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1.
PeerJ ; 11: e14606, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36643622

RESUMO

Background: Kaempferia galanga (L.) is one of the prospective therapeutic plants with an aromatic rhizome, and belongs to the Zingiberaceae family. This herb is commonly used by local practitioners in traditional Asian medicine. Methods: In the present investigation, the novel Kaempferia galanga rhizome essential oil rich in ethyl p-methoxy cinnamate (EMCKG) was evaluated using GC/MS for chemical composition analysis. EMCKG was analyzed for its possible antimicrobial, neurodegenerative inhibitory, acetylcholinesterase, anti-inflammatory, and antioxidant activities as well as for the genotoxic effects using the standard methodologies. ANOVA and post hoc was performed to test the statistical significance of the study. Results: GC/MS analysis identified ethyl p-methoxy cinnamate as the major component of EMCKG essential oil with an area percentage of 66.39%. The EMCKG exhibited moderate (DPPH assay IC50 = 15.64 ± 0.263 µg/mL; ABTS assay IC50 = 16.93 ± 0.228 µg/mL) antioxidant activity than standard ascorbic acid (DPPH assay IC50 = 21.24 ± 0.413 µg/mL; ABTS assay IC50 = 21.156 ± 0.345 µg/mL). Similarly, EMCKG showed comparable activity in albumin denaturation (IC50 = 2.93 ± 0.59 µg/mL) and protease inhibitor assay (IC50 = 17.143 ± 0.506 µg/mL) to that of standard sodium diclofenac (IC50 = 23.87 ± 0.729 µg/mL and IC50 = 19.18 ± 0.271 µg/mL, respectively). The EMCKG exhibited a dose-dependent antimicrobial activity pattern with the highest inhibitory activity at 500 µg/mL against Staphylococcus aureus and considerable anticholinesterase activities (IC50 = 21.94 ± 0.109 µg/mL) compared to the standard galanthamine (IC50 = 27.18 ± 0.511 µg/mL). EMCKG also showed strong anti-diabetic activity (IC50 = 18.503 ± 0.480 µg/mL) and anti-tyrosinase activity (IC50 = 14.756 ± 0.325 µg/mL) as compared to the standards used (acarbose IC50 = 20.39 ± 0.231 µg/mL and kojic acid IC50 = 17.73 ± 0.192 µg/mL) in the study. Genotoxicity analysis of EMCKG revealed that at 1 µg/mL concentration has no toxic effects in mitosis of Allium cepa roots (Mitotic Index MI = 13.56% and chromosomal aberration CA = 07.60%). The ANOVA confirmed that except for the anticholinesterase activity, there is insignificant difference for essential oil and standards used for all the other bioactivities thus confirming their interchangeable applicability. Conclusions: Current research provides the basis for the fact that besides being a rich source of ethyl p-methoxycinnamate, EMCKG has the potential for future formulation and development of an inexpensive skin-care agent and for the preparation of anti-diabetic drugs.


Assuntos
Alpinia , Anti-Infecciosos , Óleos Voláteis , Zingiberaceae , Inibidores da Colinesterase/farmacologia , Óleos Voláteis/farmacologia , Monofenol Mono-Oxigenase , alfa-Amilases , Acetilcolinesterase , Zingiberaceae/química , Cinamatos/farmacologia , Antioxidantes/farmacologia , Anti-Infecciosos/farmacologia
2.
Food Res Int ; 163: 112155, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36596106

RESUMO

Inhibiting carbohydrate-hydrolyzing enzymes has been considered as an effective approach for controlling starch digestion and postprandial blood glucose level. α-Amylase and amyloglucosidase (AMG) are commonly applied in analysis of starch digestion behaviour. Catechins have been shown with the inhibiting effects on α-amylase. However, the inhibitory activity of catechins against AMG needs to be further studied. Therefore, AMG inhibition of 8 catechins and the mechanisms were studied in this work through substrate depletion, inhibition kinetics, molecular docking, fluorescence quenching, differential scanning calorimetry, and isothermal titration calorimetry. The inhibitory activity of catechins with galloyl moiety (CGMs) was found to be lower than the corresponding catechins without the moiety (Cs). All catechins were anti-competitive inhibitors, indicating that they tended to bind with AMG-starch complex in the digestion system, rather than with AMG directly. Interestingly, CGMs had higher quenching effects on AMG fluorescence than Cs, due to the additional π-stacking between aromatic rings of GM and AMG fluorophores. Also, CGMs had a higher binding affinity to AMG, due to the tendency of GM to AMG active site, although the affinity was much weaker than that of starch to AMG. Besides, catechins did not affect AMG thermostability. Therefore, there was an inconsistency between catechins-AMG binding interactions and the enzyme inhibition because the predominant sites for catechins binding were the non-active sites on AMG-starch complex, rather than the enzyme active ones. Conclusively, inhibition mode should also be considered when evaluating the inhibitory activity of a polyphenol based on the polyphenol-enzyme binding affinity.


