scholarly journals Encapsulation of Freeze-dried Propolis Powder: Study of in vitro Disintegration and Dissolution

2021 ◽  
Vol 55 (2) ◽  
pp. 428-435
Author(s):  
Wan Nurul Huda Wan Zainal ◽  
Enidran Loganathan ◽  
Thavakumara Ganesan
Keyword(s):  
Freeze Dried ◽  
In Vitro Disintegration

Preparation of cultured astrocytes for x-ray microanalysis

1989 ◽  
Vol 47 ◽  
pp. 988-989
Author(s):  
N.K.R. Smith ◽  
K.E. Hunter ◽  
P. Mobley ◽  
L.P. Felpel
Keyword(s):  
Cultured Cells ◽  
Elemental Content ◽  
Elemental Distribution ◽  
Sprague Dawley ◽  
X Ray ◽  
Cultured Astrocytes ◽  
Freeze Dried ◽  
Ultimate Objective

Electron probe energy dispersive x-ray microanalysis (XRMA) offers a powerful tool for the determination of intracellular elemental content of biological tissue. However, preparation of the tissue specimen , particularly excitable central nervous system (CNS) tissue , for XRMA is rather difficult, as dissection of a sample from the intact organism frequently results in artefacts in elemental distribution. To circumvent the problems inherent in the in vivo preparation, we turned to an in vitro preparation of astrocytes grown in tissue culture. However, preparations of in vitro samples offer a new and unique set of problems. Generally, cultured cells, growing in monolayer, must be harvested by either mechanical or enzymatic procedures, resulting in variable degrees of damage to the cells and compromised intracel1ular elemental distribution. The ultimate objective is to process and analyze unperturbed cells. With the objective of sparing others from some of the same efforts, we are reporting the considerable difficulties we have encountered in attempting to prepare astrocytes for XRMA.Tissue cultures of astrocytes from newborn C57 mice or Sprague Dawley rats were prepared and cultured by standard techniques, usually in T25 flasks, except as noted differently on Cytodex beads or on gelatin. After different preparative procedures, all samples were frozen on brass pins in liquid propane, stored in liquid nitrogen, cryosectioned (0.1 μm), freeze dried, and microanalyzed as previously reported.

Freeze-Dried Clopidogrel Loaded Lyotropic Liquid Crystal: Box-Behnken Optimization, In-Vitro and In-Vivo Evaluation

2020 ◽  
Vol 17 (3) ◽  
pp. 207-217
Author(s):  
Eman A. Hakeem ◽  
Galal M. El-Mahrouk ◽  
Ghada Abdelbary ◽  
Mahmoud H. Teaima
Keyword(s):  
Statistical Analysis ◽  
Oral Bioavailability ◽  
Quadratic Model ◽  
Lipid Phase ◽  
Individual Response ◽  
In Vivo Evaluation ◽  
Freeze Dried ◽  
Suggested Formula

Background: Clopidogrel (CLP) suffers from extensive first pass metabolism results in a negative impact on its oral systemic bioavailability. Cubosomes are Lyotropic Liquid Crystalline (LLC) nano-systems comprising monoolein, a steric stabilizer and an aqueous system, it considered a promising carrier for different pharmaceutical compounds. Box-Behnken Design (BBD) is an efficient tool for process analysis and optimization skipping forceful treatment combinations. Objective: The study was designed to develop freeze-dried clopidogrel loaded LLC (cubosomes) for enhancement of its oral bioavailability. Methods: A 33 BBD was adopted, the studied independent factors were glyceryl monooleate (GMO lipid phase), Pluronic F127 (PL F127steric stabilizer) and polyvinyl alcohol powder (stabilizer). Particle Size (PS), Polydispersity Index (PDI) and Zeta Potential (ZP) were set as independent response variables. Seventeen formulae were prepared in accordance with the bottom up approach and in-vitro evaluated regarding PS, PDI and ZP. Statistical analysis and optimization were achieved using design expert software®, then the optimum suggested formula was prepared, in-vitro revaluated, freeze-dried with 3% mannitol (cryoprotectant), solid state characterized and finally packed in hard gelatin capsule for comparative in-vitro release and in-vivo evaluation to Plavix®. Results: Results of statistical analysis of each individual response revealed a quadratic model for PS and PDI where a linear model for ZP. The optimum suggested formula with desirability factor equal 0.990 consisting of (200 mg GMO, 78.15 mg PL F127 and 2% PVA). LC/MS/MS study confirmed significant higher C>max, AUC>0-24h and AUC>0-∞ than that of Plavix®. Conclusion: The results confirm the capability of developed carrier to overcome the low oral bioavailability.

