RESUMO
OBJECTIVE: Postoperative pancreatic fistula (POPF) represents a leading cause of morbidity and mortality following major pancreatic resections. This study aimed to evaluate the use of postoperative drain fluid lipase-to-amylase ratio (LAR) for the prediction of clinically relevant fistulae (CR-POPF). METHODS: Consecutive patients undergoing pancreaticoduodenectomy between 2017 and 2021 at a tertiary centre were retrospectively reviewed. Univariable and multivariable analyses were performed to identify predictors for CR-POPF (ISGPS grade B/C). Receiver operating characteristic (ROC) curve analyses were conducted to evaluate the performance of LAR and determine optimum prediction thresholds. RESULTS: Among 130 patients, 28 (21.5%) developed CR-POPF. Variables positively associated with CR-POPF included soft gland texture, acinar cell density, diagnosis other than PDAC or chronic pancreatitis, resection without neoadjuvant therapy, and postoperative drain fluid lipase, amylase, and LAR (all P <0.05). Multivariable regression analysis identified LAR as an independent predictor of CR-POPF ( P <0.05). ROC curve analysis showed that LAR had moderate ability to predict CR-POPF on POD1 (AUC,0.64; 95%CI,0.54-0.74) and excellent ability on POD3 (AUC,0.85; 95%CI,0.78-0.92) and POD 5 (AUC,0.86; 95%CI,0.79-0.92). Optimum thresholds were consistent over PODs 1 to 5 (ratio>2.6) and associated with 92% sensitivity and 46% to 71% specificity. CONCLUSIONS: Postoperative drain fluid LAR represents a reliable predictor for the development of CR-POPF. With early prognostication, the postoperative care of patients at risk of developing high-grade fistulas may be optimized.
Assuntos
Amilases , Lipase , Fístula Pancreática , Pancreaticoduodenectomia , Complicações Pós-Operatórias , Humanos , Pancreaticoduodenectomia/efeitos adversos , Fístula Pancreática/etiologia , Fístula Pancreática/diagnóstico , Feminino , Masculino , Amilases/análise , Amilases/metabolismo , Pessoa de Meia-Idade , Estudos Retrospectivos , Idoso , Lipase/análise , Lipase/metabolismo , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/etiologia , Curva ROC , Drenagem/métodos , Valor Preditivo dos Testes , Fatores de Risco , AdultoRESUMO
A quimioprevenção do câncer refere-se ao uso de compostos naturais ou sintéticos para prevenir o desenvolvimento das neoplasias antes do estabelecimento da malignidade. O ácido butirico (AB) atua como um potente quimiopreventivo na hepatocarcinogênese, reduzindo o número e o tamanho de lesões pré neoplásicas persistentes (pLPN), induzindo a apoptose e modulando mecanismos epigenéticos. Já o ácido caprílico (AC), além da sua atuação como potencializador de absorção, vem sendo investigado na área da prevenção do câncer. Neste cenário, o objetivo do trabalho visa avaliar a atividade quimiopreventiva de lipídios estruturados (EST) obtidos por interesterificação enzimática da tributirina com a tricaprilina, na fase de promoção da hepatocarcinogênese experimental. Após o processo de interesterificação, o produto final apresentou novos triacilgliceróis com composição de duas moléculas de ácido butírico para uma de ácido caprilíco. Ratos machos isogênicos da linhagem Fischer 344 foram submetidos ao modelo do hepatócito resistente, sendo distribuídos em dois grupos e tratados diariamente por via intragástrica com lipídios estruturados (EST) ou com o seu controle isocalórico, a maltodextrina (MD), durante a fase de promoção. Como esperado, não houve diferença estatística (p>0,05) em relação ao peso inicial e final dos animais dos grupos MD e EST, o que indica ausência de toxicidade dos compostos administrados. Na análise macroscópica do fígado, foi observada uma redução de 33,3% no grupo EST em relação ao número médio de nódulos macroscópicos em comparação ao grupo MD, porém essa redução não atingiu diferença estatística (p>0,05). Para a avaliação das lesões pré neoplásicas (LPN) foi utilizada a marcação imunoistoquímica para glutationa-S-transferase (GST-P). O grupo EST apresentou uma redução no número de lesões em remodelação e total GSTP-P+, quando comparado com o grupo MD (p<0,05). Quando avaliada a % de corpúsculos apoptóticos e índice de proliferação celular, não houve diferença estatística entre os grupos (p>0,05). Animais tratados com lipídios estruturados apresentaram maiores (p<0,05) concentrações de AC e AB por grama de tecido hepático em relação ao tratamento com maltodextrina. Em relação aos danos no DNA, o grupo EST resultou em cometas de comprimentos menores (p<0,05), menores níveis de γ-H2AX (p<0,05) e maiores concentrações de p53 nuclear, quando comparados aos animais que receberam maltodextrina, sugerindo uma proteção contra danos no DNA no grupo tratado com EST. Os resultados mostraram que o tratamento com EST resultou em ações efetivas na fase de promoção da hepatocarcinogênese experimental
