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1.
Afr Health Sci ; 20(2): 597-604, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33163020

RESUMO

Background: Saliva is a complex secretion produced daily by the salivary glands. Saliva consists mainly of water, enzymes, ions and amino acids and performs several important functions in oral health. Objective: The aim of this study was to investigate the flow rate and concentrations of amylase and total proteins in the saliva of hospitalized patients due to AIDS complications. Methods: Ninety-three men and women (20-64 years of age) were divided into two groups (46 HIV-infected patients and 47 controls) and had salivary flow rate and levels of amylase enzyme and total proteins evaluated. Result: The mean salivary flow rate was lower in individuals with HIV when compared to controls (P < 0.05). No significant difference between amylase enzyme levels and total proteins were observed in the saliva of patients with HIV infection when compared to controls. Conclusion: Individuals with HIV / AIDS infection (in hospital treatment) suffer no interference in levels of amylase and total salivary proteins, but they have significantly reduced salivary flow.


Assuntos
Infecções por HIV/complicações , Soropositividade para HIV/complicações , Saliva/metabolismo , Proteínas e Peptídeos Salivares/metabolismo , Salivação/fisiologia , Xerostomia/complicações , alfa-Amilases/metabolismo , Adolescente , Adulto , Idoso , Amilases/análise , Amilases/metabolismo , Fármacos Anti-HIV/uso terapêutico , Terapia Antirretroviral de Alta Atividade/efeitos adversos , Estudos de Casos e Controles , Feminino , Infecções por HIV/sangue , Infecções por HIV/tratamento farmacológico , Soropositividade para HIV/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Glândulas Salivares/metabolismo , Proteínas e Peptídeos Salivares/análise , Taxa Secretória/efeitos dos fármacos , Taxa Secretória/fisiologia , Xerostomia/etiologia , Adulto Jovem , alfa-Amilases/análise
2.
PLoS One ; 15(10): e0240805, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33104741

RESUMO

Ditylenchus destructor is a migratory plant-parasitic nematode that causes huge damage to global root and tuber production annually. The main plant hosts of D. destructor contain plenty of starch, which makes the parasitic environment of D. destructor to be different from those of most other plant-parasitic nematodes. It is speculated that D. destructor may harbor some unique pathogenesis-related genes to parasitize the starch-rich hosts. Herein, we focused on the multi-copy alpha-amylase genes in D. destructor, which encode a key starch-catalyzing enzyme. Our previously published D. destructor genome showed that it has three alpha-amylase encoding genes, Dd_02440, Dd_11154, and Dd_13225. Comparative analysis of alpha-amylases from different species demonstrated that the other plant-parasitic nematodes, even Ditylenchus dipsaci in the same genus, harbor only one or no alpha-amylase gene, and the three genes from D. destructor were closely clustered in the phylogenetic tree, indicating that there was a unique expansion of the alpha-amylase gene in D. destructor. The enzymatic activity of the three alpha-amylase proteins was verified by an enzyme assay. Quantitative real-time PCR assay showed that the expression of the three alpha-amylase genes in the post-hatching stage of D. destructor was found to be significantly higher than that in eggs. In the in situ hybridization assay, the expression of the genes was localized to the intestine, implying the association of these genes with nematode digestion. An infection assay in sweet potato demonstrated that RNA interference of any one alpha-amylase gene had no influence on the infectivity of D. destructor. Using the multi-target dsRNA cocktail method, it was found that silencing of two of the three genes inhibited nematode infection, and the infectivity of worms treated with three dsRNA simultaneously changed the most, which decreased by 76.6%. Thus, the multi-copy alpha-amylase genes in D. destructor are compensatory and crucial for nematodes to parasitize the plant host.


Assuntos
Tylenchoidea/genética , alfa-Amilases/genética , Sequência de Aminoácidos , Amilases/metabolismo , Animais , Regulação da Expressão Gênica de Plantas/genética , Interações Hospedeiro-Parasita/genética , Nematoides/genética , Parasitos/genética , Filogenia , Tubérculos/metabolismo , Tubérculos/parasitologia , Interferência de RNA , Reação em Cadeia da Polimerase em Tempo Real/métodos , Rabditídios/genética , Amido/metabolismo , Tylenchida/genética
3.
J Anim Sci ; 98(10)2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32910163