scholarly journals A High-Throughput Metabolic Microarray Assay Reveals Antibacterial Effects of Black and Red Raspberries and Blackberries against Helicobacter pylori Infection

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 845
Author(s):  
Candace Goodman ◽  
Katrina N. Lyon ◽  
Aitana Scotto ◽  
Cyra Smith ◽  
Thomas A. Sebrell ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
High Throughput ◽  
Antibacterial Properties ◽  
Treatment Strategies ◽  
Helicobacter Pylori Infection ◽  
Antibacterial Effects ◽  
Biolog System ◽  
Freeze Dried ◽  
H Pylori

Helicobacter pylori infection is commonly treated with a combination of antibiotics and proton pump inhibitors. However, since H. pylori is becoming increasingly resistant to standard antibiotic regimens, novel treatment strategies are needed. Previous studies have demonstrated that black and red berries may have antibacterial properties. Therefore, we analyzed the antibacterial effects of black and red raspberries and blackberries on H. pylori. Freeze-dried powders and organic extracts from black and red raspberries and blackberries were prepared, and high-performance liquid chromatography was used to measure the concentrations of anthocyanins, which are considered the major active ingredients. To monitor antibiotic effects of the berry preparations on H. pylori, a high-throughput metabolic growth assay based on the Biolog system was developed and validated with the antibiotic metronidazole. Biocompatibility was analyzed using human gastric organoids. All berry preparations tested had significant bactericidal effects in vitro, with MIC90 values ranging from 0.49 to 4.17%. Antimicrobial activity was higher for extracts than powders and appeared to be independent of the anthocyanin concentration. Importantly, human gastric epithelial cell viability was not negatively impacted by black raspberry extract applied at the concentration required for complete bacterial growth inhibition. Our data suggest that black and red raspberry and blackberry extracts may have potential applications in the treatment and prevention of H. pylori infection but differ widely in their MICs. Moreover, we demonstrate that the Biolog metabolic assay is suitable for high-throughput antimicrobial susceptibility screening of H. pylori.

scholarly journals Mutual influence of selenium nanoparticles and FGF2-STAB® on biocompatible properties of collagen/chitosan 3D scaffolds: in vitro and ex ovo evaluation

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Johana Muchová ◽  
Vanessa Hearnden ◽  
Lenka Michlovská ◽  
Lucie Vištejnová ◽  
Anna Zavaďáková ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Metabolic Activity ◽  
Mutual Influence ◽  
Chorioallantoic Membrane ◽  
Antagonistic Effect ◽  
Dermal Fibroblasts ◽  
Selenium Nanoparticles ◽  
Freeze Dried ◽  
Chitosan Scaffolds

AbstractIn a biological system, nanoparticles (NPs) may interact with biomolecules. Specifically, the adsorption of proteins on the nanoparticle surface may influence both the nanoparticles' and proteins' overall bio-reactivity. Nevertheless, our knowledge of the biocompatibility and risk of exposure to nanomaterials is limited. Here, in vitro and ex ovo biocompatibility of naturally based crosslinked freeze-dried 3D porous collagen/chitosan scaffolds, modified with thermostable fibroblast growth factor 2 (FGF2-STAB®), to enhance healing and selenium nanoparticles (SeNPs) to provide antibacterial activity, were evaluated. Biocompatibility and cytotoxicity were tested in vitro using normal human dermal fibroblasts (NHDF) with scaffolds and SeNPs and FGF2-STAB® solutions. Metabolic activity assays indicated an antagonistic effect of SeNPs and FGF2-STAB® at high concentrations of SeNPs. The half-maximal inhibitory concentration (IC50) of SeNPs for NHDF was 18.9 µg/ml and IC80 was 5.6 µg/ml. The angiogenic properties of the scaffolds were monitored ex ovo using a chick chorioallantoic membrane (CAM) assay and the cytotoxicity of SeNPs over IC80 value was confirmed. Furthermore, the positive effect of FGF2-STAB® at very low concentrations (0.01 µg/ml) on NHDF metabolic activity was observed. Based on detailed in vitro testing, the optimal concentrations of additives in the scaffolds were determined, specifically 1 µg/ml of FGF2-STAB® and 1 µg/ml of SeNPs. The scaffolds were further subjected to antimicrobial tests, where an increase in selenium concentration in the collagen/chitosan scaffolds increased the antibacterial activity. This work highlights the antimicrobial ability and biocompatibility of newly developed crosslinked collagen/chitosan scaffolds involving FGF2-STAB® and SeNPs. Moreover, we suggest that these sponges could be used as scaffolds for growing cells in systems with low mechanical loading in tissue engineering, especially in dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration. Due to their antimicrobial properties, these scaffolds are also highly promising for tissue replacement requiring the prevention of infection.

