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1.
Biomed Pharmacother ; 143: 112087, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34474339

RESUMO

Stress during pregnancy is not only detrimental to a woman's own physical and mental health, but can also cause changes in the intrauterine environment and even have an impact on later growth and development, this study was designed to understand the changes of gut microbiota in the maternal and offspring caused by prenatal chronic stress, and to explore the regulatory effect of LBP on gut microbiota, and then to improve the emotional damage caused by prenatal chronic stress in the offspring. A rat model of prenatal chronic stress was made and used LBP to intervene by gavage. Fresh feces of offspring were collected, the concentration of microbial metabolites were tested by ELISA. Illumina MiSeqPE300 sequencing technology was used to determine the sequence of 16S rRNA V3-V4 of microorganisms. On the PND 42, the emotional function of offspring were tested by open-field test (OFT), sucrose preference test (SPT) and tail of suspend test (TST). Results indicated that stress factors increased the plasma corticosterone level of rats during pregnancy and they appeared depressive behaviors. The body weight of offspring during prenatal chronic stress was lower than the control group, and the plasma corticosterone level was increased. Prenatal chronic stress had a significant impact on emotional performance of the offspring on OFT, SPT and TST. Alpha diversity of gut microbiota and microbiota composition in offspring of prenatal chronic stress was attenuated and some relationships existed between these parameters. LBP treatment reduced offspring's plasma corticosterone level and improved their body weight, changed the emotional function, increased the diversity of gut microbiota. Collectively, these findings disclose that prenatal chronic stress not only causes emotional injury on the offspring, but also changes the gut microbiota of the mother and offspring; LBP may regulate the intestinal flora of the mother, then reducing the influence of stress factors on the emotional injury of offspring.

2.
Int J Biol Macromol ; 188: 313-322, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34339788

RESUMO

In today's world, cancer is still the leading cause of human death. Among them, the incidence and mortality of lung cancer remain high, and have become the focus of research in the world. BEAP, a protein with anti-lung cancer activity, was isolated and purified from the edible mushroom Boletus edulis. Previous studies have shown that BEAP can inhibit the proliferation of non-small cell lung cancer A549 cells by inducing apoptosis and cell cycle arrest in vitro and in vivo. However, there are many ways in which antitumor proteins from edible and medicinal mushroom play their roles. It is worth exploring whether there are other antitumor mechanisms of BEAP, which can provide reference value for the development of new drugs targeting non-small cell lung cancer and the repurposing of existing drugs.

3.
J Food Biochem ; 45(7): e13818, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34121192

RESUMO

In this study, through the combination of AB-8 macroporous resin, Sephadex LH-20 column chromatography and semi-preparative HPLC, an antioxidant component was purified from the crude extract of Phellinus pini, thereby evaluating the cardioprotective effect of the fraction. As a result, total phenolic content of the 60% ethanol elution was increased by 4.8-fold after one run treatment on Sephadex LH-20 chromatography with gradient elution. After semi-preparative HPLC separation, the first peak (PP-S4-1) showed that inhibition ratio of erythrocyte hemolysis was 91.9%, and inhibition ratio of lipid peroxidation was also increased by 87.6%, at 50 µg/ml (p < .01). Based on the results of ESI-MS, 1 HNMR, 13 CNMR, and RP-HPLC compared to many published results, PP-S4-1was identified as catechin (MW 290.015, C15 H14 O6 ). The results showed that PP-S4-1 pretreatment made cell viability increased, and the generation of reactive oxygen species (ROS) inhibited. Meanwhile, PP-S4-1 remarkably decreased the fluorescence intensity of Ca2+ , and increased mitochondrial membrane potential (MMP; ΔΨm). In addition, PP-S4-1 could significantly inhibit the decrease of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activity as well as the increase of MDA content in H9c2 cells induced by H2 O2 . Moreover, pretreatment with PP-S4-1 significantly improved the morphological changes and prevented H2 O2 -induced DNA damage. Therefore, this study clarifies the ability of PP-S4-1 to treat H9c2 cell oxidative stress damage induced by H2 O2 through its antioxidant effect. PRACTICAL APPLICATIONS: This research is not only helpful to elaborate the cardioprotective effect of Phellinus pini but also can contribute to the development of health foods or drug supplements for heart disease in the future. This is the first report dealing with phenolic component and cardioprotective activity of a medicinal mushroom P. pini belonging to the genus Phellinus.


