scholarly journals Maternal Choline and Betaine Supplementation Modifies the Placental Response to Hyperglycemia in Mice and Human Trophoblasts

Nutrients ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 1507 ◽  
Author(s):  
Khatia Nanobashvili ◽  
Chauntelle Jack-Roberts ◽  
Rachel Bretter ◽  
Naudia Jones ◽  
Kathleen Axen ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Nutrient Transport ◽  
Human Cell Line ◽  
In Vitro Assays ◽  
High Fat ◽  
Junctional Zone ◽  
Placental Transport ◽  
Angiogenic Genes ◽  
Betaine Supplementation

Gestational diabetes mellitus (GDM) is characterized by excessive placental fat and glucose transport, resulting in fetal overgrowth. Earlier we demonstrated that maternal choline supplementation normalizes fetal growth in GDM mice at mid-gestation. In this study, we further assess how choline and its oxidation product betaine influence determinants of placental nutrient transport in GDM mice and human trophoblasts. C57BL/6J mice were fed a high-fat (HF) diet 4 weeks prior to and during pregnancy to induce GDM or fed a control normal fat (NF) diet. The HF mice also received 25 mM choline, 85 mM betaine, or control drinking water. We observed that GDM mice had an expanded placental junctional zone with an increased area of glycogen cells, while the thickness of the placental labyrinth zone was decreased at E17.5 compared to NF control mice (p < 0.05). Choline and betaine supplementation alleviated these morphological changes in GDM placentas. In parallel, both choline and betaine supplementation significantly reduced glucose accretion (p < 0.05) in in vitro assays where the human choriocarcinoma BeWo cells were cultured in high (35.5 mM) or normal (5.5 mM) glucose conditions. Expression of angiogenic genes was minimally altered by choline or betaine supplementation in either model. In conclusion, both choline and betaine modified some but not all determinants of placental transport in response to hyperglycemia in mouse and in vitro human cell line models.

scholarly journals Volatile Compounds of Endophytic Bacillus spp. have Biocontrol Activity Against Sclerotinia sclerotiorum

2018 ◽  
Vol 108 (12) ◽  
pp. 1373-1385 ◽  
Author(s):  
Venance Colman Massawe ◽  
Alvina Hanif ◽  
Ayaz Farzand ◽  
David Kibe Mburu ◽  
Sylvans Ochieng Ochola ◽  
...  
Keyword(s):  
Sclerotinia Sclerotiorum ◽  
Morphological Changes ◽  
Antagonistic Activity ◽  
In Vitro Assays ◽  
Gas Chromatography Mass Spectrometry ◽  
Bromophenol Blue ◽  
Fungal Mycelium ◽  
Negative Effects ◽  
Bacillus Spp

To develop an effective biological agent to control Sclerotinia sclerotiorum, three endophytic Bacillus spp. strains with high antagonistic activity were isolated from maize seed and characterized. In vitro assays revealed that the Bacillus endophytes could produce volatile organic compounds (VOC) that reduced sclerotial production and inhibited mycelial growth of S. sclerotiorum. Gas chromatography–mass spectrometry revealed that the selected strains produced 16 detectable VOC. Eight of the produced VOC exhibited negative effects on S. sclerotiorum, while a further four induced accumulation of reactive oxygen species in mycelial cells. A mixture of VOC produced by Bacillus velezensis VM11 caused morphological changes in the ultrastructure and organelle membranes of S. sclerotiorum mycelial cells. The bromophenol blue assay revealed a yellow color of untreated fungal mycelium, which grew faster and deeper from 24 to 72 h postinoculation, as an indication of reduced pH. The potassium permanganate (KMnO4) titration assay showed that the rate of oxalic acid accumulation was higher in minimal salt liquid medium cultures inoculated with untreated fungal plugs compared with the Bacillus VOC-treated ones. Interestingly, biological control assays using host-plant leaves challenged with treated fungal mycelial plugs produced reduced lesions compared with the control. These findings provide new viable possibilities of controlling diseases caused by S. sclerotiorum using VOC produced by Bacillus endophytes.

scholarly journals Antibodies to granulocyte precursors in selective myeloid hypoplasia and other suspected autoimmune neutropenias: use of HL-60 cells as targets