scholarly journals 127 Porcine in vitro digestibility of whole stillage predigested with multi-enzyme during different time periods

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 49-50
Author(s):  
Kevin S Jerez Bogota ◽  
Tofuko A Woyengo
Keyword(s):  
Dry Matter ◽  
In Vitro Digestion ◽  
Enzyme Treatment ◽  
Optimal Time ◽  
In Vitro Digestibility ◽  
Percentage Points ◽  
Whole Stillage ◽  
Freeze Dried ◽  
Different Sources

Abstract A study was conducted to determine the effects of the period of predigesting whole stillage (WS; slurry material that is dried into DDGS) with multi-enzyme and composition of the multi-enzyme on porcine in vitro digestibility of dry matter (IVDDM) of the WS. Four samples of whole stillage from 4 different sources were freeze-dried and divided into 13 subsamples to give 52 sub-samples. Thirteen treatments were applied to the 48 sub-samples within source. The treatments were undigested WS (control); or pre-digested with 1 of 3 multi-enzymes (MTE1, MTE2, and MTE3) at 55 °C for 6, 12, 18 or 24 h in 3 × 4 factorial arrangement. The MTE1 contained xylanase, β-glucanase, cellulase, mannanase, protease, and amylase; MTE2 contained xylanase, α-galactosidase, and cellulase; and MTE3 contained xylanase, cellulase, β-glucanase, and mannanase. The 52 subsamples were subjected to porcine in vitro digestion. The IVDDM of untreated WS was 73.3%. The IVDDM increased (P< 0.05) with an increase in the predigestion period. However, a rise in the predigestion period from 0 to 12 h resulted in greater (P< 0.05) response in mean IVDDM than an increment in the predigestion period from 12 to 24 h (11 vs. 0.83 percentage points). Predigestion period and multi-enzyme type interacted on IVDDM such that the improvement in IVDDM between 0 and 12 hours of predigestion differed (P< 0.05) among the 3 multi-enzyme types (13.3, 11.1, and 8.5 percentage points for MTE3, MTE2, and MTE1, respectively). The LS means by multi-enzyme treatment were modeled and resulted in unparallel curves (P< 0.05). The estimated maximum response of IVDDM for MTE1, MTE2 and MTE 3 were 82.4%, 84.7% and 87.1% at 15.8, 13 and 13.1 hours, respectively. In conclusion, the optimal time of predigestion of WS with multi-enzymes (with regard to improvement in its IVDDM) was approximately 14 h.

Effects of chelating agents during freeze-drying of boar spermatozoa on DNA fragmentation and on developmental ability in vitro and in vivo after intracytoplasmic sperm head injection

Zygote ◽  
2007 ◽  
Vol 15 (1) ◽  
pp. 15-24 ◽  
Author(s):  
M. Nakai ◽  
N. Kashiwazaki ◽  
A. Takizawa ◽  
N. Maedomari ◽  
M. Ozawa ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Freeze Drying ◽  
Chelating Agents ◽  
Sperm Head ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Freeze Dried ◽  
Boar Sperm

