scholarly journals Preliminary Study and Improve the Production of Metabolites with Antifungal Activity by a Bacillus Sp Strain IBA 33

2009 ◽  
Vol 2 ◽  
pp. MBI.S995 ◽  
Author(s):  
María Antonieta Gordillo ◽  
Antonio Roberto Navarro ◽  
Lidia María Benitez ◽  
Marta Inés Torres De Plaza ◽  
Maria Cristina Maldonado
Keyword(s):  
Pathogenic Fungi ◽  
Geotrichum Candidum ◽  
Culture Conditions ◽  
Penicillium Expansum ◽  
In Vitro Assays ◽  
Bacillus Sp ◽  
In Vivo Assays ◽  
Metabolites Production

Bacillus sp strain IBA 33 metabolites, isolated from decaying lemon fruits, were evaluated for the control of pathogenic and non-pathogenic fungi ( Penicillium digitatum, Geotrichum candidum, Penicillium expansum, Aspergillus clavatus, Aspergillus flavus, Aspergillus niger, and Fusarium moniliforme). These metabolites were recovered from Landy medium (LM) without aminoacids. In order to optimize metabolites production the LM was modified by adding different concentrations and sources of amino acids and carbohydrates at different culture conditions. Bacillus sp strain IBA 33 metabolites efficacy to control fungi were evaluated with in vitro and in vivo assays. A. flavus growth inhibition was 52% with the metabolites of Bacillus sp strain IBA 33 recovered from LM (MBLM) in vitro assays. MBLM supplemented with 0.5% glutamic acid, inhibited the growth of P. digitatum, G. candidum, A. clavatus, A. niger and F. moniliforme by 65%, 88.44%, 84%, 34% and 92% respectively. The highest inhibition of P. expansum was 45% with MBLM supplemented with 0.5% aspartic acid. Similar results were obtained in vivo assays. These results showed that Bacillus sp strain IBA 33 metabolites specificity against fungi depended on the composition of the LM.

scholarly journals Robust and Scalable Angiogenesis Assay of Perfused 3D Human iPSC-Derived Endothelium for Anti-Angiogenic Drug Screening

2020 ◽  
Vol 21 (13) ◽  
pp. 4804
Author(s):  
Vincent van Duinen ◽  
Wendy Stam ◽  
Eva Mulder ◽  
Farbod Famili ◽  
Arie Reijerkerk ◽  
...  
Keyword(s):  
Drug Screening ◽  
Cell Formation ◽  
Cell Types ◽  
Culture Conditions ◽  
In Vitro Assays ◽  
Vegf Receptor ◽  
Screening Assay ◽  
Angiogenesis Assay

To advance pre-clinical vascular drug research, in vitro assays are needed that closely mimic the process of angiogenesis in vivo. Such assays should combine physiological relevant culture conditions with robustness and scalability to enable drug screening. We developed a perfused 3D angiogenesis assay that includes endothelial cells (ECs) from induced pluripotent stem cells (iPSC) and assessed its performance and suitability for anti-angiogenic drug screening. Angiogenic sprouting was compared with primary ECs and showed that the microvessels from iPSC-EC exhibit similar sprouting behavior, including tip cell formation, directional sprouting and lumen formation. Inhibition with sunitinib, a clinically used vascular endothelial growth factor (VEGF) receptor type 2 inhibitor, and 3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one (3PO), a transient glycolysis inhibitor, both significantly reduced the sprouting of both iPSC-ECs and primary ECs, supporting that both cell types show VEGF gradient-driven angiogenic sprouting. The assay performance was quantified for sunitinib, yielding a minimal signal window of 11 and Z-factor of at least 0.75, both meeting the criteria to be used as screening assay. In conclusion, we have developed a robust and scalable assay that includes physiological relevant culture conditions and is amenable to screening of anti-angiogenic compounds.

