Phytochemical, Antioxidant and Anti-diabetic Potentials of the Ethanol Leave Extracts of four Medicinal Plants

For many years, medicinal plants have been a resource for healing in several local communities around the world and the phytochemicals in them such as flavonoids, alkaloids, phenolic, tannins, and terpenoids are attributed to their many medicinal values. Vernonia amydalina, Senna alata, Jatropha curcas, and Grewia pubescens are important plants with immense value. In this study, phytochemical screening, antioxidant analysis and the potential anti-hyperglycemic properties of the plants was investigated in-vitro. The ethanol leave extracts of the plants were subjected to qualitative phytochemical screening and tannin, flavonoids and phenol quantification. Ferric reducing antioxidant power and DPPH radical inhibition of the extracts was done by spectrophotometric method while the anti-diabetic potential was analyzed through the in-vitro ?-amylase and ?-glucosidase inhibition. Phytochemicals detected in the ethanol leave extracts of the four plants are tannins, flavonoids, phenolics, terpenoids, steroids, alkaloids, cardiac glycosides, and saponins. Flavonoids, phenols, and tannin content were highest in Senna alata (0.27±0.0.002 mg: mg of rutin per g of extract, 10.63±0.0.017 mg: mg of gallic acid per g of extract, and 6.72±0.06 mg/g respectively) followed by V. amygdalina (0.20±00.002 mg: mg of rutin per g of extract, 8.27±0.0.017 mg: mg of gallic acid per g of extract, and 7.98±0.03 mg/g respectively). While the least content of all was found in the extracts of Jatropha curcas. Concentration dependent and statistically significant difference was observed in both the FRAP and DPPH radical inhibition of all the extracts. Senna alata showed the strongest reducing power followed by the V. amygdalina. Both Senna alata and V. amygdalina showed DPPH radical inhibition that is not significantly (p>0.05) different from that of trolox. ?-amylase and ?-glucosidase inhibition was also demonstrated in a concentration dependent manner. In both the ?-amylase and ?-glucosidase inhibition, V. amygdalina and S. alata exhibited the most significant inhibitory properties among the plant extracts. The overall result in this study suggested that V. amygdalina, S. alata with the highest content of the phytochemicals, and antioxidant activities are potential source of antioxidant constituents and might be useful for the management of diseases such as diabetes.

Non-traumatic esophageal perforation: A case report on Boerhaave’s syndrome

Boerhaave’s syndrome is a potentially fatal condition characterized by spontaneous perforation of a previously healthy esophagus, due to severe vomiting or straining. It often presents with non-specific symptoms such as fever, pain, and vomiting and hence may go undiagnosed. The Makler’s triad, consisting of vomiting, chest pain, and subcutaneous emphysema, may be seen in only 50% of cases. Delayed diagnosis may result in complications such as sepsis, mediastinitis, pneumothorax, and multi-organ dysfunction. In general, patients presenting later than 48 h are conservatively managed with esophageal stenting. Surgical repair is usually reserved for those patients who present within 24 h, or are managed conservatively and develop complications. Mortality rises from 0% if treated within 24 h to about 29% if delayed more than 48 h. We present a case of Boerhaave’s syndrome in a 35-year-old male who presented with spontaneous respiratory distress and hemodynamic instability, about 36 h after the onset of vigorous vomiting. The case was managed initially with endoscopic insertion of a self-expanding metallic stent, followed later by surgical closure of the esophageal perforation. The patient, however, developed post-operative septic complications and died after a week

Osmoregulatory Appraisal of Some Osmoprotectants on Hydrolytic Activities of Some Enzymes on Seeds of Two Water Stressed Cultivars of Sorghum bicolor (Moench)