Assuntos
Catequina , Polifenóis , Polifenóis/farmacologia , Catequina/química , Glucana 1,4-alfa-Glucosidase , Simulação de Acoplamento Molecular , alfa-Amilases/metabolismo , Amido/química
3.
Food Res Int ; 163: 112180, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36596121

RESUMO

Tartary buckwheat was hydrolyzed with α-amylase, pullulanase, α-amylase and pullulanase double enzymes and fermented by Monascus. The fermentation products were named as enzymolysis-Monascus-fermented tartary buckwheat (EMFTB). The composition and content of volatile flavor compounds in EMFTB were investigated. The results showed that α-amylase and pullulanase hydrolysis reduced starch content and raised protein, flavonoids, Monacolin K and Monascus pigments content of EMFTB. Meanwhile, double enzyme hydrolysis significantly changed the principal components of volatile substances and affected the varieties and content of volatile organic substances in EMFTB using electronic nose and headspace gas chromatography-ion mobility chromatography (HS-GC-IMS). The volatile organic substances and main aroma components increased significantly in EMFTB, including 2-heptanone, 3-methyl-1-butanol, butan-1-ol, 2-methyl-1-propanol and other substances. These results indicate that the amylase hydrolysis plays an important role in improving the flavor quality of EMFTB.


Assuntos
Fagopyrum , Monascus , Fagopyrum/química , Hidrólise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Espectrometria de Mobilidade Iônica , alfa-Amilases
4.
Molecules ; 28(1)2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36615556

RESUMO

The leaves of Ligustrum robustum have been consumed as Ku-Ding-Cha for clearing heat and removing toxins, and they have been used as a folk medicine for curing hypertension, diabetes, and obesity in China. The phytochemical research on the leaves of L. robustum led to the isolation and identification of two new hexenol glycosides, two new butenol glycosides, and five new sugar esters, named ligurobustosides X (1a), X1 (1b), Y (2a), and Y1 (2b) and ligurobustates A (3a), B (3b), C (4b), D (5a), and E (5b), along with seven known compounds (4a and 6-10). Compounds 1-10 were tested for their inhibitory effects on fatty acid synthase (FAS), α-glucosidase, and α-amylase, as well as their antioxidant activities. Compound 2 showed strong FAS inhibitory activity (IC50 4.10 ± 0.12 µM) close to that of the positive control orlistat (IC50 4.46 ± 0.13 µM); compounds 7 and 9 revealed moderate α-glucosidase inhibitory activities; compounds 1-10 showed moderate α-amylase inhibitory activities; and compounds 1 and 10 displayed stronger 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) ammonium salt (ABTS) radical scavenging effects (IC50 3.41 ± 0.08~5.65 ± 0.19 µM) than the positive control l-(+)-ascorbic acid (IC50 10.06 ± 0.19 µM). This study provides a theoretical foundation for the leaves of L. robustum as a functional tea to prevent diabetes and its complications.


Assuntos
Ligustrum , Ligustrum/química , alfa-Glucosidases , Extratos Vegetais/química , Antioxidantes/química , Glicosídeos/química , Folhas de Planta/química , alfa-Amilases
5.
BMC Complement Med Ther ; 23(1): 3, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36604684

RESUMO

BACKGROUND: The use of conventional medical therapies has proven to have many setbacks and safety concerns that need further improvement. However, herbal medicine has been used for over 2000 years, and many studies have proven the use of herbs to be effective and safe. This article discussed the efficacy of different herbal products used in the management of obesity. To evaluate the efficacy of seven herbal-based weight loss products currently available on the Palestinian market, using in vitro assays to screen for antioxidants, anti-amylase, and anti-lipase effects for each product. METHOD: Pancreatic lipase and salivary amylase inhibitory activities, as well as antioxidant analysis, were tested in vitro on a variety of herbal products. Then the IC50 was measured for each test. RESULTS: The anti-lipase assay results, IC50 values in (µg/mL) of each of the seven products (Product A, product B, product C, product D, product E, product F, and product G) were 114.78, 532.1, 60.18, 53.33, 244.9, 38.9, and 48.97, respectively. The IC50 value for orlistat (Reference) was 12.3 µg/ml. On the other hand, the IC50 value for alpha amylase inhibition of the seven products (Product A, product B, product C, product D, product E, product F, and product F) were 345.93, 13,803.84 (Inactive), 73.79, 130.91, 165.95, 28.18, and 33.11 µg/ml respectively, while acarbose (Reference) was 23.38 µg/ml. The antioxidant activity (IC50 values) for the seven products (Product A, product B, product C, product D, product E, product F, and product F) were 1258.92, 707.94, 79.43, 186.20, 164.81, 17.53, and 10.47 µg/ml respectively. While the IC50 value for Trolox was 2.70 µg/ml. CONCLUSION: It can be concluded that the seven products showed varied anti-lipase, anti-amylase, and antioxidant effects. However, products F and G showed superiority in all categories.