Assuntos
Agaricales , Antioxidantes , Animais , Antioxidantes/farmacologia , Catalase/metabolismo , Estresse Oxidativo , Phellinus , Ratos
4.
Food Chem ; 352: 129357, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-33714165

RESUMO

In this work, we investigated whether coating fresh shiitake mushrooms with a polysaccharide isolated from Oudemansiella radicata (ORWP) would impact key quality characteristics after 18 d of storage at 4 °C. We found that ORWP-coated mushrooms had significant improvements in many qualities during storage, including reduced weight loss, improved firmness, reduced browning, decreased malondialdehyde content, and an improved physical microstructure. Further, ORWP-coated mushrooms had higher contents of nutritional and cell wall compounds compared to control samples. ORWP-coated mushrooms had reduced activities of the following enzymes: protease, polyphenol oxidase, peroxidase, phenylalanine ammonia lyase, cellulase, and chitinase, relative to control samples. However, mushrooms coated with ORWP had higher concentrations of superoxide dismutase and catalase, as well as higher contents of certain key monosodium glutamate-resembling amino acids, umami 5'-nucleotides and 1-octen-3-ol. These findings suggest that ORWP coatings have potential value as a method to improve the postharvest quality of shiitake mushrooms.


Assuntos
Agaricales/química , Armazenamento de Alimentos , Polissacarídeos Fúngicos/química , Cogumelos Shiitake/química , Paladar
5.
Biochim Biophys Acta Rev Cancer ; 1875(2): 188525, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33600824

RESUMO

Gynecological cancers are characterized by a high mortality rate when chemoresistance develops. Autophagy collaborates with apoptosis and participates in homeostasis of chemoresistance. Recent findings supported that crosstalk of necrotic, apoptotic and autophagic factors, and chemotherapy-driven hypoxia, oxidative stress and ER stress play critical roles in chemoresistance in gynecological cancers. Meanwhile, current studies have shown that autophagy could be regulated by and cooperate with metabolic regulator, survival factors, stemness factors and specific post-translation modification in chemoresistant tumor cells. Meanwhile, non-coding RNA and autophagy crosstalk also contribute to the chemoresistance. Until now, analysis of individual autophagy factors towards the clinical significance and chemoresistance in gynecological cancer is still lacking. We suggest comprehensive integrated analysis of cellular homeostasis and tumor microenvironment to clarify the role of autophagy and the associated factors in cancer progression and chemoresistance. Panel screening of pan-autophagic factors will pioneer the development of risk models for predicting efficacy of chemotherapy and guidelines for systematic treatment and precision medicine.


Assuntos
Autofagia , Resistencia a Medicamentos Antineoplásicos , Neoplasias dos Genitais Femininos/metabolismo , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Estresse Oxidativo , Transdução de Sinais , Microambiente Tumoral
6.
J Agric Food Chem ; 69(7): 2051-2061, 2021 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-33570936

RESUMO

A bacterium identified as Bacillus velezensis with a growth inhibitory effect against Fusarium solani, a pathogen that caused basal stem rot in the passion fruit Passiflora edulis, was isolated in this study. From the fermentation broth of B. velezensis, a type of antifungal peptide (named BVAP) with a molecular weight of ca. 1.5 kDa was purified and found to be fengycin. BVAP suppressed mycelial growth in F. solani with an IC50 of 5.58 µg/mL, which was superior to those of the chemical fungicides thiram (41.24 µg/mL) and hymexazol (343.31 µg/mL). The antifungal activity remained stable after exposure to 50-100 °C or following incubation with solutions at pH 1-3. Further research revealed that BVAP increased the permeability of the F. solani mycelial membrane, brought about swelling at the tips of hyphae, and elicited abnormal accumulation of nucleic acids and chitin at the sites of swelling. These findings indicate that BVAP possessed a remarkable biocontrol potential toward F. solani.