Enzymes play significant roles in metabolic processes of seeds. Therefore, this study evaluated osmoregulatory potential of some osmoprotectants on activities of some hydrolytic enzymes in the seeds of two cultivars (SOSAT.C-88 and CV. LCIC 9702) of sorghum bicolor. Matured seeds of the two cultivars were harvested and prepared for alpha, beta, total amylase and proteinase activities assay. The osmoprotectants produced significant variations on the enzymes at 10 and 14 days (DA) of 8 weeks after treatments (WAT). Seeds of well-watered SOSAT.C-88 produced higher alpha (2.10 IU/ml), beta (1.70 IU/ml) and total amylase activities (3.30 IU/ml) at 14 days (DA). Higher alpha (2.01 IU/ml and total amylase activities (2.61 IU/ml) were recorded in the seeds of CV. LCIC 9702 well-watered at 14 days DA 8WAT. Furthermore, total amylase activities (3.87 IU/ml) were recorded in the seeds produced by CV. LCIC 9702 well-watered at 14 days DA. Significant increase was noticed in beta (1.14 IU/ml) and alpha amylase (1.58 IU/ml) in the seeds of CV. LCIC 9702 treated with mycorrhiza. CV. LCIC 9702 well watered produced highest proteinase activities (1.57 U/ml) while least of the parameters were recorded in SOSAT.C-88 and CV. LCIC 9702 droughted. In conclusion, the osmoprotectants had regulatory effects on the activities of hydrolytic enzymes therefore the use of the osmoprotectants in farming should be encouraged.

Mannich bases derivatives of 2-Phenyl-5-Benzimidazole sulfonic acid; Synthesis, Characterization, Computational studies and Biological evaluation

Abstract A new series of N-Mannich bases of 2-Phenyl-5-benzimidazole sulfonic acid have been synthesized through amino methylation reaction with secondary amines. The two moieties were held together through a methylene bridge, which comes from formaldehyde (Formalin Solution 37%) used in the reaction. Chemical structures of the newly synthesized compounds have been confirmed using FT-IR, 1HNMR and 13CNMR. Different in vitro assays including Anti-oxidant, Enzyme inhibition, Anti-microbial and Cytotoxicity assay were performed to evaluate the biological potential with reference to the standard drug. Among the synthesized library, compound 3a shows maximum alpha-glucosidase inhibition with an IC50 value of 66.66 µg/ml, compound 3d was found most toxic with LC50 value of 10.17 µg/ml. ADME evaluation studies were performed with the help of Molinspiration online software. Docking calculations were also performed. Given the importance of the nucleus involved, the synthesized compound might find extensive medicinal applications as reported in the literature.

Bioactivity evaluation of least explored traditionally acclaimed medicinally potent herb Nanorrhinum ramosissimum(Wall. ) Betsche

Abstract Present study analysed the therapeutic potential of traditionally acclaimed medicinal herb Nanorrhinum ramosissimum, using plant parts extracted with different solvents (10 mg/mL). Shoot extracts exhibited comparatively better antimicrobial properties, in comparison to root extracts. Total phenolic content was estimated, to ascertain its dependency on antioxidant properties of plant extracts. Antioxidant assay revealed promising results in comparison to IC50 value of standard ascorbic acid (52.2±0.07 µg/mL), for methanolic extracts of shoot (61.07±0.53 µg/mL and 64.33±0.33 µg/mL) and root (76.705±0.12 µg/mL and 89.73±0.28 µg/ mL) for in vivo and in vitro regenerants respectively. Correlation coefficient R2 values ranged between 0.90-0.95, indicating a positive correlation between phenolic contents and antioxidant activity. Plant extracts were also able to inhibit DNA oxidative damage again indicating their antioxidative potential. Antidiabetic potential was confirmed by alpha amylase inhibition assay where shoot methanolic extracts (invivo, in vitro) exhibited the best IC50 values (54.42±0.16 µg/mL, 66.09±0.12 µg/mL) in comparison to standard metformin (41.92±0.08 µg/mL). Ethanolic extracts of roots (in vitro, invivo) exhibited the relative IC50 values (88.97±0.32µg/mL,96.63±0.44 µg/mL) indicating that shoot parts had a better alpha amylase inhibition property; thus proving the herb's bioactive potential and its prospective therapeutic source for curing various ailments.

α-amylase detection methods and applications / 生物工程学报

α-amylase is an endonucleoside hydrolase that hydrolyzes the α-1, 4-glycosidic bonds inside polysaccharides, such as starch, to generate oligosaccharides, dextrins, maltotriose, maltose and a small amount of glucose. Due to the importance of α-amylase in food industry, human health monitoring and pharmaceuticals, detection of its activity is widely required in the breeding of α-amylase producing strains, in vitro diagnosis, development of diabetes drugs, and the control of food quality. In recent years, many new α-amylase detection methods have been developed with improved speed and sensitivity. This review summarized recent processes in the development and applications of new α-amylase detection methods. The major principle of these detection methods were introduced, and their advantages and disadvantages were compared to facilitate future development and applications of α-amylase detection methods.