RESUMO

Developed initially for use in fuel ethanol production, Enogen Feed Corn (EFC; Syngenta Crop Protection) is genetically modified to express high concentrations of α-amylase in the corn kernel. Experiments were conducted to evaluate processing characteristics of EFC, in vitro digestion, and effects on feedlot performance, carcass characteristics, and liver abscess incidence. Experiment 1 used a randomized complete block design (3 × 3 × 5 factorial) to evaluate starch availability, in situ dry matter disappearance (ISDMD), in vitro gas production (IVGP), and volatile fatty acid (VFA) profiles of in vitro cultures. Grains (EFC or mill-run control [CON]) were flaked to a density of 360 g/L, and mixtures with 0%, 25%, 50%, 75%, or 100% EFC were prepared. Grains were tempered with added moisture (0%, 3%, or 6%) prior to steam conditioning for 15, 30, or 45 min. No two- or three-way interactions were observed. Adding moisture improved starch availability (linear; P < 0.01), and tended to improve ISDMD (linear, P = 0.06). Steam conditioning for 30 min improved starch availability, IVGP, and production of acetate, propionate, butyrate, valerate, and total VFA (P < 0.01) compared with conditioning for 15 or 45 min. Starch availability, ISDMD, IVGP, acetate, propionate, valerate, and total VFA production increased with an increasing proportion of EFC (linear, P < 0.01). Experiment 2 used 700 beef heifers (394 ± 8.5 kg initial body weight [BW]) fed finishing diets with steam-flaked corn as CON or EFC for 136 d. Targeting similar starch availabilities, grains were processed to 360 g/L (CON) and 390 g/L for CON and EFC, respectively. Heifers were blocked by BW, stratified, and then randomly assigned to 28 dirt-surfaced pens (25 animals per pen). Dry matter intakes were similar between treatments (P = 0.78), but cattle fed EFC had greater average daily gain (P < 0.01), improving feed efficiency by 5% (P < 0.01). Hot carcass weight was 6 kg greater for EFC cattle (P <0.01) than CON. No differences were observed for longissimus muscle area (P = 0.89), 12th-rib fat thickness (P = 0.21), or USDA yield grade (P = 0.13). Cattle fed CON had greater marbling scores than EFC (P = 0.04), but this did not affect the USDA quality grade (P > 0.33). Cattle fed EFC had 23% fewer abscessed livers than CON (P = 0.03). High-amylase corn may be used to improve microbial digestion, mill-throughput, and cattle performance, and it may mitigate liver abscesses.


Assuntos
Amilases/farmacologia , Ração Animal/análise , Bovinos/fisiologia , Dieta/veterinária , Zea mays/enzimologia , Amilases/administração & dosagem , Amilases/química , Amilases/metabolismo , Fenômenos Fisiológicos da Nutrição Animal , Animais , Composição Corporal , Digestão/fisiologia , Ácidos Graxos Voláteis/farmacologia , Feminino , Amido/farmacologia , Vapor
4.
Ann Surg ; 272(5): 863-870, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32833754

RESUMO

OBJECTIVE: We investigated the activation of pancreatic proenzymes and signs of peripancreatic inflammation in patients with clinically relevant postoperative pancreatic fistulas (POPFs). SUMMARY BACKGROUND DATA: An increase of systemic amylase concentration was associated with POPFs. This suggested parallels in the pathomechanisms between the development of POPFs and pancreatitis. METHODS: Trypsinogen, procathepsin B, and IL-6 concentrations as well as cathepsin B, myeloperoxidase and trypsin activities were determined throughout the first 7 postoperative days in drain fluids of 128 consecutive patients after pancreas resection. Histology and immunohistochemistry were performed in pancreatic specimens after total pancreatectomy due to complications and after placing experimental pancreatic sutures in the pancreatic tail of C57/Bl6 mice. RESULTS: Trypsin activity, cathepsin B activity and myeloperoxidase activity on the first postoperative day were elevated and predictive for clinically relevant pancreatic fistulas. Drain fluid stabilized trypsin activity and prevented the activation of the cascade of digestive enzymes. Leukocytes were the source of cathepsin B in drain fluid. Findings differed between fistulas after distal pancreatectomy and pancreatoduodenectomy. Immunohistochemistry of the pancreatic remnant revealed an inflammatory infiltrate expressing cathepsin B, independent of the presence of pancreatic fistulas. The infiltrate could be reproduced experimentally by sutures placed in the pancreatic tail of C57/Bl6 mice. CONCLUSIONS: Trypsinogen activation, increased cathepsin B activity and inflammation around the pancreato-enteric anastomosis on post operative day 1 are associated with subsequent clinically relevant POPFs after pancreatoduodenectomy. The parenchymal damage seems to be induced by placing sutures in the pancreatic parenchyma during pancreatic surgery.


Assuntos
Pancreatectomia , Fístula Pancreática/enzimologia , Complicações Pós-Operatórias/enzimologia , Amilases/metabolismo , Animais , Catepsina B/metabolismo , Precursores Enzimáticos/metabolismo , Feminino , Humanos , Inflamação/enzimologia , Interleucina-6/metabolismo , Masculino , Camundongos , Peroxidase/metabolismo , Estudos Prospectivos , Tripsina/metabolismo , Tripsinogênio/metabolismo
5.
Res Vet Sci ; 132: 54-56, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32485464

RESUMO

The European eel has recently been included on the Red List of the International Union for Conservation of Nature (IUCN) as a critically endangered species. The rearing of Anguilla larvae is seen as a key bottleneck to the mass production of glass eels since very little ecological information is available regarding their natural nutrition. Studies of digestive physiology and ontogenetic development in eel larvae could provide useful information for solving some of the puzzles regarding larval fish culture. The aim of this study was to characterize the ontogeny of pancreatic enzymes (trypsin, lipase and amylase) and a peptide hormone regulator of pancreatic secretion (cholecystokinin) in terms of gene expression in European eel larvae from day 0 (P0) of hatching to 5, 10, 15 and 20 days post hatching during fasting. The results in the present study showed that all the genes selected were present, with different levels of expression and increasing trends, during larval development. At P0, the increase in the gene expression of lipase and amylase was higher than that of trypsin and cholecystokinin, confirming that enzymatic activity began before mouth opening and that larvae, provided with a complete enzymatic set, might have the capacity of digesting and absorbing various nutrients.