Polystachyasaponin with Adjuvant Activity from Entada polystachya

2010 ◽  
Vol 65 (5) ◽  
pp. 628-634 ◽  
Author(s):  
Bernadete P. da Silva ◽  
José P. Parente
Keyword(s):  
2D Nmr ◽  
In Vitro Assays ◽  
Triterpenoid Saponin ◽  
Spectroscopic Techniques ◽  
Adjuvant Activity ◽  
In Vivo Assays ◽  
Cellular Immune ◽  
C Nmr

A new complex triterpenoid saponin, polystachyasaponin, was isolated from leaves of Entada polystachya (L.) DC. (Leguminosae) by using chromatographic methods. Its structure was established as 15,16-dihydroxy-3-[[O-β -D-xylopyranosyl-(1→2)-O-α-L-arabinopyranosyl-(1→6)-2- (acetylamino)-2-deoxy-β -D-glucopyranosyl]oxy]-(3β ,15α,16α)-olean-12-en-28-oic acid O-D-apio- β -D-furanosyl-(1→3)-O-β -D-xylopyranosyl-(1→2)-O-[β -D-glucopyranosyl-(1→4)]-6-O-[(2E,6R)- 6-hydroxy-2,6-dimethyl-1-oxo-2,7-octadienyl]-β -D-glucopyranosyl ester. Structural elucidation was performed using detailed analyses of 1H and 13C NMR spectra including 2D NMR spectroscopic techniques and chemical conversions. The hemolytic activity of the saponin was evaluated using in vitro assays, and its adjuvant potential on the cellular immune response against ovalbumin antigen was investigated using in vivo assays.

Renewing potency assays: Moving forward from traditional methods

2020 ◽  
Author(s):  
Francesco Nevelli
Keyword(s):  
Growth Hormone ◽  
Physiological Mechanism ◽  
Global Market ◽  
Hormone Deficiency ◽  
In Vitro Assays ◽  
In Vitro Methods ◽  
In Vivo Assays ◽  
Market Leader

Merck is global market leader in the fertility and growth hormone deficiency treatment. The quality control analytical panels for each new produced batch envisage the potency quantification that is estimated using a dedicated in vivo assay. Indeed, no in vitro methods for gonadotropin potency quantification are available in any pharmacopoeia. Merck Ivrea started a project to replace the in vivo assays with in vitro assays able to mimic the physiological mechanism of action of each gonadotropin and growth hormone.

scholarly journals The Extracellular Matrix Influences Ovarian Carcinoma Cells’ Sensitivity to Cisplatinum: A First Step towards Personalized Medicine

Cancers ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 1175 ◽  
Author(s):  
Andrea Balduit ◽  
Chiara Agostinis ◽  
Alessandro Mangogna ◽  
Veronica Maggi ◽  
Gabriella Zito ◽  
...  
Keyword(s):  
Standardized Testing ◽  
Ovarian Carcinoma ◽  
Tumor Cells ◽  
Culture Conditions ◽  
Carcinoma Cells ◽  
In Vitro Assays ◽  
Set Up ◽  
Platinum Chemotherapy

The development of personalized therapies for ovarian carcinoma patients is still hampered by several limitations, mainly the difficulty of predicting patients’ responses to chemotherapy in tumor cells isolated from peritoneal fluids. The main reason for the low predictive power of in vitro assays is related to the modification of the cancer cells’ phenotype induced by the culture conditions, which results in changes to the activation state and drug sensitivity of tumor cells compared to their in vivo properties. We have defined the optimal culture conditions to set up a prognostic test to predict high-grade serous ovarian carcinoma (HGSOC) patients’ responses to platinum chemotherapy. We evaluated the effects of hyaluronic acid (HA) and fibronectin matrices and the contribution of freezing/thawing processes to the cell response to platinum-based treatment, collecting spheroids from the ascitic fluids of 13 patients with stage II or III HGSOC. Our findings indicated that an efficient model used to generate predictive data for in vivo sensitivity to platinum is culturing fresh spheroids on HA, avoiding the use of previously frozen primary tumor cells. The establishment of this easy, reproducible and standardized testing method can significantly contribute to an improvement in therapeutic effectiveness, thus bringing the prospect of personalized therapy closer for ovarian carcinoma patients.

scholarly journals Recent Reduction in Thrombogenic Enzyme Content of Prothrombin Complex Concentrates

1977 ◽  
Author(s):  
H.S. Kingdon
Keyword(s):  
Calcium Ions ◽  
Calcium Ion ◽  
Deae Cellulose ◽  
In Vitro Assays ◽  
Prothrombin Complex Concentrates ◽  
In Vivo Assays ◽  
Prolonged Contact ◽  
Plasma Fractions

