scholarly journals Gliotoxin Is a Virulence Factor of Aspergillus fumigatus: gliP Deletion Attenuates Virulence in Mice Immunosuppressed with Hydrocortisone

2007 ◽  
Vol 6 (9) ◽  
pp. 1562-1569 ◽  
Author(s):  
Janyce A. Sugui ◽  
Julian Pardo ◽  
Yun C. Chang ◽  
Kol A. Zarember ◽  
Glenn Nardone ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Virulence Factor ◽  
High Performance ◽  
Superoxide Production ◽  
In Vitro Assays ◽  
Mouse Strains ◽  
Human Neutrophils ◽  
Pathogenic Potential ◽  
Chromatography Analysis

ABSTRACT Gliotoxin is an immunosuppressive mycotoxin long suspected to be a potential virulence factor of Aspergillus fumigatus. Recent studies using mutants lacking gliotoxin production, however, suggested that the mycotoxin is not important for pathogenesis of A. fumigatus in neutropenic mice resulting from treatment with cyclophosphomide and hydrocortisone. In this study, we report on the pathobiological role of gliotoxin in two different mouse strains, 129/Sv and BALB/c, that were immunosuppressed by hydrocortisone alone to avoid neutropenia. These strains of mice were infected using the isogenic set of a wild type strain (B-5233) and its mutant strain (gliPΔ) and the the glip reconstituted strain (gliP R). The gliP gene encodes a nonribosomal peptide synthase that catalyzes the first step in gliotoxin biosynthesis. The gliPΔ strain was significantly less virulent than strain B-5233 or gliP R in both mouse models. In vitro assays with culture filtrates (CFs) of B-5233, gliPΔ, and gliP R strains showed the following: (i) deletion of gliP abrogated gliotoxin production, as determined by high-performance liquid chromatography analysis; (ii) unlike the CFs from strains B-5233 and gliP R, gliPΔ CFs failed to induce proapoptotic processes in EL4 thymoma cells, as tested by Bak conformational change, mitochondrial-membrane potential disruption, superoxide production, caspase 3 activation, and phosphatidylserine translocation. Furthermore, superoxide production in human neutrophils was strongly inhibited by CFs from strain B-5233 and the gliP R strain, but not the gliPΔ strain. Our study confirms that gliotoxin is an important virulence determinant of A. fumigatus and that the type of immunosuppression regimen used is important to reveal the pathogenic potential of gliotoxin.

scholarly journals Olive Fruit and Leaf Wastes as Bioactive Ingredients for Cosmetics—A Preliminary Study

Antioxidants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 245
Author(s):  
María de la Luz Cádiz-Gurrea ◽  
Diana Pinto ◽  
Cristina Delerue-Matos ◽  
Francisca Rodrigues
Keyword(s):  
Olive Oil ◽  
High Performance ◽  
Value Added ◽  
Bioactive Compound ◽  
In Vitro Assays ◽  
Oil Consumption ◽  
Cosmetic Ingredients ◽  
Bioactive Ingredients ◽  
Olive Industry

Olea europaea cultivar, native in the Mediterranean basin, has expanded worldwide, mainly due to the olive oil industry. This expansion is attributed to the benefits of olive oil consumption, since this product is rich in nutritional and bioactive compounds. However, the olive industry generates high amounts of wastes, which could be related to polluting effects on soil and water. To minimize the environmental impact, different strategies of revalorization have been proposed. In this sense, the aim of this work was to develop high cosmetic value added oleuropein-enriched extracts (O20 and O30), a bioactive compound from olive byproducts, performing a comprehensive characterization using high performance liquid chromatography coupled to mass spectrometry and evaluate their bioactivity by in vitro assays. A total of 49 compounds were detected, with oleuropein and its derivatives widely found in O30 extract, whereas iridoids were mainly detected in O20 extract. Moreover, 10 compounds were detected for the first time in olive leaves. Both extracts demonstrated strong antioxidant and antiradical activities, although O30 showed higher values. In addition, radical oxygen and nitrogen species scavenging and enzyme inhibition values were higher in O30, with the exception of HOCl and hyaluronidase inhibition assays. Regarding cell viability, olive byproduct extracts did not lead to a decrease in keratinocytes viability until 100 µg/mL. All data reported by the present study reflect the potential of industrial byproducts as cosmetic ingredients.