SUMMARYSuccessful offspring production after intracytoplasmic injection of freeze-dried sperm has been reported in laboratory animals but not in domesticated livestock, including pigs. The integrity of the DNA in the freeze-dried sperm is reported to affect embryogenesis. Release of endonucleases from the sperm is one of the causes of induction of sperm DNA fragmentation. We examined the effects of chelating agents, which inhibit the activation of such enzymes, on DNA fragmentation in freeze-dried sperm and on the in vitro and in vivo developmental ability of porcine oocytes following boar sperm head injection. Boar ejaculated sperm were sonicated, suspended in buffer supplemented with (1) 50 mM EGTA, (2) 50 mM EDTA, (3) 10 mM EDTA, or (4) no chelating agent and freeze-dried. A fertilization medium (Pig-FM) was used as a control. The rehydrated spermatozoa in each group were then incubated in Pig-FM at room temperature. The rate of DNA fragmentation in the control group, as assessed by the TUNEL method, increased gradually as time after rehydration elapsed (2.8% at 0 min to 12.2% at 180 min). However, the rates in all experimental groups (1–4) did not increase, even at 180 min (0.7–4.1%), which were all significantly lower (p < 0.05) than that of the control group. The rate of blastocyst formation after the injection in the control group (6.0%) was significantly lower (p < 0.05) than those in the 50 mM EGTA (23.1%) and 10 mM EDTA (22.6%) groups incubated for 120–180 min. The average number of blastocyst cells in the 50 mM EGTA group (33.1 cells) was significantly higher (p < 0.05) than that in the 10 mM EDTA group (17.8 cells). Finally, we transferred oocytes from 50 mM EGTA or control groups incubated for 0–60 min into estrous-synchronized recipients. The two recipients of the control oocytes became pregnant and one miscarried two fetuses on day 39.The results suggested that fragmentation of DNA in freeze-dried boar sperm is one of the causes of decreased in vitro developmental ability of injected oocytes to the blastocyst stage. Supplementation with EGTA in a freeze-drying buffer improves this ability.

PSXIII-2 Effect of varying proportions of cereal rye and turnip on ruminal fermentation and methane output through in vitro batch culture

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 461-461
Author(s):  
Jordan L Cox-O’Neill ◽  
Vivek Fellner ◽  
Alan J Franluebbers ◽  
Deidre D Harmon ◽  
Matt H Poore ◽  
...  
Keyword(s):  
Methane Production ◽  
Dry Matter ◽  
Ruminal Fermentation ◽  
Variable Response ◽  
Cool Season ◽  
Cereal Rye ◽  
Freeze Dried ◽  
Main Effect ◽  
Cool Season Grass

Abstract Ruminant animal performance has been variable in studies grazing annual cool-season grass and brassica monocultures and mixtures. There is little understanding of the fermentation mechanisms causing variation. The aim of this study was to determine apparent dry matter (DM) digestibility, methane, and volatile fatty acid (VFA) concentration from different proportions of cereal rye (Secale cereal; R) and turnip (Brassica rapa L.; T) (0R:100T, 40R:60T, 60R:40T, and 100R:0T) via in vitro batch fermentation. Freeze-dried forage samples from an integrated crop-livestock study was assembled into the four treatments with a 50:50 leaf to root ratio for turnip. Measurements were made following a 48 hr fermentation with 2:1 buffer and ruminal fluid inoculum. Data were analyzed using Mixed Procedure of SAS with batch (replicate) and treatment (main effect) in the model; differences were declared at P ≤ 0.05, with tendencies declared at &gt; 0.05 but &lt; 0.10. Rumen apparent DM digestibility (26.8%; overall mean) was not different among treatments. Methane production was less (P &lt; 0.01) with inclusion of turnip ranging from 774 nmol/ml for 0R:100T to 1416 nmol/ml for 100R:0T. Total VFA production, acetate to propionate ratio, acetate, and valerate were not affected by forage treatments (117 mM, 1.45, 39.84 mol/100 mol, and 7.86 mol/100 mol, respectively; overall mean). Propionate, isobutyrate, and isovalerate concentrations were greater and butyrate concentration less with greater (P &lt; 0.01) proportions of rye in the mixture. No effect of R:T ratio on digestibility or total VFA production along with the observed differences in individual VFA concentration do not explain variable response in grazing animals. Additionally, methane production results indicate that grazing turnips could potentially reduce methane production and thus reduce ruminant livestock’s contribution to greenhouse gas emissions.

scholarly journals Citrus limon Extract Loaded in Vesicular Systems for the Protection of Oral Cavity

Medicines ◽  
2018 ◽  
Vol 5 (4) ◽  
pp. 108 ◽  
Author(s):  
Maria Manconi ◽  
Maria Manca ◽  
Carla Caddeo ◽  
Giorgia Sarais ◽  
Alessandra Palmieri ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Lactobacillus Acidophilus ◽  
Freeze Drying ◽  
Citrus Limon ◽  
Antibacterial Effects ◽  
Bacterial Proliferation ◽  
Freeze Dried ◽  
Ethanol Evaporation ◽  
Dried Extract

