scholarly journals Pinocembrin-7-Glucoside (P7G) Reduced Postharvest Blue Mold of Navel Orange by Suppressing Penicillium italicum Growth

2020 ◽  
Vol 8 (4) ◽  
pp. 536
Author(s):  
Chuying Chen ◽  
Jinyin Chen ◽  
Chunpeng Wan
Keyword(s):  
Electron Microscopy ◽  
Cell Membrane ◽  
Mycelial Growth ◽  
Cell Membrane Permeability ◽  
Wall Structure ◽  
Navel Orange ◽  
Penicillium Italicum ◽  
Blue Mold

The current study aimed to examine the in vitro and in vivo antifungal potential of pinocembrin-7-glucoside (P7G). P7G is an antifungal flavanone glycoside isolated from Ficus hirta Vahl. fruit against Penicillium italicum, a causative pathogen of blue mold disease in citrus fruit, and this study elucidates its possible action mechanism. P7G had a prominent mycelial growth inhibitory activity against P. italicum, with an observed half maximal effective concentration, minimum inhibitory concentration and minimum fungicidal concentration of 0.08, 0.2, and 0.8 g/L, respectively. The data from the in vivo test show that P7G significantly reduced blue mold symptoms and disease development of P. italicum in artificially inoculated “Newhall” navel orange. Compared to the control, increases in the cell membrane permeability of P. italicum supernatant and decreases in the intracellular constituent (e.g., soluble protein, reducing sugar, and total lipid) contents of P. italicum mycelia were identified, supporting scanning electron microscopy and transmission electron microscopy observations. Furthermore, a marked decline in both chitin and glucanase contents of P. italicum mycelia treated with P7G was induced by increasing its related degrading enzyme activities, suggesting that the cell wall structure was destroyed. The current study indicated that P7G may be a novel alternative for reducing blue mold by suppressing mycelial growth of P. italicum via a cell membrane/wall-targeting mechanism.

Ultrasound Mediated Enhancement of Nanoparticle Uptake in PC-3 Cancer Cells

2013 ◽  
Author(s):  
Antony Thomas ◽  
Paige Baldwin ◽  
Yaling Liu
Keyword(s):  
Drug Delivery ◽  
Cell Membrane ◽  
Contrast Agents ◽  
Cellular Uptake ◽  
Cancer Detection ◽  
Inert Gas ◽  
Cell Membrane Permeability ◽  
In Vivo Gene Delivery

Ultrasound in the presence of microbubbles brings in transient increase in cell membrane permeability, which allows the entry of foreign molecules into cells. This platform has been applied in in vitro and in vivo gene delivery studies in recent years[1–2]. The frequently used microbubbles are air or inert gas encapsulated in a protein, lipid or polymer which is commonly used as FDA approved contrast agents in diagnostic ultrasound. On exposure to ultrasound the microbubbles lead to formation of small pores on the cell membrane. This work concentrates on application of this platform to enhance cellular uptake of nanoparticles and thereby achieve enhanced drug delivery. Nanoparticles can be manipulated at the nano level and have been applied in the realm of cancer detection and treatment for imaging, targeting tumors, and drug delivery purposes [2].

scholarly journals Glyceraldehyde-3-Phosphate Dehydrogenase Increases the Adhesion of Lactobacillus reuteri to Host Mucin to Enhance Probiotic Effects

2020 ◽  
Vol 21 (24) ◽  
pp. 9756
Author(s):  
Zhaoxi Deng ◽  
Tian Dai ◽  
Wenming Zhang ◽  
Junli Zhu ◽  
Xin M. Luo ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Membrane Permeability ◽  
Bacterial Cell ◽  
Lactobacillus Reuteri ◽  
Surface Proteins ◽  
Cell Membrane Permeability ◽  
In Vivo Studies ◽  
Bacterial Surface

The ability to adhere to the intestinal mucus layer is an important property of probiotic bacteria. Lactobacillus reuteri strains ZJ615 and ZJ617 show low and high adhesion, respectively, to intestinal epithelial cells. In this study, we quantified bacterial cell wall-associated glyceraldehyde-3-phosphate dehydrogenases (cw-GAPDH) and bacterial cell membrane permeability in both strains using immunoblotting and flow cytometry, respectively. Highly adhesive L. reuteri ZJ617 possessed significantly more cw-GAPDH, higher cell membrane permeability, and significantly higher adhesive ability toward mucin compared with low-adhesive L. reuteri ZJ615. In vitro adhesion studies and analysis of interaction kinetics using the Octet, the system revealed significantly decreased interaction between L. reuteri and mucin when mucin was oxidized when bacterial surface proteins were removed when bacteria were heat-inactivated at 80 °C for 30 min, and when the interaction was blocked with an anti-GAPDH antibody. SWISS-MODEL analysis suggested intensive interactions between mucin glycans (GalNAcα1-O-Ser, GalNAcαSer, and Galβ3GalNAc) and GAPDH. Furthermore, in vivo studies revealed significantly higher numbers of bacteria adhering to the jejunum, ileum, and colon of piglets orally inoculated with L. reuteri ZJ617 compared with those inoculated with L. reuteri ZJ615; this led to a significantly decreased rate of diarrhea in piglets inoculated with L. reuteri ZJ617. In conclusion, there are strong correlations among the abundance of cw-GAPDH in L. reuteri, the ability of the bacterium to adhere to the host, and the health benefits of this probiotic.