Fabrication of 1,4-alpha-D-glucan glucanohydrolase holding Gel-Scaffolds using Agar-Agar, a natural polysaccharide and Polyacrylamide, a synthetic organic polymer for continuous liquefaction of starch

1,4-alpha-D-glucan glucanohydrolase is among the most widely used commercial hydrolytic enzymes acting randomly on the glycosidic linkages of starch resulting in its saccharification and liquefaction. Its applicability in different industries can be improved by enhancing its stability and reusability. Therefore, in the present study attempts have been made to enhance the industrial applicability of 1,4-alpha-D-glucan glucanohydrolase from Bacillus subtilis KIBGE-HAR by adapting immobilization technology. The study developed mechanically stable, enzyme containing gel-frameworks using two support matrices including agar-agar, a natural polysaccharide and polyacrylamide gel, a synthetic organic polymer. These catalytic gel-scaffolds were compared with each other in terms of kinetics and stability of entrapped 1,4-α-D-glucan glucanohydrolase. In case of polyacrylamide gel, Km value for immobilized enzyme increased to 7.95 mg/mL, while immobilization in agar-agar resulted in decreased Km value i.e 0.277 mg/mL as compared to free enzyme. It was found that immobilized enzyme showed maximum activity at 70 °C in both the supports as compared to free enzyme having maximum activity at 60 °C. Immobilized 1,4-α-D-glucan glucanohydrolase exhibited no change in optimal pH 7.0 before and after entrapment in polyacrylamide gel and agar-agar. The enzyme containing gel-scaffold was found suitable for repeated batches of starch liquefaction in industrial processes. Agar-agar entrapped 1,4-α-D-glucanglucanohydrolase was capable to degrade starch up to seven repeated operational cycles whereas polyacrylamide entrapped enzyme conserved its activity up to sixth operational cycle.

Extract Development of sorghum Grains during Malting and Utilization of Bitter Leaf Extract for Beer Production Using Saccharomyces cerevisiae

Aims: The aim of this study was to evaluate extract development of sorghum grains during malting and utilization of bitter leaf for production of beer using S. cerevisiae. Study Design: Randomized experimental design was used to achieve the aim of the study. Place and Duration of Study: Study was conducted at the Food and Industrial Microbiology Laboratory, Department of Microbiology, Faculty of Science, University of Port Harcourt, Choba, Rivers State between June 2021 and September 2021. Methodology: The grains were bought at a local market, steeped for 24h, germinated for 5days at 30°C and kilned at 45°C for 24h. Alpha and beta amylases were extracted from the malted sorghum and their activities were determined by measuring the maltose produced. Mashing was done using infusion method, bitter leaf extract was used in place of hops, and 500ml of the wort was pitched with 50ml inoculum of Saccharomyces cerevisiae isolated from fresh palm wine. Fermentation lasted for 14 days. Results: The results indicated that ?-amylase activities were higher with a peak of 1.5mg/ml maltose as against 0.9mg/ml maltose for alpha amylase. Wort properties such as diastatic power and hot water extract were measured as 40°l and 415°l/kg respectively. The resulting beer with bitter leave as substitute for hops gave alcohol content (%) of 3.6 and 3.4, bitterness was 100.38 IBU (International Bitterness Units) and 101.82 IBU while colour was 19.15 EBC (European Brewery Convention) and 21.45 EBC for sorghum wort having 5ml and 10ml of bitter leaf extract respectively. Conclusion: The results obtained in this study depicts sorghum malt as a source of enzymes and further reveals the brewing potentials of sorghum grains in beer production with bitter leaf as a potential substitute for hops. The bitter leaf which offered a good degree of bitterness to the beer can potentially serve as a good substitute for hops in brewing industry.