Assuntos
Fusarium , Passiflora , Antifúngicos/farmacologia , Bacillus , Frutas , Doenças das Plantas
7.
Int J Biol Macromol ; 174: 77-88, 2021 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-33508361

RESUMO

A new anti-tumor protein (designated as Boletus edulis or in short BEAP) was isolated from dried fruit bodies of the edible bolete mushroom Boletus edulis. The purification protocol employed comprised fast ion exchange chromatography on a Hitrap Q column and ion exchange chromatography on a DEAE-52 cellulose column. Superdex G75 gel filtration and SDS-PAGE analysis revealed that BEAP was a protein with a molecular weight of 16.7 KD. The protein exhibited potent anti-cancer activity on A549 cells both in vitro and in vivo. With the use of AO/EB staining, annexin V-FITC/PI, and Western blotting, it was demonstrated in vitro that the cytotoxicity of BEAP was mediated by induction of apoptosis and arrest of A549 cells in the G1 phase of the cell cycle. BEAP significantly suppressed the growth of A549 solid tumors in vivo. These results prove that BEAP is a new multifunctional protein with anti-tumor and anti-metastasis capabilities.


Assuntos
Basidiomycota/enzimologia , Basidiomycota/metabolismo , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Células A549/efeitos dos fármacos , Agaricales/química , Agaricales/enzimologia , Animais , Apoptose/efeitos dos fármacos , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Proliferação de Células/efeitos dos fármacos , Cromatografia em Gel/métodos , Cromatografia por Troca Iônica/métodos , Eletroforese em Gel de Poliacrilamida/métodos , Feminino , Carpóforos/química , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas/análise , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
8.
Environ Health Prev Med ; 26(1): 8, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33451279

RESUMO

BACKGROUND: Prenatal stress can cause neurobiological and behavioral defects in offspring; environmental factors play a crucial role in regulating the development of brain and behavioral; this study was designed to test and verify whether an enriched environment can repair learning and memory impairment in offspring rats induced by prenatal stress and to explore its mechanism involving the expression of insulin-like growth factor-2 (IGF-2) and activity-regulated cytoskeletal-associated protein (Arc) in the hippocampus of the offspring. METHODS: Rats were selected to establish a chronic unpredictable mild stress (CUMS) model during pregnancy. Offspring were weaned on 21st day and housed under either standard or an enriched environment. The learning and memory ability were tested using Morris water maze and Y-maze. The expression of IGF-2 and Arc mRNA and protein were respectively measured by using RT-PCR and Western blotting. RESULTS: There was an elevation in the plasma corticosterone level of rat model of maternal chronic stress during pregnancy. Maternal stress's offspring exposed to an enriched environment could decrease their plasma corticosterone level and improve their weight. The offspring of maternal stress during pregnancy exhibited abnormalities in Morris water maze and Y-maze, which were improved in an enriched environment. The expression of IGF-2, Arc mRNA, and protein in offspring of maternal stress during pregnancy was boosted and some relationships existed between these parameters after being exposed enriched environment. CONCLUSIONS: The learning and memory impairment in offspring of prenatal stress can be rectified by the enriched environment, the mechanism of which is related to the decreasing plasma corticosterone and increasing hippocampal IGF-2 and Arc of offspring rats following maternal chronic stress during pregnancy.


Assuntos
Proteínas do Citoesqueleto/genética , Regulação da Expressão Gênica , Fator de Crescimento Insulin-Like II/genética , Deficiências da Aprendizagem/genética , Transtornos da Memória/genética , Proteínas do Tecido Nervoso/genética , Efeitos Tardios da Exposição Pré-Natal/genética , Meio Social , Estresse Psicológico/genética , Animais , Proteínas do Citoesqueleto/metabolismo , Feminino , Hipocampo/metabolismo , Fator de Crescimento Insulin-Like II/metabolismo , Aprendizagem , Deficiências da Aprendizagem/psicologia , Masculino , Transtornos da Memória/psicologia , Proteínas do Tecido Nervoso/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/psicologia , Distribuição Aleatória , Ratos , Ratos Wistar
9.
BMC Cancer ; 20(1): 964, 2020 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-33023525