Cancer chemoprevention refers to the use of natural or synthetic compounds to prevent the development of neoplasms before the establishment of malignancy. Butyric acid (AB) acts as a potent chemopreventive in hepatocarcinogenesis, reducing the number and size of persistent preneoplastic lesions (pLPN), inducing apoptosis and modulating epigenetic mechanisms. Caprylic acid (CA), in addition to its role as an absorption enhancer, has been investigated in the area of cancer prevention. In this scenario, the objective of this work was to evaluate the chemopreventive activity of structured lipids (EST) obtained by enzymatic interesterification of tributyrin with tricaprylin, in the phase of promotion experimental hepatocarcinogenesis. After the interesterification process, the final product presented new triacylglycerols with a composition of two molecules of butyric acid to one of caprylic acid. Isogenic male Fischer 344 rats were submitted to the resistant hepatocyte model, divided into two groups and treated daily intragastrically with structured lipids (EST) or with its isocaloric control, maltodextrin (MD), during the promotion phase. As expected, there was no statistical difference (p>0.05) in relation to the initial and final weight of the animals in the MD and EST groups, which indicates the absence of toxicity of the administered compounds. In the macroscopic analysis of the liver, a reduction of 33.3% was observed in the EST group in relation to the mean number of macroscopic nodules compared to the MD group, but this reduction did not reach a statistical difference (p>0.05). For the evaluation of pre-neoplastic lesions (PNL) immunohistochemical staining for glutathione-Stransferase (GST-P) was used. The EST group showed a reduction in the number of remodeling lesions and total GSTP-P+, when compared to the MD group (p<0.05). Animals treated with structured lipids had higher (p<0.05) concentrations of AC and AB per gram of liver tissue compared to treatment with maltodextrin. Regarding DNA damage, the EST group resulted in comets of shorter lengths (p<0.05), lower levels of γ-H2AX (p<0.05) and high concentration of nuclear p53, when compared to animals that received maltodextrin, suggesting protection against DNA damage in the EST treated group. The results showed that EST treatment resulted in effective actions in the promotion phase of experimental hepatocarcinogenesis
Assuntos
Animais , Masculino , Ratos , Quimioprevenção , Lipase/análise , Neoplasias/patologia , Ferimentos e Lesões/complicações , Biotecnologia/classificação , Carcinoma Hepatocelular/patologia , AbsenteísmoRESUMO
Membrane-based processes are increasingly used to clarify and concentrate thermo-sensitive fruit juices and plant extracts as alternatives to conventional processes. This work aimed to evaluate the quality of red fruit juices clarified and concentrated by an integrated membrane process with special regard to the preservation of valuable compounds. A red fruit juice obtained from a blend of pomegranate, cactus pear, and red orange juices of Sicilian origin was clarified by microfiltration (MF) and then pre-concentrated up to 33 °Brix by nanofiltration (NF). The pre-concentrated juice was finally concentrated by osmotic distillation (OD) up to 50 and 60 °Brix. Samples of clarified, pre-concentrated, and concentrated juice were analyzed for their physico-chemical composition and in terms of the antioxidant activity and inhibitory activity against α-amylase and lipase. The results clearly confirmed the assumption of a mild fruit juice processing method, allowing us to preserve the original nutritional and functional properties of the fresh juice. In particular, the OD retentate at 60 °Brix resulted the most active sample against pancreatic lipase and α-amylase inhibitory activity with IC50 values of 44.36 and 214.65 µg/mL, respectively.
