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1.
Int J Cancer ; 148(12): 3111-3118, 2021 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-33594666

RESUMO

It is known that core-type fucosylation is higher in prostate cancer cells than in other cancer cell types and is associated with high-risk prostate cancer. Here, we developed an automated microcapillary electrophoresis-based immunoassay system for measuring serum core-type fucosylated prostate-specific antigen (PSA) and evaluated whether the serum fucosylated PSA index (FPI) can detect high-risk prostate cancer. Core-type fucosylated-free PSA was measured by our automated microcapillary electrophoresis-based immunoassay system with Pholiota squarrosa lectin. The FPI was calculated from total PSA and the percentage of fucosylated-free PSA. The optimum model to predict Gleason grade (GG) ≥2 was constructed by multivariate logistic regression analysis. Discrimination was assessed by determining the area under the receiver operator characteristic curve (AUC). The study included 252 men who underwent prostate needle biopsy due to elevated serum PSA levels (4-20 ng/mL), including 138 with GG ≥2. A higher FPI was significantly associated with GG (P < .0001). Multivariate logistic regression analysis showed that age, prostate volume and FPI were significant predictors of GG ≥2. The AUC of FPI and the model were 0.729 (95% confidence interval [CI]: 0.668-0.790) and 0.837 (95% CI: 0.788-0.886), respectively, compared to 0.629 (95% CI: 0.561-0.698) for PSA. Decision curve analysis showed the superior benefit of FPI and the model when compared to PSA. In a cohort with serum PSA levels <20 ng/mL, FPI could differentiate high-risk prostate cancer from biopsy-negative or low-risk prostate cancer. Therefore, FPI could be a useful adjunct in prostate biopsy counseling for men with abnormal PSA levels.


Assuntos
Lectinas/química , Pholiota/metabolismo , Antígeno Prostático Específico/análise , Neoplasias da Próstata/diagnóstico , Fatores Etários , Idoso , Biópsia por Agulha , Detecção Precoce de Câncer , Fucose/química , Proteínas Fúngicas/química , Glicosilação , Humanos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Gradação de Tumores , Antígeno Prostático Específico/química , Neoplasias da Próstata/sangue , Neoplasias da Próstata/patologia
2.
Mycologia ; 113(1): 146-167, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33175650

RESUMO

Multigene data sets were assembled to evaluate the phylogeny of species attributed to the genus Pholiota sensu A.H. Sm. & Hesler. This effort included generation of just more than 200 new sequences from 19 type collections of Pholiota and recent samples from East Asia. Phylogenetic analyses reinforced the autonomous phylogenetic positions of pholiotoid taxa in the genera Flammula (Hymenogastraceae) and Kuehneromyces (Strophariaceae). Samples of Pholiota astragalina from diverse geographic regions split into two species-level lineages but occupied an isolated phylogenetic position apart from Pholiota sensu stricto. The new genus Pyrrhulomyces is described to accommodate P. astragalina and a new cryptic species from the Southern Appalachians, Pyrrhulomyces amariceps. Pyrrhulomyces is distinguished from other genera of Strophariaceae by the blackening basidiomata with a bitter taste, smooth basidiospores without a germ pore under light microscopy, presence of pleurochrysocystidia, an ixocutis, rugulose spore ornamentation under scanning electron microscope (SEM), and association with late stages of conifer wood decay. Pholiota subochracea was found to be sister to a clade containing samples of Hypholoma and Bogbodia, but this portion of the Strophariaceae will require further taxon and gene sampling to resolve relationships between these three taxa. Pholiota sensu stricto comprised at least two major groups, but several residual poorly placed lineages were also noted depending on the data set analyzed. New combinations are made in the genera Flammula, Kuehneromyces, and Stropharia for three species of Pholiota-P. abieticola, P. obscura, and P. scabella, respectively, based on molecular annotation of type collections. Overall, 20 new synonymies are proposed, mostly in Pholiota. Illustrations of Pyrrhulomyces are provided along with a key to genera of Strophariaceae and Hymenogastraceae.