RESUMO

BACKGROUND: Previous studies have shown a major green tea polyphenol (-)-epigallocatechin-3-gallate ((-)-EGCG) as a powerful anti-cancer agent. However, its poor bioavailability and requirement of a high dosage to manifest activity have restricted its clinical application. Recently, our team synthesized a peracetate-protected derivative of EGCG, which can act as a prodrug of (-)-EGCG (ProEGCG) with enhanced stability and improved bioavailability in vitro and in vivo. Herein, we tested the therapeutic efficacy of this novel ProEGCG, in comparison to EGCG, toward human endometrial cancer (EC). METHODS: In this study, the effects of ProEGCG and EGCG treatments on cell growth, cell survival and modulation of intracellular signaling pathways in RL95-2 and AN3 CA EC cells were compared. The antiproliferative effect was evaluated by cell viability assay. Apoptosis was measured by annexin/propidium iodide staining. Expression of mitogen-activated protein kinases, markers of proliferation and apoptosis were measured by immunoblot analysis. In addition, the effects of ProEGCG and EGCG on tumor growth, vessel formation and gene expression profiles on xenograft models of the EC cells were investigated. RESULTS: We found that treatment with ProEGCG, but not EGCG, inhibited, in a time- and dose-dependent manner, the proliferation and increased apoptosis of EC cells. Treatment with low-dose ProEGCG significantly enhanced phosphorylation of JNK and p38 MAPK and inhibited phosphorylation of Akt and ERK which are critical mediators of apoptosis. ProEGCG, but not EGCG, elicited a significant decrease in the growth of the EC xenografts, promoted apoptotic activity of tumour cells in the EC xenografts, and decreased microvessel formation, by differentially suppressing anti-apoptotic molecules, NOD1 and NAIP. Notably, no obvious adverse effects were detected. CONCLUSIONS: Taken together, ProEGCG at a low dose exhibited anticancer activity in EC cells through its anti-proliferative, pro-apoptotic and anti-tumor actions on endometrial cancer in vitro and in vivo. In contrast, a low dose of EGCG did not bring about similar effects. Importantly, our data demonstrated the efficacy and safety of ProEGCG which manifests the potential of a novel anticancer agent for the management of endometrial cancer.


Assuntos
Catequina/análogos & derivados , Neoplasias do Endométrio/tratamento farmacológico , Pró-Fármacos/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Chá/química , Animais , Apoptose , Catequina/química , Proliferação de Células , Feminino , Humanos , Camundongos , Camundongos Nus , Pró-Fármacos/farmacologia , Transdução de Sinais
10.
Artigo em Inglês | MEDLINE | ID: mdl-32924196

RESUMO

Chitinases play an important role in many industrial processes, including the preparation of oligosaccharides with potential applications. In the present study, a 1,713 bp gene of Chi1602, derived from a marine bacterium Microbulbifer sp. BN3, encoding a GH18 family chitinase, was expressed at high levels in Pichia pastoris. Distinct from most of the marine chitinases, the recombinant chitinase 1602 exhibited maximal activity at 60 °C and over a broad pH range between 5.0 and 9.0, and was stable at 50 °C and over the pH range 4.0-9.0. The hydrolytic products derived from colloidal chitins comprised mainly (GlcNAc)2 and GlcNAc, indicating that rChi1602 is a GH18 processive chitinase in conformity with its hypothetical structure. However, rChi1602 showed traces of ß-N-acetylglucosaminidase activity on substrates such as powder chitin, chitosan, and ethylene glycol chitin. The thermophilic rChi1602, which manifests adaptation to a wide pH range and can be expressed at a high level in P. pastoris, is advantageous for applications in industrial processes.