In vitro ?-amylase and ?-glucosidase inhibitory potential of Pleurotus ostreatus cv. Florida extract

Pleurotus ostreatus cv. Florida is one of the widely used edible mushroom. The polysaccharides from this mushrooms have been studied for antidiabetic potential; however, no efforts have been made to explore the potential of this mushroom to influence carbohydrate metabolizing enzymes viz. ?-amylase and ?-glucosidase. The present work was undertaken to investigate the inhibitory potential of Pleurotus ostreatus cv. Florida on enzymes ?-amylase and ?-glucosidase. Several concentrations of extracts were used to study inhibition of enzymatic activity of ?-amylase and ?-glucosidase. A dose dependent inhibitory effect on enzymes was observed. The current study, for the first time, uncovered ?-amylase and ?-glucosidase inhibitory potential of Pleurotus ostreatus cv. Florida. The study could be helpful to isolate and characterize compounds responsible for it.

Chemical characterization, antioxidant activity, -amylase and acetylcholinesterase inhibitory potential of Angelica pancicii Vandas ex Velen / Caracterización química, actividad antioxidante, potencial inhibidor de -amilasa y acetilcolinesterasa de Angelica pancicii Vandas ex Velen

Abstract: Comparative study GC - FID /M S of essential oils of fruits, leaves and roots of the endemic plant Angelica pancicii Vandas ex Velen. revealed a significant difference in their chemical composition. The enantiomeric purity of the main component in the fruit oil (+) - ß - phellandrene was a lso confirmed. In addition, imperatorin, isoimperatorin, oxypeucedanin, oxypeucedanin hydrate, angeloylpangelin and umbelliprenin were isolated from the fruit hexane extract. The content of these coumarins in the hexane extracts from different plant parts was further determined by HPLC. The essential oils and hexane extracts were assessed for their antioxidant potential and inhibitory effect towards  - amylase and acetylcholinesterase enzymes. The fruit and leaf essential oils (> 80%) as well as the fruit he xane extract (> 62%) significantly inhibited acetylcholinesterase enzyme. Distinguish free radical scavenging properties were detected for the leaf (Inh. 95.0 ± 2.2 %) and the root (Inh. 66.0 ± 2.4 %) extracts.

Resumen: Estudio comparativo GC - FID / MS de aceites esenciales de frutas, hojas y raíces de la planta endémica Angelica pancicii Vandas ex Velen revelaron una dife rencia significativa en su composición química. También se confirmó la pureza enantiomérica del componente principal del aceite de fruta (+) - ß - felandreno. Además, se aislaron imperatorina, isoimperatorina, oxipeucedanina, hidrato de oxipeucedanina, angeloi lpangelina y umbeliprenina del extracto de hexano del fruto. El contenido de estas cumarinas en los extractos de hexano de diferentes partes de la planta se determinó adicionalmente mediante HPLC. Los aceites esenciales y extractos de hexano se evaluaron p or su potencial antioxidante efecto inhibidor de las enzimas -  - amilasa y acetilcolinesterasa. Los aceites esenciales de frutas y hojas (> 80%), así como el extracto de hexano de frutas (> 62%) inhibieron significativamente la enzima acetilcolinesterasa. Se detectaron propiedades de captación de radicales libres diferenciadas para los extractos de hoja (Inh. 95,0 ± 2,2%) y de raíz (Inh. 66,0 ± 2,4%).

Metabolites in Conocarpus erectus leaves attenuate α-amylase activity by modulating amino acid residues of α-amylase: an in vitro and docking study / Los metabolitos en las hojas de Conocarpus erectus atenúan la actividad de la α-amilasa al modular los residuos de aminoácidos de la α-amilasa: un estudio in vitro y de acoplamiento

The antioxidant activity and the inhibitory potential of α-amylase of lyophilized hydroethanolic extracts of Conocarpus erectus leaves obtained by ultrasonication were determined. The most potent extract was subjected to ultra-high performance liquid chromatography system equipped with mass spectrometer for metabolite identification. The identified metabolites were docked in α-glucosidase to assess their binding mode. The results revealed that 60% ethanolic extract exhibited highest ferric reducing antioxidant power (4.08 ± 0.187 mg TE/g DE) and α-amylase inhibition (IC50 58.20 ± 1.25 µg/mL. The metabolites like ellagic acid, 3-O-methyl ellagic acid, ferujol, 5, 2 ́-dihydroxy-6,7,8-trimethyl flavone and kaempferol glucoside were identified in the extract and subjected to molecular docking studies regarding α-amylase inhibition. The comparison of binding affinities revealed 3-O-methyl ellagic acid as most effective inhibitor of α-amylase with binding energy of -14.5911 kcal/mol comparable to that of acarbose (-15.7815 kcal/mol). The secondary metabolites identified in the study may be extended further for functional food development with antidiabetic properties.