Assuntos
Fármacos Antiobesidade , Extratos Vegetais , Plantas Medicinais , alfa-Amilases/química , Antioxidantes/química , Antioxidantes/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Oriente Médio , Fármacos Antiobesidade/química , Fármacos Antiobesidade/farmacologia
6.
Cells ; 12(1)2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36611986

RESUMO

Feeding is crucial for the growth and survival of animals, including humans, but relatively little is known about how it is regulated. Here, we show that larval feeding in Ostrinia furnacalis is regulated by neuropeptide F (NPF, the homologous peptide of mammalian NPY) via the insulin signalling pathway in the midgut. Furthermore, the genes pi3k and mtor in the insulin pathway positively regulate α-amylase and lipase of the midgut by recruiting the transcription factors c-Myc and PPARγ for binding to the promotors of these two enzymes. Importantly, we find that the feeding behaviour and the digestive system of midgut in O. furnacalis larvae are closely related and interactive in that knocking down α-amylase or lipase induces a reduction in larval feeding, while food-deprived larvae lead to fewer expressions of α-amylase and lipase. Importantly, it is the gut NPF that regulates the α-amylase and lipase, while variations of α-amylase and lipase may feed back to the brain NPF. This current study reveals a molecular feedback mechanism between feeding behaviour and the digestive system that is regulated by the conserved NPF via insulin signalling systems in the midgut of O. furnacalis larvae.


Assuntos
Insulinas , Mariposas , Animais , Humanos , Larva/genética , Lipase , Digestão , alfa-Amilases/genética , Mamíferos
7.
Talanta ; 255: 124215, 2023 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-36603441

RESUMO

Protein biosensors hold a promise to transform the way we collect physiological data by enabling quantification of biomarkers outside of specialized laboratory environment. However, achieving high specificity and sensitivity in homogeneous assay format remains challenging. Here we report construction of fluorescent biosensor arrays based on artificial allosteric α-amylase-activated PQQ-dependent glucose dehydrogenase (Amy-GDH). Amy-GDH was covalently immobilized on silica nanoparticles that were then arrayed on fiberglass sheets. The activity of the biosensor was monitored using a smartphone camera via emergence of bright fluorescence (λex 365 nm) originating from reduced phenazine methosulfate upon glucose oxidation by Amy-GDH. We show that such biosensor arrays demonstrate an apparent Kd of 115 pM for α-amylase with a detection limit of 2 pM. Using the developed biosensor arrays, we were able to specifically and accurately quantify the concentration of α-amylase in biological fluids such as serum and saliva. We propose that the presented approach can enable construction of ultrasensitive point-of-care diagnostic arrays.


Assuntos
Técnicas Biossensoriais , alfa-Amilases , Glucose , Saliva , Oxirredução
8.
ACS Synth Biol ; 12(1): 196-202, 2023 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-36580286

RESUMO

Developing effective bacterial autolytic systems for fast release of intracellular bioproducts could simplify purification procedures and help with the high throughput screening of mutant libraries in protein engineering. Here, we developed a fast and tightly regulated E. coli autolytic system, named the FhuD-lysozyme-SsrA mediated autolytic (FLSA) system, by integrating the secretion signal peptide, T7 lysozyme, and E. coli ClpX/P-SsrA protein degradation machinery. To decrease the cytotoxicity of leaky T7 lysozymes, the SsrA tag was fused to the C-terminus of T7 lysozyme to confer a tight regulation of its production. Using sfGFP as a reporter, we demonstrated that anchoring the Sec-Tat dual pathway signal peptide FhuD to the N-terminus of T7 lysozyme-SsrA could give the highest cell lysing efficiency. The optimization of the FLSA system indicated that weak alkaline conditions (pH 8.0) and 0.5% Triton X-100 could further increase the lysing efficiency by about 24%. The FLSA system was validated by efficient production of sfGFP and human growth hormone 1 (hGH1) in a shake flask, with a cell lytic efficiency of approximately 82% and 80%, respectively. Besides, the FLSA system was applied for large-scale fermentation, in which approximately 90% sGFP was released with a cell density OD600 of 110. Moreover, the FLSA system was also tested for α-amylase mutant library screening in microplates, and the results showed that intracellular α-amylase can be efficiently released out of cells for activity quantitation. In all, the FLSA system can facilitate the release of intracellular recombinant proteins into the cell culture medium, which has the potential to serve as an integrated system for large-scale production of recombinant targets and high throughput enzyme engineering in synthetic biology.