Background: The nanoincorporation of the extract of Citrus limon (L.) Osbeck var. pompia into liposomes was aimed at improving its antioxidant and antibacterial effects. Methods: The extract of the rind of Citrus limon (L.) Osbeck var. pompia was obtained by maceration in ethanol, evaporation, and freeze-drying. The extract phytochemical fingerprint was obtained by HPLC and mass spectrometry, and it was determined that gallic acid, neohesperidin, eriocitrin, and neoeriocitrin were the most abundant components. The freeze-dried extract was loaded in liposomes, glycerosomes, and penetration-enhancer-containing vesicles prepared with propylene glycol (PG-PEVs). Results: Capability of the vesicles of improving efficacy of the extract in counteracting oxidative stress was studied in vitro in keratinocytes, along with antimicrobial activity against planktonic cultures of Streptococcus mutans, Lactobacillus acidophilus, and Streptococcus sanguinis. Conclusion: Results showed that the vesicles, especially glycerosomes and PG-PEVs, prevented oxidative damage and cell death, and inhibited bacterial proliferation.

scholarly journals Selection of phytotoxin producing rhizobacteria

2011 ◽  
Vol 83 (3) ◽  
pp. 1091-1096 ◽  
Author(s):  
Daniel D.C. Carvalho ◽  
Denilson F. Oliveira ◽  
Vicente P. Campos ◽  
Moacir Pasqual
Keyword(s):  
Seed Germination ◽  
Sucrose Solution ◽  
Triticum Aestivum L ◽  
In Vitro Assays ◽  
Lettuce Seed ◽  
Bacterial Strains ◽  
Coleoptile Growth ◽  
Liquid Phases ◽  
Freeze Dried

In order to select phytotoxin producing rhizobacteria to control weed plants, twenty five bacterial strains previously isolated from the rhizospheres of various plants were grown in a liquid medium and, after cell removal by centrifugation, the liquid phases were freeze-dried and the products were extracted with ethyl acetate/methanol. The extracts were concentrated to dryness under vacuum and dissolved in water and sucrose solution to be submitted to in vitro assays of lettuce (Lactuca sativa L.) seed germination and wheat (Triticum aestivum L.) coleoptile growth. Although most samples affected coleoptile growth, only those from four strains reduced lettuce seed germination. Two strains of Bacillus cereus, one strain of B. pumilus and one of Stenotrophoonas altophilia were the most promising microorganisms for producing phytotoxin and, consequently, for the development of new weed control products.

scholarly journals Toxicity Evaluation of Selected Plant Water Extracts on a Honey Bee (Apis mellifera L.) Larvae Model

Animals ◽  
2022 ◽  
Vol 12 (2) ◽  
pp. 178
Author(s):  
Roksana Kruszakin ◽  
Paweł Migdal
Keyword(s):  
Apis Mellifera ◽  
Honey Bee ◽  
Honey Bees ◽  
Larval Rearing ◽  
Distilled Water ◽  
Chelidonium Majus ◽  
Freeze Dried ◽  
Study Laboratory ◽  
The Impact

So far, larval rearing in vitro has been an important method in the assessment of bee toxicology, particularly in pesticide risk assessment. However, natural products are increasingly used to control honey bee pathogens or to enhance bee immunity, but their effects on honey bee larvae are mostly unknown. In this study, laboratory studies were conducted to determine the effects of including selected aqueous plant infusions in the diet of honey bee (Apis mellifera L.) larvae in vitro. The toxicity of infusions from three different plant species considered to be medicinal plants was evaluated: tansy (Tanacetum vulgare L.), greater celandine (Chelidonium majus L.), and coriander (Coriandrum sativum L.). The impact of each on the survival of the larvae of honey bees was also evaluated. One-day-old larvae were fed a basal diet consisting of distilled water, sugars (glucose and fructose), yeast extract, and freeze-dried royal jelly or test diets in which distilled water was replaced by plant infusions. The proportion of the diet components was adjusted to the age of the larvae. The larvae were fed twice a day. The experiment lasted seven days. Significant statistical differences in survival rates were found between groups of larvae (exposed or not to the infusions of tansy, greater celandine, and coriander). A significant decrease (p < 0.05) in the survival rate was observed in the group with the addition of a coriander herb infusion compared to the control. These results indicate that plant extracts intended to be used in beekeeping should be tested on all development stages of honey bees.

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