In previous studies of prothrombin complex concentrates, it was demonstrated that there was a high content of potentially thrombogenic enzymes in the products from certain manufacturers, and that the enzyme conteat correlated closely with in vivo assays for thrombogenicity and with the observed incidence of thrombotic episodes following infusion (Thromb. Diath. Haemorrh. 33, 617, 1 975\Subsequently, it was shown that the thrombogenic enzymes could be generated by prolonged contact with DEAE-cellulose or calcium ions during preparation or later handling (Blood, 49, 159, 1977). In view of. these observations, efforts have been made to reduce the contact time of plasma fractions with either DEAE-cellulose or calcium ion during purification of these fractions for therapeutic use. In vitro assays for thrombogenic enzymes using the nonactivated partial thromboplastin time (NAPTT) were recently repeated on some of the currently available therapeutic materials. Assays were standardized and compared with previous results by using a provisional standard provided by Dr. David Aronson, Bureau of Biologies, USFDA. In the 1975 study, one manufacturer was identified as making a concentrate virtually free of thrombogenic enzyme. The concentrate currently being made by this manufacturer still does not significantly shorten the NAPTT. In the 1975 study, 2 manufacturers were shown to be making concentrates with high titers of thrombogenic enzyme. The current products of these two manufacturers contain detectable but significantly lower levels of thrombogenic enzymes. Thus it appears that for these two manufacturers, minor changes in production procedures have led to a product containing less potentially thrombogenic material.

scholarly journals In vitro assays misrepresent in vivo lineage potentials of murine lymphoid progenitors

Blood ◽  
2011 ◽  
Vol 117 (9) ◽  
pp. 2618-2624 ◽  
Author(s):  
Lauren I. Richie Ehrlich ◽  
Thomas Serwold ◽  
Irving L. Weissman
Keyword(s):  
T Cell ◽  
The Other ◽  
In Vitro Assays ◽  
In Vivo Assays ◽  
Other Hand ◽  
Multipotent Progenitors ◽  
Lymphoid Progenitors

Abstract The identity of T-cell progenitors that seed the thymus has remained controversial, largely because many studies differ over whether these progenitors retain myeloid potential. Contradictory reports diverge in their use of various in vitro and in vivo assays. To consolidate these discordant findings, we compared the myeloid potential of 2 putative thymus seeding populations, common lymphoid progenitors (CLPs) and multipotent progenitors (MPPs), and the earliest intrathymic progenitor (DN1), using 2 in vitro assays and in vivo readouts. These assays gave contradictory results: CLP and DN1 displayed surprisingly robust myeloid potential on OP9-DL1 in vitro stromal cocultures but displayed little myeloid potential in vivo, as well as in methylcellulose cultures. MPP, on the other hand, displayed robust myeloid potential in all settings. We conclude that stromal cocultures reveal cryptic, but nonphysiologic, myeloid potentials of lymphoid progenitors, providing an explanation for contradictory findings in the field and underscoring the importance of using in vivo assays for the determination of physiologic lineage potentials.

Evaluation of antioxidant activity and toxicity of sulfur- or selenium-containing 4-(arylchalcogenyl)-1H-pyrazoles

2020 ◽  
Vol 98 (7) ◽  
pp. 441-448
Author(s):  
Daniela Hartwig de Oliveira ◽  
Fernanda Severo Sabedra Sousa ◽  
Paloma Taborda Birmann ◽  
Ana Paula Pesarico ◽  
Diego Alves ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Mouse Brain ◽  
Aminolevulinic Acid ◽  
In Vitro Assays ◽  
Scavenging Activity ◽  
In Vivo Assays ◽  
Reducing Ability ◽  
Liver And Kidney