Assuntos
Escherichia coli , Muramidase , Humanos , alfa-Amilases/metabolismo , Escherichia coli/metabolismo , Muramidase/genética , N-Acetil-Muramil-L-Alanina Amidase/metabolismo , Sinais Direcionadores de Proteínas , Histidina Quinase/metabolismo
9.
Mar Environ Res ; 183: 105849, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36565507

RESUMO

In the marine environment, plastic pollution may occur simultaneously with hypoxia. However, current ecological risk assessments of nanoplastics have rarely considered the impact of additional environmental factors, such as hypoxia. In this study, we investigated the effect of polystyrene nanospheres (PS-NPs) on the digestive performance (antioxidant system and digestive enzymes) of mussels Mytilus galloprovincialis under different patterns of hypoxia (normoxia, constant hypoxia, and fluctuating hypoxia). The result showed that PS-NPs caused oxidative damage in the digestive glands of mussels, while all patterns of hypoxia exacerbated this oxidative damage. Activities of four digestive enzymes (α-amylase, cellulase, trypsin, and lipase) were examined. Among these, the activity of the α-amylase was inhibited by PS-NPs, and the inhibition was aggravated by all the hypoxia patterns. The cellulase activity and trypsin activity was enhanced by PS-NPs, and the increase was further stimulated by hypoxia. Lipase activity was not affected by PS-NPs alone, but significant inhibition was detected after the coexposure to PS-NPs and hypoxia. Conclusively, the combined stress of hypoxia and nanoplastics can significantly affect the digestive performance of mussels and may alter the mussel nutrient uptake strategy. Our work has provided new insight into the ecological risk assessment of plastics under global climate change.


Assuntos
Celulases , Mytilus , Poluentes Químicos da Água , Animais , Antioxidantes , Microplásticos , Tripsina/farmacologia , Proteínas , Plásticos/toxicidade , Poliestirenos/toxicidade , Hipóxia , Lipase/farmacologia , alfa-Amilases/farmacologia , Celulases/farmacologia , Poluentes Químicos da Água/toxicidade
10.
Arch Oral Biol ; 145: 105574, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36395562

RESUMO

OBJECTIVES: Salivary statherin and alpha-amylase play significant roles in biofilm formation and pathogenic bacteria adhesion. Examination of these proteins may provide information on their roles in periodontal diseases. The present study was based on the hypothesis that; the salivary proteins -statherin and alpha-amylase- effective on biofilm formation, may play important roles in the etiology of periodontal disease. Therefore, we aimed to analyze the differences in periodontal diseases compared to periodontal health in order to search their roles in periodontal disease. METHODS: Patients with gingivitis (n = 26) and periodontitis (n = 20), and periodontally healthy individuals (n = 21) were included in this study. Unstimulated whole saliva samples were obtained from a total of 67 individuals. Salivary statherin level and alpha-amylase activity were determined using ELISA and enzymatic methods, respectively. RESULTS: Statherin levels in saliva were significantly higher in the periodontitis group compared to the gingivitis group (p = 0.014), while alpha-amylase activities and total protein levels were slightly higher in the periodontitis and gingivitis groups compared to controls, without significant differences among the groups (p = 0.295 and p = 0.019, respectively). Statherin levels showed positive correlations with gingival and plaque indices in the disease groups. CONCLUSIONS: The results suggest that statherin level in saliva increase to provide a protective effect against periodontitis, and higher salivary statherin level is related to the degree of gingival inflammation and plaque accumulation.


Assuntos
Gengivite , Doenças Periodontais , Periodontite , Humanos , alfa-Amilases/metabolismo , Aderência Bacteriana , Gengivite/metabolismo , Doenças Periodontais/metabolismo , Periodontite/metabolismo , Saliva/metabolismo
11.
Sci Total Environ ; 858(Pt 3): 159581, 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36397605

RESUMO

Efficient substrate metabolism is the premise for stable operation of aerobic granular sludge and can be regulated by quorum sensing (QS). In this study, starch and acetate were selected to represent complex and simple substrates to provide comparable amount of metabolic energy for granules cultivation. Starch-fed granules were larger in size and contained higher EPS content than acetate-fed granules, though both granules exhibited similar substrate-degradation rates during sequencing batch reactor (SBR) cycle. Three N-acylhomoserine lactones (AHLs), including C8-HSL, 3OHC8-HSL and 3OHC12-HSL, were detected as dominant autoinducers in granules. They accumulated more in starch-fed granules than acetate-fed granules. The batch experiments were implemented to investigate QS regulation for granular stability in terms of substrate hydrolysis and transformation. The addition of three AHLs increased the activity of α-amylase, the main starch hydrolase, 4-6 times, significantly (p < 0.01) higher than the control treatment without AHLs amendment. While activity of dehydrogenase, the main simple substrate degradation enzyme, was increased only 1-2 times. Higher enzyme activity, especially α-amylase, significantly (p < 0.05) promoted the substrate-degradation rate (65 % than control group) and EPS yield in starch-fed system. Overall, QS can facilitate complex substrate uptake via hydrolysis enhancement and EPS secretion, which together promote sludge granulation and stability.


