM) are similar to those on the PNIPAM homopolymer. As the mole fraction of PNIPAM decreases to a certain value, a V-shaped anion series can be observed in terms of the anion-specific cloud point temperature of the copolymer, as induced by the interplay between different anion-polymer interactions. Our study also suggests that both the direct and the indirect anion-polymer interactions contribute to the anion specificities of the copolymers. This work would improve our understanding of the relationship between the ion specificities and the ion-macromolecule interactions for naturally occurring macromolecules.

4.
Med Sci Monit ; 27: e928374, 2021 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-33388740

RESUMO

BACKGROUND Although influenza primarily affects the respiratory system, it can cause severe neurological complications, especially in younger children, but knowledge about the early indicators of acute necrotizing encephalopathy (ANE) is limited. The main purpose of this article is to summarize the clinical characteristics, diagnosis, and treatment of neurological complications of influenza in children, and to identify factors associated with ANE. MATERIAL AND METHODS This was a retrospective study of children with confirmed influenza with neurological complications treated between 01/2014 and 12/2019 at Guangzhou Women and Children's Medical Center. A receiver operating characteristics curve analysis was performed to determine the prognostic value of selected variables. RESULTS Sixty-three children with IAE (n=33) and ANE (n=30) were included. Compared with the IAE group, the ANE group showed higher proportions of fever and acute disturbance of consciousness, higher alanine aminotransferase, higher aspartate aminotransferase, higher creatinine kinase, higher procalcitonin, higher cerebrospinal fluid (CSF) protein, and lower CSF white blood cells (all P<0.05). The areas under the curve (AUCs) for procalcitonin and CSF proteins, used to differentiate IAE and ANE, were 0.790 and 0.736, respectively. The sensitivity and specificity of PCT >4.25 ng/ml to predict ANE were 73.3% and 100.0%, respectively. The sensitivity and specificity of CSF protein >0.48 g/L to predict ANE were 76.7% and 69.7%, respectively. Thirteen (43.3%) children with ANE and none with IAE died (P<0.0001). CONCLUSIONS High levels of CSF protein and serum procalcitonin might be used as early indicators for ANE. All children admitted with neurological findings, especially during the influenza season, should be evaluated for influenza-related neurological complications.

5.
J Agric Food Chem ; 69(1): 428-436, 2021 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-33377774

RESUMO

In order to reduce the immunoreactivity of sarcoplasmic calcium-binding protein (SCP), site-directed mutations were used to replace key amino acids in the conformational epitopes and calcium-binding sites. The mutant SCPs (mSCPs) were expressed in Escherichia coli, and their immunoreactivities were analyzed using iELISA and basophil activation assays. Furthermore, the structural changes of mSCPs were determined from the circular dichroism spectra. The iELISA results showed that mSCPs could effectively inhibit the binding of wild-type SCP (wtSCP) to sensitive serum, with inhibition rates that reached 90%. Moreover, mSCPs could downregulate the expression levels of CD63 and CD203c on the basophil surface. Compared with wtSCP, the peak values were significantly changed, and the calcium binding ability was impaired, which explained the decline in immunoreactivities of the mSCPs. All of the data confirmed that this approach was effective in reducing the immunoreactivity of SCP and could be applied to other shellfish allergens.

6.
Food Chem ; : 128735, 2020 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-33279350

RESUMO

Mud crab (Scylla paramamosain) is widely consumed after thermal processing. It is necessary to comprehensively evaluate of the allergenic potential and epitopes of allergens in high temperature-pressure (HTP) treated S. paramamosain. Tropomyosin and arginine kinase presented higher prevalence (30.77% and 42.13%) than the other three important crab allergens by component-resolved diagnosis. The surface expression of basophils CD63 and CD203c were decreased in HTP treated crab, an effect that was even more evident after digestion and absorption by the intestinal Caco-2 cell model. Of the 35 stable epitope, six were for the first time identified in shellfish. Seven heat/digested stable peptides of tropomyosin retained IgE-binding capacity and were shown to interact with MHC-II. Five epitopes (amino acids 19-29, 99-109, 153-162, 170-188 and 211-221) were the first identified in crab. The study provides insight into prevention and therapy of crab allergy, as well as helps to reduce crab allergenicity during thermal processing.