Assuntos
Sucos de Frutas e Vegetais , Frutas , Frutas/química , Destilação/métodos , alfa-Amilases , Lipase/análiseRESUMO
Phenolic compounds from the flower of Clitoria ternatea L. (PCFCTL) were extracted using a high-speed shearing extraction technique and purified by AB-8 macroporous resins, and the phytochemical composition of the purified phenolic compounds from the flower of Clitoria ternatea L. (PPCFCTL) was then analyzed. Subsequently, its bioactivities including antioxidant properties, enzyme inhibitory activities, and antiproliferative activities against several tumor cell lines were evaluated. Results indicated that the contents of total phenolics, flavonoids, flavonols, flavanols, and phenolic acids in PPCFCTL were increased by 3.29, 4.11, 2.74, 2.43, and 2.96-fold, respectively, compared with those before being purified by AB-8 macroporous resins. The results showed PPCFCTL have significant antioxidant ability (measured by reducing power, RP, and ferric reducing antioxidant power method, FRAP) and good DPPH, ABTS+, and superoxide anion radical scavenging activities. They can also significantly inhibit lipase, α-amylase, and α-glucosidase. In addition, morphological changes of HeLa, HepG2, and NCI-H460 tumor cells demonstrated the superior antitumor performance of PPCFCTL. However, the acetylcholinesterase inhibitory activity was relatively weak. These findings suggest that PPCFCTL have important potential as natural antioxidant, antilipidemic, anti-glycemic and antineoplastic agents in health-promoting foods.
Assuntos
Clitoria , Acetilcolinesterase , Antioxidantes/química , Clitoria/química , Flavonoides/análise , Flavonoides/farmacologia , Flavonóis/análise , Flores/química , Lipase/análise , Fenóis/análise , Fenóis/farmacologia , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/análise , Extratos Vegetais/farmacologia , Superóxidos/análise , alfa-Amilases , alfa-GlucosidasesRESUMO
Purpose: Effectively controlling the accumulation of adipose tissue can be a therapeutic strategy for treating obesity, which is a global problem. The present study was designed for comparative assessment of in vitro antiobesity activities of the Psoralea corylifolia-dichloromethane seed extract (DCME) and the isolated phytochemicals, bakuchiol, isopsoralen, and psoralen, through antiadipogenesis and pancreatic lipase (PL) inhibition assays. Material and Methods. In vitro pancreatic lipase activity was determined spectrophotometrically by measuring the hydrolysis of p-nitrophenyl butyrate (p-NPB) to p-nitrophenol at 405 nm, and adipogenesis was assayed in 3 T3-L1 adipocytes (by using Oil Red O staining) using P. corylifolia-dichloromethane seed extract (DCME) and individual compounds, isolated from the extract. Result: Antilipase as well as antiadipogenesis activity was displayed by both the DCME and the compounds. Maximum antilipase property was recorded in DCME (26.02 ± .041%) at 100 µg/ml, while, among the isolated compounds, bakuchiol exhibited a higher activity (24.2 ± 0.037%) at 100 µg/ml concentration, compared to other isolates. DCME was found to exhibit antiadipogenesis property, 75 ± 0.003% lipid accumulation, compared to the control at 100 µg/ml dose. Bakuchiol, isopsoralen, and psoralen inhibited the lipid accumulation in 3T3-L1 preadipocytes, 78.06 ± 0.002%, 80.91 ± 0.004%, and 80.91 ± 0.001%, respectively, lipid accumulation in comparison to control at 25 µM dose. Conclusion: The present study highlights the antiobesity potential of P. corylifolia and its active constituents. Thus, it can be concluded that P. corylifolia has the potential to treat obesity and related diseases; however, further research on dose standardization and clinical trials are required.
Assuntos
Fabaceae , Furocumarinas , Psoralea , Ficusina/farmacologia , Lipase/análise , Lipídeos/análise , Cloreto de Metileno , Obesidade/tratamento farmacológico , Extratos Vegetais/química , Psoralea/química , Sementes/químicaRESUMO
Du-Zhong tea is a health beverage produced from Eucommia ulmoides leaves (EUL) as raw materials using traditional tea processing and Chinese herbal pieces processing methods. To evaluate the differences between tender leaves and mature leaves of Du-Zhong tea, UPLC-Q-TOF MS was used to analyze the constituents in EUL collected in April and August. A total of 52 compounds, including iridoids, phenylpropanoids, flavonoids, lignans, and other types of compounds were identified. The contents of nine ingredients in the tender and mature leaves of E. ulmoides were determined by HPLC-DAD analysis. The results show that the average contents of the compounds in tender leaves were significantly higher than those in mature leaves. Lastly, the antioxidant and antipancreatic lipase activities of commercial Du-Zhong tea made by leaves of different maturity were evaluated. Du-Zhong tea with d-grade exhibited relatively higher antioxidant, while C-grade exhibited greater lipase inhibitory activities.