Assuntos
Anguilla/metabolismo , Fenômenos Fisiológicos do Sistema Digestório , Proteínas de Peixes/metabolismo , Privação de Alimentos/fisiologia , Amilases/metabolismo , Anguilla/crescimento & desenvolvimento , Fenômenos Fisiológicos da Nutrição Animal , Animais , Aquicultura , Colecistocinina/metabolismo , Feminino , Lipase/metabolismo , Tripsina/metabolismo
6.
Food Microbiol ; 91: 103507, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32539950

RESUMO

Bacillus spp. cause ropy bread spoilage of bread, which is characterized by a rotten fruity odor, followed by discoloration and degradation of the crumb. Bacillus spp. are wheat grain endophytes and form heat resistant endospores, therefore, process hygiene and heating during baking do not prevent ropy spoilage. This study used 8 strains of Bacillus subtilis and Bacillus amyloliquefaciens to determine whether the presence and the copy number of spoVA2mob operon influences survival after baking; in addition, the spoilage phenotype was correlated with the presence of amylolytic enzymes in genomes of Bacillus spp.. The presence and copy number of the spoVA2mob operon had only a minor effect on survival of Bacillus endospores. Strains of B. amyloliquefaciens caused ropy spoilage faster than strains of B. subtilis, this difference correlated to the number and type of extracellular amylases encoded in the genomes of the strains of B. amyloliquefaciens and B. subtilis. The inhibitory effect of sourdough on ropy spoilage was determined by addition of 3-24% sourdough fermented with L. reuteri TMW1.656. Addition of 12% and 24% sourdough, corresponding to a bread pH of 5.93 ± 0.041 and 5.53 ± 0.040, respectively, delayed ropy spoilage for 2 and more than 5 d, respectively. The comparison of addition of 12% sourdough fermented with the reutericyclin producing L. reuteri TMW1.656 and the isogenic reutericyclin-negative strain L. reuteri TMW1.656ΔgtfAΔrtcN demonstrated that reutericyclin produced in sourdough inhibits growth of Bacillus in bread. In conclusion, sourdough inhibits germination of Bacillus spores in bread and the effect of sourdough is enhanced by reutericyclin.


Assuntos
Bacillus/metabolismo , Proteínas de Bactérias/genética , Pão/microbiologia , Ácido Tenuazônico/análogos & derivados , Amilases/genética , Amilases/metabolismo , Bacillus/classificação , Bacillus/genética , Bacillus/crescimento & desenvolvimento , Proteínas de Bactérias/metabolismo , Variações do Número de Cópias de DNA , Fermentação , Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Lactobacillus reuteri/genética , Lactobacillus reuteri/metabolismo , Viabilidade Microbiana , Óperon , Esporos Bacterianos/classificação , Esporos Bacterianos/genética , Esporos Bacterianos/crescimento & desenvolvimento , Temperatura , Ácido Tenuazônico/metabolismo
7.
Am J Physiol Gastrointest Liver Physiol ; 319(1): G74-G86, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32538138

RESUMO

The mechanism for segregation of cargo proteins into the regulated and constitutive secretory pathways in exocrine cells remains to be elucidated. We examined the transport of HaloTag proteins fused with full-length cystatin D (fCst5-Halo) or only its signal peptide (ssCst5-Halo) in parotid acinar cells. Although both fusion proteins were observed to be colocalized with amylase in the secretory granules, the coefficients for overlapping and correlation of fCst5-Halo with amylase were higher than those of ssCst5-Halo. The secretion of both the proteins was enhanced by the addition of the ß-adrenergic receptor agonist isoproterenol as well as endogenous amylase. In contrast, unstimulated secretion of ssCst5-Halo without isoproterenol was significantly higher than that of fCst5-Halo and amylase. Simulation analysis using a mathematical model revealed that a large proportion of ssCst5-Halo was secreted through the constitutive pathway, whereas fCst5-Halo was transported into the secretory granules more efficiently. Precipitation of fCst5-Halo from cell lysates was increased at a low pH, which may mimic the milieu of the trans-Golgi networks. These data suggest that the addition of a full-length sequence of cystatin D facilitates efficient selective transport into the regulated pathway by aggregation at low pH in the trans-Golgi network.NEW & NOTEWORTHY The mechanism underlying the segregation of cargo proteins to the regulated and constitutive secretory pathways in exocrine cells remains to be solved. We analyzed unstimulated secretion in salivary acinar cells by performing double-labeling experiments using HaloTag technology and computer simulation. It revealed that the majority of HaloTag with only signal peptide sequence was secreted through the constitutive pathway and that the addition of a full-length cystatin D sequence changed its sorting to the regulated pathway.