scholarly journals In Vitro Propagation, Huperzine A Content and Antioxidant Activity of Three Genotypic Huperzia serrata

Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1112
Author(s):  
Yan Yang ◽  
Liangfang Dai ◽  
Decai Wu ◽  
Limin Dong ◽  
Yisheng Tu ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
High Performance ◽  
In Vitro Propagation ◽  
Huperzine A ◽  
Induction Medium ◽  
Naphthalene Acetic Acid ◽  
Regeneration Rate ◽  
Huperzia Serrata ◽  
Chromatography Analysis

Huperzia serrata is a traditional herb and endangered Chinese medicinal material, which has attracted much attention due to its production of Huperzine A (HupA). In vitro propagation of H. serrata is considered a new way to relieve the resource pressure of H. serrata. In this study, three different genotypic wild H. serrata were used for in vitro propagation. Then, the antioxidant activity and the content of HupA in the regenerated H. serrata were investigated. The results showed the survival rate of the explant was increased to 25.37% when using multiple sterilization processes. The best induction medium for H. serrata was the Schenk and Hildebrandt (SH) medium supplemented with 0.5 mg·L−1 Naphthalene acetic acid (NAA) and 0.1 mg·L−1 2,4-Dichlorophenoxyacetic acid (2,4-D), where the regeneration rate of the explant was to 57.04%. The best proliferation medium was the SH medium with NAA (1.0 mg·L−1), as the biomass of in vitro tissue increased 164.17 ± 0.41 times. High-performance liquid chromatography analysis showed that the in vitro culture of three genotypes could produce HupA and the content of HupA was 53.90–87.17 µg·g−1. The antioxidant experiment showed that the methanol extract of in vitro H. serrata had higher antioxidant activity than that of wild H. serrata. This study provides a reliable in vitro H. serrata culture protocol and laid an important foundation for the antioxidant capacity of the thallus and the content of HupA.

scholarly journals Abnormal physiological properties and altered cell wall composition in Streptococcus pneumoniae grown in the presence of clavulanic acid.

1997 ◽  
Vol 41 (3) ◽  
pp. 504-510 ◽  
Author(s):  
A Severin ◽  
E Severina ◽  
A Tomasz
Keyword(s):  
Streptococcus Pneumoniae ◽  
High Performance ◽  
Biochemical Properties ◽  
Beta Lactam ◽  
Chromatography Analysis ◽  
Peptide Composition ◽  
Increased Sensitivity ◽  
Altered Cell ◽  
Quantitative Changes

Subinhibitory concentrations of clavulanate caused premature induction of stationary-phase autolysis, sensitization to lysozyme, and reductions in the MICs of deoxycholate and penicillin for Streptococcus pneumoniae. In the range of clavulanate concentrations producing these effects, this beta-lactam compound was selectively bound to PBP 3. Cell walls isolated from pneumococci grown in the presence of clavulanate showed increased sensitivity to the hydrolytic action of purified pneumococcal autolysin in vitro. High-performance liquid chromatography analysis of the peptidoglycan isolated from the clavulanate-grown cells showed major qualitative and quantitative changes in stem peptide composition, the most striking feature of which was the accumulation of peptide species carrying intact D-alanyl-D-alanine residues at the carboxy termini. The altered biological and biochemical properties of the clavulanate-grown pneumococci appear to be the consequences of suppressed D,D-carboxypeptidase activity.

scholarly journals A Comparative Study of the Antioxidative Effects of Helichrysum italicum and Helichrysum arenarium Infusions

Antioxidants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 380
Author(s):  
Katja Kramberger ◽  
Zala Jenko Pražnikar ◽  
Alenka Baruca Arbeiter ◽  
Ana Petelin ◽  
Dunja Bandelj ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
High Performance ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Morphological Type ◽  
Mass Spectrometry Analysis ◽  
In Vitro Assays ◽  
Spectrometry Analysis ◽  
Stress Related Genes