Assuntos
Eucommiaceae , Antioxidantes/química , Eucommiaceae/química , Lipase/análise , Folhas de Planta/química , Chá/químicaRESUMO
The aim of the present study is to investigate the chemical profile, antioxidant activity, carbohydrate-hydrolysing enzyme inhibition, and hypolipidemic effect of essential oils (EOs) extracted from Sicilian Citrus maxima (pomelo) flavedo. Using gas-chromatography-mass spectrometry analysis (GC-MS) we analysed the Eos of five cultivars of C. maxima, namely, 'Chadock', 'Maxima', 'Pyriformis', 'Terracciani', and 'Todarii', and their blends. The antioxidant activity was performed by using a multi-target approach using 2,2'-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic acid (ABTS), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), ferric reducing ability power (FRAP), and ß-carotene bleaching tests. The α-amylase, α-glucosidase, and lipase-inhibitory activities were also assessed. GC-MS analyses revealed D-limonene as the main monoterpene hydrocarbon in all cultivars, albeit with different percentages in the range of 21.72-71.13%. A good content of oxygenated monoterpenes was detected for all cultivars, especially for 'Todarii'. The analysis of the principal components (PCA), and related clusters (HCA), was performed to find chemo-diversity among the analysed samples. EOs from 'Chadock' and 'Maxima' were statistically similar to each other, and they differed from P3 in the smaller amount of sesquiterpene hydrocarbons, while the oils from 'Terracciani' and 'Todarii' were found to be chemically and statistically different. 'Chadock' EO was the most active to scavenge radicals (IC50 values of 22.24 and 27.23 µg/mL in ABTS and DPPH tests, respectively). 'Terracciani' EO was the most active against both lipase and α-amylase, whereas the blends obtained by the combination (1:1 v/v) of C. maxima 'Maxima' + 'Todarii' were the most active against α-glucosidase. Generally, the blends did not exert a unique behaviour in potentiating or reducing the bioactivity of the pomelo EOs.
Assuntos
Citrus , Óleos Voláteis , Eliminação de Resíduos , Antioxidantes/química , Citrus/química , Frutas/química , Lipase/análise , Monoterpenos/análise , Óleos Voláteis/química , alfa-Amilases , alfa-GlucosidasesRESUMO
The effect-directed detection (EDD) of Schisandra rubriflora fruit and leaves extracts was performed to assess their pharmacological properties. The EDD comprised TLC-direct bioautography against Bacillus subtilis, a DPPH assay, as well as α-glucosidase, lipase, tyrosinase, and acetylcholinesterase (AChE) inhibition assays. The leaf extracts showed stronger antioxidant activity than the fruit extract as well as inhibition of tyrosinase and lipase. The fruit extract was found to be extremely active against B. subtilis and to inhibit α-glucosidase and AChE slightly more than the leaf extracts. UHPLC-MS/MS analysis was carried out for the bioactive fractions and pointed to the possible anti-dementia properties of the dibenzocyclooctadiene lignans found in the upper TLC fractions. Gomisin N (518 mg/100 g DW), schisanhenol (454 mg/100 g DW), gomisin G (197 mg/100 g DW), schisandrin A (167 mg/100 g DW), and gomisin O (150 mg/100 g DW) were the quantitatively dominant compounds in the fruit extract. In total, twenty-one lignans were found in the bioactive fractions.
Assuntos
Lignanas , Schisandra , Acetilcolinesterase , Ciclo-Octanos/química , Frutas/química , Lignanas/química , Lipase/análise , Monofenol Mono-Oxigenase , Extratos Vegetais/química , Schisandra/química , Espectrometria de Massas em Tandem , alfa-GlucosidasesRESUMO
Bee pollen (BP) is full of nutrients and phytochemicals, and so it is widely used as a health food and alternative medicine. Its composition and bioactivity mainly depend on the floral pollens. In this work, BP collected by Apis mellifera with different monoculture flowering crops (BP1-6) were used. The types of floral pollen in each BP were initially identified by morphology, and subsequently confirmed using molecular phylogenetic analysis. Data from both approaches were consistent and revealed each BP to be monofloral and derived from the flowers of Camellia sinensis L., Helianthus annuus L., Mimosa diplotricha, Nelumbo nucifera, Xyris complanata, and Ageratum conyzoides for BP1 to BP6, respectively. The crude extracts of all six BPs were prepared by sequential partition with methanol, dichloromethane (DCM), and hexane. The crude extracts were then tested for the in vitro (i) α-amylase inhibitory, (ii) acetylcholinesterase inhibitory (AChEI), and (iii) porcine pancreatic lipase inhibitory (PPLI) activities in terms of the percentage enzyme inhibition and half maximum inhibitory concentration (IC50). The DCM partitioned extract of X. complanata BP (DCMXBP) had the highest active α-amylase inhibitory activity with an IC50 value of 1,792.48 ± 50.56 µg/mL. The DCM partitioned extracts of C. sinensis L. BP (DCMCBP) and M. diplotricha BP (DCMMBP) had the highest PPLI activities with an IC50 value of 458.5 ± 13.4 and 500.8 ± 24.8 µg/mL, respectively), while no crude extract showed any marked AChEI activity. Here, the in vitro PPLI activity was focused on. Unlike C. sinensis L. BP, there has been no previous report of M. diplotricha BP having PPLI activity. Hence, DCMMBP was further fractionated by silica gel 60 column chromatography, pooling fractions with the same thin layer chromatography profile. The pooled fraction of DCMMBP2-1 was found to be the most active (IC50 of 52.6 ± 3.5 µg/mL), while nuclear magnetic resonance analysis revealed the presence of unsaturated free fatty acids. Gas chromatography with flame-ionization detection analysis revealed the major fatty acids included one saturated acid (palmitic acid) and two polyunsaturated acids (linoleic and linolenic acids). In contrast, the pooled fraction of DCMMBP2-2 was inactive but pure, and was identified as naringenin, which has previously been reported to be present in M. pigra L. Thus, it can be concluded that naringenin was compound marker for Mimosa BP. The fatty acids in BP are nutritional and pose potent PPLI activity.