Blood ◽  
1987 ◽  
Vol 69 (2) ◽  
pp. 529-536 ◽  
Author(s):  
MS Currie ◽  
JB Weinberg ◽  
PK Rustagi ◽  
GL Logue
Keyword(s):  
Cell Line ◽  
Protein A ◽  
Human Cell Line ◽  
Normal Serum ◽  
Antibody Binding ◽  
In Vitro Assays ◽  
Staphylococcal Protein A ◽  
In Vitro Differentiation ◽  
Igg Binding

Abstract Patients with syndromes of autoantibody-mediated hematocytopenias may manifest signs of increased cell destruction and/or decreased cell production, depending on the maturity of the target cell and the effects of antibody binding. The purpose of this study was to use a cultured human cell line of hematopoietic origin for in vitro assays of antibody binding to overcome the relative inaccessibility of natural human marrow progenitor cells. This report describes the detection, using radioiodinated staphylococcal protein A (SPA), of antibodies binding to a human promyelocytic cell line (HL-60) in sera from three patients with chronic idiopathic granulocytic hypoplasia (“pure white cell aplasia,” PWCA) and 22 patients with other syndromes of suspected immune neutropenia. Bone marrow from patients with increased IgG binding to HL-60 cells showed less than 15% granulocytic lineage cellularity in 11 of 17 cases. In vitro differentiation of HL-60 cells by retinoic acid resulted in increased IgG binding for sera that had shown increased IgG binding to mature granulocytes but not undifferentiated HL-60 cells; in contrast, for sera with antibodies to untreated HL-60 cells and for normal serum, in vitro differentiation had little effect on IgG binding. Antibodies eluted from mature granulocytes were similar to the parent serum regarding the ratio of IgG binding to mature cells v HL-60 cells. No sera from 19 patients with febrile transfusion reactions showed increased IgG binding to HL- 60 cells in the absence of increased IgG binding to mature granulocytes, although two sera had antibodies to both cell types. The use of HL-60 cells as targets may permit measurement of serum antibodies associated with granulocytic hypoplasia. In combination with assays to detect antibody binding to mature granulocytes, these techniques may discriminate among autoantibody specificities for antigens that are gained, conserved, or lost during myeloid maturation.

scholarly journals Cupric Oxide Nanoparticles Induce Cellular Toxicity in Liver and Intestine Cell Lines

2020 ◽  
Vol 10 (2) ◽  
pp. 213-220 ◽  
Author(s):  
Mahmoud Abudayyak ◽  
Elif Guzel ◽  
Gül Özhan
Keyword(s):  
Cell Lines ◽  
Cupric Oxide ◽  
Morphological Changes ◽  
Neutral Red ◽  
In Vitro Assays ◽  
Oxide Nanoparticles ◽  
Cuo Nps ◽  
Cell Death Pathway ◽  
Cupric Oxide Nanoparticles

Purpose: The wide application of cupric oxide nanoparticles (copper (II) oxide, CuO-NPs) in various fields has increased exposure to the kind of active nanomaterials, which can cause negative effects on human and environment health. Although CuO-NPs were reported to be harmful to human, there is still a lack information related to their toxic potentials. In the present study, the toxic potentials of CuO-NPs were evaluated in the liver (HepG2 hepatocarcinoma) and intestine (Caco-2 colorectal adenocarcinoma) cells. Methods: After the characterization of particles, cellular uptake and morphological changes were determined. The potential of cytotoxic, genotoxic, oxidative and apoptotic damage was investigated with several in vitro assays. Results: The average size of the nanoparticles was 34.9 nm, about 2%-5% of the exposure dose was detected in the cells and mainly accumulated in different organelles, causing oxidative stress, cell damages, and death. The IC50 values were 10.90 and 10.04 µg/mL by MTT assay, and 12.19 and 12.06 µg/mL by neutral red uptake (NRU) assay, in HepG2 and Caco-2 cells respectively. Apoptosis assumes to the main cell death pathway; the apoptosis percentages were 52.9% in HepG2 and 45.5% in Caco-2 cells. Comet assay result shows that the highest exposure concentration (20 µg/mL) causes tail intensities about 9.6 and 41.8%, in HepG2 and Caco-2 cells, respectively. Conclusion: CuO-NPs were found to cause significant cytotoxicity, genotoxicity, and oxidative and apoptotic effects in both cell lines. Indeed, CuO-NPs could be dangerous to human health even if their toxic mechanisms should be elucidated with further studies.

scholarly journals Characterization of ctx gene Negative Vibrio fluvialis Organisms Isolated from the Environment