Assuntos
Classificação/métodos , Pholiota , Filogenia , Agaricales/classificação , Agaricales/genética , Genes Fúngicos , Pholiota/classificação , Pholiota/citologia , Pholiota/genética , Pholiota/ultraestrutura
3.
Int J Biol Macromol ; 166: 1220-1229, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33157137

RESUMO

The aim of this study was to investigate sulfuric acid degradation of the Pholiota nameko polysaccharide (AIPS-1). Three stepwise degraded polysaccharides (AIPS-2, AIPS-3, and AIPS-4) were obtained by sequentially increasing the strength of sulfuric acid treatment. Structural characterization showed that sulfuric acid treatment significantly decreased molecular weight, increased the content of uronic acid and changed the molar ratio of monosaccharide composition, while the major functional groups and the triple helical conformation of polysaccharides did not change significantly. In vitro experiments proved that the antioxidation ability of the stepwise degraded polysaccharides gradually increased (AIPS-1 < AIPS-2 < AIPS-3 < AIPS-4). An oxidative stress zebrafish model was established, which demonstrated that the ability of AIPS-3 and AIPS-4 to scavenge free radicals in zebrafish was significantly improved compared to AIPS-1. In conclusion, sulfuric acid treatment is an effective method for improving the antioxidant activity of polysaccharides, and increased antioxidant activity was closely related to the changes in their structural characteristics.


Assuntos
Antioxidantes/farmacologia , Pholiota/química , Polissacarídeos/química , Polissacarídeos/farmacologia , Ácidos Sulfúricos/química , Vermelho Congo/química , Sequestradores de Radicais Livres/farmacologia , Conformação Molecular , Peso Molecular , Monossacarídeos/análise , Polissacarídeos/isolamento & purificação , Espectroscopia de Infravermelho com Transformada de Fourier , Sulfatos/análise , Ácidos Urônicos/análise
4.
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
5.
Nat Prod Res ; 34(14): 1957-1961, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30724606

RESUMO

Pholiotone A (1), a new polyketide derivative, with tetrahydrobenzofuran-4(2H)-one skeleton, together with four known compounds, trichodermatides A (2) and B (3) and koninginins B (4) and E (5), were isolated from the crude extract of Pholiota sp. The structures of all the isolated compounds were determined mainly by NMR experiments, the modified Mosher method and electronic circular dichroism (ECD) calculations. The antifungal and cytotoxicity of all isolates were evaluated.


Assuntos
Pholiota/química , Policetídeos/isolamento & purificação , Antifúngicos/química , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Dicroísmo Circular , Citotoxinas/química , Citotoxinas/farmacologia , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Policetídeos/química
6.
Nat Prod Res ; 34(19): 2754-2759, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30887855

RESUMO

One new sterpurane sesquiterpene (1), named (3R,6S,7S,8R,10S)-3,7,14-trihydroxy-1-sterpurene was isolated from cultures of the basidiomycete Pholiota nameko. The structure of new compound was elucidated by extensive spectroscopic. Additionally, a single crystal X-ray diffraction not only confirmed the structure, but also determined the absolute configuration of the new compound. The compound was evaluated for cytotoxicity against five human cancer cell lines, but no significant cytotoxicity were found (IC50 values > 40 µM).


Assuntos
Pholiota/metabolismo , Sesquiterpenos/química , Sesquiterpenos/farmacologia , Linhagem Celular Tumoral , Cristalografia por Raios X , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Pholiota/crescimento & desenvolvimento , Espectrometria de Massas por Ionização por Electrospray
7.
Int J Biol Macromol ; 135: 1020-1027, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31152832

RESUMO

The effect of the ultrasonic treatment on the physicochemical properties, oil-holding capacities, foaming capacities, and TLR2-affinity of the polysaccharides from Pholiota nameko (PNPS), was evaluated. Compared with the protein content of PNPS before ultrasonic treatment, the protein content of PNPS all presented a decrease under different ultrasonic conditions. The viscosity of PNPS showed a decrease when the ultrasonic intensity was strong enough, as well as the molecular weight. The oil-holding capacity and the foaming capacity of PNPS showed a continuous increasing trend with the increase of the ultrasonic treatment time under a set ultrasonic power of 400 W. Further, the ultrasonic operation could induce the decrease of the affinity binding between PNPS and the receptor proteins TLR2. These data confirmed that applying the ultrasonic treatment could obtain PNPS with high carbohydrate contents, low viscosity and low TLR2-affinity under proper ultrasonic condition.