Assuntos
Acil-Butirolactonas , Homosserina , Esgotos , alfa-Amilases , Amido
12.
J Dairy Sci ; 106(2): 1002-1012, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36543642

RESUMO

The objective of this study was to determine the effects of including exogenous amylolytic or fibrolytic enzymes in a diet for high-producing dairy cows on in vitro ruminal fermentation. Eight dual-flow continuous-culture fermentors were used in a replicated 4 × 4 Latin square. The treatments were control (CON), a xylanase and glucanase mixture (T1), an α-amylase mixture (T2), or a xylanase, glucanase, and α-amylase mixture (T3). Treatments were included at a rate of 0.008% of diet dry matter (DM) for T1 and T2 and at 0.02% for T3. All treatments replaced the equivalent amount of soybean meal in the diet compared with CON. All diets were balanced to have the same nutrient composition [30.2% neutral detergent fiber (NDF), 16.1% crude protein (CP), and 30% starch; DM basis], and fermentors were fed 106 g/d divided into 2 feedings. At each feeding, T2 was pipetted into the respective fermentor and an equivalent amount of deionized water was added to each fermentor to eliminate potential variation. Experimental periods were 10 d (7 d for adaptation and 3 d for sample collection). Composite samples of daily effluent were collected and analyzed for volatile fatty acids (VFA), NH3-N, and lactate concentrations, degradability of DM, organic matter, NDF, CP, and starch, and flow and metabolism of N. Samples of fermentor contents were collected from each fermentor at 0, 1, 2, 4, 6, and 8 h after feeding to determine kinetics of pH, NH3-N, lactate, and VFA concentrations over time. All data were analyzed using PROC GLIMMIX of SAS (SAS Institute Inc.), and the repeated variable of time was included for kinetics measurements. Treatment did not affect mean pH, degradability, N flow and metabolism, or the concentrations of VFA, NH3-N, or lactate in the effluent samples. Treatment did not affect pH, acetate:propionate ratio, or the concentrations of lactate, NH3-N, total VFA, acetate, propionate, butyrate, isobutyrate, valerate, or caproate. However, the concentration of total VFA tended to change at each time point depending upon the treatment, and T2 tended to have a greater proportion of 2-methylbutyrate and isovalerate than CON, T1, or T3. As 2-methylbutyrate and isovalerate are branched-chain VFA that are synthesized from branched-chain amino acids, T2 may have an increased fermentation of branched-chain amino acids or decreased uptake by fibrolytic microorganisms. Although we did not observe changes in N metabolism due to the enzymes, there could be changes in microbial populations that utilize branched-chain VFA. Overall, the tested enzymes did not improve in vitro ruminal fermentation in the diet of high-producing dairy cows.


Assuntos
Lactação , Propionatos , Animais , Bovinos , Feminino , alfa-Amilases/metabolismo , Ração Animal/análise , Dieta/veterinária , Digestão , Ácidos Graxos Voláteis/metabolismo , Fermentação , Lactatos/metabolismo , Propionatos/metabolismo , Rúmen/metabolismo , Amido/metabolismo
13.
IUBMB Life ; 75(2): 161-180, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36565478

RESUMO

This study was designed to screen novel thiourea derivatives against different enzymes, such as α-amylase, α-glucosidase, protein tyrosine phosphatase 1 B, and advanced glycated end product (AGEs). A cytotoxicity analysis was performed using rat L6 myotubes and molecular docking analysis was performed to map the binding interactions between the active compounds and α-amylase and α-glucosidase. The data revealed the potency of five compounds, including E (1-(2,4-difluorophenyl)-3-(3,4-dimethyl phenyl) thiourea), AG (1-(2-methoxy-5-(trifluoromethyl) phenyl)-3-(3-methoxy phenyl) thiourea), AF (1-(2,4-dichlorophenyl)-3-(4-ethylphenyl) thiourea), AD (1-(2,4-dichlorophenyl)-3-(4-ethylphenyl) thiourea), and AH (1-(2,4-difluorophenyl)-3-(2-iodophenyl) thiourea), showed activity against α-amylase. The corresponding percentage inhibitions were found to be 85 ± 1.9, 82 ± 0.7, 75 ± 1.2, 72 ± 0.4, and 65 ± 1.1%, respectively. These compounds were then screened using in vitro assays. Among them, AH showed the highest activity against α-glucosidase, AGEs, and PTP1B, with percentage inhibitions of 86 ± 0.4% (IC50  = 47.9 µM), 85 ± 0.7% (IC50  = 49.51 µM), and 85 ± 0.5% (IC50  = 79.74 µM), respectively. Compound AH showed an increased glucose uptake at a concentration of 100 µM. Finally, an in vivo study was conducted using a streptozotocin-induced diabetic mouse model and PTP1B expression was assessed using real-time PCR. Additionally, we examined the hypoglycemic effect of compound AH in diabetic rats compared to the standard drug glibenclamide.