11.
Pharmacol Res ; 161: 105157, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32814169

RESUMO

Increasing evidence shows that Traditional Chinese Medicine (TCM) has an obvious appeal for cancer treatment, but there is still a lack of scientific investigation of its underlying molecular mechanisms. Bitter melon or bitter gourd (Momordica charantia) is an edible fruit that is commonly consumed, and it is used to cure different diseases in various ancient folk medical practices. We report that a bioactive protein, MAP30, isolated from bitter melon seeds exhibited potent anticancer and anti-chemoresistant effects on ovarian cancer cells. Functional studies revealed that MAP30 inhibited cancer cell migration, cell invasion, and cell proliferation in various ovarian cancer cells but not normal immortalized ovarian epithelial cells. When administered with cisplatin, MAP30 produced a synergistic effect on cisplatin-induced cell cytotoxicity in ovarian cancer cells. When low doses of cisplatin and MAP30 were co-injected intraperitoneally, a remarkable reduction of tumor dissemination and tumor growth was observed in an ovarian cancer ascites mouse model. Notably, blood tests confirmed that MAP30 did not cause any adverse effects on liver and kidney functions in the treated mice. MAP30 activated AMP-activated protein kinase (AMPK) signaling via CaMKKß and induced cell cycle arrest in the S-phase. MAP30 modulated cell metabolism of ovarian cancer cells via suppression of GLUT-1/-3-mediated glucose uptake, adipogenesis, and lipid droplet formation in tumor development and progression. MAP30 also induced an increase in intracellular Ca2+ ion concentration, which triggered ROS-mediated cancer cell death via apoptosis and ferroptosis. Collectively, these findings suggest that natural MAP30 is a non-toxic supplement that may enhance chemotherapeutic outcomes and benefit ovarian cancer patients with peritoneal metastases.

12.
Int J Antimicrob Agents ; 56(3): 106118, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32738305

RESUMO

The coronavirus disease 2019 (COVID-19) pandemic is rapidly advancing across the globe despite drastic public and personal health measures. Antivirals and nutritional supplements have been proposed as potentially useful against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the novel coronavirus that causes COVID-19, but few have been clinically established. Lactoferrin (Lf) is a naturally occurring, non-toxic glycoprotein that is orally available as a nutritional supplement and has established in vitro antiviral efficacy against a wide range of viruses, including SARS-CoV, a closely related coronavirus to SARS-CoV-2. Furthermore, Lf possesses unique immunomodulatory and anti-inflammatory effects that may be especially relevant to the pathophysiology of severe COVID-19 cases. Here we review the underlying biological mechanisms of Lf as an antiviral and immune regulator, and propose its unique potential as a preventative and adjunct treatment for COVID-19. We hope that further research and development of Lf nutritional supplementation would establish its role for COVID-19.


Assuntos
Anti-Inflamatórios/uso terapêutico , Antivirais/uso terapêutico , Infecções por Coronavirus/tratamento farmacológico , Infecções por Coronavirus/epidemiologia , Fatores Imunológicos/uso terapêutico , Lactoferrina/uso terapêutico , Pandemias , Pneumonia Viral/tratamento farmacológico , Pneumonia Viral/epidemiologia , Betacoronavirus/efeitos dos fármacos , Betacoronavirus/imunologia , COVID-19 , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/virologia , Proteoglicanas de Heparan Sulfato/antagonistas & inibidores , Proteoglicanas de Heparan Sulfato/metabolismo , Humanos , Interferons/agonistas , Interferons/biossíntese , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Pneumonia Viral/virologia , SARS-CoV-2 , Índice de Gravidade de Doença , Internalização do Vírus/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos
13.
Int J Biol Macromol ; 164: 3171-3178, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32858105

RESUMO

Breast cancer is the most common malignant tumor in women and one of the three most common cancers worldwide. It is a life-threatening disease among women and the leading cause of death among women. New drugs or new drug translations and laboratory clinical studies are ongoing. A new antitumor protein (PNAP) purified from edible fungus Pholiota nameko has potential for treating breast cancer. We have previously found that PNAP exhibits anti-proliferative and apoptosis-inducing activities in a human breast cancer cell line (MCF-7). In this study, we constructed a BALB/c mouse model of MCF-7 tumor xenografts. In vivo experiments show that PNAP can effectively inhibit the malignant proliferation of MCF-7 solid tumors. This is because PNAP can successfully activate the death receptor pathway and mitochondrial apoptosis pathway of MCF-7 tumor cells in vivo, and induce tumor cells to wither. It is estimated that PNAP may also have an immunoregulatory ability to indirectly inhibit malignant proliferation of tumors. We also found that PNAP may also have the immunomodulatory ability to indirectly inhibit the malignant proliferation of tumors, which can shift the balance of Th1/Th2 to Th1 and eventually inhibit the growth of tumors. The study reveals a new therapeutic approach for breast cancer patient.