Assuntos
Fenômenos Químicos , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/isolamento & purificação , Extração Líquido-Líquido , Pholiota/química , Receptor 2 Toll-Like/química , Ondas Ultrassônicas , Ligação Proteica , Reologia , Receptor 2 Toll-Like/metabolismo
8.
Glycobiology ; 29(7): 576-587, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30913288

RESUMO

Fucosylation of the N-glycan core via the α1-6 linkage (core fucosylation) is detected in specific types of cancers and related diseases, and thereby serves for a relevant biomarker. The lectin from a mushroom Pholiota squarrosa (PhoSL) shows a clear specificity to core fucosylation, without recognizing those with other types of fucosylation, such as the H type via the α1-2 linkage or the Lewis type via the α1-3 or α1-4 linkage. Here we determined the crystal structure of the PhoSL trimer in complex with a disaccharide fucose(α1-6)N-acetylglucosamine (GlcNAc). In the three sugar-binding pockets of PhoSL, extensive hydrophobic and hydrogen-bonding contacts were formed with the fucose moiety. In contrast, the GlcNAc moiety showed only a few hydrophobic and hydrogen-bonding contacts. To elucidate the mechanism for the specificity, we performed molecular dynamics simulations on this disaccharide and a trisaccharide fucose(α1-6)[GlcNAc(ß1-4)]GlcNAc in complex with PhoSL. It was observed that the GlcNAc corresponding to the outer one of the N-glycan core entered the sugar-binding pocket with the N-acetyl group placed stably at the bottom, forming extensive hydrophobic and hydrogen-bonding interactions. In addition, these glycans adopted unstressed favorable conformations when bound to PhoSL. In contrast, H- and Lewis-types of fucosylated trisaccharides adopting favorable conformations caused inevitable steric hindrance with the steep edge of the binding pocket, when docked with PhoSL. Therefore, the specificity to core fucosylation of PhoSL was achieved by a combination of these preferential and exclusive mechanisms.


Assuntos
Fucose/metabolismo , Pholiota/metabolismo , Polissacarídeos/metabolismo , Configuração de Carboidratos , Fucose/química , Simulação de Dinâmica Molecular , Polissacarídeos/química
9.
Int J Biol Macromol ; 131: 1021-1031, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30910671

RESUMO

This study explored Pholiota nameko (P. nameko) polysaccharide fractions, PNP-40, PNP-60, and PNP-80, purified by gradient concentrations of ethanol (40%, 60%, and 80% (v/v)). The physicochemical properties, functional group composition, moisture-preserving, and antioxidant ability were determined. The results indicate that the polysaccharide contents of PNP-40, PNP-60, and PNP-80 are 45.12%, 78.04%, and 72.22%, respectively, while the ß-glucan, protein, and uronic acid contents are 20.20%, 12.20%, and 10.15%, respectively; 11.24%, 14.53%, and 26.94%; and 5.99%, 7.73%, and 3.78%. Furthermore, PNP-60 has better moisture absorption, while PNP-80 has better antioxidant ability and H2O2-injury resistance activity. Monosaccharide composition analysis shows that P. nameko belongs to heteropolysaccharides, which consists of galactose, glucose, and mannose with different types and ratios, and the molecular weight are distributed at 4.40-333.49kDa. It was found that different polysaccharide fractions have the potential to be a moisturizer and an antioxidant, and their active ingredients could be used in the development of cosmetic ingredients.


Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Pholiota/química , Absorção Fisico-Química , Antioxidantes/isolamento & purificação , Fenômenos Químicos , Precipitação Fracionada , Polissacarídeos Fúngicos/isolamento & purificação , Quelantes de Ferro/química , Quelantes de Ferro/isolamento & purificação , Quelantes de Ferro/farmacologia , Peso Molecular , Monossacarídeos , Espectroscopia de Infravermelho com Transformada de Fourier , beta-Glucanas
10.
Sci Rep ; 9(1): 1724, 2019 02 11.
Artigo em Inglês | MEDLINE | ID: mdl-30741980