Se determinó la actividad antioxidante y el potencial inhibidor de la α-amilasa de extractos hidroetanólicos liofilizados de hojas de Conocarpus erectus obtenidos por ultrasónicación. El extracto más potente se sometió a un sistema de cromatografía líquida de ultra alto rendimiento equipado con un espectrómetro de masas para la identificación de metabolitos. Los metabolitos identificados se acoplaron en α-glucosidasa para evaluar su modo de unión. Los resultados revelaron que el extracto etanólico al 60% exhibió el mayor poder antioxidante reductor férrico (4.08 ± 0.187 mg TE/g DE) e inhibición de la α-amilasa (IC50 58.20 ± 1.25 µg/mL. Los metabolitos como el ácido elágico, 3-O-metil elágico ácido, ferujol, 5, 2 ́-dihidroxi-6,7,8-trimetil flavona y kaempferol glucósido se identificaron en el extracto y se sometieron a estudios de acoplamiento molecular con respecto a la inhibición de la α-amilasa. La comparación de las afinidades de unión reveló 3-O-metil El ácido elágico como inhibidor más eficaz de la α-amilasa con una energía de unión de -14,5911 kcal/mol comparable a la de la acarbosa (-15,7815 kcal/mol). Los metabolitos secundarios identificados en el estudio pueden ampliarse aún más para el desarrollo funcional de alimentos con propiedades antidiabéticas.

Alpha (?)-amylase Inhibitory Property of Anthocleista nobilis Leaf Extract

Aims: Alpha (?)-amylase inhibitors from plants are preferable for type 2 diabetes treatment because of their relative potency and safety. This study examined, in vitro, the inhibitory effect of Anthocleista nobilis leaf extract on starch hydrolysis catalyzed by ? -amylase (extracted from moated sorghum). Place and Duration of Study: Department of Biochemistry (School of Biological Sciences) and Department of Chemistry, School of Physical Sciences, University of Cape Coast, Ghana, between June 2021 and August 2021. Methodology: Leaves of A. nobilis were air-dried, pulverized, and macerated in 70% aqueous ethanol for 72 hrs. The filtrate was concentrated and reconstituted in 0.02M Sodium phosphate buffer (pH 6.9) for further analysis including Phytoconstituents screening. In vitro analysis of ? -amylase activity as well as inhibitory effect of A. nobilis extract on ? -amylase was performed. The Lineweaver-Burk plot was employed in the inhibition analysis to determine the inhibition type, maximum initial reaction rate (Vmax), as well as the Michaelis constant (KM). Results: The percentage inhibition of ?-amylase ranged from 25.0 ± 0.46% - 85.7 ± 2.17% for 0.1mg/mL and 0.9mg/mL of the A. nobilis leaf extract respectively. The mode of ?-amylase inhibition was found from the Lineweaver-Burk plot as mixed noncompetitive. The KM and Vmax were determined as 0.2043 mM and 0.1282 mM/min respectively. In contrast, KM for the control were 0.1537mM and Vmax of 0.09750 mM/min. The inhibition property of A. nobilis could be attributed to its phytochemicals such as flavonoids, saponins, alkaloids, tannins, and terpenoids that were present. Conclusion: Anthocleista Nobilis leaf extract contains certain naturally occurring anti-diabetic compounds and could be explored for treating type 2 diabetic patients. These findings, however, need further work to validate the exact bioactive constituents responsible for the inhibitory effect.