Assuntos
Células Acinares/metabolismo , Movimento Celular/fisiologia , Transporte Proteico/fisiologia , Via Secretória/fisiologia , Amilases/metabolismo , Animais , Células Cultivadas , Exocitose/fisiologia , Glândula Parótida/metabolismo
8.
Int J Nanomedicine ; 15: 3405-3414, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32523340

RESUMO

Background: Fluorescent metallic nanodots (NDs) have become a promising nanoprobe for a wide range of biomedical applications. Because Ag NDs have a high tendency to be oxidized, their synthesis and storage are a big challenge. Thus, the method for preparing stable Ag NDs is urgently needed. Surface modification and functionalization can enrich the capability of Ag NDs. Methods: In this work, fluorescent Ag NDs were synthesized in deoxygenated water by using porcine pancreatic α-amylase (PPA) as the stabilizing/capping agent. The absorption and fluorescence of PPA-protected Ag NDs (PPA@AgNDs) were measured with a spectrophotometer and a spectrofluorometer, respectively. The morphology of PPA@AgNDs was characterized by high-angle annular dark-field (HAADF) scanning transmission electron microscopy (STEM). The biocompatibility of PPA@AgNDs was evaluated by tetrazolium (MTT)-based assay. PolyLys-Cys-SH (sequence: KKKKKKC) peptides were conjugated to PPA@AgNDs via heterobifunctional crosslinkers. PolyLys-Cys-linked PPA@AgNDs absorbed 5-aminolevulinic acid (ALA) by electrostatic interaction at physiological pH. The capability of tumor targeting was evaluated by intravenously injecting PPA@AgND-ALA into 4T1 breast cancer xenograft mouse models. Photodynamic therapy (PDT) against tumors was performed under 635 nm laser irradiation. Results: PPA@AgNDs emitted at 640 nm with quantum yield of 2.1%. The Ag NDs exhibited strong photostability over a long period and a fluorescence lifetime of 5.1 ns. PPA@AgNDs easily entered the cells to stain the nuclei, showing the capabilities of living cell imaging with negligible cytotoxicity. ALA-loaded PPA@AgNDs (PPA@AgND-ALA) presented the superiority of passive tumor targeting via the enhanced permeability and retention (EPR) effect. Tumors were visualized in the near-infrared (NIR) region with reduced background noise. ALA molecules released from PPA@AgND-ALA was converted into the photosensitizer (PS) of protoporphyrin IX (PpIX) intracellularly and intratumorally, which greatly improved the PDT efficacy. Conclusion: Our approach opens a new way to design a novel theranostic nanoplatform of PPA@AgND-ALA for effective tumor targeting and fluorescence image-guided PDT.


Assuntos
Amilases/metabolismo , Nanopartículas/química , Neoplasias/tratamento farmacológico , Imagem Óptica , Fotoquimioterapia , Prata/química , Animais , Linhagem Celular Tumoral , Fluorescência , Humanos , Ácidos Levulínicos/farmacologia , Camundongos , Tamanho da Partícula , Espectrofotometria Ultravioleta , Suínos , Nanomedicina Teranóstica , Ensaios Antitumorais Modelo de Xenoenxerto
9.
Nutrients ; 12(4)2020 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-32260553

RESUMO

It is widely recognized that smelling food results in a mouth-watering feeling and influences appetite. However, besides changes in volume, little is known about the effects that food odours have on the composition of saliva. The aim of the present study was to access the effects that smelling bread has on saliva proteome and to compare such effects with those of chewing and ingesting it. Besides a significant increase in saliva flow rate, together with a decrease in total protein concentration, bread odour induced changes in the proportion of different salivary proteins. The expression levels of two spots of cystatins and two spots of amylase increased due to olfactory stimulation, similar to what happened with bread mastication, suggesting that odour can allow anticipation of the type of food eaten and consequently the physiological oral changes necessary to that ingestion. An interesting finding was that bread odour increased the expression levels of several protein spots of immunoglobulin chains, which were decreased by both bread or rice mastication. This may be of clinical relevance since food olfactory stimulation of salivary immunoglobulins can be used to potentiate the oral immune function of saliva. Moreover, the effects of bread odour in the levels of salivary proteins, previously observed to be involved in oral food processing led to the hypothesis of an influence of this odour in the sensory perception of foods further ingested. Further studies are needed to elucidate this point, as well as whether the changes observed for bread odour are specific, or if different food odours lead to similar salivary proteome responses.


Assuntos
Pão , Odorantes , Saliva/química , Saliva/metabolismo , Adulto , Amilases/genética , Amilases/metabolismo , Apetite , Feminino , Regulação da Expressão Gênica , Humanos , Oryza , Proteínas/química , Proteínas/genética , Proteínas/metabolismo , Adulto Jovem
10.
Med Sci Monit ; 26: e920684, 2020 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-32283546