Assuntos
Adenocarcinoma/tratamento farmacológico , Antineoplásicos/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Citocinas/genética , Proteínas Fúngicas/administração & dosagem , Pholiota/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Feminino , Proteínas Fúngicas/química , Proteínas Fúngicas/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Células MCF-7 , Camundongos , Camundongos Endogâmicos BALB C , Ensaios Antitumorais Modelo de Xenoenxerto
14.
Toxins (Basel) ; 12(8)2020 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-32727016

RESUMO

Aflatoxin B1 (AFB1) is a known toxic human carcinogen and can be detoxified by laccases, which are multicopper oxidases that convert several environmental pollutants and toxins. In this study, a new laccase that could catalyze AFB1 degradation was purified and identified from the white-rot fungus Cerrena unicolor 6884. The laccase was purified using (NH4)2SO4 precipitation and anion exchange chromatography, and then identified as Lac 2 through zymogram and UHPLC-MS/MS based on the Illumina transcriptome analysis of C. unicolor 6884. Six putative laccase protein sequences were obtained via functional annotation. The lac 2 cDNA encoding a full-length protein of 512 amino acids was cloned and sequenced to expand the fungus laccase gene library for AFB1 detoxification. AFB1 degradation by Lac 2 was conducted in vitro at pH 7.0 and 45 °C for 24 h. The half-life of AFB1 degradation catalyzed by Lac 2 was 5.16 h. Acetosyringone (AS), Syrinagaldehyde (SA) and [2,2' -azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)] (ABTS) at 1 mM concentration seemed to be similar mediators for strongly enhancing AFB1 degradation by Lac 2. The product of AFB1 degradation catalyzed by Lac 2 was traced and identified to be Aflatoxin Q1 (AFQ1) based on mass spectrometry data. These findings are promising for a possible application of Lac 2 as a new aflatoxin oxidase in degrading AFB1 present in food and feeds.


Assuntos
Aflatoxina B1/química , Lacase/química , Polyporales/enzimologia , Sequência de Aminoácidos , Contaminação de Alimentos/prevenção & controle , Lacase/genética , Lacase/isolamento & purificação , Filogenia , Polyporales/genética
15.
Int J Biol Macromol ; 158: 1125-1134, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32360969

RESUMO

Chitinases play an important role in the process of chitin bioavailability. In this study, we cloned a new chitinase gene and characterized its recombinant protein. The new 1251 bp gene of chitinase (ChiT-7) was cloned from the metagenome of the mangrove tidal flat soil in the city of Zhangzhou in Fujian Province (China) by genome walking. The gene encoded a mature protein with 381 amino acids, which manifested certain sequence similarity (59% identity) to characterized GH18 chitinases. The mature protein of ChiT-7 was successfully expressed in E. coli BL21 (DE3). After purification, the specific activity of the recombinant enzyme was 0.63 U/mg at the optimal pH of 6.0 and the optimal temperature of 45 °C. The rChiT-7 was active over a wide pH range, and the residual enzyme activity reached 80% or higher at 30 °C-50 °C. rChiT-7 hydrolyzed colloidal chitin with (GlcNAc)2 and GlcNAc as the main final products. Structural analysis of ChiT-7 indicated that ChiT-7 could be a processive chitinase. rChiT-7 manifested characteristics analogous to those of fungi and actinomycetes and exhibited sequence homology.