scholarly journals Limonin Enhances the Antifungal Activity of Eugenol Nanoemulsion against Penicillium Italicum In Vitro and In Vivo Tests

2021 ◽  
Vol 9 (5) ◽  
pp. 969
Author(s):  
Yi Li ◽  
Runan Zhao ◽  
Yan Li ◽  
Zhiqin Zhou
Keyword(s):  
Antifungal Activity ◽  
Pathogenic Fungus ◽  
Germination Rate ◽  
Challenge Test ◽  
Postharvest Quality ◽  
Citrus Fruits ◽  
Penicillium Italicum ◽  
Blue Mold

Penicillium italicum, the cause of citrus blue mold, is a pathogenic fungus that seriously affects the postharvest quality of citrus fruit and causes serious economic loss. In this study, a eugenol nanoemulsion containing limonin, an antimicrobial component from citrus seeds, was prepared using a high-pressure microfluidizer and the antifungal activity of the nanoemulsions against P. italicum was evaluated based on the conidial germination rate, mycelial growth, and scanning electron microscopy analysis. The results showed that the minimum inhibitory concentration and the inhibition rate of limonin-loaded eugenol nanoemulsion was 160 μg/mL and 59.21%, respectively, which was more potent than that of the limonin-free eugenol emulsion. After treatment with the nanoemulsions, the integrity of the P. italicum cell membrane was disrupted, the cell morphology was abnormal, and the leakage of nucleic acid and protein was observed. In addition, the challenge test on citrus fruits revealed that the limonin-loaded eugenol emulsion inhibited citrus infection for longer periods, with an infection rate of 29.2% after 5 days. The current research shows that nanoemulsions containing limonin and eugenol have effective antifungal activity against P. italicum, and may be used as a substitute for inhibiting blue mold in citrus fruits.

scholarly journals Use of Food Additives to Control Postharvest Citrus Blue Mold Disease

2017 ◽  
Vol 2 (3) ◽  
pp. 147-153 ◽  
Author(s):  
Askarne L. ◽  
H. Boubaker ◽  
E. H. Boudyach ◽  
A. Ait Ben Aoumar
Keyword(s):  
Boric Acid ◽  
Food Additives ◽  
Citrus Fruit ◽  
Colony Growth ◽  
Sodium Metabisulfite ◽  
Penicillium Italicum ◽  
Postharvest Diseases ◽  
Blue Mold

The aim of this study was to find an alternative to the chemical fungicide currently used in the control of postharvest citrus diseases. The antifungal activity of 10 salt compounds, considered as common food additives was assayed in in vitro and in vivo trials against Penicillium italicum, causal agent of citrus blue mold. Among the 10 tested salt compounds, sodium carbonate, ammonium carbonate, copper sulfate, sodium EDTA and sodium metabisulfite completely inhibited mycelial growth of Penicillium italicum at 20 mM. Colony growth of P. italicum on pH adjusted medium was evaluated. Results indicate that P. italicum can grow on both acidic and alkaline pH, with the optimum growth occurred in the range of 4.0 and 8.0. Results of the in vivo trials with tested salt compounds indicate that sodium metabisulfite (100 and 200 mM), boric acid (400 mM), sodium salicylate and sodium sulfite (200, 300 and 400 mM) completely inhibited blue mold development on citrus fruit. Boric acid (400 mM) and sodium metabisulfite (100 mM) gave the best results as they completely inhibited the fungus development without damaging fruit rind. Such healthy products therefore may represent a sustainable alternative to the use of chemical fungicides for controlling postharvest diseases of citrus fruit.