Assuntos
Acetilcolinesterase , Ácidos Graxos , Abelhas , Animais , Suínos , Ácidos Graxos/análise , Acetilcolinesterase/análise , Filogenia , Pólen/química , Lipase/análise , alfa-Amilases/análiseRESUMO
Vegetable oil is one of the most important components of human nutrition. Soybean (Glycine max) is an important oil crop worldwide and contains rich unsaturated fatty acids. Diacylglycerol acyltransferase (DGAT) is a key rate-limiting enzyme in the Kennedy pathway from diacylglycerol (DAG) to triacylglycerol (TAG). In this study, we conducted further research using T3 AhDGAT3 transgenic soybean. A high-performance gas chromatography flame ionization detector showed that oleic acid (18:1) content and total fatty acid content of transgenic soybean were significantly higher than those of the wild type (WT). However, linoleic acid (18:2) was much lower than that in the WT. For further mechanistic studies, 20 differentially expressed proteins (DEPs) and 119 differentially expressed metabolites (DEMs) were identified between WT (JACK) and AhDGAT3 transgenic soybean mature seeds using proteomic and lipidomics analyses. Combined proteomic and lipidomics analyses showed that the upregulation of the key DEP (lipase GDSL domain-containing protein) in lipid transport and metabolic process induced an increase in the total fatty acid and 18:1 composition, but a decrease in the 18:2 composition of fatty acids. Our study provides new insights into the deep study of molecular mechanism underlying the enhancement of fatty acids in transgenic soybeans, especially oleic acid and total fatty acid, which are enhanced by over-expression of AhDGAT3.
Assuntos
Ácidos Graxos , Glycine max , Ácidos Graxos/metabolismo , Humanos , Lipase/análise , Lipase/metabolismo , Lipidômica , Plantas Geneticamente Modificadas/genética , Proteômica , Sementes/genética , Glycine max/genética , Glycine max/metabolismoRESUMO
PURPOSE: Pancreatic enzyme replacement therapy (PERT) involves exogenous enzyme supplementation and is used in the treatment of pancreatic exocrine insufficiency. Clinical efficacy of PERT preparations is a function of physical properties and release kinetics that vary between commercially available products. In this study, we evaluated the physical properties, in vitro dissolution, and release kinetics of commercially available pancreatic enzyme preparations available in the Indian market. METHODS: Physical properties such as particle size distribution and water content of the capsules were measured by dynamic light scattering and Karl-Fischer titration method, respectively. An analytical procedure based on the European pharmacopoeia (EP) method was used to determine lipase activity, and a modified United States pharmacopoeia (USP)-based method was used for dissolution studies. Enzyme release was ascertained under gastroduodenal conditions in buffered media. RESULTS: Considerable variations in physical properties such as particle size and water content were observed between pancreatic enzyme preparations. Some preparations failed to meet the labeled lipase content as per USP standards (>90% label claim) and showed inconsistent release behavior (>5% relative standard deviation). CONCLUSION: Differences exist between pancreatic enzyme preparations in terms of physical properties, dissolution, and release behavior that can affect their clinical efficacy. The present study suggests, therefore, that these preparations should not be used interchangeably.