16.
Appl Microbiol Biotechnol ; 104(11): 4675-4703, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32274562

RESUMO

This article reviews mushrooms with anti-breast cancer activity. The mushrooms covered which are better known include the following: button mushroom Agaricus bisporus, Brazilian mushroom Agaricus blazei, Amauroderma rugosum, stout camphor fungus Antrodia camphorata, Jew's ear (black) fungus or black wood ear fungus Auricularia auricula-judae, reishi mushroom or Lingzhi Ganoderma lucidum, Ganoderma sinense, maitake mushroom or sheep's head mushroom Grifola frondosa, lion's mane mushroom or monkey head mushroom Hericium erinaceum, brown beech mushroom Hypsizigus marmoreus, sulfur polypore mushroom Laetiporus sulphureus, Lentinula edodes (shiitake mushroom), Phellinus linteus (Japanese "meshimakobu," Chinese "song gen," Korean "sanghwang," American "black hoof mushroom"), abalone mushroom Pleurotus abalonus, king oyster mushroom Pleurotus eryngii, oyster mushroom Pleurotus ostreatus, tuckahoe or Fu Ling Poria cocos, and split gill mushroom Schizophyllum commune. Antineoplastic effectiveness in human clinical trials and mechanism of anticancer action have been reported for Antrodia camphorata, Cordyceps sinensis, Coriolus versicolor, Ganoderma lucidum, Grifola frondosa, and Lentinula edodes.


Assuntos
Agaricales/química , Agaricales/classificação , Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Animais , Linhagem Celular Tumoral , Ensaios Clínicos como Assunto , Misturas Complexas/química , Misturas Complexas/farmacologia , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , Ratos
17.
Appl Microbiol Biotechnol ; 104(10): 4211-4226, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32193575

RESUMO

Ribosome-inactivating proteins (RIPs) consist of three varieties. Type 1 RIPs are single-chained and approximately 30-kDa in molecular weight. Type 2 RIPs are double-chained and composed of a type 1 RIP chain and a lectin chain. Type III RIPs, such as maize b-32 barley and JIP60 which are produced as single-domain proenzymes, possess an N-terminal domain corresponding to the A domain of RIPs and fused to a C-terminal domain. In addition to the aforementioned three types of RIPs originating from flowering plants, there are recently discovered proteins and peptides with ribosome-inactivating and protein synthesis inhibitory activities but which are endowed with characteristics such as molecular weights distinctive from those of the regular RIPs. These new/unusual RIPs discussed in the present review encompass metazoan RIPs from Anopheles and Culex mosquitos, antimicrobial peptides derived from RIP of the pokeweed Phytolacca dioica, maize RIP (a type III RIP derived from a precursor form), RIPs from the garden pea and the kelp. In addition, RIPs with a molecular weight smaller than those of regular type 1 RIPs are produced by plants in the Cucurbitaceae family including the bitter gourd, bottle gourd, sponge gourd, ridge gourd, wax gourd, hairy gourd, pumpkin, and Chinese cucumber. A small type II RIP from camphor tree (Cinnamomum camphora) seeds and a snake gourd type II RIP with its catalytic chain cleaved into two have been reported. RIPs produced from mushrooms including the golden needle mushroom, king tuber mushroom, straw mushroom, and puffball mushroom are also discussed in addition to a type II RIP from the mushroom Polyporus umbellatus. Bacterial (Spiroplasma) RIPs associated with the fruitfly, Shiga toxin, and Streptomyces coelicolor RIP are also dealt with. The aforementioned proteins display a diversity of molecular weights, amino acid sequences, and mechanisms of action. Some of them are endowed with exploitable antipathogenic activities.