7.
Med Sci Monit ; 26: e928835, 2020 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-33335084

RESUMO

BACKGROUND This study summarizes the characteristics of children screened for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and reports the case of 1 child who was diagnosed with SARS-CoV-2 infection in Guangzhou Women and Children's Medical Center and the cases of his family members. MATERIAL AND METHODS The medical records of 159 children who were admitted to our hospital from January 23 to March 20, 2020, were retrospectively analyzed. Samples from pharyngeal or/and anal swabs were subjected to reverse-transcription polymerase chain reaction (RT-PCR) testing for SARS-CoV-2 within 12 h of patient admission; a second RT-PCR test was done 24 h after the first test. RESULTS Of the 159 patients, 151 patients had epidemiological histories, 14 patients had cluster onset, and 8 patients had no epidemiological history but had symptoms similar to coronavirus disease 2019 (COVID-19). The most common symptom was fever (n=125), followed by respiratory and gastrointestinal symptoms. A 7-year-old boy in a cluster family from Wuhan was confirmed with asymptomatic SARS-CoV-2 infection with ground-glass opacity shadows on his lung computed tomography scan, and his swab RT-PCR test had not turned negative until day 19 of his hospitalization. In patients who did not test positive for SARS-CoV-2, influenza, respiratory syncytial virus, and adenovirus were observed. A total of 158 patients recovered, were discharged, and experienced no abnormalities during follow-up. CONCLUSIONS For SARS-CoV-2 nosocomial infections, taking a "standard prevention & contact isolation & droplet isolation & air isolation" strategy can prevent infection effectively. Children with clustered disease need close monitoring.


Assuntos
/diagnóstico , /epidemiologia , /métodos , Criança , Pré-Escolar , China/epidemiologia , Coronavirus/metabolismo , Coronavirus/patogenicidade , Infecção Hospitalar/epidemiologia , Feminino , Febre , Hospitalização , Hospitais , Humanos , Masculino , Registros Médicos , Alta do Paciente , Estudos Retrospectivos , /patogenicidade
8.
Food Funct ; 2020 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-33226057

RESUMO

Nevadensin (NEV), a natural flavonoid compound derived from Lysionotus pauciflorus Maxim, has numerous biological activities. However, few researchers have examined its potential impact on alleviating allergies. In the present study, NEV was found to upregulate rectal temperature, suppress the development of diarrhea, and decrease the levels of serum specific immunoglobulin E, histamine and mouse MC protease-1 in ovalbumin-allergic mice. Moreover, NEV also alleviated passive cutaneous anaphylaxis reactions and inhibited the release of ß-hexosaminidase and histamine in bone marrow-derived mast cells. Furthermore, we provide the first demonstration that NEV decreases the expression of c-Kit and suppresses the proliferation of bone marrow-derived mast cells and accelerates their apoptosis. These findings indicated that L. pauciflorus-derived NEV might have the potential to alleviate food hypersensitivity.

9.
Int J Biol Macromol ; 2020 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-33144260

RESUMO

The ameliorative effect of depolymerized sulfated polysaccharides from Eucheuma serra (DESP) on ovalbumin (OVA)-caused induced food allergy was investigated in this work. Results showed that OVA stimulated the secretion of allergy-related cytokines (OVA-specific IgE, mMCP-1, IgA, TNF-α) and led to diarrhea, intestinal epithelial damage, and intestinal microflora dysbiosis in sensitized mice. After the administration of DESP, however, the anaphylactic symptoms (shortness of breath, hypothermia, diarrhea), along with the allergy-related cytokines, were effectively suppressed. Moreover, the reduced intestinal inflammation was discovered in the DESP-treated group. Additionally, 16S rRNA sequencing of fecal samples was performed, and gene count and α-diversity analysis revealed that DESP improved microbial community richness. Taxonomic composition analysis showed that DESP modulated the proportion of Firmicutes and Bacteroidetes/Proteobacteria. Particularly, DESP increased probiotics (Lactobacillaceae, Bifidobacteriaceae and Prevotellaceae) and decreased pathogenic bacteria (Helicobacteraceae and Desulfovibrionaceae). These findings, therefore, suggest that DESP may ameliorate food allergy through the regulation of intestinal microbiota.

