scholarly journals In-vitro Analysis of Trypsin and Alpha - Amylase Inhibitory Activities in Selected Legume Varieties in Sri Lanka

2021 ◽  
pp. 1-9
Author(s):  
A. M. C. N. Abeykoon ◽  
H. M. T. Herath ◽  
M. A. Jayasinghe

Aim: To quantify the Trypsin Inhibitory Activity (TIA) and Alpha - Amylase Inhibitory Activity (AIA) in legume varieties with effect of cooking. Study Design: Seeds of twelve legume varieties grown in Complete Randomized Block Design (CRBD) in experimental field conditions were used and data analysis was performed using one-way ANOVA at 95% confidence interval using MINITAB statistical software.                                                 Place and Duration of Study: Grain Legumes and Oil Crops Research and Development Centre (GLOCRDC), Angunakolapalessa and Industrial Technology Institute (ITI), Colombo, Sri Lanka between June 2019 and Dec 2019. Methods: Ethanolic (80%) extracts of raw and cooked grain legumes were used. In determining TIA, N-α-benzoyl-DL-arginine-p-nitroanilide hydrochloride (BAPA) is used as a synthetic substrate for trypsin enzyme and the rate of hydrolysis was measured by intensity of colour released by p-nitroaniline. AIA was carried out determining the maltose content which was released by hydrolysis of starch in the presence of amylase enzyme using reduction of 3, 5-dinitrosalicylic acid. Results: TIA in raw legumes ranged from 0.65±0.02 mg/g (ANK- Brown) to 1.52±0.01 mg/g (ANKCP1) while in cooked legumes ranged from -0.11±0.1 mg/g (ANK-Black) to 0.61±0.02 mg/g (MI 5). In pressure cooking (120oC, 10 min) considerable reduction in TIA of 53.74% (MI6) to 100% (ANK-Black, Bombay) was observed. A significant difference (p<0.05) in TIA among the varieties as well as among cooked form of varieties were observed (one- way ANOVA). Further a significant difference (p<0.05) in TIA was observed between cooked and raw form in each legume variety (Paired T-Test). AIA of cooked samples was ranged from - 11.61% (MI 6) - 23.05% (MISB1) and there was no significant difference (p<0.05) in AIA among the most of the legume varieties.   Conclusion: A significant reduction of TIA among the legumes varieties was observed in the pressure cooking process while a significant activity of alpha- amylase was not seen in cooked legumes.

2016 ◽  
Vol 3 ◽  
pp. 160-163
Author(s):  
Muhammad Amir Masruhim ◽  
Wisnu Cahyo Prabowo ◽  
Dita Paramitha

Hyperuricemia is a condition in which increased levels of uric acid in the blood. Xanthine oxidase role in the oxidation of hypoxanthine and xanthine to uric acid. One treatment of hyperuricemia is inhibiting xanthine oxidase in the process of formation of uric acid. The purpose of this study to determine the inhibitory activity of xanthine oxidase in the ethanol extract of black betel leaf (Piper sp). Xanthine oxidase inhibitory activity test using UV-Vis spectrophotometry in vitro with a concentration of 5 ppm, 10 ppm and 20 ppm. The data obtained were analyzed using one-way ANOVA. The result is the ethanol extract of black betel leaf has a different activity significantly and IC50 values obtained is 65.96 ppm.


Author(s):  
Jirawat Riyaphan ◽  
Chien-Hung Jhong ◽  
May-Jwan Tsai ◽  
Der-Nan Lee ◽  
Max K. Leong ◽  
...  