RESUMO

BACKGROUND Acute pancreatitis (AP) is a symptom of sudden pancreas inflammation, which causes patients severe suffering. In general, fibroblast growth factor (FGF) levels are increased and amylase and lipase activities are elevated during AP pathogenesis, but protein concentration are low. However, the mechanism through which FGF signaling regulates AP pathogenesis remains elusive. MATERIAL AND METHODS The concentrations of PGE2, TNF-alpha, sCRP, FGF1, and FGF2 in the serum samples of the AP group and healthy control group were detected by enzyme-linked immunosorbent assay. In addition, IkappaBalpha and p-IkappaBalpha levels were analyzed in the serum samples. Subsequently, the AP rat model was established, and FGF1, FGF2, anti-FGF1, and anti-FGF2 antibodies and Bay11-7082 were injected into AP rats. TNF-alpha, PAI-1 JNK, p-JNK, IkappaBalpha, and p-IkappaBalpha levels were also examined. RESULTS Results showed that levels of PGE2, TNF-alpha, sCRP, p-IkappaBalpha, FGF1, and FGF2, as well as amylase and lipase activity were increased in patients with AP compared with those in healthy people. In addition, protein concentrations were lower in patients with AP than in the healthy group. Activation of FGF signaling by injecting FGF1 or FGF2 also inhibited AP-induced inflammation response in the pancreas and increased amylase and lipase activities, as well as protein concentration. However, the injection of FGF1 and FGF2 antibodies accelerated AP-mediated inflammation responses in the serum. In addition, Bay11-7082 injection inhibited AP activation of inflammation response and amylase and lipase activities. Protein concentration were also increased in AP rats. CONCLUSIONS FGF signaling protects against AP-mediated damage by inhibition of AP-activating inflammatory responses.


Assuntos
Fator 1 de Crescimento de Fibroblastos/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , Inflamação/metabolismo , Pancreatite/patologia , Transdução de Sinais , Doença Aguda , Adulto , Amilases/metabolismo , Animais , Proteína C-Reativa/análise , Estudos de Casos e Controles , Dinoprostona/sangue , Feminino , Fator 1 de Crescimento de Fibroblastos/sangue , Fator 2 de Crescimento de Fibroblastos/sangue , Humanos , Inflamação/patologia , Lipase/metabolismo , Masculino , Pessoa de Meia-Idade , Inibidor de NF-kappaB alfa/sangue , Nitrilos/farmacologia , Ratos , Ratos Sprague-Dawley , Sulfonas/farmacologia , Fator de Necrose Tumoral alfa/sangue
11.
Toxicon ; 180: 43-48, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32298663

RESUMO

Zearalenone, an oestogenic mycotoxin produced by Fusarium sp., occurs naturally in agricultural commodities. Economic losses and health concerns associated to mycotoxins has attracted research interest towards exploring novel approaches to detoxify mycotoxin-contaminated food and feed. The aim of the present work was to study the ability of 11 aflatoxin-degrading Bacillus strains to degrade ZEA. In addition, a qualitative assessment of protease, amylase and cellulase activity of the studied Bacillus strains was made. All strains were able to degrade 58-96.9% ZEA after 72 h. Toxicity towards Artemia salina was significantly reduced (P < 0.0001). Degradation extracts fluorescence decreased 50% indicating a probable cleavage of the lactone ring. Strains RC1A, RC3A and RC6A showed a remarkable enzymatic activity, showing potential to be used as feed additives.


Assuntos
Aflatoxinas/metabolismo , Amilases/metabolismo , Bacillus/metabolismo , Celulases/metabolismo , Peptídeo Hidrolases/metabolismo , Zearalenona/metabolismo , Agricultura , Inativação Metabólica
12.
Biochem Biophys Res Commun ; 526(3): 692-698, 2020 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-32248979

RESUMO

BACKGROUND: We reported that the pancreas of the interferon-regulatory factor (IRF) 2 knock-out (KO) mouse represents an early phase of acute pancreatitis, including defective regulatory exocytosis, intracellular activation of trypsin, and disturbance of autophagy. The significantly upregulated and downregulated genes in the IRF2 KO pancreas have been reported. The catalogue of gene transcripts included two types of calcium-binding proteins (S100 calcium binding protein G [S100g] and Annexin A10 [Anxa10]), which were highly upregulated in the IRF2 KO pancreas. As the intracellular calcium signal plays a pivotal role in regulatory exocytosis and its disturbance is related to pancreatitis, we then evaluated the role of S100g and Anxa10 in acute pancreatitis. METHOD: We induced cerulein-pancreatitis in wild-type mice and examined the changes in the expression of these genes by qPCR and immunohistochemistry. We constructed S100g-overexpressing or Anxa10-overexpressing AR42J cells (AR42J-S100g, AR42J-Anxa10). We examined the changes in amylase secretion, intracellular calcium ([Ca2+]i), and cell viability in these cells, when incubated with cholecystokinin (CCK). RESULTS: The expression of S100g and Anxa10 was increased in cerulean-induced pancreatitis. The acini were patchily stained for S100g and the cytosol of acini was evenly but weakly stained for Anxa10. Stimulation with 100pM CCK-8, decreased amylase secretion and inhibited the [Ca2+]i increase in AR42J-S100g cells. These effects were weak in AR42J-Anxa10 cells. Cell viability was not changed by incubation with cerulein. CONCLUSION: In cerulean pancreatitis, the expression of S100g and Anxa10 was induced in the acini. S100g may work as a Ca2+ buffer in acute pancreatitis.