10.
Langmuir ; 36(43): 13051-13059, 2020 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-33094611

RESUMO

In this work, a negatively charged poly(3-sulfopropyl methacrylate potassium) (PSPMA) brush has been employed as a model system to demonstrate the tuning of the light response of strong polyelectrolyte brushes (SPBs) with counterions. The substitution of K+ counterions by azobenzene-containing counterions (Azo-N+) renders the PSPMA brush light-responsive in aqueous solutions. Nevertheless, the strength of the light response of the PSPMA brush is weak due to the inefficient disassembly of the micelle-like aggregates in the brush upon irradiation with ultraviolet light. Counterion mixtures of Azo-N+ and K+ are employed to realize a strong light response of the PSPMA brush by incorporating a reasonable amount of Azo-N+ counterions into the brush. The strength of the light response of the PSPMA brush can be tuned by the mole ratio of Azo-N+ to K+. Furthermore, properties including the hydration and conformation of the PSPMA brush can be reversibly switched via alternating ultraviolet and visible light irradiation. This work opens up the opportunities available for the use of counterions to tune the light response of SPBs.

11.
Cell Cycle ; 19(22): 3182-3194, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33121314

RESUMO

Tuberculosis is one of the most important infectious diseases worldwide and macrophage apoptosis is the major host defense mechanism against TB. We attempted to characterize the role of miRNA (miR)-125b-5p on mycobacterium tuberculosis (Mtb) infection and macrophages behaviors in vitro. According to fluorescence-activated cell separation (FACS), primary monocytes (CD14+) in TB patients were accumulated, and apoptotic monocytes were decreased. Peripheral blood mononuclear cells (PBMCs)-derived macrophages (MDMs) and monocytic cells THP-1-derived macrophage-like cells (TDMs) in vitro were used to be infected with H37Rv. After infection, colony-forming units assay revealed the increase of bacterial activity, FACS demonstrated the decrease of apoptosis rate of MDMs and TDMs, as well as promoted levels of IL-6, TNF-α, Bax, and Bim and suppressed levels of IL-10 and Bcl-2, examined by enzyme-linked immunosorbent assay (ELISA) and western blot assay. Expression of miR-125b-5p and DNA damage-regulated autophagy modulator 2 (DRAM2) was examined, and real-time PCR and western blot assay showed that miR-125b-5p was upregulated, whereas DRAM2 was downregulated in primary monocytes and H37Rv-infected macrophages (MDMs and TDMs). Moreover, blocking miR-125b-5p could attenuated H37Rv-induced bacterial activity and inflammatory response of MDMs and TDMs, accompanied with apoptosis inhibition. Whereas these effects of miR-125b-5p knockdown were abolished by downregulating DRAM2. In mechanism, DRAM2 was a downstream target of miR-125b-5p, as evidenced by dual-luciferase reporter assay. Collectively, silencing miR-125b-5p could protect human macrophages against Mtb infection through promoting apoptosis and inhibiting inflammatory response via targeting DRAM2, suggesting a novel target for Mtb eliminating. Abbreviations: TB: tuberculosis; PBMCs: peripheral blood mononuclear cells; Mtb: mycobacterium tuberculosis; AFB: acid fast bacilli; FITC: fluorescein isothiocyanate; MDMs: monocytes-derived macrophages; TDMs: THP-1-derived macrophage-like cells; ERFP: Mtb-enhanced red fluorescent protein; CFU: colony-forming units; ELISA: enzyme-linked immunosorbent assay; FACS: fluorescence-activated cell separation; PI: propidium iodide; DRAM2: DNA damage-regulated autophagy modulator 2; Real-time PCR: real-time polymerase chain reaction; in-miR-125b-5p: miR-125b-5p inhibitor; si-DRAM2: siRNA against DRAM2.