The inhibition of alpha-glucosidase and alpha-amylase is one of clinic strategies for remedy the type II diabetes. Herbal medicines are reported to alleviate hyperglycemia. However, the constituents from those sources whether are targeted to the alpha-glucosidase and alpha-amylase still unexplored. This study attempted to select the compounds for efficacy of hypoglycemia via cellular and mouse levels. The results illustrated that the cytotoxicity in all tested compounds at various concentrations except the concentration of 16-hydroxy-cleroda-3,13-dine-16,15-olide (HCD) at 30 &micro;M were not significant difference (p &gt; 0.05) when compared with the untreated control. Acarbose (reference drug), Antroquinonol, Catechin, Quercetin, Actinodaphnine, Curcumin, HCD, Docosanol, Tetracosanol, Berberine, and Rutin could effectively inhibit the alpha-glucosidase activity of Caco-2 cells when compared with the control (maltose). The compounds (Curcumin, HCD, Tetracosanol, Antroquinonol, Berberine, Catechin, Actinodaphnine, and Rutin) could reduce blood sugar level at 30 min in tested mice. The effects of tested compounds on area under curve (AUC) were significant (p &lt; 0.05) among Acarbose, Tetracosanol, Antroquinonol, Catechin, Actinodaphnine, and Rutin along with Berberine and Quercetin. In in vitro (alpha-glucosidase) with in vivo (alpha-amylase) experiments suggest that bioactive compounds can be a potential inhibitor candidate of alpha-glucosidase and alpha-amylase for the alleviation of type II diabetes.


2021 ◽  
Vol 3 (1) ◽  
pp. 22
Author(s):  
Pubudu Kumara ◽  
Kandiah Pakeerathan ◽  
Liyanage P. P. Deepani

Green gram (Vigna radiata (L.) R. Wilczek) is one of the most economically important grain legumes of the traditional farming systems of Sri Lanka because it is a cheap source of protein and animal feed, and sustains soil fertility by fixing atmospheric nitrogen. Weeds are one of the major problems in green gram cultivation, reducing the yield through competition, interference with harvest and harboring pests and diseases. Controlling of weeds by applying herbicides would definitely cause unexpected damage to human health and the abundant biodiversity of Sri Lanka. Therefore, an investigation was planned to evaluate the yield loss due to weeds and to determine the optimum weed free period to minimize the yield losses. Two experiments were performed. The first experiment was conducted to determine the effects of different weed functional groups on the yield of green gram. In the second experiment, weeds were continuously hand weeded and areas kept weed free. In the third, weeds were allowed to compete with green gram until 2, 3, 4, 5 or 6 weeks after cultivation. All the treatments were conducted in randomized complete block design with three replicates. The data collected on types of weed, number of pods and pod weight at 3–6 weeks after planting (WAP) were analyzed using the SAS 9.4 statistical package, and DMRT was performed to determine the best treatment combination. The results from the first experiment showed that average yield loss due to total weed populations was 54.77%. Yield loss due to grasses alone was 46.56%, far worse than broad leaves (16.49%) and sedges (18.01%) at p < 0.05. Crop stand count at 3–4 WAP was not significantly different among treatments. However, biomass weight of 50 plants, number of pods in 50 plants and grain weight of 10 plants were found to be significantly different after 3–4 WAP in weed free conditions at p < 0.05%. When the weed free period increased, the yield was increased until 3 WAP; thereafter, not significant yield increment was observed. In contrast, yield steeply declined in plots that had weeds until 3 WAP. According to the results of the present study, it can be concluded that the critical weed free period from the planting of green gram is 3 WAP. Maintaining a weed free period for 3–4 weeks is recommended to minimize the yield loss of green gram at minimal weed management cost.


2005 ◽  
Vol 17 (2) ◽  
pp. 190
Author(s):  
W.C. Chang ◽  
J. Xu ◽  
S. Jiang ◽  
X.C. Tian ◽  
X. Yang ◽  
...  