RESUMO

PAP80-2a, purified from Pholiota adiposa mycelia, is a polysaccharide exhibiting prominent antitumor effects. However, the yield of PAP80-2a was low and its structure has not been characterized, impeding the exploration of its structure-function relationship, thus influencing the development of oral drugs for antitumor therapy and immunomodulation. In order to improve the yield of PAP80-2a, response surface methodology along with Box-Behnken design was applied to optimize the ultrasonic-assisted extraction conditions for polysaccharides. Then, the structure of PAP80-2a exhibiting antitumor activity was determined from different angles. The results showed that the extraction yield of P. adiposa polysaccharides increased by 11.5% under optimized ultrasonic extraction conditions. Structural analysis showed that PAP80-2a was mainly composed of glucose, rhamnose, xylose, and galactose in a ratio of 10.00: 2.09: 4.09: 1.13. The total amino acid content in the sugar chain was 69.92 µg/mL. The sugar chain structure was [α-Rha (1 → 3)-]n, and rhamnose was located at the non-reducing end of the sugar chain, while glucose was located at the non-reducing end or in the sugar chain in 1,2,6- and 1,3,6-linked forms. Our study clearly illuminates the primary structure of PAP80-2a, but 3D structure of PAP80-2a and its structure-function relationship is a future challenge.


Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Micélio/química , Pholiota/química , Animais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Espectroscopia de Ressonância Magnética , Camundongos , Estrutura Molecular , Análise Espectral , Relação Estrutura-Atividade , Ensaios Antitumorais Modelo de Xenoenxerto
11.
Nat Prod Res ; 33(14): 1992-1996, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29911899

RESUMO

Two new sesquiterpenes (1-2), along with one known sesquiterpene (3), were isolated from cultures of the higher fungus Pholiota nameko. The structures of new compounds were elucidated by extensive spectroscopic methods. The known compound was identified by comparing its spectroscopic data with those reported in the literature. All compounds were evaluated for their cytotoxicities against five human cancer cell lines.


Assuntos
Pholiota/química , Sesquiterpenos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Pholiota/citologia , Sesquiterpenos/isolamento & purificação , Sesquiterpenos/toxicidade , Análise Espectral
12.
Nat Prod Res ; 33(11): 1563-1569, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29308664

RESUMO

A polysaccharide named PNP was extracted and purified from Pholiota nameko. The total sugar content of PNP was 95.29% and the molecular weight was 1.89 × 103 kDa. The structural features of PNP were investigated by the combination of chemical and instrumental analysis such as UV spectrophotometer, specific rotation determination, FT-IR, methylisation analysis and Congo red. The results showed that the optical rotation of PNP was +120° and that it had a triple-helical structure. Besides, PNP was mainly composed of glucose and mannose at the molar ratio of 4.24:1.00. The backbone of PNP was composed of (1→3)-linked-Glc and (1→3)-linked-Man whereas the branches of (1→3,6)-linked- Glc, (1→3,6)-linked-Man and T- Glc. Consistenting with the results of UV-Vis spectra, FT-IR spectroscopy and 1H NMR, indicated that PNP was a complex of polysaccharides and polyphenols. In vitro antioxidant results suggested that PNP was processed with certain scavenging capacity.


Assuntos
Antioxidantes/farmacologia , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/farmacologia , Pholiota/química , Antioxidantes/química , Vermelho Congo/química , Carpóforos/química , Polissacarídeos Fúngicos/isolamento & purificação , Glucose/análise , Espectroscopia de Ressonância Magnética , Manose/análise , Metilação , Peso Molecular , Ácido Periódico/química , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier
13.
Int J Biol Macromol ; 122: 1128-1135, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30218731

RESUMO

A ß-D-glucan was obtained from the edible mushroom Pholiota nameko by hot aqueous extraction and purification. NMR and methylation analyses of the purified fraction (GHW-PN, 1.46% yield) indicated the presence of a (1 → 3)-linked ß-D-glucan, highly substituted (~27%) at O-6 by single units of ß-D-Glcp or by (1 → 6)-ß-D-Glcp fragments. The ß-glucan (at 0.5, 1, and 2%) showed shear thinning behavior and when the concentration of the solution increased, there was an increase in apparent viscosity. The ß-D-glucan presented gel-like behavior and thermal stability under a simulated pasteurization process, suggesting its potential as a thickening and gelling agent in products submitted to temperature variations. The ß-D-glucan at 0.3, 1.0 and 3.0 mg kg-1 significantly inhibited the inflammatory pain in 24.8, 56.9 and 82.3%, respectively, in the formalin-induced nociception in mice. The results pointed out that the ß-D-glucan (GHW-PN) isolated from P. nameko presents potential application for the food industry or for medical purposes.