Assuntos
Diabetes Mellitus Experimental , alfa-Glucosidases , Camundongos , Ratos , Animais , alfa-Glucosidases/genética , alfa-Glucosidases/química , alfa-Glucosidases/metabolismo , Diabetes Mellitus Experimental/tratamento farmacológico , Simulação de Acoplamento Molecular , Reação de Maillard , Hipoglicemiantes/farmacologia , Produtos Finais de Glicação Avançada/genética , alfa-Amilases , Tioureia/farmacologia
14.
Poult Sci ; 102(2): 102366, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36565629

RESUMO

Two experiments (Exp.) were conducted to evaluate the effects of exogenous carbohydrases on nutrient and energy utilization of corn with different compositions by broilers. In Exp. 1, a total of 448 Cobb 500 male chicks were distributed in a 2 × 4 factorial arrangement (corn from regions geographically located in the North or South of Brazil × 4 carbohydrases supplementation), with 8 replicate cages of 7 birds each. In Exp. 2, 672 Cobb 500 male chicks were fed 12 experimental feeds, in a 3 × 4 factorial arrangement [3 corn endosperm compositions (waxy, semi-dent, or semi-flint) × 4 carbohydrases], with 8 replicate cages of 7 birds. Birds were fed semi-purified test diets with 95.9% corn from d 14 to 24 in both studies. In Exp. 1, α-amylase, ß-xylanase, or carbohydrase complex (cellulase, glucanase, and xylanase) were added to the diet. In Exp. 2, α-amylase, ß-xylanase, or α-amylase + ß-xylanase were supplemented. Digestibility of DM, N, ether extract (EE), Ca, and P as well as AME, AMEn, and IDE were determined. In Exp. 2, jejunal starch digestibility was determined on d 24. Data were subjected to analysis of variance and means were compared by Tukey test (P ≤ 0.05). Corn from the North origin had higher AME, AMEn, and digestibility of DM and N compared to the South (P ≤ 0.05). Amylase supplementation led to increases of 3% in AME and 2% in N digestibility when compared to the non-supplemented feeds (P ≤ 0.01). In Exp. 2, the highest ME values and EE digestibility were observed in the semi-flint corn compared to the waxy, whereas the semi-dent presented the highest digestibility of N and starch. Corn diets supplemented with amylase + xylanase had improvements of 2.5% AMEn and 3% starch digestibility. In conclusion, energy and nutrient utilization of corn by broilers depend on the region where it was grown. Corn genetics, expressed by the endosperm composition, and carbohydrase supplementation influenced energy and nutrient utilization by broilers.


Assuntos
Galinhas , Zea mays , Animais , Masculino , Ração Animal/análise , Dieta/veterinária , Suplementos Nutricionais/análise , Endo-1,4-beta-Xilanases/farmacologia , Nutrientes , Amido , alfa-Amilases/farmacologia , Fenômenos Fisiológicos da Nutrição Animal , Digestão
15.
Addict Behav ; 136: 107489, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36181746

RESUMO

BACKGROUND: To advance our understanding of the health-related consequences of chronic cannabis use, this study examined hypothalamic-pituitary-adrenal (HPA) axis and sympathetic nervous system (SNS) reactivity and regulation in response to a well-characterized, acute, social evaluative stress task among cannabis users and non-users. We also explored differences in HPA-SNS coordination across the stress task in cannabis users and non-users. METHOD: Participants were 75 adults (53% female) who reported no use of tobacco/nicotine products. Cannabis use was measured using self-report and salivary/urinary THC levels. Participants were classified as cannabis users (n = 33) if they reported using cannabis at least twice per week in the prior year and had a positive salivary/urinary THC test or as non-users (n = 42) if they reported no use in the prior year and had a negative THC test. During a laboratory visit, participants completed the standard Trier Social Stress Test (TSST) and provided saliva samples before, and 5, 20, and 40 min after the task. Samples were assayed for salivary cortisol and alpha-amylase (sAA) as indices of HPA axis and SNS activity, respectively. RESULTS: Multilevel piecewise growth models revealed that, relative to non-users, cannabis users showed (a) blunted cortisol reactivity and recovery to the TSST, and (b) greater reductions in sAA concentrations following the TSST. Chronic cannabis users may exhibit blunted HPA axis responses and greater SNS recovery to acute psychosocial stress. Implications of individual differences in stress reactivity and regulation for the biobehavioral health of chronic cannabis users are discussed.