An Attempt at Biological Control of Blossom Blight of Rose Caused by Botrytis cinerea Using some Local Trichoderma spp. Strains

2020 ◽  
Vol 55 (1) ◽  
pp. 27-34
Author(s):  
G. Zadehdabagh ◽  
K. Karimi ◽  
M. Rezabaigi ◽  
F. Ajamgard
Keyword(s):  
Botrytis Cinerea ◽  
Mycelial Growth ◽  
Limiting Factor ◽  
Dual Culture ◽  
Trichoderma Spp ◽  
Khuzestan Province ◽  
Inhibitory Potential ◽  
Cut Rose

The northern of Khuzestan province in Iran is mainly considered as one of the major areas of miniature rose production. Blossom blight caused by Botrytis cinerea has recently become a serious limiting factor in rose production in pre and post-harvest. In current study, an attempt was made to evaluate the inhibitory potential of some local Trichoderma spp. strains against B. cinerea under in vitro and in vivo conditions. The in vitro results showed that all Trichoderma spp. strains were significantly able to reduce the mycelial growth of the pathogen in dual culture, volatile and non-volatile compounds tests compared with control, with superiority of T. atroviride Tsafi than others. Under in vivo condition, the selected strain of T. atroviride Tsafi had much better performance than T. harzianum IRAN 523C in reduction of disease severity compared with the untreated control. Overall, the findings of this study showed that the application of Trichoderma-based biocontrol agents such as T. atroviride Tsafi can be effective to protect cut rose flowers against blossom blight.

Light Chain LC and TAT-EGFP-HCS of Botulinum Toxin Expression and Biological Function in vitro and in vivo

2019 ◽  
Vol 16 (3) ◽  
pp. 175-180
Author(s):  
Fengjin Hao ◽  
Yueqin Feng ◽  
Yifu Guan
Keyword(s):  
Biological Activity ◽  
Botulinum Toxin ◽  
Cell Membrane ◽  
Western Blot ◽  
Biological Function ◽  
C6 Cells ◽  
Green Fluorescence ◽  
Bv2 Cells

Objective: To verify whether the botulinum toxin heavy chain HCS has specific neuronal targeting function and to confirm whether TAT-EGFP-LC has hydrolyzable SNAP-25 and has transmembrane biological activity. Methods: We constructed the pET-28a-TAT-EGFP-HCS/LC plasmid. After the plasmid is expressed and purified, we co-cultured it with nerve cells or tumors. In addition, we used Western-Blot to identify whether protein LC and TAT-EGFP-LC can digest the protein SNAP-25. Results: Fluorescence imaging showed that PC12, BV2, C6 and HeLa cells all showed green fluorescence, and TAT-EGFP-HCS had the strongest fluorescence. Moreover, TAT-EGFP-LC can hydrolyze intracellular SNAP-25 in PC12 cells, C6 cells, BV2 cells and HeLa, whereas LC alone cannot. In addition, the in vivo protein TAT-EGFP-HCS can penetrate the blood-brain barrier and enter mouse brain tissue. Conclusion: TAT-EGFP-HSC expressed in vitro has neural guidance function and can carry large proteins across the cell membrane without influencing the biological activity.

scholarly journals Improving the Biocontrol Potential of Bacterial Antagonists with Salicylic Acid against Brown Rot Disease and Impact on Nectarine Fruits Quality

Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 209
Author(s):  
Nadia Lyousfi ◽  
Rachid Lahlali ◽  
Chaimaa Letrib ◽  
Zineb Belabess ◽  
Rachida Ouaabou ◽  
...  
Keyword(s):  
Salicylic Acid ◽  
Disease Severity ◽  
Mycelial Growth ◽  
Brown Rot ◽  
Antagonistic Bacteria ◽  
Biological Treatments ◽  
Rot Disease ◽  
The Impact

The main objective of this study was to evaluate the ability of both antagonistic bacteria Bacillus amyloliquefaciens (SF14) and Alcaligenes faecalis (ACBC1) used in combination with salicylic acid (SA) to effectively control brown rot disease caused by Monilinia fructigena. Four concentrations of salicylic acid (0.5%, 2%, 3.5%, and 5%) were tested under in vitro and in vivo conditions. Furthermore, the impact of biological treatments on nectarine fruit parameters’ quality, in particular, weight loss, titratable acidity, and soluble solids content, was evaluated. Regardless of the bacterium, the results indicated that all combined treatments displayed a strong inhibitory effect on the mycelial growth of M. fructigena and disease severity. Interestingly, all SA concentrations significantly improved the biocontrol activity of each antagonist. The mycelial growth inhibition rate ranged from 9.79% to 88.02% with the highest reduction rate recorded for bacterial antagonists in combination with SA at both concentrations of 0.5% and 3.5%. The in vivo results confirmed the in vitro results with a disease severity varying from 0.00% to 51.91%. A significant biocontrol improvement was obtained with both antagonistic bacteria when used in combination with SA at concentrations of 0.5% and 2%. The lowest disease severity observed with ACBC1 compared with SF14 is likely due to a rapid adaptation and increase of antagonistic bacteria population in wounded sites. The impact of all biological treatments revealed moderate significant changes in the fruit quality parameters with weight loss for several treatments. These results suggest that the improved disease control of both antagonistic bacteria was more likely directly linked to both the inhibitory effects of SA on pathogen growth and induced fruit resistance.