Helichrysum arenarium (L.) Moench (abbrev. as HA) has a long tradition in European ethnomedicine and its inflorescences are approved as a herbal medicinal product. In the Mediterranean part of Europe, Helichrysum italicum (Roth) G. Don (abbrev. as HI) is more common. Since infusions from both plants are traditionally used, we aimed to compare their antioxidative potential using in vitro assays. Two morphologically distinct HI plants, HIa and HIb, were compared to a commercially available HA product. Genetic analysis using microsatellites confirmed a clear differentiation between HI and HA and suggested that HIb was a hybrid resulting from spontaneous hybridization from unknown HI subspecies. High-performance liquid chromatography–mass spectrometry analysis showed the highest amounts of hydroxycinnamic acids and total arzanol derivatives in HIa, whereas HIb was richest in monohydroxybenzoic acids, caffeic acids, and coumarins, and HA contained the highest amounts of flavonoids, especially flavanones. HIa exhibited the highest radical scavenging activity; it was more efficient in protecting different cell lines from induced oxidative stress and in inducing oxidative stress-related genes superoxide dismutase 1, catalase, and glutathione reductase 1. The antioxidative potential of HI was not only dependent on the morphological type of the plant but also on the harvest date, revealing important information for obtaining the best possible product. Considering the superior properties of HI compared to HA, the evaluation of HI as a medicinal plant could be recommended.

scholarly journals Monoclonal antibody AHN-1 inhibits phagocytosis by human neutrophils

Blood ◽  
1985 ◽  
Vol 65 (2) ◽  
pp. 333-339
Author(s):  
KM Skubitz ◽  
DJ Weisdorf ◽  
PK Peterson
Keyword(s):  
Monoclonal Antibody ◽  
Lysosomal Enzyme ◽  
Surface Proteins ◽  
Superoxide Production ◽  
Human Neutrophils ◽  
Enzyme Release ◽  
Specific Monoclonal Antibody ◽  
Normal Human ◽  
Major Surface

The granulocyte-specific monoclonal antibody, AHN-1, immunoprecipitates two major surface-iodinated proteins of 105,000 and 145,000 to 150,000 daltons from normal human neutrophils. In this study, the effect of AHN- 1 on a number of neutrophil functions was evaluated in vitro. Both complement- and antibody-mediated phagocytosis were inhibited when human neutrophils were pretreated with AHN-1 and opsonized bacteria were used as targets. The inhibition of phagocytosis was specific, in that lysosomal enzyme release and chemotaxis were not altered by treatment with AHN-1. AHN-1 did inhibit superoxide production by neutrophils in response to particulate stimuli, but not in response to the soluble stimulus, 12-O-tetradecanoylphorbol-13-acetate. The data indicate that one or both of these surface proteins may be important in the process of phagocytosis. AHN-1 should be useful in isolating and further characterizing the nature of these molecules.

scholarly journals CHARACTERIZATION OF MOLLUSCICIDAL COMPONENT OF Moringa oleifera LEAF AND Momordica charantia FRUITS AND THEIR MODES OF ACTION IN SNAIL Lymnaea acuminata

2013 ◽  
Vol 55 (4) ◽  
pp. 251-259 ◽  
Author(s):  
Aparna Upadhyay ◽  
Vinay K. Singh ◽  
Dinesh K. Singh
Keyword(s):  
High Performance ◽  
Moringa Oleifera ◽  
Momordica Charantia ◽  
Chromatography Analysis ◽  
Leaf Powder ◽  
Fruit Powder ◽  
Lymnaea Acuminata ◽  
Molluscicidal Activity

SUMMARY The molluscicidal activity of the leaf powder of Moringa oleifera and lyophilized fruit powder of Momordica charantia against the snail Lymnaea acuminata was time and concentration dependent. M. oleifera leaf powder (96 h LC50: 197.59 ppm) was more toxic than M. charantia lyophilized fruit powder (96 h LC50: 318.29 ppm). The ethanolic extracts of M. oleifera leaf powder and Momordica charantia lyophilized fruit powder were more toxic than other organic solvent extracts. The 96 h LC50 of the column purified fraction of M. oleifera leaf powder was 22.52 ppm, while that of M. charantia lyophilized fruit powder was 6.21 ppm. Column, thin layer and high performance liquid chromatography analysis show that the active molluscicidal components in M. oleifera leaf powder and lyophilized fruit of M. charantia are benzylamine (96 h LC50: 2.3 ppm) and momordicine (96 h LC50: 1.2 ppm), respectively. Benzylamine and momordicine significantly inhibited, in vivo and in vitro, the acetylcholinesterase (AChE), acid and alkaline phosphatase (ACP/ALP) activities in the nervous tissues of L. acuminata. Inhibition of AChE, ACP and ALP activity in the nervous tissues of L. acuminata by benzylamine and momordicine may be responsible for the molluscicidal activity of M. oleifera and M. charantia fruits, respectively.