Assuntos
Anexinas/metabolismo , Cálcio/metabolismo , Pancreatite/metabolismo , Proteína G de Ligação ao Cálcio S100/metabolismo , Células Acinares/citologia , Células Acinares/metabolismo , Amilases/metabolismo , Animais , Anexinas/genética , Autofagia , Sobrevivência Celular , Ceruletídeo/metabolismo , Colecistocinina/metabolismo , Exocitose , Fator Regulador 2 de Interferon/metabolismo , Camundongos Knockout , Pâncreas/efeitos dos fármacos , Fragmentos de Peptídeos/metabolismo , Proteína G de Ligação ao Cálcio S100/genética , Transdução de Sinais , Regulação para Cima
13.
J Sci Food Agric ; 100(8): 3544-3553, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32242927

RESUMO

BACKGROUND: Jiuqu are vital saccharifying and fermenting agents for Chinese fermented foods. Natural ventilation during Jiuqu fermentation causes changes in temperature, oxygen and moisture content, resulting in mass and heat gradients from the outer to inner areas of Jiuqu blocks. In the present study, microbiota stratification in Jiuqu was investigated by single molecule real-time sequencing and culture isolation. The contributors of Bacillus to amylase activity of Jiuqu and the dynamics of their biomass during Jiuqu fermentation were also analyzed. RESULTS: The dominant orders, genera and species between the inner and outer layers of Huangjiu qu (HJQ) were similar, although they displayed greater variance in two layers of Baijiu qu (BJQ). Bacillus possessed the highest diversity (including 27 species) in Jiuqu. Bacillus licheniformis, Bacillus altitudinis, Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus megaterium were most prevalent in HJQ, whereas B. licheniformis, B. amyloliquefaciens and Bacillus cereus were dominant in BJQ. Isolates of B. amyloliquefaciens, B. subtilis and B. cereus exhibited high activities of amylase and glucoamylase. Quantification of Bacillus members possessing genes of α-amylase revealed that B. cereus and B. licheniformis were the most dominant microbes to secret α-amylase in Jiuqu and their biomass were increasing during Jiuqu fermentation. CONCLUSION: The present study demonstrates the microbial distribution in different layers of Jiuqu and clarifies the Bacillus species processing the activity of α-amylase. These results will help industries control the quality of Jiuqu by rationally selecting starters and optimizing their microbiota. © 2020 Society of Chemical Industry.


Assuntos
Bacillus/metabolismo , Proteínas de Bactérias/metabolismo , Microbiota , Oryza/microbiologia , Amilases/genética , Amilases/metabolismo , Bacillus/classificação , Bacillus/enzimologia , Bacillus/genética , Proteínas de Bactérias/genética , Fermentação , Alimentos e Bebidas Fermentados/microbiologia , Microbiologia de Alimentos
14.
Biochem Pharmacol ; 177: 113992, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32335141

RESUMO

IL-17A combined with TNF-α plays a vital role in inflammatory response and interference of the synergistic effect is an effective strategy for treating inflammatory diseases. Ellipticine, a natural alkaloid, has biological activities on anti-tumor and anti-HIV. However, it is still unknown whether ellipticine can inhibit IL-17A and TNF-α-mediated signaling and has treatment effect on PALI. Here, we reported that ellipticine significantly inhibited the production of pro-inflammatory cytokines and chemokines in pulmonary epithelial cell BEAS-2B treated with IL-17A and TNF-α, but not IL-17A or TNF-α alone. Meanwhile, ellipticine attenuated NF-κB and MAPKs activation in response to IL-17A and TNF-α treatment, inhibited Act1 and TRAF6-mediated NF-κB activation, and blocked the interaction of Act1 with TRAF6. Furthermore, we found that ellipticine significantly alleviated CAE and LPS-induced SAP/PALI. Ellipticine treatment dramatically reduced inflammatory cells infiltration, MPO activity, serum amylase and lipase activity and the protein concentration of BALF. Collectively, our findings indicate that ellipticine inhibits the synergistic effect of IL-17A and TNF-α by targeting on Act1 and TRAF6 interaction and is a potential therapeutic agent for the treatment of SAP/PALI.


Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Anti-Inflamatórios/farmacologia , Elipticinas/farmacologia , Interleucina-17/antagonistas & inibidores , Pancreatite/tratamento farmacológico , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/complicações , Lesão Pulmonar Aguda/genética , Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Amilases/antagonistas & inibidores , Amilases/genética , Amilases/metabolismo , Animais , Linhagem Celular Transformada , Ceruletídeo/administração & dosagem , Modelos Animais de Doenças , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Regulação da Expressão Gênica , Células HEK293 , Humanos , Interleucina-17/farmacologia , Lipase/antagonistas & inibidores , Lipase/genética , Lipase/metabolismo , Lipopolissacarídeos/administração & dosagem , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , NF-kappa B/metabolismo , Pancreatite/induzido quimicamente , Pancreatite/complicações , Pancreatite/genética , Peroxidase/antagonistas & inibidores , Peroxidase/genética , Peroxidase/metabolismo , Transdução de Sinais , Fator 6 Associado a Receptor de TNF/antagonistas & inibidores , Fator 6 Associado a Receptor de TNF/genética , Fator 6 Associado a Receptor de TNF/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
15.
Cell Host Microbe ; 27(4): 571-584.e7, 2020 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-32220647

RESUMO

Legionella pneumophila has co-evolved with amoebae, their natural hosts. Upon transmission to humans, the bacteria proliferate within alveolar macrophages causing pneumonia. Here, we show L. pneumophila injects the effector LamA, an amylase, into the cytosol of human macrophage (hMDMs) and amoebae to rapidly degrade glycogen to generate cytosolic hyper-glucose. In response, hMDMs shift their metabolism to aerobic glycolysis, which directly triggers an M1-like pro-inflammatory differentiation and nutritional innate immunity through enhanced tryptophan degradation. This leads to a modest restriction of bacterial proliferation in hMDMs. In contrast, LamA-mediated glycogenolysis in amoebae deprives the natural host from the main building blocks for synthesis of the cellulose-rich cyst wall, leading to subversion of amoeba encystation. This is non-permissive for bacterial proliferation. Therefore, LamA of L. pneumophila is an amoebae host-adapted effector that subverts encystation of the amoebae natural host, and the paradoxical hMDMs' pro-inflammatory response is likely an evolutionary accident.