scholarly journals Assessment of SnFe2O4 Nanoparticles for Potential Application in Theranostics: Synthesis, Characterization, In Vitro, and In Vivo Toxicity

Materials ◽  
2021 ◽  
Vol 14 (4) ◽  
pp. 825
Author(s):  
Saman Sargazi ◽  
Mohammad Reza Hajinezhad ◽  
Abbas Rahdar ◽  
Muhammad Nadeem Zafar ◽  
Aneesa Awan ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
A549 Cells ◽  
In Vitro Cytotoxicity ◽  
Hek293 Cells ◽  
Hydrothermal Route ◽  
X Ray ◽  
Tin Chloride ◽  
Bet Analysis

In this research, tin ferrite (SnFe2O4) NPs were synthesized via hydrothermal route using ferric chloride and tin chloride as precursors and were then characterized in terms of morphology and structure using Fourier-transform infrared spectroscopy (FTIR), Ultraviolet–visible spectroscopy (UV-Vis), X-ray power diffraction (XRD), Scanning electron microscopy (SEM), Transmission electron microscopy (TEM), and Brunauer–Emmett–Teller (BET) method. The obtained UV-Vis spectra was used to measure band gap energy of as-prepared SnFe2O4 NPs. XRD confirmed the spinel structure of NPs, while SEM and TEM analyses disclosed the size of NPs in the range of 15–50 nm and revealed the spherical shape of NPs. Moreover, energy dispersive X-ray spectroscopy (EDS) and BET analysis was carried out to estimate elemental composition and specific surface area, respectively. In vitro cytotoxicity of the synthesized NPs were studied on normal (HUVEC, HEK293) and cancerous (A549) human cell lines. HUVEC cells were resistant to SnFe2O4 NPs; while a significant decrease in the viability of HEK293 cells was observed when treated with higher concentrations of SnFe2O4 NPs. Furthermore, SnFe2O4 NPs induced dramatic cytotoxicity against A549 cells. For in vivo study, rats received SnFe2O4 NPs at dosages of 0, 0.1, 1, and 10 mg/kg. The 10 mg/kg dose increased serum blood urea nitrogen and creatinine compared to the controls (P < 0.05). The pathology showed necrosis in the liver, heart, and lungs, and the greatest damages were related to the kidneys. Overall, the in vivo and in vitro experiments showed that SnFe2O4 NPs at high doses had toxic effects on lung, liver and kidney cells without inducing toxicity to HUVECs. Further studies are warranted to fully elucidate the side effects of SnFe2O4 NPs for their application in theranostics.

scholarly journals Cemp1-p3 Peptide Promotes the Transformation of Octacalcium Phosphate into Hydroxyapatite Crystals

Crystals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1131
Author(s):  
Maricela Santana ◽  
Gonzalo Montoya ◽  
Raúl Herrera ◽  
Lía Hoz ◽  
Enrique Romo ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Octacalcium Phosphate ◽  
Sequence Motifs ◽  
Simple Method ◽  
Transmission Electron ◽  
Precursor Phase ◽  
Mineralized Tissues ◽  
Potent Growth

Dental cementum contains unique molecules that regulate the mineralization process in vitro and in vivo, such as cementum protein 1 (CEMP1). This protein possesses amino acid sequence motifs like the human recombinant CEMP1 with biological activity. This novel cementum protein 1-derived peptide (CEMP1-p3, from the CEMP1’s N-terminal domain: (QPLPKGCAAVKAEVGIPAPH), consists of 20 amino acids. Hydroxyapatite (HA) crystals could be obtained through the combination of the amorphous precursor phase and macromolecules such as proteins and peptides. We used a simple method to synthesize peptide/hydroxyapatite nanocomposites using OCP and CEMP1-p3. The characterization of the crystals through scanning electron microscopy (SEM), powder X-ray diffraction (XRD), high--resolution transmission electron microscopy (HRTEM), and Raman spectroscopy revealed that CEMP1-p3 transformed OCP into hydroxyapatite (HA) under constant ionic strength and in a buffered solution. CEMP1-p3 binds and highly adsorbs to OCP and is a potent growth stimulator of OCP crystals. CEMP1-p3 fosters the transformation of OCP into HA crystals with crystalline planes (300) and (004) that correspond to the cell of hexagonal HA. Octacalcium phosphate crystals treated with CEMP1-p3 grown in simulated physiological buffer acquired hexagonal arrangement corresponding to HA. These findings provide new insights into the potential application of CEMP1-p3 on possible biomimetic approaches to generate materials for the repair and regeneration of mineralized tissues, or restorative materials in the orthopedic field.

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