Piper nigrum and Morinda lucida Possess Antioxidant Capacities and Regulate the Activities of Key Carbohydrate and Lipid Digestive Enzymes

Introduction: Carbohydrate and lipid digestive enzymes are instrumental in the absorbability of nutrients associated to diabetes and obesity. This study evaluated hydroethanolic extracts of Piper nigrum leaf and Morinda lucida stem bark for antioxidant capacity and enzymes (carbohydrate and lipid digestive) inhibition. Methods: Colorimetric assays determined enzyme (?-amylase, ?-glucosidase, lipase and cholesterol esterase) inhibition and antioxidant capacity (total phenolic (TPC) and flavonoid (TFC) content, radical scavenging activity (DPPH, ABTS), and ferric reducing antioxidant power (FRAP)) of hydroethanolic ethanolic extracts, ethyl acetate and hexane fractions. Results: At 1 mg/ml extracts of P nigrum and M lucida inhibited ?- amylase (9.82±1.05 - 36.63±0.69 %) and ?-glucosidase (22.47±0.34 - 67.77±0.58 %) activities. At 100 µg/ml extracts and fractions inhibited lipase (56.72±1.11 - 81.61±0.71 %) and cholesterol esterase (18.14 ±0.79 - 36.84±0.70 %) activities. IC50 for ?- amylase (2.20±0.02 - 7.8±1.42 mg/ml), ?-glucosidase (0.16±0.01 - 3.74±0.01 mg/ml), lipase (8.58±2.57 - 53.03±5.20 µg/ml) and cholesterol esterase (172.20±5.12 - 419.80±4.55 µg/ml) were registered. At 4 mg/ml, P. nigrum presented a higher TPC (153.78±8.31 - 354.63±6.33 mg/ml), TFC (21.65±1.14 -33.86±0.00 mg/ml) than M lucida TPC (10.21±0.11 - 169.89±6.54 mg/ml), TFC (ND - 87.32±6.14 mg/ml). P nigrum presented radical scavenging (DPPH and ABTS) activity with IC50 0.12±0.00 - 1.27±0.01 mg/ml compared to 1.31±0.02 - 3.44±0.12 mg/ml of M lucida. The FRAP IC50 values were better for P nigrum (3.38±0.14- 4.48±1.05 mg/ml) than M lucida (3.34±1.32 - 15.4±2.03 mg/ml). Conclusion: P nigrum presented better antioxidant capacity and more effective on lipid digestive enzymes while M lucida was more effective on carbohydrate digestive enzymes.

Evaluation of trans-cinnamaldehyde as an anti-hyperglycemic compound through inhibition of ?- amylase

The present study aims to evaluate the effectiveness of Cinnamon (Cinnamomum verum) derive bioactive compound viz.trans-cinnamaldehyde, cinnamyl alcohol, and cinnamic acid on inhibition of Bacillus licheniformis ?-amylase (BLA) and pancreatic porcine ?-amylase (PPA) activity. The inhibition extent of each of the compounds was determined along with their inhibition kinetics and compared with standard inhibitor-acarbose (Synthetic anti-diabetic agent). The IC50 values for trans-cinnamaldehyde with respect to BLA and PPAwere observed to 5.38 ?g mL?1 and 3.76 ?g mL?1, respectively. The IC50 value of acarbose was estimated to be 6.2 ?g mL?1 for both the amylases. The maximum percent enzyme inhibition of 75.8 (at 10.75 µg mL?1) and 71.6 (5.38 µg mL?1) were observed in case of BLA and PPA, respectively, using trans-cinnamaldehyde. Cinnamyl alcohol and cinnamic acid on the other hand were observed to show no specific inhibitory effect on the both the ?-amylases even at high concentrations. Catalytic efficiency (Vmax/Km) of both the amylases was observed to decrease significantly in presence of trans-cinnamaldehyde compared to acarbose. Overall, trans-cinnamaldehyde was observed as a better inhibitor of ?-amylase compared to known synthetic inhibitor-acarbose. Thus, trans-cinnamaldehyde could effectively be used for controlling hyperglycemia and diabetes mellitus.