Assuntos
Biossíntese de Proteínas/efeitos dos fármacos , Proteínas Inativadoras de Ribossomos/metabolismo , Sequência de Aminoácidos , Animais , Culicidae/química , Proteínas de Insetos/metabolismo , Proteínas de Plantas/metabolismo , Proteínas Inativadoras de Ribossomos/classificação , Proteínas Inativadoras de Ribossomos/farmacologia , Sementes/química
18.
Int J Med Mushrooms ; 22(10): 991-1000, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33426828

RESUMO

A ribonuclease was purified from dry fruiting bodies of the wild edible mushroom Lepista personata (LPR) to 259-fold with a specific activity of 280 U/mg. The purification protocol involved ion-exchange chromatography on DEAE-cellulose, CM-cellulose and SP-sepharose, followed by size exclusion chromatography on Superdex 75. LPR is a homodimeric protein with a molecular weight of 27.8 kDa as determined by SDS-PAGE and by gel filtration. Three inner peptide sequences for LPR were obtained by LC-MS-MS analysis. It demonstrated the optimum pH of 4.0 and temperature optimum of 60°C. The specificity ribonuclease potencies order toward polyhomoribonucleotides was poly C > poly A > poly G > poly U. The ribonuclease inhibited HIV-1 reverse transcriptase with an IC50 of 0.53 µM.


Assuntos
Agaricales/enzimologia , Proteínas Fúngicas/isolamento & purificação , Transcriptase Reversa do HIV/antagonistas & inibidores , Ribonucleases/isolamento & purificação , Agaricales/química , Agaricales/metabolismo , Estabilidade Enzimática , Carpóforos/química , Carpóforos/enzimologia , Proteínas Fúngicas/química , Transcriptase Reversa do HIV/química , Concentração de Íons de Hidrogênio , Peso Molecular , Ribonucleases/química
19.
AMB Express ; 9(1): 194, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31797089

RESUMO

In this study, a single component paclitaxel was obtained from fermentation broth by molecular imprinting technique, and its antiviral, antitumor and immunomodulatory activities were studied. The results showed that paclitaxel had a good inhibitory activity on human breast cancer MCF-7 cells and showed a concentration- dependent relationship with an IC50 of about 15 µg/mL in the sulforhodamine B assay. At the same time, paclitaxel exerted a weak inhibitory activity on cervical cancer Hela cells. In addition, paclitaxel not only inhibited the invasion of HIV-1 pseudovirus into cells, but also exhibited inhibitory activity to a certain extent after viral invasion of the cells. At a paclitaxel concentration of 20 µg/mL, the inhibition of HIV-1 pseudovirus reached about 66%. The inhibition of HIV-1 protease activity was concentration-dependent. At a concentration of 20 µg/mL, the inhibitory effect of paclitaxel on HIV-1 protease was similar to that of the positive control pepstatin A, being 15.8%. The HIV-1 integrase inhibiting activity of paclitaxel was relatively weak. Paclitaxel significantly up-regulated the expression of interleukin-6.

20.
Sci Rep ; 9(1): 17128, 2019 11 20.
Artigo em Inglês | MEDLINE | ID: mdl-31748652

RESUMO

Adolescent idiopathic scoliosis (AIS) is a complex three-dimensional structural deformity of the spine with unknown etiology. Although leptin has been postulated as one of the etiologic factors in AIS, its effects on osteoblastic activity remain unknown. Herein, we conducted this study to investigate whether there are abnormal functional responses to leptin and abnormal expression of leptin receptor in AIS osteoblasts. In vitro assays were performed with osteoblasts isolated from 12 severe AIS girls and 6 non-AIS controls. The osteoblasts were exposed to different concentrations of leptin (0, 10, 100, 1000 ng/mL). The effects of leptin on cell proliferation, differentiation and mineralization were determined. Protein expressions of leptin receptor (LEP-R) under basal and osteogenic conditions were also evaluated by Western blot. Our results showed that leptin significantly stimulated osteoblasts from non-AIS subjects to proliferate, differentiate and mineralized. However, in the AIS group, the stimulatory effects of leptin on cell proliferation, differentiation, and mineralization were not observed. In addition, no statistically significant difference in the expression of leptin receptor under both basal and osteogenic conditions was found between AIS and control group. In conclusion, these findings might help to explain the low bone mass and deranged bone quality that is clinically associated with AIS girls.


Assuntos
Leptina/metabolismo , Osteoblastos/metabolismo , Escoliose/metabolismo , Adolescente , Adulto , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Receptores para Leptina/metabolismo , Coluna Vertebral/metabolismo , Adulto Jovem
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