scholarly journals UV-visible photodegradation of naproxen in water – Structural elucidation of photoproducts and potential toxicity

2020 ◽  
Vol 26 (6) ◽  
pp. 400-408
Author(s):  
Noemi Cazzaniga ◽  
Zsuzsanna Varga ◽  
Edith Nicol ◽  
Stéphane Bouchonnet
Keyword(s):  
Mass Spectrometry ◽  
High Performance ◽  
Vibrio Fischeri ◽  
Software Tool ◽  
In Vitro Assays ◽  
Naphthaleneacetic Acid ◽  
Ion Cyclotron Resonance ◽  
Potential Toxicity ◽  
Uv Visible

The UV-visible photodegradation of Naproxen (6-methoxy-α-methyl-2-naphthaleneacetic acid, CAS: 22204-53-1), one of the most used and detected non-steroidal anti-inflammatory drugs (NSAIDs) in the world, and its ecotoxicological consequences were investigated in an aqueous medium. The photo-transformation products were analyzed and the structures of photoproducts were elucidated using gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) and high-performance liquid chromatography coupled with ultrahigh-resolution Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR-MS). Seven photoproducts were detected and characterized, photo-transformation mechanisms have been postulated to rationalize their formation under irradiation. In silico Q.S.A.R. (Quantitative Structure-Activity Relationship) toxicity predictions were performed with the Toxicity Estimation Software Tool (T.E.S.T.) and in vitro assays were carried out on Vibrio fischeri bacteria. Some of the obtained photoproducts exhibit higher potential toxicity than Naproxen itself but the whole toxicity of the irradiated solution is not of major concern.

A method for high-purity isolation of neutrophil granulocytes for functional cell migration assays

2019 ◽  
Vol 44 (6) ◽  
pp. 810-821
Author(s):  
Edibe Avci ◽  
Yeliz Z. Akkaya-Ulum ◽  
Digdem Yoyen-Ermis ◽  
Gunes Esendagli ◽  
Banu Balci-Peynircioglu
Keyword(s):  
Flow Cytometry ◽  
Cell Migration ◽  
Neutrophil Migration ◽  
Cost Effective ◽  
In Vitro Assays ◽  
Human Neutrophils ◽  
Neutrophil Granulocytes ◽  
Inflammatory Conditions ◽  
Migration Analysis

Abstract Background Neutrophil-mediated killing of pathogens is one of the most significant functions of the primary defense of the host. Neutrophil activity and migration play a key role in inflammatory conditions. To gain insights into the interactions between neutrophils and neutrophil migration-related disorders, a large number of sophisticated methods have been developed. The technical limitations of isolating highly purified neutrophil populations, minimizing both cell death and activation during the isolation process, and the short lifespan of neutrophils present challenges for studying specific functions of neutrophils in vitro. In this study, we aimed to evaluate a separation medium-based density gradient method to obtain highly purified neutrophil populations and combined this protocol with a model for studying neutrophil migration in-vitro. Materials and methods Human granulocytes were isolated using Lympholyte-poly solution. The purity and viability of isolated neutrophils were assessed by flow cytometry and morphological analysis. Neutrophil activation was confirmed by immunocytochemistry. Lastly, filter assay was performed to measure neutrophil chemotaxis. Results and discussion All validation experiments revealed that this method was capable of generating a highly purified neutrophil population for further functional in-vitro assays. Consequently, this study demonstrates a quick, cost effective, and easy-to-follow model, and may be a significant alternative to isolation methods that need extra subsequent steps such as flow cytometry-based cell sorting for reaching highly purified neutrophil population. Conclusion The suggested combination of methods for the isolation and cell migration analysis of human neutrophils is highly recommended to use for disease models involving neutrophil migration such as autoinflammatory disorders.