Assuntos
Analgésicos/química , Analgésicos/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Glucanos/química , Glucanos/farmacologia , Pholiota/química , Animais , Formaldeído/farmacologia , Géis , Masculino , Camundongos , Nociceptividade/efeitos dos fármacos , Reologia
14.
Mycologia ; 110(6): 997-1016, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30481131

RESUMO

A systematic reevaluation of North American pyrophilous or "burn-loving" species of Pholiota is presented based on molecular and morphological examination of type and historical collections. Confusion surrounds application of the names P. brunnescens, P. carbonaria, P. castanea, P. fulvozonata, P. highlandensis, P. molesta, and P. subsaponacea, with multiple names applied to a single species and multiple species described more than once. Molecular annotations using nuc rDNA ITS1-5.8S-ITS2 (internal transcribed spacer [ITS] barcode) and RPB2 (RNA polymerase II second largest subunit) are used to aid in application of these names in a phylogenetic context. Based on ITS molecular annotations of 13 types, the following heterotypic synonymies are proposed: P. highlandensis (syn. P. carbonaria and P. fulvozonata); P. molesta (syn. P. subsaponacea); and P. brunnescens (syn. P. luteobadia). In addition, we observed that the species P. castanea, known previously only from the type collection in Tennessee, is found commonly on burned sites near the Gulf Coast and other southeast regions of the United States. Overall, the pyrophilous trait is evolutionarily derived in Pholiota. Endophytic and endolichenic stages were deduced for P. highlandensis, the most widely distributed of the pyrophilous Pholiota. As a result, we introduce the "body snatchers" hypothesis that explains the maintenance of some pyrophilous fungi in ecosystems as endophytes and/or endolichenic fungi. Photographs, taxonomic descriptions, and a dichotomous key to pyrophilous species of Pholiota that occur in North America are presented.


Assuntos
Pholiota/classificação , Filogenia , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Técnicas de Tipagem Micológica , América do Norte , RNA Polimerase II/genética , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
15.
Sci Rep ; 8(1): 7740, 2018 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-29773815

RESUMO

The core α1-6 fucosylation-specific lectin from a mushroom Pholiota squarrosa (PhoSL) is a potential tool for precise diagnosis of cancers. This lectin consists of only 40 amino acids and can be chemically synthesized. We showed here that a synthesized PhoSL peptide formed a trimer by gel filtration and chemical cross-linking assays, and determined a structure of the PhoSL trimer by NMR. The structure possesses a ß-prism motif with a three-fold rotational symmetry, where three antiparallel ß-sheets are tightly connected by swapping of ß-strands. A triad of Trp residues comprises the structural core, forming NH-π electrostatic interactions among the indole rings. NMR analysis with an excess amount of fucose revealed the structural basis for the molecular recognition. Namely, fucose deeply enters a pocket formed at a junction of ß-sheet edges, with the methyl group placed at the bottom. It forms a number of hydrophobic and hydrogen-bonding interactions with PhoSL residues. In spite of partial similarities to the structures of other functionally related lectins, the arrangement of the antiparallel ß-sheets in the PhoSL trimer is novel as a structural scaffold, and thus defines a novel type of lectin structure.


Assuntos
Fucose/metabolismo , Lectina de Ligação a Manose/química , Lectina de Ligação a Manose/metabolismo , Fragmentos de Peptídeos/química , Pholiota/metabolismo , Multimerização Proteica , Sequência de Aminoácidos , Sítios de Ligação , Fucose/química , Ligação de Hidrogênio , Espectroscopia de Ressonância Magnética , Manose/química , Manose/metabolismo , Modelos Moleculares , Fragmentos de Peptídeos/metabolismo , Conformação Proteica
16.
Carbohydr Polym ; 169: 1-8, 2017 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-28504124

RESUMO

A crude fraction (SCW) was extracted with cold water from Pholiota nameko and showed mannose (24.1%), galactose (44.9%) and glucose (31%). Purification procedures resulted in a polysaccharide fraction (bG-PN), that showed only glucose. NMR and methylation analyses of bG-PN indicated a ß-d-glucan-(1→3)-linked, substituted at O-6 by ß-d-Glcp or (1→6)-linked ß-d-Glcp side chains. Rheological studies of crude and purified fractions at the same concentration showed similar shear-thinning behavior and gel-like structure which indicates no need to isolate the polymer to achieve some desirable rheological properties. SCW (at 1% and 2%) and bG-PN (at 2%) presented thermal stability during heating and cooling, suggesting that the physical structure of gels (SCW and bG-PN at 2%) and viscoelastic fluid (SCW at 1%) formed were not altered in the tested temperature range. Our results suggest that P. nameko ß-d-glucans can be applied in different food preparations as thickener or gelling agents modifying their rheological properties.