Study of anti-diabetic, beta-carotene-bleaching inhibiting and iron chelating properties of Carissa opaca root extracts

Abstract Degenerative diseases diabetes and oxidative stress constitute a major health concern worldwide. Medicinal plants are expected to provide effective and affordable remedies. The present research explored antidiabetic and antioxidant potential of extracts of Carissa opaca roots. Methanolic extract (ME) was prepared through maceration. Its fractions were obtained, sequentially, in hexane, chloroform, ethyl acetate and n-butanol. An aqueous decoction (AD) of the finely ground roots was obtained by boiling in distilled water. The leftover biomass with methanol was boiled in water to obtain biomass aqueous decoction (BAD). The extracts and fractions showed considerable porcine pancreatic α-amylase inhibitory activity with IC50 in the range of 5.38-7.12 mg/mL while acarbose had 0.31 mg/mL. The iron chelating activity in terms of EC50 was 0.2939, 0.3429, 0.1876, and 0.1099 mg/mL for AD, BAD, ME, and EDTA, respectively. The EC50 of beta-carotene bleaching activity for AD, BAD, ME, and standard BHA were 4.10, 4.71, 3.48, and 2.79 mg/mL, respectively. The total phenolic content (TPC) and total flavonoid content (TFC) of AD and BAD were also considerable. In general, ethyl acetate fraction proved to be the most potent. Thus, the C. opaca roots had excellent antioxidant activity while having moderate α-amylase inhibitory potentia

Escherichia coli expression and characterization of alfa-amylase from Geobacillus thermodenitrificans DSM-465 / Expressão e caracterização de Escherichia coli de alfa-amilase de Geobacillus thermodenitrificans DSM-465

Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM-465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% α-helices, 15% β-sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (∆G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.

A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (∆G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.

Escherichia coli expression and characterization of -amylase from Geobacillus thermodenitrificans DSM-465

Abstract Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM-465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% -helices, 15% -sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.

Resumo A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.

Escherichia coli expression and characterization of α-amylase from Geobacillus thermodenitrificans DSM-465 / Expressão e caracterização de Escherichia coli de alfa-amilase de Geobacillus thermodenitrificans DSM-465

Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% α-helices, 15% ß-sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (∆G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.

A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (∆G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.

Eurytrema coelomaticum infection: correlation between parasite burden and impairment of pancreatic exocrine enzyme secretion / Eurytrema coelomaticum: correlação entre carga parasitária e insuficiência pancreática exócrina

Eurytrema coelomaticum is a trematode reported in the pancreatic ducts of ruminants. It is conjectured that may cause disorders in the pancreas, as well as digestive and metabolic processes dependent on them. This study, determined if there is an impairment of exocrine pancreatic function, and correlated it with parasite burden. Pancreas, blood, and fecal samples were collected from 119 bovines at a abattoir. Stool samples were subjected to the gelatin and x-ray film digestion tests (to detect the presence of trypsin in feces). Using blood samples, the following biochemical tests were performed: amylase, lipase, glucose, fructosamine, cholesterol, triglycerides, total protein, albumin, and globulins. Analyses were correlated with pancreatic parasite burden. Cattle with a high parasitic load presented higher incidence of negative tests in both gelatin digestion and x-ray film digestion tests (P < 0.001) when compared to non-parasitized animals and those with a low parasitic load. Changes in those tests only occurred if the parasitemia was moderate or severe. The activity of the amylase and lipase enzymes was significantly higher in animals with low parasitemia (P < 0.05), compared to non-parasitized animals and with a high parasitic burden. In this study, in cases of high parasitemia, negative results were observed in both gelatin and x-ray film in the feces digestion tests. However, the low infection of E. coelomaticum, higher levels of serum amylase and lipase that also indicated loss of pancreatic exocrine functions were reported.

Eurytrema coelomaticum, um trematódeo de ductos pancreáticos de ruminantes. Conjectura-se que possa ocasionar transtornos nas funções pancreáticas, mais especificamente nos processos digestivos e metabólicos dependentes destas. Neste estudo, o objetivo foi determinar se há comprometimento da função pancreática exócrina, correlacionado-a a carga parasitária. Foram utilizados pâncreas e respectivas amostras de sangue e fezes de 119 bovinos. As amostras de fezes foram submetidas aos testes de digestão da gelatina em tubo e digestão de filme radiográfico, ambos para detecção de tripsina nas fezes. Foram realizados os seguintes exames bioquímicos em amostras de sangue: amilase, lipase, glicemia, frutosamina, colesterol, triglicerídeos, proteínas totais, albumina e globulinas. Após isto, as análises bioquímicas foram correlacionadas com a quantidade numérica de parasitas encontrados no pâncreas (post-mortem). Houve maior quantidade de testes negativos (digestão do filme radiográfico e prova de digestão da gelatina) nos animais com alta carga parasitária (P < 0.001), quando comparados aos animais não parasitados e com baixa carga parasitária. Portanto, os exames supracitados se alteram somente se a quantidade de parasitas for moderada ou severa. As atividades das enzimas amilase e lipase foram significativamente maiores nos animais que apresentavam baixa parasitemia (P < 0.05), em comparação com os animais com alta carga parasitária e não parasitados. Conclui-se que em quadros de alta parasitemia há alteração significativa nos testes de digestão nas fezes, e que em quadros de baixa parasitemia há alterações significativas nos valores de amilase e lipase séricas, ambos comprovando alterações pancreáticas importantes, de acordo com o quadro de parasitemia.

