scholarly journals Identification of Bioactive Phytochemicals from Six Plants: Mechanistic Insights into the Inhibition of Rumen Protozoa, Ammoniagenesis, and α-Glucosidase

Biology ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1055
Author(s):  
Aurele Gnetegha Ayemele ◽  
Lu Ma ◽  
Xiumei Li ◽  
Peilong Yang ◽  
Jianchu Xu ◽  
...  

Rumen protozoa prey on feed-degrading bacteria synthesizing microbial protein, lowering nitrogen utilization efficiency in ruminants. In this in vitro study, we evaluated six plants (Adansonia digitata, Flemingia macrophylla, Kalimeris indica,Brassica rapa subsp. chinensis, Portulaca oleracea, and Calotropis gigantea) for their potential to inhibit rumen protozoa and identified the phytochemicals potentially responsible for protozoa inhibition. Rumen protozoa were anaerobically cultured in vitro in the presence of each plant at four doses. All of the tested plants reduced total rumen protozoa (p ≤ 0.05), but C. gigantea and B. rapa were the most inhibitory, inhibiting rumen protozoa by 45.6 and 65.7%, respectively, at the dose of 1.1 mg/mL. Scanning electron microscopy revealed a disruption of the extracellular structure of protozoa cells. Only C. gigantea also decreased the wasteful ammoniagenesis (p ≤ 0.05). Moreover, the A. digitata extract inhibited α-glucosidase activity by about 70% at 100 µg/mL. Reversed-phase high-performance liquid chromatography analysis detected quercetin, anthraquinone, 3-hydroxybenzoic acid, astragaloside, and myricetin in the tested plant leaves. These plants may hold potential as feed additives to reduce rumen protozoa and α- glucosidase activity. Future research is needed to identify the specific anti-protozoal compound(s), the effects on the rumen microbiome, and its fermentation characteristics.

2021 ◽  
Author(s):  
Aurele Gnetegha Ayemele ◽  
Lu Ma ◽  
Xiumei Li ◽  
Peilong Yang ◽  
Jianchu Xu ◽  
...  

Abstract Rumen protozoa have a little contribution to the feed digestibility but Entodinium, the most predominant genus, is unfortunately culprit of the nitrogen utilization inefficiency. To bridge the gap, antibiotics have been used to inhibit the rumen protozoa but unfortunately, due to the health concern, this could not be really applied at the farm level, especially in the organic farms where the use of natural plants is an utmost priority. Therefore, our study aimed at investigating the nutritional and functional properties of six forest plants for their potential as feed additives in animal husbandry. The plants were analyzed for major phytochemicals using reversed phase-HPLC analysis and then evaluated for their in-vitro suppressing effect on rumen protozoa, ammoniagenesis, and microbial α-glucosidase activity. For each plant, four doses (0, 0.7, 0.9, and 1.1 mg/mL culture fluid) displayed in a complete randomized design were used. Quercetin, anthraquinone, 3-hydroxybenzoic acid, astragaloside, and myricetin were found to different extent (P ≤ 0.05) in the plant leaves. All the tested plants reduced total rumen protozoa counts but C. gigantea and B. rapa had the most inhibitory effect (P ≤ 0.05), inhibiting the rumen protozoa by 45.6 and 65.7%, respectively, at the dose of 1.1 mg/mL. Moreover, the scanning electron microscopy revealed the mechanistic disruption of the extracellular membrane of the protozoa, indicating their metabolic death pathway. Only C. gigantea inhibited the rumen protozoa in a proportion that also led to the reduction of the wasteful ammonia production (P ≤ 0.05). Besides, A. digitata and F. macrophylla have the higher inhibition rate (70%) of the microbial α-glucosidase activity at 100 µg/mL of crude extract. Overall, the plants showed promising results as functional feed additives although future research on bio-guided fractionation is needed to accurately identify the pure anti-protozoal bioactive compound(s).