2020 ◽  
Vol 36 (2) ◽  
pp. 91-97
Author(s):  
Juthi Biswas ◽  
Tahsina Jainab ◽  
Mahmud Hossain ◽  
Mahmuda Yasmin ◽  
Jamalun Nessa ◽  
...  
Keyword(s):  
Mdck Cells ◽  
Morphological Changes ◽  
Neuronal Cell ◽  
In Vitro Assays ◽  
Ileal Loop ◽  
Vibrio Fluvialis ◽  
Cell Culture Assay ◽  
Hind Limb Paralysis

A total of five Vibrio fluvialis organisms were isolated from the different environmental samples collected from Dhaka, Satkhira and Khulna. All these isolates were confirmed following API 20NE tests. Molecular analysis showed the absence of ctx, toxR, tdh, trh, stx1, and stx2 genes in these organisms. However, culture filtrates and crude proteins prepared from these organisms showed fluid accumulation in rabbit ileal loop assay, haemolysis of sheep red blood cess, rounding of BHK-21, HeLa and MDCK cells in cell culture assay, hind limb paralysis and death of mice in mice lethality assay and morphological changes in mouse neuronal cell assay. All these results indicated that the environmental V. fluvialis organisms, may not contain different virulence genes, including the ctx gene. However, the other in vivo and in vitro assays indicate that the toxins produced by the V. fluvialis organisms may contain enterotoxin, haemolysin, cytotoxin and neurotoxin. Bangladesh J Microbiol, Volume 36 Number 2 December 2019, pp 91-97

scholarly journals Antibodies to granulocyte precursors in selective myeloid hypoplasia and other suspected autoimmune neutropenias: use of HL-60 cells as targets

Blood ◽  
1987 ◽  
Vol 69 (2) ◽  
pp. 529-536
Author(s):  
MS Currie ◽  
JB Weinberg ◽  
PK Rustagi ◽  
GL Logue
Keyword(s):  
Cell Line ◽  
Protein A ◽  
Human Cell Line ◽  
Normal Serum ◽  
Antibody Binding ◽  
In Vitro Assays ◽  
Staphylococcal Protein A ◽  
In Vitro Differentiation ◽  
Igg Binding

Patients with syndromes of autoantibody-mediated hematocytopenias may manifest signs of increased cell destruction and/or decreased cell production, depending on the maturity of the target cell and the effects of antibody binding. The purpose of this study was to use a cultured human cell line of hematopoietic origin for in vitro assays of antibody binding to overcome the relative inaccessibility of natural human marrow progenitor cells. This report describes the detection, using radioiodinated staphylococcal protein A (SPA), of antibodies binding to a human promyelocytic cell line (HL-60) in sera from three patients with chronic idiopathic granulocytic hypoplasia (“pure white cell aplasia,” PWCA) and 22 patients with other syndromes of suspected immune neutropenia. Bone marrow from patients with increased IgG binding to HL-60 cells showed less than 15% granulocytic lineage cellularity in 11 of 17 cases. In vitro differentiation of HL-60 cells by retinoic acid resulted in increased IgG binding for sera that had shown increased IgG binding to mature granulocytes but not undifferentiated HL-60 cells; in contrast, for sera with antibodies to untreated HL-60 cells and for normal serum, in vitro differentiation had little effect on IgG binding. Antibodies eluted from mature granulocytes were similar to the parent serum regarding the ratio of IgG binding to mature cells v HL-60 cells. No sera from 19 patients with febrile transfusion reactions showed increased IgG binding to HL- 60 cells in the absence of increased IgG binding to mature granulocytes, although two sera had antibodies to both cell types. The use of HL-60 cells as targets may permit measurement of serum antibodies associated with granulocytic hypoplasia. In combination with assays to detect antibody binding to mature granulocytes, these techniques may discriminate among autoantibody specificities for antigens that are gained, conserved, or lost during myeloid maturation.