12.
Mar Drugs ; 18(10)2020 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-33081290

RESUMO

Viridicatol is a quinoline alkaloid isolated from the deep-sea-derived fungus Penicillium griseofulvum. The structure of viridicatol was unambiguously established by X-ray diffraction analysis. In this study, a mouse model of ovalbumin-induced food allergy and the rat basophil leukemia (RBL)-2H3 cell model were established to explore the anti-allergic properties of viridicatol. On the basis of the mouse model, we found viridicatol to alleviate the allergy symptoms; decrease the levels of specific immunoglobulin E, mast cell protease-1, histamine, and tumor necrosis factor-α; and promote the production of interleukin-10 in the serum. The treatment of viridicatol also downregulated the population of B cells and mast cells (MCs), as well as upregulated the population of regulatory T cells in the spleen. Moreover, viridicatol alleviated intestinal villi injury and inhibited the degranulation of intestinal MCs to promote intestinal barrier repair in mice. Furthermore, the accumulation of Ca2+ in RBL-2H3 cells was significantly suppressed by viridicatol, which could block the activation of MCs. Taken together, these data indicated that deep-sea viridicatol may represent a novel therapeutic for allergic diseases.

13.
Cell Transplant ; 29: 963689720963948, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33028110

RESUMO

Circular RNAs (circRNAs) could sponge micro-RNAs (miRNAs) to regulate tumor progression of hepatocellular carcinoma (HCC). Hsa_circ_104566 contributes to papillary thyroid carcinoma progression. However, the tumorigenic mechanism of hsa_circ_104566 on HCC remains enigmatic. The role of hsa_circ_104566 on HCC was therefore evaluated in this study. First, the high expression of hsa_circ_104566 was found in HCC tissues, which was significantly associated with poor prognosis in HCC patients. Second, Hsa_circ_104566 promoted HCC progression by decreasing apoptosis and E-cadherin, while increasing cell viability, proliferation, migration, invasion, and N-cadherin. On the other hand, HCC progression was suppressed by knockdown of hsa_circ_104566. Hsa_circ_104566 could target miR-338-3p, and its expression was negatively correlated with miR-338-3p in HCC patients. Moreover, miR-338-3p suppressed protein expression of Forkhead box protein 1 (FOXP1) and had a negative correlation with FOXP1 in HCC patients. Functional assay further indicated that the promotion of HCC progression by hsa_circ_104566 was reversed by miR-338-3p, and miR-338-3p inhibitor could counteract the effect of hsa_circ_104566 knockdown on the suppression of HCC progression. In vivo assay indicated that hsa_circ_104566 knockdown suppressed HCC tumor growth and metastasis. In conclusion, hsa_circ_104566 sponged miR-338-3p to promote HCC progression, providing a potential therapeutic target for cancer intervention.

14.
Int J Biol Sci ; 16(15): 2938-2950, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33061807

RESUMO

Large amounts of long non-coding RNAs (lncRNAs) have been annotated whereas most of them have not been functionally characterized. Here we identified lncRNA ENST00000441932 as an oncogenic lncRNA in esophageal squamous cell carcinoma (ESCC) and named lnc-MCEI (lncRNA mediated the chemosensitivity of ESCC by regulating IGF2). What's more, the effect of lnc-MCEI on the chemosensitivity of ESCC was further evaluated. Bioinformatics analysis demonstrated that lnc-MCEI was involved in the tumorigenesis of ESCC and lnc-MCEI levels were significantly increased in ESCC cells and tissues. Additionally, lnc-MCEI knockdown retarded cell proliferation, colony formation of ESCC cells, but induced cell apoptosis. Moreover, lnc-MCEI knockdown significantly improved the chemosensitivity of ESCC to cisplatin (DDP) both in vivo and in vitro. Further mechanisms disclosed that lnc-MCEI functioned as a competing endogenous RNA (ceRNA) via sponging miR-6759-5p and IGF2 was a target of miR-6759-5p. Meanwhile, we found that IGF2 suppressed chemosensitivity of ESCC cells via PI3K/AKT pathway. These data suggested that lnc-MCEI was an oncogenic lncRNA and lnc-MCEI knockdown enhanced chemosensitivity of ESCC cells to cisplatin by targeting miR-6759-5p /IGF2/PI3K/AKT axis.