The aim of this experiment was to determine the effect of the sucrose concentration (0 to 0.33 M) in the dilution medium on the viability, fertilizability, and development of vitrified bovine oocytes. Bovine oocyte-cumulus complexes were collected from slaughterhouse ovaries and in vitro-matured as reported previously. After 24-h maturation in TCM199-based medium under 5% CO2 humidified air at 39°C, these were exposed to hyaluronidase and carefully pipetted to remove all except the 3–5 innermost layers of cumulus. Oocytes were put into the pre-equilibration medium for 3 min and then into vitrification solution containing HEPES-buffered TCM199 supplemented with 20% FBS, ethylene glycol, and dimethylsulphoxide for 25–30 s; they were then vitrified by modified solid surface vitrification (Dinnyes et al. 2000 Biol. Reprod. 63, 513–518).The oocytes were warmed at 39°C by placing them in holding medium with 0, 0.08, 0.17, 0.25, or 0.33 M sucrose. Non-vitrified oocytes were used as controls. Oocytes were inseminated 30 min after warming, and the presumptive zygotes were cultured in CR1-aa medium supplemented with 6 mg/mL BSA at 39°C in a humidified atmosphere of 5% CO2, 5% O2, and 90% N2 for eight days. Data were analyzed by one-way ANOVA. As shown in Table 1, there was no significant difference in survival rate (P > 0.05) of the vitrified oocytes that were placed in dilution solution containing 0.17, 0.25, or 0.33 M sucrose and the non-treated controls. On Day 2 (fertilized on Day 0), cleavage to the 8-cell stage was similar for the 0.17, 0.25, and 0.33 M dilution groups, but the rates for all three were significantly lower (P < 0.05) than for the control group. The blastocyst rate on Day 8 was significantly higher for the 0.25 M group than for any other experimental group but still significantly lower than for the control. In conclusion, this study suggests that with this vitrification/warming procedure the optimum concentration of sucrose in the dilution solution is 0.25 M. Table 1. Oocyte survival after vitrification/warming and subsequent embryo development The authors would like to thank Ms Colleen Shaffer for the preparation of bovine oocytes.


2012 ◽  
Vol 10 (1) ◽  
pp. 17-22
Author(s):  
SITA AULIA SARI ◽  
RUBEN DHARMAWAN ◽  
PARAMASARI DIRGAHAYU

Sari SA, Dharmawan R, Dirgahayu P. 2012. The antifungal effect of roselle calyx extract on Trichophyton rubrum growth in vitro. Biofarmasi 10: 17-22. Dermatophytosis is a fungal infection on skin that one of them caused by Trichophyton rubrum. Dermatophytosis treatment by using chemical drugs has many shortcomings, such as a high cost and a drug resistance. Roselle (Hibiscus sabdariffa L.) calyx content was flavonoid, which have an antifungal effect. Flavonoids on roselle calyx include anthocyanin, gossypeptin (hexahydroxyflavone) 3-glucoside, flavonol glucoside hibiscritin, flavonoid gossypeptin, delphinidine 3-monoglucoside, cyanidin 3-monoglucoside. The objective of this study was to determine the effect of roselle calyx on Trichophyton rubrum growth in vitro. The study was performed as an experimental laboratory. The object of study was T. rubrum. The sample of T. rubrum colonies in this study was taken by a random sampling. The study used T. rubrum colonies on seven Sabouraud Dextrose Agar plates. Each plate had four holes. Each hole was filled by aquadest as a negative control, fluconazole 25 µg/mL as a positive control, and various roselle calyx extract concentrations (10%, 20%, 30%, 40% and 50%). The plates were incubated in an incubator with a temperature of 25oC for 7 days and measured for the diameter of roselle calyx extract inhibition effect. The data were collected and analyzed by One-way ANOVA and Least Significance Difference (LSD) tests on SPSS 16.0 for Windows. The result of One-way ANOVA test showed that there was a difference of inhibition diameter mean among all of the various roselle calyx extract concentration groups (p<0.05). The diameter of roselle calyx extract inhibition effect increased for each concentration up to 50%. The inhibition diameter of positive control compared to 20% roselle calyx extract concentration had no a significant difference. The study was concluded that roselle calyx extract has an antifungal effect to T. rubrum growth in vitro.