Assuntos
Amoeba/microbiologia , Amilases/metabolismo , Legionella pneumophila , Macrófagos Alveolares/microbiologia , Amoeba/metabolismo , Evolução Biológica , Citocinas/metabolismo , Glicogenólise , Interações Hospedeiro-Parasita , Humanos , Imunidade Inata , Legionella pneumophila/imunologia , Legionella pneumophila/metabolismo , Macrófagos Alveolares/metabolismo , Encistamento de Parasitas
16.
Appl Microbiol Biotechnol ; 104(9): 3745-3756, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32170387

RESUMO

Diastatic strains of Saccharomyces cerevisiae possess the unique ability to hydrolyze and ferment long-chain oligosaccharides like dextrin and starch. They have long been regarded as important spoilage microbes in beer, but recent studies have inspired a re-evaluation of the significance of the group. Rather than being merely wild-yeast contaminants, they are highly specialized, domesticated yeasts belonging to a major brewing yeast lineage. In fact, many diastatic strains have unknowingly been used as production strains for decades. These yeasts are used in the production of traditional beer styles, like saison, but also show potential for creation of new beers with novel chemical and physical properties. Herein, we review results of the most recent studies and provide a detailed account of the structure, regulation, and functional role of the glucoamylase-encoding STA1 gene in relation to brewing and other fermentation industries. The state of the art in detecting diastatic yeast in the brewery is also summarized. In summary, these latest results highlight that having diastatic S. cerevisiae in your brewery is not necessarily a bad thing. KEY POINTS: •Diastatic S. cerevisiae strains are important spoilage microbes in brewery fermentations. •These strains belong to the 'Beer 2' or 'Mosaic beer' brewing yeast lineage. •Diastatic strains have unknowingly been used as production strains in breweries. •The STA1-encoded glucoamylase enables efficient maltotriose use.


Assuntos
Amilases/genética , Amilases/metabolismo , Fermentação , Saccharomyces cerevisiae/enzimologia , Cerveja/microbiologia , Dextrinas/metabolismo , Saccharomyces cerevisiae/classificação , Amido/metabolismo
17.
Electron. j. biotechnol ; 44: 41-46, Mar. 2020. tab, ilus
Artigo em Inglês | LILACS | ID: biblio-1087698

RESUMO

Background: The main objective of this study was to isolate fungi associated with Anthopleura xanthogrammica and measure their antimicrobial and enzymatic activities. A total of 93 fungal strains associated with A. xanthogrammica were isolated in this study, of which 32 isolates were identified using both morphological characteristics and internal transcribed spacer (ITS) sequence analysis. The antibacterial activities of 32 fungal isolates were tested against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Edwardsiella tarda, Vibrio harveyi, Fusarium oxysporum, and Pyricularia oryzae by agar diffusion assay. Extracellular hydrolytic enzyme activities of the fungal isolates were determined by agar diffusion assays. Enzyme activities were detected from clear halo size. Results: The isolated fungi belonged to 18 genera within 7 taxonomic orders of 1 phylum. The genera Aspergillaceae were the most diverse and common. The antimicrobial activities of 32 isolates were evaluated, and 19 (59.4%) of fungi isolate displayed unique antimicrobial activities. All fungal strains displayed at least one enzyme activity. The most common enzyme activities in the fungi isolates were amylase and protease, while the least common were pectinase and xylanase. Conclusions: This is first report on the sea anemone-derived fungi with antimicrobial and enzyme activities. Results indicated that sea anemone is a hot spot of fungal diversity and a rich resource of bioactive natural products.


Assuntos
Aspergillus/isolamento & purificação , Anêmonas-do-Mar/microbiologia , Antibacterianos/isolamento & purificação , Peptídeo Hidrolases/metabolismo , Filogenia , Poligalacturonase/metabolismo , Aspergillus/enzimologia , Aspergillus/genética , Bactérias/efeitos dos fármacos , DNA Espaçador Ribossômico , Biodiversidade , Fungos/isolamento & purificação , Fungos/genética , Amilases/metabolismo , Antibacterianos/farmacologia
18.
Yeast ; 37(5-6): 336-347, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32065695