Diagnostic value of endoscopic ultrasonography for pancreatic cystic lesions / 中华胰腺病杂志

Objective:To explore the diagnostic value of endoscopic ultrasonography (EUS) for pancreatic cystic lesions (PCLs).Methods:Clinical data of 211 patients with PCLs, who underwent EUS at least once and were pathologically confirmed in First Affiliated of Naval Medical University from January 2011 to December 2021 was retrospectively analyzed. EUS imaging characteristics, biochemical analysis of cystic fluid and pathological data were recorded. The pathological diagnosis results of intraductal papillary mucinous neoplasms and mucinous cystic neoplasms were included in the mucinous lesion group, while pancreatic pseudocyst, serous cystic neoplasms, solid pseudopapillary neoplasms and pancreatic neuroendocrine tumors were included as non-mucinous lesions group; those with pancreatic ductal adenocarcinoma, adenocarcinoma or with atypical or cancer cells were included as malignant lesion group, and the else were included as benign lesions group. The level of CEA in cyst fluid between mucinous and non-mucinous lesions and the level of amylase in cyst fluid between benign and malignant lesion groups were compared, and the area under the curve (AUC) was calculated by drawing receiver operating characteristic curve (ROC), which was used to analyze the differential diagnosis efficiency of cyst fluid CEA and amylase test indexes. The basic characteristics and EUS imaging characteristics, and the diagnostic efficiency of EUS and liquid-based cytology and histopathology between benign and malignant lesions were studied and analyzed.Results:Among the 211 PCL patients, cyst fluid was obtained in 201 patients, of which 188 patients (93.5%) underwent cytological examination, and 33 patients were diagnosed with an accuracy rate of 17.6%; 41 cases were obtained for histological examination, and 23 cases were confirmed, with an accuracy rate of 56.1%. Among all confirmed cases, 45 cases had benign lesions, including 22 cases of mucinous lesions and 23 cases of non-mucinous lesions, with the cyst fluid CEA of 1458.16(19.80, 1500.00), 4.4(0.50, 341.14)ng/ml respectively, and the difference of cyst fluid CEA level between mucinous and non-mucinous lesions was statistically significant( P<0.05). The cyst fluid CEA<10.15 ng/ml could be used to diagnose non-mucinous PCLs with the sensitivity of 89.5%(95% CI0.686-0.981), and the specificity of 66.7%(95% CI0.438-0.837). The cyst fluid amylase levels in benign and malignant lesions were 379.00(50.00, 18405.50), 42.00(13.50, 340.00)U/L, and the difference was statistically significant ( P<0.05). The cyst fluid amylase>747.50 U/L might help to identify benign PCLs with the sensitivity of 91.7%(95% CI0.646-0.996), and the specificity of 48.0%(95% CI0.300-0.665). EUS showed that the proportion of cyst wall thickening, main duct dilatation and cystic solid components in patients with malignant lesions was significantly higher than that in patients with benign lesions, and the differences were statistically significant ( P<0.05), while there was no significant difference in the proportion of cyst wall nodules and cystic septum between the two groups. The accuracy of EUS combined with liquid-based cytology or histopathology in malignant lesions was over 80%. Conclusions:The cyst fluid CEA level can help to differentiate non-mucinous PCLs from mucinous PCLs, and the cystic amylase level could be useful to identify the benign and malignant PCLs. EUS combined with cytology or histology had high diagnostic value for malignant or potentially malignant PCLs, and EUS-FNA examination can be recommended as soon as possible for those with high-risk factors.