Assuntos
Fármacos Gastrointestinais/análise , Lipase/análise , Pancreatina/análise , Cápsulas , Liberação Controlada de Fármacos , Terapia de Reposição de Enzimas , Fármacos Gastrointestinais/química , Fármacos Gastrointestinais/normas , Humanos , Índia , Lipase/química , Lipase/normas , Pancreatina/química , Pancreatina/normas , Tamanho da PartículaRESUMO
Water content is an important factor in lipase-catalyzed reactions in organic media but is frequently ignored in the study of lipases by molecular dynamics (MD) simulation. In this study, Candida antarctica lipase B, Candida rugosa lipase and Rhizopus chinensis lipase were used as research models to explore the mechanisms of lipase in micro-aqueous organic solvent (MAOS) media. MD simulations indicated that lipases in MAOS systems showed unique conformations distinguished from those seen in non-aqueous organic solvent systems. The position of water molecules aggregated on the protein surface in MAOS media is the major determinant of the unique conformations of lipases and particularly impacts the distribution of hydrophilic and hydrophobic amino acids on the lipase surface. Additionally, two maxima were observed in the water-lipase radial distribution function in MAOS systems, implying the formation of two water shells around lipase in these systems. The energy landscapes of lipases along solvent accessible areas of catalytic residues and the minimum energy path indicated the dynamic open states of lipases in MAOS systems differ from those in other solvent environments. This study confirmed the necessity of considering the influence of the microenvironment on MD simulations of lipase-catalyzed reactions in organic media.
Assuntos
Microambiente Celular/fisiologia , Biologia Computacional/métodos , Lipase/química , Simulação de Dinâmica Molecular , Água/química , Basidiomycota/química , Basidiomycota/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Lipase/análise , Lipase/metabolismo , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Saccharomycetales/química , Saccharomycetales/metabolismo , Água/metabolismoRESUMO
OBJECTIVES: To determine the influence of pH on recovery of analytes in body fluids (BFs), investigate the mechanism of pH interference, measure the frequency of abnormal-pH BFs received, and compare pH measured by meter and paper. METHODS: We performed pH titration in residual BFs. A low-pH BF was spiked and neutralized to investigate pH interference. We measured analytes on a Roche cobas c501 analyzer (Roche Diagnostics) and calculated the percent recovery. Measurement of pH using a meter and paper was conducted on 122 BF samples received in the laboratory. RESULTS: Enzyme activity in BFs was unaffected when pHâ =â 7.4-8.5 lactate dehydrogenase, pHâ =â 7.3-10.2 amylase, pHâ =â 6.0-9.9 lipase, and pHâ =â 1.3-11.7 all other analytes. BFs had mean (range) pH of 8.0 (5.1-8.9), with a mean (range) difference (paperâ ââ meter) of â0.4 (â0.6 to 1.1). CONCLUSIONS: Irreversible loss of enzyme activity occurs in BFs at low pH. Few clinical BFs have pHâ <â 7.0, but laboratories should incorporate pH measurement in BF workflows.
Assuntos
Líquidos Corporais/química , Testes Diagnósticos de Rotina , Ensaios Enzimáticos/métodos , Concentração de Íons de Hidrogênio , Amilases/análise , Testes Diagnósticos de Rotina/instrumentação , Testes Diagnósticos de Rotina/métodos , Humanos , L-Lactato Desidrogenase/análise , Lipase/análiseRESUMO
Lipases and esterases are important catalysts with wide varieties of industrial applications. Although many methods have been established for detecting their activities, a simple and sensitive approach for picogram detection of lipolytic enzyme quantity is still highly desirable. Here we report a lipase detection assay which is 1000-fold more sensitive than previously reported methods. Our assay enables the detection of as low as 5 pg and 180 pg of lipolytic activity by direct spotting and zymography, respectively. Furthermore, we demonstrated that the detection sensitivity was adjustable by varying the buffering capacity, which allows for screening of both high and low abundance lipolytic enzymes. Coupled with liquid chromatography-mass spectrometry, our method provides a useful tool for sensitive detection and identification of lipolytic enzymes.
Assuntos
Ensaios Enzimáticos/métodos , Esterases/análise , Lipase/análise , Cromatografia Líquida/métodos , Compostos Cromogênicos/química , Eletroforese em Gel de Poliacrilamida/métodos , Esterases/química , Esterases/metabolismo , Lipase/química , Lipase/metabolismo , Lipólise , Espectrometria de Massas/métodos , Especificidade por SubstratoRESUMO
BACKGROUND/AIM: Hepatocellular carcinoma (HCC) remains one of the biggest medical issues. Pigment epithelial-derived factor (PEDF) is a glycoprotein that belongs to the superfamily of serine protease inhibitors. PEDF interacts with its two receptors, adipose triglyceride lipase (ATGL) and laminin receptor (LR). MATERIALS AND METHODS: We conducted immunohistochemical staining for PEDF, LR and ATGL in 151 resected HCCs and their background liver tissues. RESULTS: High expression of LR in HCC was associated with high histological grade and portal vein invasion, while high expression of PEDF in HCC was associated with absence of portal vein invasion. High LR expression in background liver was statistically associated with low serum albumin levels and was an independent prognostic factor of worse outcomes. No cases with more than 5% fatty degeneration in the background liver tissue showed high PEDF expression. CONCLUSION: PEDF/LR/ATGL could be potential biomarkers in HCC and various chronic hepatic disorders.