2012 ◽  
Vol 19 (1) ◽  
pp. 25
Author(s):  
Yulita Kristanti ◽  
D. Dessy

Latar Belakang. Penelitian seputar ozonazed water akhir-akhir ini cukup banyak diminati karena bahan ini mempunyai potensi antibakteria yang menjanjikan. Tujuan Penelitian ini adalah untuk mengetahui pengaruh ozonated water pada pertumbuhan bakteri Staphylococcus Aureus. Metode Penelitian. Penelitian ini dilakukan dengan cara mencampurkan 1 ml larutan bakteri 106 CFU/ml dengan 10 ml ozonated water konsentrasi 4 ppm selama 10 detik (kelompok I), 20 detik (kelompok II), 30 detik  (kelompok III), dan 40 detik (kelompok IV), akuades (kontrol negatif) kemudian semua diencerkan dengan akuabides 10ml. selanjutnya dari masing-masing kelompok diambil 0,01 ml untuk ditanam pada MHA, inkubasi 24 jam pada suhu 370 C dalam anaerobic jar kemudian dilakikan perhitungan koloni. Data dianalisis dengan Avana satu jalur dilanjutkan dengan uji LSD. Hasil. Avana satu jalur memperlihatkan adanya perbedaan yang bermakna antar kelompok perlakuan. Uji LSD menunjukkan perbedaan rerarta yang bermakna (p<0,05) antar kelompok I, II, III, IV dengan kelompok V (akuedes) terdapat perbedaan rerata yang bermakna antara kelompok I dan kelompok VI (iod). Tidak terdapat perbedaan bermakna antara kelompok II, III, IV terdapat kelompok VI (oid). Uji korelasi Produk Momen Pearson menunjukkan adanya korelasi negatif yang sangat kuat. Kesimpulan. Ozonated Water 4 ppm memiliki daya antibakreri dalam menghambat pertumbuhan Staphylococcus Aureus. Semakin lama waktu kontak, daya anti bakterinya semakain besar. Background. Some previous research show that the antibacterial effect of ozonated water is very promi-sing. The aim of this study was to know the antibacterial effect of ozonated water on staphylococcus aureus. Method. One ml of 106 CFU/ ml S. aureus suspension was mixed with 10 ml ozonated water for 10 second (group I), 20 second (group II), 30 second (group III), 40 second (group IV)As negative control S. aureus was mixed with aquadest (group V), and as positive control S. aureus was mixed with iod (groupVI). Furthermore, 0,01 ml from each group was cultivated on MHA, incubated for 24 hours-37C followed by colony caunting. Data was analyzed using one way anova followed by LSD. Result. One way anova show significant difference among the group and LSD test show significant mean different between group I, II, III, IV, and V. Significant difference can also be seen between group I and V. No significant difference between group II, III, IV and VI. Product momen test show strong negative correlation. Conclusion. Ozonated water 4 ppm has antibacterial effect on S. aureus. The longer the contact time, the stronger the antibacterial effect.


2020 ◽  
Vol 19 ◽  
pp. e207286
Author(s):  
Kamila Aguiar Figueiredo Alves ◽  
Janaina Emanuela Damasceno ◽  
Viviane Maia Barreto de Oliveira ◽  
Luiz Gustavo Cavalcanti Bastos ◽  
Andrea Nóbrega Cavalcanti

Aim: This study evaluated the precision of a CAD/CAM system by measuring marginal, internal and proximal fits in implantsupported single-crown restorations. Methods: Ten models of the upper arch were made in which implants replaced the upper left premolars. For fabrication of the zirconia infrastructures, titanium bases (TiBase) were coded and scanned using a scan body. A second digital impression was made for the fabrication of prostheses. Silicone impression material was used to determine the internal clearance between the TiBase and infrastructure and between the infrastructure and crown, whose thickness was measured at three points [P1 (cervical), P2 (middle) and P3 (occlusal)] with a stereoscopic microscope at 70x and 100x magnification. One-way ANOVA for repeated measures and the Student t-test were used for the analysis of internal and marginal adaptation. Proximal contacts were analyzed qualitatively. Results: There was no significant difference between the teeth evaluated (Student’s t-test; p>0.05) or between the corresponding points evaluated in either tooth (one-way ANOVA; p>0.05). Analysis of the internal clearance between the infrastructure and crown demonstrated that all points were significantly different compared to the reference standardized at 100 μm (Student’s t-test p<0.0001). There was no significant difference between P1 and P2, with the thickness at these two points being lower than that obtained at P3 (one-way ANOVA, p<0.05). The proximal contacts did not coincide with the quality defined by the device. Conclusion: The system tested was unable to produce implantsupported single-crown ceramic restorations with marginal, internal and proximal fits matching the digital workflow, with the inferior fits requiring adjustment prior to cementation.