Assuntos
Cannabis , Sistema Hipófise-Suprarrenal , Adulto , Dronabinol , Feminino , Humanos , Hidrocortisona , Sistema Hipotálamo-Hipofisário/metabolismo , Masculino , Nicotina , Sistema Hipófise-Suprarrenal/metabolismo , Saliva/metabolismo , Estresse Psicológico/psicologia , Sistema Nervoso Simpático/metabolismo , alfa-Amilases/metabolismo
16.
Biomater Sci ; 11(2): 630-640, 2023 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-36484349

RESUMO

Effective treatment of bacterial biofilm-related infections is a great challenge for the medical community. During the formation of biofilms, bacteria excrete extracellular polymeric substances (EPS), including polysaccharides, proteins, nucleic acids, etc., to encapsulate themselves and form a "fort-like" structure, which greatly reduces the efficiency of therapeutic agents. Herein, we prepared a nanoagent (MnO2-amylase-PEG-ICG nanosheets, MAPI NSs) with biofilm degradation capability for efficient photothermal therapy and fluorescence imaging of methicillin-resistant Staphylococcus aureus (MRSA) biofilm infections. MAPI NSs were constructed by sequentially modifying α-amylase, polyethylene glycol (PEG), and indocyanine green (ICG) on manganese dioxide nanosheets (MnO2 NSs). Experimental results exhibited that MAPI NSs could accumulate in infected tissues after intravenous injection, degrade in the acidic biofilm microenvironment, and release the loaded ICG for near-infrared (NIR) fluorescence imaging of the infected tissues. Importantly, MAPI NSs could efficiently eliminate MRSA biofilm infections in mice by α-amylase enhanced photothermal therapy. In addition, MAPI NSs exhibited neglectable toxicity towards mice. Given the superior properties of MAPI NSs, the enzyme-degradation enhanced therapeutic strategy presented in this work offers a promising solution for effectively combating biofilm infectious diseases.


Assuntos
Infecções Bacterianas , Staphylococcus aureus Resistente à Meticilina , Camundongos , Animais , Terapia Fototérmica , Compostos de Manganês , Amilases , Óxidos , Polietilenoglicóis/química , Verde de Indocianina/química , Biofilmes , alfa-Amilases , Imagem Óptica , Antibacterianos/farmacologia , Antibacterianos/química
17.
Spectrochim Acta A Mol Biomol Spectrosc ; 289: 122251, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36542921

RESUMO

Developing naturally active components to control α-amylase/α-glucosidase activity is highly desired for preventing and managing type 2 diabetes. Rapeseed oil is rich in active phenolic compounds and seed oil is a major source of liposoluble inhibitors to these enzymes. However, it remains unclear about the interaction of phenolic compounds in rapeseed oil with α-amylase/α-glucosidase. This study found that the important phenolic compounds from rapeseed oil (Sinapic acid, SA; canolol, CAO; canolol dimer, CAO dimer) possessed effective inhibition performance against α-amylase and α-glucosidase. CAO showed the lowest and highest inhibitory effect, respectively. In the kinetics studies, the inhibition mechanism of SA/CAO/CAO dimer against α-glucosidase was non-competitive, exhibiting a different way from α-amylase. Fluorescence quenching spectra implied that the static processes were responsible for the spontaneous binding between the compounds and enzymes. Fourier-transform infrared spectroscopy (FT-IR) displayed these compounds-induced conformation alterations of α-amylase/α-glucosidase. Molecular docking revealed that SA/CAO/CAO dimer decreased the catalytic efficiency of α-amylase/α-glucosidase through hydrogen bonds, hydrophobic force, or π-π interaction. Molecular dynamics matched well with the experimental and docking results regarding the inhibitory behaviors and interactions toward α-amylase/α-glucosidase. These results demonstrated the potential benefits of phenolic compounds from rapeseed oil in antidiabetic-related activities.


Assuntos
Diabetes Mellitus Tipo 2 , Simulação de Dinâmica Molecular , Humanos , Simulação de Acoplamento Molecular , Inibidores de Glicosídeo Hidrolases/química , alfa-Glucosidases/metabolismo , Óleo de Brassica napus , alfa-Amilases/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier
18.
Chem Biol Interact ; 370: 110312, 2023 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-36535312

RESUMO

In this study, eight new compounds (7a-h) based on triazole compounds containing ester groups were synthesized with high yields. The structures of the synthesized compounds (7a-h) were elucidated by various spectroscopic methods (element analysis, FT-IR, 1H-(13C) NMR). Antioxidant, anticancer, and α-amylase enzyme inhibition activities of synthesized new triazole derivatives were carried out, and the effects of different groups on the activity were investigated. When the determined antioxidant properties of the compounds were examined, all synthesized compounds showed a moderate radical scavenging effect against radicals depending on the concentration (6.25-200 g/mL). All compounds except the three derivatives were found to have higher IC50 values than the standard drug acarbose (IC50: 891 µg/mL) according to the α-amylase enzyme inhibition results. Compound 7g (IC50: 50 g/mL) was discovered to have nearly eighteen (18) times the activity of the conventional medication acarbose (IC50: 891 µg/mL). Compounds synthesized for anticancer activity studies were screened against the Hela cell line, and the results were compared with standard cis-platinum (IC50: 16.30 µg/mL). Compound 7g (IC50: 19.78 µg/mL) was found to have almost the same activity as cis-platinum. Using Qikprop, the compounds were thoroughly tested for ADME qualities, and none violated any drug similarity standards. According to ADME data, whole physicochemical drug-likeness parameters of molecules remained within defined ranges as stipulated in the Lipinski rules (RO5) and revealed a high bioavailability profile. The molecular docking results with 2QV4 and 4GQR alpha-amylase enzymes demonstrated that all molecules have a high affinity, indicating polar and apolar interaction with critical amino acids in the α-amylase binding pocket.