Assuntos
Carcinoma Hepatocelular/química , Proteínas do Olho/análise , Lipase/análise , Neoplasias Hepáticas/química , Fígado/química , Fatores de Crescimento Neural/análise , Receptores de Laminina/análise , Receptores de Neuropeptídeos/análise , Serpinas/análise , Carcinoma Hepatocelular/patologia , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/patologia , Masculino , Invasividade Neoplásica , Prognóstico , Albumina Sérica/análiseRESUMO
Simultaneous reduction in activity of fat destabilizing enzymes (lipase and lipoxygenase), contaminants heavy metals (As, Cd, Pb, and Hg), antinutrient phytic acid and hazardous coliforms in rice bran was investigated. Application of washing, soaking the washed sample at different pH values (2, 6 and 9) alone or in combination with ultrasonication were examined. While washing was beneficial, its low efficiency acquired further treatment, which was prevailed by application of acidic pH and ultrasound (28 kHz) treatments. Free fatty acids and peroxide value, as indicators of enzymes activity, implied the effectiveness of treatments with adverse impact of sonication on peroxide value. Remarkably, reduction of dominant heavy metals (As, Pb and Zn) and phytic acid were synergistically facilitated by sonication. Coliforms growth was inhibited at pH 2 even at the absence of ultrasonic treatment. Evidently, combination of acidic pH and ultrasound is a practical approach to improve rice bran stability and safety.
Assuntos
Análise de Alimentos , Lipase/análise , Lipoxigenase/análise , Metais Pesados/análise , Oryza/química , Ácido Fítico/análise , Concentração de Íons de Hidrogênio , Proteínas de Plantas/análiseRESUMO
Baliospermum montanum (Willd.) Muell. Arg, a medicinal plant distributed throughout India from Kashmir to peninsular-Indian region is extensively used to treat jaundice, asthma, and constipation. In the current study, 203 endophytic fungi representing twenty-nine species were isolated from tissues of B. montanum. The colonization and isolation rate of endophytes were higher in stem followed by seed, root, leaf and flower. The phytochemical analysis revealed 70% endophytic isolates showed alkaloids and flavonoids, 13% were positive for phenols, saponins and terpenoids. Further, these endophytes produced remarkable extracellular enzymes such as amylase, cellulase, phosphates, protease and lipase. The most promisive three endophytic fungi were identified by ITS region and secreted metabolites were identified by gas chromatography-mass spectrometry (GC-MS/MS). The GC-MS profile detected twenty-five bioactive compounds from ethyl acetate extracts. Among endophytic fungi, Trichoderma reesei isolated from flower exhibited nine bioactive compounds namely, 2-Cyclopentenone, 2-(4-chloroanilino)-4-piperidino, Oxime-methoxy-Phenyl, Methanamine N-hydroxy-N-methyl, Strychane, Cyclotetrasiloxane, Octamethyl and 1-Acetyl-20a-hydroxy-16-methylene. The endophyte, Aspergillus brasiliensis isolated from root and Fusarium oxysporum isolated from seed produced nine and seven bioactive compounds, respectively. Overall, a significant contribution of bioactive compounds was noticed from the diverse endophytic fungi associated with B. montanum and could be explored for development of novel drug with commercial values.