1998 ◽  
Vol 66 (12) ◽  
pp. 5826-5832 ◽  
Author(s):  
Spencer R. Hedges ◽  
Matthew S. Mayo ◽  
Lisa Kallman ◽  
Jiri Mestecky ◽  
Edward W. Hook ◽  
...  

ABSTRACT Immunoglobulin A1 (IgA1) protease, an enzyme that selectively cleaves human IgA1, may be a virulence factor for pathogenic organisms such as Neisseria gonorrhoeae. Host protection from the effects of IgA1 protease includes antibody-mediated inhibition of IgA1 protease activity, and it is believed that the relative balance between IgA1 protease and inhibitory antibodies contributes to the pathogenesis of disease caused by IgA1 protease-producing organisms. We have examined the levels of these two opposing factors in genital tract secretions and sera from women with uncomplicated infection withN. gonorrhoeae. When IgA1 in cervical mucus was examined by Western blotting, no evidence of cleavage fragments characteristic of IgA1 protease activity was seen in gonococcus-infected or control patients. Cleavage fragments typical of IgA1 protease were detected, however, after the addition of exogenous IgA1 protease to cervical mucus. Degraded IgA1 was detected in some vaginal wash samples, but the fragment pattern was not typical of IgA1 protease activity. AllN. gonorrhoeae isolates from the infected patients produced IgA1 protease in vitro. All but two serum samples and 16 of 65 cervical mucus samples displayed inhibitory activity against gonococcal IgA1 protease, but there was no significant difference in the level of inhibitory activity between gonococcus-infected and noninfected patients in either cervical mucus or serum. There was no difference in the levels of IgA1 protease-inhibitory activity in serum or cervical mucus collected from patients at recruitment and 2 weeks later. These results suggest that cleavage of IgA1 by gonococcal IgA1 protease within the lumen of the female lower genital tract is unlikely to be a significant factor in the pathogenesis of infections by N. gonorrhoeae.


Author(s):  
Shahram Mosharrafian ◽  
Maryam Shafizadeh ◽  
Zeinab Sharifi

Objectives: This study aimed to compare the fracture resistance of a bulk-fill and a conventional composite and a combination of both for coronal restoration of severely damaged primary anterior teeth. Materials and Methods: In this in vitro experimental study, 45 primary anterior teeth were randomly divided into three groups. After root canal preparation, the canals were filled with Metapex paste such that after the application of 1 mm of light-cure liner, 3 mm of the coronal third of the canal remained empty for composite post fabrication. Filtek Z250 conventional composite was used in group 1, Sonic-Fill bulk-fill composite was used in group 2 and Sonic-Fill with one layer of Filtek Z250 as the veneering were used in group 3. Adper Single Bond 2 was used in all groups. The teeth were thermocycled, and fracture resistance was measured by a universal testing machine. The mode of fracture was categorized as repairable or irreparable. Data were analyzed using one-way ANOVA. Results: The mean fracture resistance was 307.00±74.72, 323.31±84.28 and 333.30±63.96 N in groups 1 to 3, respectively (P=0.55). The mean fracture strength was 14.53±2.98, 15.08±2.82 and 15.26±3.02 MPa in groups 1 to 3, respectively (P=0.77). The frequency of repairable mode of failure was 80% for the conventional, 73.6% for the bulk-fill and 80% for the bulk-fill plus conventional group, with no significant difference (P>0.05). Conclusions: Bulk-fill composites can be used for coronal reconstruction of severely damaged primary anterior teeth similar to conventional composites to decrease the treatment time in pediatric patients.


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