Pyrazoles represent a significant class of heterocyclic compounds that exhibit pharmacological properties. The present study aimed to investigate the antioxidant potential of pyrazol derivative compounds in brain of mice in vitro and the effect of pyrazol derivative compounds in the oxidative damage and toxicity parameters in mouse brain and plasma of mice. The compounds tested were 3,5-dimethyl-1-phenyl-4-(phenylselanyl)-1H-pyrazol (1a), 3,5-dimethyl-4-(phenylselanyl)-1H-pyrazole (2a), 4-((4-methoxyphenyl)selanyl)-3,5-dimethyl-1-phenyl-1H-pyrazole (3a), 4-((4-chlorophenyl)selanyl)-3,5-dimethyl-1-phenyl-1H-pyrazole (4a), 3,5-dimethyl-1-phenyl-4-(phenylthio)-1H-pyrazole (1b), 3,5-dimethyl-4-(phenylthio)-1H-pyrazole (2b), 4-((4-methoxyphenyl)thio)-3,5-dimethyl-1-phenyl-1H-pyrazole (3b), 4-((4-chlorophenyl)thio)-3,5-dimethyl-1-phenyl-1H-pyrazole (4b), and 3,5-dimethyl-1-phenyl-1H-pyrazole (1c). In vitro, 4-(arylcalcogenyl)-1H-pyrazoles, at low molecular range, reduced lipid peroxidation and reactive species in mouse brain homogenates. The compounds also presented ferric-reducing ability as well nitric oxide-scavenging activity. Especially compounds 1a, 1b, and 1c presented efficiency to 1,1-diphenyl-2-picryl-hydrazyl-scavenging activity. Compounds 1b and 1c presented 2,20 -azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)-scavenging activity. In vivo assays demonstrated that compounds 1a, 1b, and 1c (300 mg/kg, intragastric, a single administration) did not cause alteration in the of δ-aminolevulinic acid dehydratase activity, an enzyme that exhibits high sensibility to prooxidants situations, in the brain, liver, and kidney of mice. Compound 1c reduced per se the lipid peroxidation in liver and brain of mice. Toxicological assays demonstrate that compounds 1a, 1b, and 1c did not present toxicity in the aspartate aminotransferase, alanine aminotransferase, urea, and creatinine levels in the plasma. In conclusion, the results demonstrated the antioxidant action of pyrazol derivative compounds in in vitro assays. Furthermore, the results showed low toxicity of compounds in in vivo assays.

scholarly journals Cytochrome P450-mediated warfarin metabolic ability is not a critical determinant of warfarin sensitivity in avian species: In vitro assays in several birds and in vivo assays in chicken

2015 ◽  
Vol 34 (10) ◽  
pp. 2328-2334 ◽  
Author(s):  
Kensuke P. Watanabe ◽  
Minami Kawata ◽  
Yoshinori Ikenaka ◽  
Shouta M. M. Nakayama ◽  
Chihiro Ishii ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Avian Species ◽  
In Vitro Assays ◽  
Critical Determinant ◽  
In Vivo Assays

scholarly journals BIOCONTROL OF TEAK CANKER CAUSED BY Lasiodiplodia theobromae

2018 ◽  
Vol 42 (3) ◽  
Author(s):  
Rafaela Cristina Ferreira Borges ◽  
Eder Marques ◽  
Monica Alves Macedo ◽  
Irene Martins ◽  
José Getulio da Silva Filho ◽  
...  
Keyword(s):  
Pathogenic Fungus ◽  
In Vitro Assays ◽  
Timber Species ◽  
Bacillus Sp ◽  
Trichoderma Spp ◽  
Vitro Assay ◽  
Lasiodiplodia Theobromae ◽  
In Vivo Tests

ABSTRACT Teak is a forest species that has assumed great importance in Brazil, where it has found excellent conditions for development since its introduction into the country in the 1960s. However, phytosanitary problems are beginning to threaten the production of this timber species. An example is teak canker, caused by the fungus Lasiodiplodia theobromae (Lt), which has only recently been reported in Brazil, and for which, therefore, there are no recommended control methods. Thus, this study evaluated the control of this pathogen, investigating the potential of the biocontrol agents (BCAs) Trichoderma spp., Bacillus sp. and Enterobacter sp., initially through in vitro assays and, subsequently, with in vivo tests. According to the in vitro assay results, the Trichoderma isolates CEN162 and CEN1153 and the strain of Bacillus sp. (UnB1366) were the treatments that stood out, as they were able to completely inhibit mycelial growth of some isolates of Lt. When these isolates were tested in a preventive way, the control levels varied depending on the Lt isolate and the antagonist-clone interaction, where CEN162 (T. asperellum) and UnB166 (Bacillus sp.) showed 100% control. Thus, there is a positive correlation between the in vitro and in vivo tests, since the same BCAs stood out. Although good levels of control have been obtained with the BCAs used, it can be concluded that there is a variation in the antagonism to different Lt isolates or even in the antagonist-clone interaction, corroborating the information available in the scientific literature on this plant-pathogenic fungus.

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