Assuntos
Acarbose , Antioxidantes , Humanos , Simulação de Acoplamento Molecular , Relação Estrutura-Atividade , Antioxidantes/farmacologia , Células HeLa , Cisplatino , Triazóis/farmacologia , Triazóis/química , Espectroscopia de Infravermelho com Transformada de Fourier , alfa-Amilases/metabolismo , Estrutura Molecular
19.
Food Funct ; 14(1): 262-276, 2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36484426

RESUMO

The scientific interest in the medicinal properties of Kombucha beverages, a carbonated drink with live microorganisms, has increased recently. Hence, the aim of this study was to determine the chemical profile and to examine the antioxidant, antidiabetic and antineurodegenerative potential of unfermented and also Kombucha fermented Camellia sinensis (green tea), Coffea arabica (coffee), and Ganoderma lucidum (Reishi) extracts. The extracts were prepared as follows: the first (unfermented) set contained 1 L of water, 50 g of sucrose and 20 g of dried and ground green tea, coffee, or Reishi basidiocarp, while the second (fermented) set contained all of the aforementioned ingredients individually inoculated with Kombucha and fermented for 21 days. The chemical analysis was conducted using liquid chromatography-mass spectrometry (LC-MS). The antioxidant activity was assessed by DPPH, total reducing power (TRP), and ß-carotene bleaching assays. The inhibition of α-amylase and α-glucosidase activity was used to estimate the antidiabetic potential, while the level of inhibition of acetylcholinesterase (AChE) and tyrosinase (TYR) was used to evaluate the antineurodegenerative activity. The results suggested that the fermented extracts of green tea, coffee, and Reishi exert significant antioxidant effects, although they were lower compared to the unfermented extracts. The unfermented green tea extract exhibited the highest DPPH-scavenging activity (87.46%) and the highest preservation of ß-carotene (92.41%), while the fermented coffee extract showed the highest TRP (120.14 mg AAE per g) at 10 mg mL-1. Although the extracts did not inhibit the activity of α-amylase, they were quite effective at inhibiting α-glucosidase, especially the unfermented Reishi extract, inhibiting 95.16% (at a concentration of 10 mg mL-1) of α-glucosidase activity, which was slightly higher than the positive control at the same concentration. The most effective AChE inhibitor was unfermented green tea extract (68.51%), while the fermented coffee extract inhibited 34.66% of TYR activity at 10 mg mL-1. Altogether, these results are in accordance with the differences found in the extracts' chemical composition. Finally, this is the first report that highlights the differences in the chemical profile between the unfermented and Kombucha fermented green tea, coffee and Reishi extracts, while it also reveals, for the first time, the antineurodegenerative potential of Kombucha fermented Reishi extract. The examined extracts represent potent functional foods, while their more detailed mechanisms of action are expected to be revealed in future research.


Assuntos
Camellia sinensis , Coffea , Reishi , Antioxidantes/farmacologia , Antioxidantes/análise , Camellia sinensis/química , Hipoglicemiantes/farmacologia , Hipoglicemiantes/análise , alfa-Glucosidases , Acetilcolinesterase , beta Caroteno/análise , Cromatografia Gasosa-Espectrometria de Massas , Chá/química , alfa-Amilases , Extratos Vegetais/farmacologia , Extratos Vegetais/química
20.
Int J Biol Macromol ; 226: 102-110, 2023 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-36495986

RESUMO

Starch microspherical aerogel (SMA) prepared by enzymatic hydrolysis of starch with α-amylase was demonstrated to be higher adsorption capacity for methylene blue. Proper cleavage of α-1,4 glycosidic bonds could enhance the adsorption capacity of SMA, while the cleavage of α-1,6 glycosidic bonds showed an opposite effect. Compared with tapioca starch (TS), α-amylase hydrolyzed starch exhibited a 9.46 % decrease in amylose content, a 25.40 % increase in adsorbability, and significant decreases in weight-average molecular weight (Mw) of different amylases. When the Mw of enzymolysis starch was 6.39 × 106 g/mol, it was suitable for the preparation of SMA, and could significantly increase its adsorption capacity. The adsorbability of the crosslinked starch microspherical aerogel (CSMA) was 1.816 ± 0.026 mg/g, which was increased by 100.60 % relative to that of native starch microspherical aerogel (NSMA). CSMA had the best adsorption effect on oil and could be applied to the adsorption and removal of vegetable oil.


Assuntos
Amido , alfa-Amilases , Amido/química , Adsorção , alfa-Amilases/química , Hidrólise , Amilose
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