RESUMO

Saccharomyces cerevisiae cells grown in a small volume of chemically defined media neither reach the desired cell density nor grow at a fast enough rate to scale down the volume and increase the sample number of classical biochemical assays, as the detection limit of the readout often requires a high number of cells as an input. To ameliorate this problem, we developed and optimised a new high cell density (HCD) medium for S. cerevisiae. Starting from a widely used synthetic medium composition, we systematically varied the concentrations of all components without the addition of other compounds. We used response surface methodology to develop and optimise the five components of the medium: glucose, yeast nitrogen base, amino acids, monosodium glutamate, and inositol. We monitored growth, cell number, and cell size to ensure that the optimisation was towards a greater density of cells rather than just towards an increase in biomass (i.e., larger cells). Cells grown in the final medium, HCD, exhibit growth more similar to the complex medium yeast extract peptone dextrose (YPD) than to the synthetic defined (SD) medium. Whereas the final cell density of HCD prior to the diauxic shift is increased compared with YPD and SD about threefold and tenfold, respectively. We found normal cell-cycle behaviour throughout the growth phases by monitoring DNA content and protein expression using fluorescent reporters. We also ensured that HCD media could be used with a variety of strains and that they allow selection for all common yeast auxotrophic markers.


Assuntos
Meios de Cultura/química , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/genética , Aminoácidos/metabolismo , Amilases/metabolismo , Biomassa , Ciclo Celular , Tamanho Celular , Proteínas Fúngicas
19.
Ann Biol Clin (Paris) ; 78(1): 93-107, 2020 02 01.
Artigo em Francês | MEDLINE | ID: mdl-32108587

RESUMO

The measurement performance of 13 biochemistry parameters (CEA, CA 19-9, amylase, lipase, sodium, potassium, chloride, creatinine, glucose, protein, albumin, LDH, triglycerides) was tested in a panel of biological fluids other than blood and urine (peritoneal, pleural, pancreatic fluids ...). Our protocol, based on a risk analysis, allowed us to justify our choices and compare the performance obtained with those of the serum or plasma matrix already validated. Thus, the coefficients of variation obtained in body fluids are comparable. The assessment of accuracy (spiking and dilution tests) shows the absence of bias, which is consistent with the absence of matrix effect. The linearity studied by dilution tests shows that the upper limits of the measurement interval communicated by the supplier are applicable to body fluids. The absence of contamination and stability have been also confirmed. All analytes are stable for 3 days at room temperature, 7 days between 2 and 8̊C, and 6 months at -20̊C; except LDH and lipase. For most analytes, at least one interference (hemolysis, icterus, lipemia) was found. Finally, a bibliographical study, confronted with the experience of prescribers, led us to define optimal thresholds to help interpret patients' results. In conclusion, this work has allowed us to validate analytical methods for body fluids testing after relying on their comparability to the blood matrix. We have also been able to adapt our practices and finally be accredited according to the standard NF IN ISO 15189.


Assuntos
Biomarcadores/análise , Líquidos Corporais/química , Técnicas de Laboratório Clínico , Albuminas/análise , Albuminas/metabolismo , Amilases/análise , Amilases/metabolismo , Biomarcadores/metabolismo , Líquidos Corporais/metabolismo , Antígeno CA-19-9/análise , Antígeno CA-19-9/metabolismo , Antígeno Carcinoembrionário/análise , Antígeno Carcinoembrionário/metabolismo , Cloretos/análise , Cloretos/metabolismo , Técnicas de Laboratório Clínico/métodos , Técnicas de Laboratório Clínico/normas , Creatinina/análise , Creatinina/metabolismo , Glucose/análise , Glucose/metabolismo , Humanos , L-Lactato Desidrogenase/análise , L-Lactato Desidrogenase/metabolismo , Lipase/análise , Lipase/metabolismo , Potássio/análise , Potássio/metabolismo , Proteínas/análise , Proteínas/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Sódio/análise , Sódio/metabolismo , Temperatura , Triglicerídeos/análise , Triglicerídeos/metabolismo
20.
J Agric Food Chem ; 68(3): 838-844, 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31896254

RESUMO

Maltooligosaccharide-forming amylases (MFAses) are promising tools for a variety of food industry applications because they convert starch into functional maltooligosaccharides. The MFAse from Bacillus stearothermophilus STB04 (BstMFAse) is a thermostable enzyme that preferentially produces maltopentaose and maltohexaose. An X-ray crystal structure of the BstMFAse-acarbose complex suggested that mutation of glycine 109 would increase its maltohexaose specificity. Using site-directed mutagenesis, glycine 109 was replaced with several different amino acids. Mutant-containing asparagine (G109N), aspartic acid (G109D), and phenylalanine (G109F) produced 36.1, 42.4, and 39.0% maltohexaose from starch, respectively, which was greater than that produced by the wild-type (32.9%). These mutants also exhibited substantially altered oligosaccharide hydrolysis patterns in favor of maltohexaose production. Homology models suggested that the mutants form extra interactions with the substrate at subsite -6, which were responsible for the enhanced maltohexaose specificity of BstMFAse. The results of this study support the proposition that binding of the substrate's nonreducing end in the nonreducing end-subsite of the MFAse active center plays a crucial role in its product specificity.


Assuntos
Amilases/genética , Amilases/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Geobacillus stearothermophilus/enzimologia , Oligossacarídeos/metabolismo , Amilases/química , Proteínas de Bactérias/química , Sítios de Ligação , Geobacillus stearothermophilus/genética , Mutagênese Sítio-Dirigida , Oligossacarídeos/química , Engenharia de Proteínas , Amido/química , Amido/metabolismo , Especificidade por Substrato
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