2020 ◽  
Vol 11 (4) ◽  
pp. 5373-5381
Author(s):  
Iskandarsyah ◽  
Camelia Dwi Putri Masrijal ◽  
Harmita

A hormonal contraception progestin such as medroxyprogesterone acetate (MPA) is used to helps regulate ovulation thus as a part of contraception hormone therapy as a method of birth control. This study aimed to formulate, characterized, evaluated transfersomal gel containing medroxyprogesterone acetate and to increased subcutaneous penetration of medroxyprogesterone acetate. In this research, three transfersomes formulas were prepared and optimized, e.g. F1, F2 and F3 with phosphatidylcholine: tween 80 concentration were 90:10; 85:15; and 75:25, respectively. F2 was the best formula with the highest entrapment efficiency 81.20±0.42 %, Average 81.35 ±0.78 nm, morphology of vesicles were spheres, indeks polidispersity 0.198±0.012 and zeta potential was -34.83±0.64 mV. The transpersonal gel (FGT) containing F2, and non-transpersonal gel containing MPA in methanol(FG) were prepared. In vitro penetration test were conducted to both of them using Franz Diffusion cells. Analysis of medroxyprogesterone acetate used a high performance liquid chromatographic (HPLC) method with an ultraviolet detector on reversed-phase C18, 5µm; 150 x 4.6 mmcolumn; using acetonitrile-0.1% formic acid (60:40/v:v) and was detected at a wavelength of 240 nm with flow rate at 1.0 mL/min. Gel stability evaluation results showed that FGT was better than FG on pH stability, viscosity and rheological properties. Based on in vitro penetration study, cumulative subcutaneous penetration of medroxyprogesterone acetate from FGT was 2356.45 ± 197.73 ng.cm-2 and from FG 359.15 ± 13.60 ng.cm-2, respectively. Flux value for FGT and FG were 112.77 ± 6,47 ng.cm-2.hr-1and 17.99 ± 4.81 ng.cm-2.hr-1, respectively. It could be concluded that transfersomal gel medroxyprogesterone acetate for transdermal drug delivery increased cumulative transdermal penetration of medroxyprogesterone acetate by six times more than non-transfersomal gel dosage form.


2021 ◽  
Vol 104 (3) ◽  
pp. 36-46
Author(s):  
Vitaly A Ryazanov ◽  
◽  
Georgy I Levakhin ◽  
Galimzhan K Duskaev ◽  
Elena V Sheyda ◽  
...  

Author(s):  
John Fanikos ◽  
Kathleen Marquis ◽  
Leo Francis Buckley ◽  
Lena Kim Tran ◽  
Kevin C McLaughlin ◽  
...  

Introduction Ultrasound-facilitated catheter-directed thrombolysis is used with low-dose alteplase to treat pulmonary embolism. This reduces the bleeding risk that accompanies systemic administration of higher alteplase doses. While studies suggest that alteplase given over 2 to 6 hours is safe and effective, few data exist to support alteplase stability under these conditions. Therefore, we undertook this in vitro study to determine the duration of alteplase stability. Methods Alteplase was prepared in solutions of 8 mg in 100 mL, 6 mg in 150 mL, and 8 mg in 200 mL. Solutions were administered through the EkoSonicTM Endovascular System with and without ultrasound, to simulate administration over 2, 4, and 6 hours. Alteplase was assessed with reversed-phase high-performance liquid chromatography (RP-HPLC). Assays were performed at time 0 and at 30-minute intervals during simulated infusion. An enzyme-linked immunosorbent assay (ELISA) assay was used to measure alteplase concentrations that were at time 0 and at 15-minute intervals during simulated infusion. Results Using RP-HPLC, in the absence of ultrasound, the alteplase concentration remained within 1% of the original concentration through 120, 240, and 360 minutes of infusion. Using RP-HPLC for measurement, alteplase, in the presence of ultrasound, degraded steadily over time to approximately 90%, 80%, and 70% of its original amounts in 120, 240, and 360 minutes, respectively. Alteplase that remained was available for enzymatic activity. Conclusions Alteplase solutions of 0.04 and 0.08 mg/mL degraded steadily over time during simulated ultrasound-facilitated catheter-directed administration. Alteplase that did not degrade remained available for enzymatic activity.


Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 2008
Author(s):  
Shin Ja Lee ◽  
Hyun Sang Kim ◽  
Jun Sik Eom ◽  
You Young Choi ◽  
Seong Uk Jo ◽  
...  