Assuntos
Aspergillus/isolamento & purificação , Endófitos/isolamento & purificação , Enzimas/análise , Euphorbiaceae/microbiologia , Fusarium/isolamento & purificação , Hypocreales/isolamento & purificação , Alcaloides/análise , Amilases/análise , Aspergillus/química , Celulase/análise , Endófitos/química , Flavonoides/análise , Fusarium/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Hypocreales/química , Índia , Lipase/análise , Peptídeo Hidrolases/análise , Folhas de Planta/microbiologia , Plantas Medicinais/microbiologiaRESUMO
In many different cell types neutral lipids can be stored in lipid droplets (LDs). Nowadays, LDs are viewed as dynamic organelles, which store and release fatty acids depending on energy demand (LD dynamics). Proteins like perilipin 2 (PLIN2) and PLIN5 decorate the LD membrane and are determinants of LD lipolysis and fat oxidation, thus affecting LD dynamics. Trained athletes and type 2 diabetes (T2D) patients both have high levels of intramyocellular lipid (IMCL). While IMCL content scales negatively with insulin resistance, athletes are highly insulin sensitive in contrast to T2D patients, the so-called athlete's paradox. Differences in LD dynamics may be an underlying factor explaining the athlete's paradox. We aimed to quantify PLIN2 and PLIN5 content at individual LDs as a reflection of the ability to switch between fatty acid release and storage depending on energy demand. Thus, we developed a novel fluorescent super-resolution microscopy approach and found that PLIN2 protein abundance at the LD surface was higher in T2D patients than in athletes. Localization of adipocyte triglyceride lipase (ATGL) to the LD surface was lower in LDs abundantly decorated with PLIN2. While PLIN5 abundance at the LD surface was similar in athletes and T2D patients, we have observed previously that the number of PLIN5 decorated LDs was higher in athletes, indicating more LDs in close association with mitochondria. Thus, in athletes interaction of LDs with mitochondria was more pronounced and LDs have the protein machinery to be more dynamic, while in T2D patients the LD pool is more inert. This observation contributes to our understanding of the athlete's paradox.
Assuntos
Diabetes Mellitus Tipo 2/metabolismo , Treino Aeróbico , Lipase/metabolismo , Gotículas Lipídicas/metabolismo , Perilipina-2/metabolismo , Perilipina-5/metabolismo , Adulto , Idoso , Atletas , Biomarcadores/análise , Biomarcadores/metabolismo , Biópsia , Diabetes Mellitus Tipo 2/patologia , Ácidos Graxos/metabolismo , Humanos , Insulina/metabolismo , Resistência à Insulina , Lipase/análise , Lipólise , Masculino , Microscopia Confocal/métodos , Pessoa de Meia-Idade , Mitocôndrias/metabolismo , Fibras Musculares Esqueléticas/citologia , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/patologia , Oxirredução , Perilipina-2/análise , Perilipina-5/análise , Adulto JovemRESUMO
BACKGROUND: Pancreatic enzyme-replacement therapy (PERT), provided as pancreatin to patients with pancreatic exocrine insufficiency (PEI), is considered an essential substitute for the pivotal physiological function the pancreas fulfills in digestion. PEI involves a reduction in the synthesis and secretion of pancreatic enzymes (lipase, protease, amylase), which leads to an inadequate enzymatic response to a meal and consequently to maldigestion and malabsorption of nutrients. The efficacy of PERT is strongly dependent on enzyme activity, dissolution, and pancreatin particle size. OBJECTIVE: The physiological properties of eight pancreatin preparations (nine batches; five different brands) available in Russia and CIS (Commonwealth of Independent States: Armenia, Azerbaijan, Belarus, Kazakhstan, Kyrgyzstan, Moldova, Russia, Tajikistan, Uzbekistan) were investigated. METHODS: The lipase activity, dissolution, and particle size distribution of samples from multiple batches of pancreatin of different strengths were measured. RESULTS: Regarding lipase activities, all pancreatin preparations except Micrazim® matched the labeled content. Considerable differences were observed in particle size and dissolution. CONCLUSION: Pancreatin preparations available in Russia and CIS demonstrate product-to-product and batch-to-batch variability regarding the measured properties of lipase activity, dissolution, and particle size. This may impact the efficacy of PERT and therefore clinical outcomes.
Assuntos
Fármacos Gastrointestinais/química , Fármacos Gastrointestinais/metabolismo , Lipase/análise , Lipase/metabolismo , Pancreatina/química , Pancreatina/metabolismo , Comunidade dos Estados Independentes , Liberação Controlada de Fármacos , Insuficiência Pancreática Exócrina/tratamento farmacológico , Fármacos Gastrointestinais/uso terapêutico , Humanos , Pancreatina/uso terapêutico , Tamanho da Partícula , Preparações Farmacêuticas/análise , Preparações Farmacêuticas/química , Federação RussaRESUMO
Acute pancreatitis is uncommon in children and adolescents, and when it does occur, it is rarely fatal. The cause of death may be misdiagnosed as a death due to diabetic ketoacidosis or an infectious disease if an autopsy is not performed. Because these deaths are often unexpected and may occur before medical care, the forensic pathologist may be contacted to determine the cause of death. Even deaths that occur after hospitalization may be referred to the medical examiner because of the concern for drug involvement, the unusual age of the decedent, or the decreased availability for a hospital autopsy. We report a fatal case of acute pancreatitis in a 14-year-old adolescent girl who was brought unresponsive to the emergency room of a university hospital and died after unsuccessful resuscitation efforts.