scholarly journals Acantholippia salsoloides: Phytochemical Composition and Biological Potential of a Thujonic Population

2019 ◽  
Vol 14 (6) ◽  
pp. 1934578X1985854
Author(s):  
Liliana Celaya ◽  
Carmen Viturro ◽  
Luís R. Silva
Keyword(s):  
Essential Oil ◽  
High Performance ◽  
Micrococcus Luteus ◽  
Pharmaceutical Formulations ◽  
In Vitro Assays ◽  
Array Detector ◽  
Biological Potential ◽  
Polar Extracts ◽  
Hydroethanolic Extract

Acantholippia salsoloides (Verbenaceae) is an aromatic plant widespread in the Andean region. The infusion (leaves and flowers) is widely used as a digestive stimulant as well as for the treatment of various diseases in traditional medicine. A. salsoloides attributes its common name “rica-rica” to the fresh and sweet fragrance of the plant. In this work, 2 different polar extracts and the essential oil of a selected rica-rica population were studied. The phenolic composition was analyzed by high-performance liquid chromatography diode array detector; the essential oil profile was determined by gas-chromatography ion-trap mass spectrometry/flame ionization detection. For all extracts, the antibacterial potential was performed by in vitro assays; the antioxidant and α-glucosidase inhibition were determined in decoction and hydroethanolic extracts. The volatile profile allowed the identification of 26 volatile compounds, β-thujone (84%) being the major one in this rica-rica population. Eighteen phenolic compounds were identified; isoferulic acid (16%-18%) and cynaroside (45%-47%) were the larger ones. In a general way, the hydroethanolic extract was more active against Staphylococcus aureus and Micrococcus luteus (minimum inhibitory concentrations= 0.3- 1.3 mg/mL). Both polar extracts have strong antiradical activities although decoction extract proved to be more active against DPPH· (half-maximal inhibitory concentration [IC50] =36 µg/mL) and O2•− (IC50 =28 µg/mL) while hydroethanolic extract shows higher action over α-glucosidase (IC50 =217 µg/mL). The results suggest that A. salsoloides leaves and flowers may be an interesting source of natural antioxidants, antidiabetics, or antimicrobials, and could be used in dietary supplements, medicinal products and pharmaceutical formulations.

Effects of homœopathic preparations of organic acids and minerals on the oxidative metabolism of human neutrophils

1993 ◽  
Vol 82 (04) ◽  
pp. 237-244 ◽  
Author(s):  
Salvatore Chirumbolo ◽  
Andrea Signorini ◽  
Ivo Bianchi ◽  
Guiseppe Lippi ◽  
Paolo Bellavite
Keyword(s):  
Organic Acids ◽  
Oxidative Metabolism ◽  
Saline Solution ◽  
Superoxide Production ◽  
Human Neutrophils ◽  
Critical Factors ◽  
Inhibitory Effects ◽  
Series Of Experiments

AbstractA number of different potencies of commercially available homœopathic preparations in saline solution were tested for their ability to regulate the oxidative metabolism (superoxide production) and adhesion function of human neutrophils in vitro. 15% to 30% inhibition of oxidative metabolism was caused by Sulphur 6x, Manganum phosphoricum 6x and 8x, and Magnesium phosphoricum 6x and 8x. Phosphorus slightly reduced superoxide production, with varying results in a series of experiments. Using Magnesium phosphoricum and Phosphorus, small inhibitory effects (8–11%) were noted event at high potencies. Among the organic acids, a group (Acidum malicum 4x and Acidum fumaricum 4x) enhanced superoxide production, while others either inhibited the response (Acidum citricum and Acidum succinicum, 3x and 4x) or had no effect (Acidum α-ketoglutaricum and Acidum cis-aconitum). Attempts to reproduce these effects using solutions prepared in the laboratory confirmed the inhibitory effects of Manganum phosphoricum 6x and of organic acids in the 3x, while other data indicated that critical factors in the methodology of preparation may affect the results.

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