We evaluated whether olive leaves (OLs) are effective as feed additives and supplements for ruminants and the potential methane reduction effects during in vitro fermentation. Two Hanwoo cows (460 ± 20 kg) equipped with cannula were fed Timothy hay and corn-based feed 3% of the body weight at a ratio of 6:4 (8:30 a.m. and 5:00 p.m.). Ruminal fluid from the cows was collected and mixed before morning feeding. In vitro batch fermentation was monitored after 12 and 24 h of incubation at 39 °C, and OLs were used as supplements to achieve the concentration of 5% in the basal diet. At 12 h of fermentation, methane production decreased in the 5% OLs group compared to that in the control group, but not at 24 h. The proportion of cellulose-degrading bacteria, Fibrobacter succinogenes, Ruminococcus albus, and Ruminococcus flavefaciens, tended to increase in the 5% OLs group at 12 h. The amount of ammonia produced was the same as the polymerase chain reaction result for Prevotella ruminicola. At 12 h, the proportion of Prevotella ruminicola was significantly higher in the 5% OLs group. OLs may be used incorporated with protein byproducts or other methane-reducing agents in animal feed.


Author(s):  
Kristian Kniha ◽  
Frank Hölzle ◽  
Faruk Al-Sibai ◽  
Johannes Jörg ◽  
Reinhold Kneer ◽  
...  

Thermal treatment may reverse the osseointegration of implants and could become an atraumatic controlled method for implant removal in the future. The aim of this non-random in vitro study was to empirically identify suitable sources for a controlled heating process, in order to generate a homogenous temperature distribution at a threshold level of 47°C for future in vivo research. Two different set-ups evaluating four different sources (water, laser, monopolar and an electrical joule heater device) were used to carry out infrared measurements and numerical calculations at 47°C along the implant axis and along the periimplant area at the axial plane. Furthermore, required time intervals to heat up the implant tip from 33°C to 47°C were determined. The monopolar electric device led to the most uneven and unpredictable implant heating and was therefore excluded. The thermal analysis suggested identical thermal distributions without any significant differences for water and electrical joule sources with a heat maximum at the implant shoulder (p > 0.05). On the other hand, the laser device may produce the temperature maximum in the middle of the implant without any afterglow effect (p < 0.01). When the implant was heated from 33°C up to 47°C, the water device indicated the fastest approach. Thermal distributions of water and laser sources may be suitable for clinical applications. For future research the numerical analysis may suggests an ideal time interval of 120s to 180s for a homogenous implant temperature of 47°C.


2019 ◽  
Vol 30 (2) ◽  
Author(s):  
Juan Camilo Duque-Aristizabal ◽  
Liza Maria Isaza-Areiza ◽  
Diego Tobon-Calle ◽  
Martha Elena Londoño Lopez

Introduction: the aim of this study is to evaluate the antimicrobial capacity of silver nanoparticles immobilized in a zinc oxide-eugenol (ZOE) cement against Enterococcus faecalis for potential use in endodontic treatments. Method: experimental in vitro study, performing synthesis of silver nanoparticles (AgNPs) and UV-visible spectroscopy to confirm presence of AgNPs in the prepared solutions. The ZOE mixture was standardized, producing the integrated AgNPs/ZOE material. Scanning electron microscopy (SEM) and Fourier-Transform Infrared Spectroscopy (FTIR) were used to characterize the integrated material; a KirbyBauer assay was finally run to measure the inhibition halos produced by the compound-microorganism interaction. Results: the UV-visible spectroscopy showed presence of AgNPs in the created solution; both SEM and FTIR show that the AgNPs are integrated into the ZOE system, not altering their properties when performed under conditions like those found in the mouth. The Kirby-Bauer assay shows that all samples had inhibition halos. The AgNPs in guava extract had statistically significant differences with the halos of the other samples (p < 0.05). Conclusions: the obtained AgNPs show bactericidal activity against Enterococcus faecalis, as a statistically significant difference was found between the AgNPs suspended in guava extract and the other groups; this will be the starting point for future research


2019 ◽  
Vol 244 (8) ◽  
pp. 709-720 ◽  
Author(s):  
Maxwell Allen-Benton ◽  
Heather E Findlay ◽  
Paula J Booth