Assuntos
beta-Glucanas/química , Pholiota , Reologia , Substâncias Viscoelásticas/química , Substâncias Viscoelásticas/isolamento & purificação , beta-Glucanas/isolamento & purificação
17.
Glycoconj J ; 34(4): 537-544, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28455724

RESUMO

An increase in Lewis- and core-type fucosylation of haptoglobin has been reported in patients with pancreatic cancer (PC), suggesting that fucosylated haptoglobin is a candidate PC biomarker. Previously, we developed a Pholiota squarrosa lectin antibody enzyme-linked immunosorbent assay (PhoSL-ELISA) system for the detection of core-fucosylated haptoglobin. However, with this methodology, positive results were only obtained for some patients with PC, demonstrating the need for a more sensitive detection system. In the current study, we developed an improved PhoSL-ELISA system with higher sensitivity to detect core-fucosylated haptoglobin using high-concentration urea as a denaturing agent with lectin to facilitate detection. We then reevaluated the performance of PhoSL reactive-core-fucosylated haptoglobin (PhoSL-HP) as a PC biomarker using the improved PhoSL-ELISA system. PhoSL-HP levels in the sera of patients with PC were significantly higher than those in healthy volunteers, with an area under the curve (AUC) value of 0.753. Furthermore, the AUC value of CA19-9 improved from 0.793 to 0.907 when combined with PhoSL-HP. Additionally, several CA19-9-negative cases among the patients with PC were diagnosed as positive for PhoSL-HP. In conclusion, PhoSL-HP detection using our improved ELISA system might allow PhoSL-HP to serve as a potential biomarker for PC and thus might be useful to complement the detection of CA19-9 in PC diagnosis.


Assuntos
Biomarcadores Tumorais/metabolismo , Ensaio de Imunoadsorção Enzimática/métodos , Haptoglobinas/metabolismo , Lectinas/metabolismo , Neoplasias Pancreáticas/metabolismo , Pholiota/metabolismo , Adulto , Idoso , Antígenos Glicosídicos Associados a Tumores/metabolismo , Biomarcadores Tumorais/sangue , Biotinilação , Feminino , Células Hep G2 , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/sangue , Desnaturação Proteica
18.
Food Chem ; 216: 176-85, 2017 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-27596407

RESUMO

Edible mushrooms including Pholiota nameko are excellent sources of extractable bioactive compounds with much to explore. Enzymatic extractions with Cellulase and Viscozyme were responsible for highest extraction yields (67-77%). No strong antioxidant activity was observed although extracts were able to scavenge ABTS(+) and OH(+). Potential prebiotic activity was observed in all extracts, some increasing 1.4-2 Log cycles of Lactobacillus acidophilus La-5 and Bifidobacterium animalis BB12. 30-50% α-glucosidase inhibition was observed in ultrasound, Flavourzyme and Cellulase extracts. Flavourzyme and Cellulase extracts are richer in macro (Mg, K and P) and micro elements (Zn, Mn and Fe) in comparison to mushroom. Monosaccharides content and profile varied slightly among both extracts with predominance of glucose, galactose and mannose with no uronic acids detection; Flavourzyme extract reported higher free amino acids content. Presence of α and ß-glycosidic structures such as glucans and glucan-protein complexes are among the polysaccharides found in both extracts.