15.
Int J Food Microbiol ; 335: 108891, 2020 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-32977153

RESUMO

Antimicrobial peptides are being explored for use as food preservatives to prevent foodborne diseases. In this study, bioinformatics tools were used to screen potential antimicrobial amino acid sequences from the whey acidic protein (WAP) of large yellow croaker (Larimichthys crocea). A novel antimicrobial peptide, designated as LCWAP, was identified and its antimicrobial effect and mechanism of action on Staphylococcus aureus was explored. The minimal inhibitory concentration (MIC) of LCWAP on S. aureus was 15.6 µg/mL. Transmission electron microscopy and laser confocal microscopy revealed that LCWAP kills bacteria by aggregating on the cell surface, destroying the integrity of bacterial cell membrane and resulting in the leakage of intracellular solutes. Moreover, peptide LCWAP inhibit biofilm formation, at concentrations of 1-1/16 × MIC, with biofilm formation found to decrease by 94.3%-13.7% upon LCWAP treatment. The ability of peptide LCWAP to bind bacteria DNA was revealed using electrophoresis analysis and ultraviolet absorption spectroscopy, with peptide LCWAP/DNA weight ratios of 125/1, and 17.3% decrease in the absorption peak of LCWAP. Furthermore, LCWAP had no cytotoxic effects on normal human hepatocytes, although it had strong inhibitory effect on S. aureus growth in milk.


Assuntos
Antibacterianos/farmacologia , Proteínas do Leite/farmacologia , Leite/microbiologia , Perciformes , Staphylococcus aureus/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Antibacterianos/química , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Membrana Celular/efeitos dos fármacos , Sobrevivência Celular , DNA Bacteriano/efeitos dos fármacos , DNA Bacteriano/metabolismo , Conservantes de Alimentos/química , Conservantes de Alimentos/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Proteínas do Leite/química , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Fragmentos de Peptídeos/farmacologia , Staphylococcus aureus/crescimento & desenvolvimento
16.
Chem Commun (Camb) ; 56(74): 10930-10933, 2020 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-32940269

RESUMO

Herein, we have demonstrated that the properties of both strong and weak polyelectrolyte brushes including hydration, stiffness, conformation, and wettability can be tuned by the hydrogen bonding between the bound counterions and the grafted chains beyond the hydronium and hydroxide ions. This will greatly extend the application of ionic hydrogen bond effects in polymer systems.

17.
Mar Drugs ; 18(8)2020 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-32751049

RESUMO

Seaweed sulfated polysaccharides have attracted significant attention due to their antibacterial activity. This work investigated the antibacterial activity and mechanism of depolymerized sulfated galactans from Eucheuma serra (E. serra) and Gracilaria verrucosa (G. verrucosa) against enterotoxigenic Escherichia coli (ETEC) K88. The results show that removing the metal ions improves the anti-ETEC K88 activity of the galactans. The fluorescence labeling study confirmed that the sulfated galactans penetrated the cell walls and eventually reached the interior of the ETEC K88. Nucleic acid staining and intracellular protein leakage were also observed, indicating the destruction of permeability and integrity of the cell membrane. Interestingly, the two polysaccharides exhibited no effect on the proliferation of the selected Gram-positive bacteria and yeast. This indicates that the cell wall structure of the microorganisms could influence the bacteriostatic activity of the sulfated polysaccharides, as well. These results suggest that the sulfated seaweed polysaccharides might have potential application value in antibacterial diarrhea.