Integral membrane proteins comprise a large proportion of drug targets, yet are challenging to study in vitro due to their amphiphilic nature. Conducting useful functional in vitro studies requires an artificial membrane that can mimic the lipid environment of the biogenic membrane. Droplet interface bilayer technology provides a method to form artificial bilayers with a robustness and physicochemical complexity that has not previously been possible, facilitating more sophisticated in vitro studies of membrane proteins. This mini-review examines functional studies of membrane proteins that utilize droplet interface bilayers to date and comments on possible directions of future research. Observations from our own laboratory regarding the study of a flippase protein in droplet interface bilayers are also presented. Impact statement The paper presents a comprehensive review of integral membrane protein studies utilizing droplet interface bilayers. Droplet interface bilayers are a novel method of constructing artificial lipid bilayers with enhanced stability and physicochemical complexity compared to existing methods. Their unique morphology also suggests applications in the construction of synthetic biological systems and protocells. As well as serving as a guide to in vitro membrane protein functional studies using droplet interface bilayers in the literature to date, a novel in vitro study of a flippase protein in a droplet interface bilayer is presented.


2021 ◽  
Vol 9 (5) ◽  
pp. 966
Author(s):  
Barbara Rossi ◽  
Maria Angéles Esteban ◽  
José María García-Beltran ◽  
Giulia Giovagnoni ◽  
Alberto Cuesta ◽  
...  

Vibrosis is one of the major threats in aquaculture farming, and due to the increasing antimicrobial resistance of different Vibrio species, there is an urgent need to replace conventional treatments with more sustainable solutions. Antimicrobial molecules such as organic acids (OA) and nature-identical compounds (NIC) are currently finding a central role in the infection management of terrestrial livestock, but little is known about their usage in aquaculture. The aim of this study was to perform a preliminary screening of the in vitro antimicrobial activity of a wide panel of OA and NIC against 2 Vibrio species characteristic of the Mediterranean area, Vibrio harveyi and Vibrio anguillarum, through minimal inhibitory/bactericidal concentration tests. The active principles that showed the best effective pathogen control were medium-chain fatty acids, sorbic and benzoic acid among OA and phenolic monoterpenoids (thymol, carvacrol and eugenol) and aromatic monoterpene aldehydes (vanillin and cinnamaldehyde) among NIC. These results showed how the usage of OA and NIC can open promising perspectives in terms of Vibrio spp. load control in aquaculture. Natural or nature-identical feed additives can make aquaculture production not only more efficient by reducing the need for medicated treatments, but also more sustainable.


Author(s):  
Pranjali P. Dhawal ◽  
Siddhivinayak S. Barve ◽  
Dyotona Sen-Roy

Background: Hyper-acidity is excessive formation of acid (pH=1.5-3.5) in the stomach by parietal cells which causes a burning sensation in the chest. The preservation of gastric acid insult is crucial because of the implications of hyperacidity in gastroesophageal reflux disease (GERD), peptic ulcers and duodenal ulcers. Acidity is controlled by use of some over-the-counter (OTC) antacid formulations containing magnesium or aluminum hydroxides.Methods: In the present study, the preliminary antacid test (PAT), the pH acid neutralizing capacity (ANC), acid neutralizing potential (ANP) along with buffering capacity of two well-known quick release formulations (F1 [Digene Ultra Fizz] and F2 [a standard, commercially available product]) were determined. Results: According to US pharmacopeia USP, both the antacid formulations passed the PAT test. PAT results revealed that the pH of the acid-antacid solution was higher in F1 (8.20±0.02) as compared to F2, (6.53±0.01). The ANC results revealed that F1 (46.89±0.6 mEq/dosage) had higher neutralizing capacity as compared to F2(30.12±1.3 mEq/dosage). Higher ANP was observed for F1 (245 mins), and it was 2.7 times that of F2 (90 min). The onset of action for both the antacids was <2 seconds. Additionally, buffering capacity was evidently observed during ANP analysis in the case of F1. Independent T test performed for all the tests revealed that the data obtained was highly significant (p<0.01).Conclusions: F1 showed high antacid and buffering properties when tested in vitro. The present study highlights the need for future research on specific OTC non-prescribed antacid formulations with respect to their price, efficacy and side effects.


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