Assuntos
Pholiota/química , Celulase/metabolismo , Glucanos/análise , Inibidores de Glicosídeo Hidrolases/farmacologia , Espectroscopia de Ressonância Magnética , Monossacarídeos/análise , Polissacarídeos/análise
19.
Int J Med Mushrooms ; 18(11): 977-989, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28008810

RESUMO

A polysaccharide purified from Pholiota nameko (PNPS-1) was found to have anticancer and anti-inflammatory activity. This study investigated the effect of PNPS-1 on the nuclear factor (NF)-κB signaling pathway of TLR2 small interfering RNA-silenced murine bone marrow-derived dendritic cells (BMDCs) and relevant mechanisms. The expression of messenger RNA of 4 NF-κB-related genes, including MyD88, IKBKB, RelA(p65), and CCL2, was determined by real-time polymerase chain reaction; the expression of the phenotype molecule intercellular adhesion molecule-1 (ICAM-1) by flow cytometry; the protein expression of IKKß and p65 by Western blot; the production of p65 by enzyme-linked immunosorbent assay; and the expression of p65 by immunocytochemistry. The results showed that TLR2-specific small interfering RNA could effectively inhibit the decrease in the expression of MyD88, IKBKB, CCL2, p65, and ICAM-1 in BMDCs induced by PNPS-1, and thus the transcription inactivation of NF-κB, which obviously suggests that PNPS-1 could downregulate the NF-κB signaling pathway via the TLR2 receptor.


Assuntos
Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Imunossupressores/isolamento & purificação , NF-kappa B/antagonistas & inibidores , Pholiota/química , Polissacarídeos/isolamento & purificação , Receptor 2 Toll-Like/agonistas , Animais , Western Blotting , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Perfilação da Expressão Gênica , Imunossupressores/farmacologia , Masculino , Camundongos Endogâmicos C57BL , Polissacarídeos/farmacologia , Reação em Cadeia da Polimerase em Tempo Real
20.
Acc Chem Res ; 49(11): 2372-2380, 2016 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-27696815

RESUMO

Bioluminescence, the ability of a living organism to produce light through a chemical reaction, is one of Nature's most amazing phenomena widely spread among marine and terrestrial species. There are various different mechanisms underlying the emission of "cold light", but all involve a small molecule, luciferin, that provides energy for light-generation upon oxidation, and a protein, luciferase, that catalyzes the reaction. Different species often use different proteins and substrates in the process, which suggests that the ability to produce light evolved independently several times throughout evolution. Currently, it is estimated that there are more than 30 different mechanisms of bioluminescence. Even though the chemical foundation underlying the bioluminescence phenomenon is by now generally understood, only a handful of luciferins have been isolated and characterized. Today, the known bioluminescence reactions are used as indispensable analytical tools in various fields of science and technology. A pressing need for new bioluminescent analytical techniques with a wider range of practical applications stimulates the search and chemical studies of new bioluminescent systems. In the past few years two such systems were unraveled: those of the earthworms Fridericia heliota and the higher fungi. The luciferins of these two systems do not share structural similarity with the previously known ones. This Account will survey structure elucidation of the novel luciferins and identification of their mechanisms of action. Fridericia luciferin is a key component of a novel ATP-dependent bioluminescence system. Structural studies were performed on 0.005 mg of natural substance and revealed its unusual extensively modified peptidic nature. Elucidation of Fridericia oxyluciferin revealed that oxidative decarboxylation of a lysine fragment of luciferin supplies energy for light generation, while a fluorescent CompX moiety remains intact and serves as a light emitter. Along with luciferin, a number of its natural analogs were found in the extracts of worm biomass. They occurred to be highly unusual modified peptides comprising a set of amino acids, including threonine, aminobutyric acid, homoarginine, unsymmetrical N,N-dimethylarginine and extensively modified tyrosine. These natural compounds represent a unique peptide chemistry found in terrestrial animals and raise novel questions concerning their biosynthetic origin. Also in this Account we discuss identification of the luciferin of higher fungi 3-hydroxyhispidin which is biosynthesized by oxidation of the precursor hispidin, a known fungal and plant secondary metabolite. Furthermore, it was shown that 3-hydroxyhispidin leads to bioluminescence in extracts from four diverse genera of luminous fungi, thus suggesting a common biochemical mechanism for fungal bioluminescence.


Assuntos
Proteínas Fúngicas/química , Proteínas de Helminto/química , Substâncias Luminescentes/química , Peptídeos/química , Pironas/química , Animais , Ácidos Cumáricos/química , Luciferases/química , Luminescência , Lisina/análogos & derivados , Lisina/química , Modelos Químicos , Oligoquetos , Oxirredução , Pholiota
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