18.
J Colloid Interface Sci ; 579: 369-378, 2020 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-32615480

RESUMO

HYPOTHESIS: A detailed understanding of the influence of electrolytes on the conformation of polyelectrolyte chains is an important goal made challenging by the strong coupling between electrostatic interactions and chain conformation. This challenge is particularly evident at moderate to high salt concentrations where mean-field theories of electrolytes are no longer applicable and are therefore unable to predict the interactions between neutral or like charged surfaces that leads to re-entrant swelling of DNA and other polyelectrolytes at high salt concentrations. Recent developments arising from studies of surface forces in ionic liquids that have been extended to include a wide variety of monovalent electrolytes reveal a hitherto unknown increase in the electrostatic decay length at high electrolyte concentrations. We hypothesise that the re-entrant behaviour of polyelectrolytes is driven by an increasing electrostatic decay length with increasing electrolyte concentration. EXPERIMENTS: We survey numerous experiments in the literature on re-entrant swelling and calculate the effect of ion pairing on the electrostatic decay length in concentrated electrolytes. FINDINGS: Re-entrant solubility is driven by an increasing electrostatic decay length at high salt concentrations and is universal across all polyelectrolytes.

19.
Food Chem ; 333: 127452, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-32673951

RESUMO

Aimed to study the characteristics of prolyl endopeptidase (PEP, EC 3.4.21.26) and its possible role in the degradation of collagen, we cloned the full-length cDNA sequence of PEP from abalone (Haliotis discus hannai) (Hdh-PEP). Recombinant Hdh-PEP (rHdh-PEP) was expressed in vitro, its enzymatic properties were detected, and its secondary structure was analyzed by Circular Dichroism (CD). We for the first time determined the 1.5 Å crystal structure of rHdh-PEP. The decomposition effect of rHdh-PEP on collagen peptides was analyzed. Our data revealed that the molecular weight of rHdh-PEP is 85 kDa, consisting of a catalytic domain and a ß-propeller domain. The optimal pH and temperature of rHdh-PEP were pH 6.0 and 20 °C, respectively. Using small collagen peptides as substrates, HPLC-ESI-MS analysis confirmed that rHdh-PEP specifically cleaved at the carboxyl side of proline residues, suggesting its role in the degradation of collagen peptides during autolysis.


Assuntos
Colágeno/metabolismo , Gastrópodes/enzimologia , Serina Endopeptidases/química , Serina Endopeptidases/metabolismo , Animais , Dicroísmo Circular , Cristalografia por Raios X , DNA Complementar/genética , Concentração de Íons de Hidrogênio , Prolina/metabolismo , Estrutura Secundária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Serina Endopeptidases/genética , Espectrometria de Massas por Ionização por Electrospray , Temperatura
20.
Microb Pathog ; 147: 104302, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32504846

RESUMO

Vibrio parahaemolyticus is a Gram-negative bacterium and the one of leading causal agent of human foodborne diseases such as gastroenteritis upon consumption of raw, or contaminated marine products. There is an increased interest in the use of antimicrobial peptides (AMPs) as alternative food preservatives to prevent foodborne diseases. In this study, bioinformatics tools were used to predict and screen AMPs derived from hemoglobin of blood clam (Tegillarca granosa). A novel AMP, T. granosa hemoglobin-derived peptide (TGH1), was identified and its antimicrobial effect and mechanism of action on V. parahaemolyticus was explored. The minimal inhibitory concentration (MIC) of TGH1 on V. parahaemolyticus was 12.5 µg/mL. Transmission electron microscopy (TEM) revealed that TGH1 kills bacteria by perforating the cell wall perforation and destroying integrity of the cell membrane. Similarly, laser confocal microscopy confirmed that TGH1 entered bacterial cells by aggregating on the cell surface to destroy the cell. In addition, TGH1 increased the inner-membrane permeability of V. parahaemolyticus in a concentration-dependent manner, as well as prevented biofilm formation. Moreover, TGH1 has 55.6% ß-sheet (antiparallel) structure and has no cytotoxic effects on normal human hepatocytes. Thus, peptide TGH1 has good potential use and application in antimicrobial control of foodborne pathogens.

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