Terpenoids from Leaves of Guarea macrophylla Display In Vitro Cytotoxic Activity and Induce Apoptosis In Melanoma Cells

Planta Medica ◽  
2017 ◽  
Vol 83 (16) ◽  
pp. 1289-1296 ◽  
Author(s):  
Geanne Conserva ◽  
Natalia Girola ◽  
Carlos Figueiredo ◽  
Ricardo Azevedo ◽  
Sasha Mousdell ◽  
...  
Keyword(s):  
Cytotoxic Activity ◽  
Tumor Cells ◽  
Nuclear Dna ◽  
Morphological Changes ◽  
Folk Medicine ◽  
Melanoma Cells ◽  
In Vitro Assays ◽  
Apoptotic Pathway ◽  
Isolation And Characterization

Abstract Guarea macrophylla is a Brazilian plant species that has been used in folk medicine to treat a range of diseases. Our ongoing work focuses on the discovery of new bioactive natural products derived from Brazilian flora. The current study describes the identification of cytotoxic compounds from the EtOH extract of leaves from G. macrophylla using bioactivity-guided fractionation. This approach resulted in the isolation and characterization of four compounds: cycloart-23E-ene-3β,25-diol (1), (23S*,24S*)-dihydroxycicloart-25-en-3-one (2), isopimara-7,15-diene-2α,3β-diol (3), and isopimara-7,15-dien-3β-ol (4), in which 2 and 3 are identified as new derivatives. In vitro assays were conducted to evaluate the cytotoxic activity of compounds 1–4 against a panel of cancer cell lines and to determine the possible mechanism(s) related to the activity of the compounds on B16F10Nex2 cells. The most active compound 1 induced cytotoxic effects on tumor cells, with IC50 values of 18.3, 52.1, and 58.9 µM against HL-60, HeLa, and B16F10-Nex2 tumor cells, respectively. Furthermore, it was observed in melanoma cells that compound 1 induced several specific apoptotic hallmarks, such as morphological changes in the cell shape structure, nuclear DNA condensation, specific chromatin fragmentation, and disruption in the mitochondrial membrane potential, which are related to the intrinsic apoptotic pathway.

scholarly journals Genotoxic and Antitumor Activity of Pollen Grains against Prostate Cancer Cell Line

2021 ◽  
Vol 11 (3) ◽  
Author(s):  
Magdah Ganash
Keyword(s):  
Prostate Cancer ◽  
Cancer Cell ◽  
Prostate Cancer Cell ◽  
Antioxidant Activities ◽  
Morphological Changes ◽  
Pollen Grains ◽  
In Vitro Assays ◽  
Maize Pollen ◽  
Pc3 Cells

Since the use of engineered antioxidants and antitumor is under investigation, inferable from its likely poisonousness, scientists have deflected their thoughtfulness regarding the quest for characteristic sources to meet the human medication and diet requests. Therefore the study aimed to evaluate the antitumor and antioxidant activities of maize pollen grains against the Prostate Cancer Cell (Pc3) line. Maize pollen grains were collected by Bee through a pollen trap, and then subjected for flavonoids and alkaloids analysis by HPLC method. an in vitro assays, were used to test the antitumor properties, against Pc3 cells. Furthermore, its antioxidant potential was also evaluated by DPPH. The detected flavonoids were identified to be quercetin, luteolin kaempferol, rutin, apigenin and naringin and the alkaloids were quinolone, hydroxyindolenine and conofoline. The antitumor efficacy of pollen grains extract increased with concentration and reached to 94.92 % that similar to the toxicity % of adriamycin at 1000 µg/mL, however, the IC50 (339.81 µg) of pollen grains extract was highest than IC50 (58.07 µg) of adriamycin. At 500 μg/mL of pollen grains extract, morphological changes of Pc3 were recorded. These changes deformed more at 1000 μg/mL. DPPH scavenging activity was found to be 92.26 % at 1280 µg/mL of pollen grains extracted with IC50 425.4 µg/mL compared with IC50 (13.9 µg/mL) of the ascorbic acid. DNA fragmentation and quantitative RT-PCR examinations of Bax and Bcl-2 genes demonstrated that pollen grains extract induced cellular apoptosis of Pc3 cells. This study concluded that the maize pollen grains may applied as natural safe source for inhibit Pc3 Cells proliferation as well as applied as antioxidant.

Correlation Between Cellular Functions and Morphological Changes of Human Trophoblast in Vitro

1975 ◽  
Vol 33 ◽  
pp. 600-601
Author(s):  
John C. Garancis ◽  
Robert O. Hussa ◽  
Michael T. Story ◽  
Donald Yorde ◽  
Roland A. Pattillo
Keyword(s):  
Electron Microscopy ◽  
Cellular Proliferation ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Cellular Functions ◽  
Human Trophoblast ◽  
Transmission Electron ◽  
Marked Activity ◽  
Malignant Trophoblast

Human malignant trophoblast cells in continuous culture were incubated for 3 days in medium containing 1 mM N6-O2'-dibutyryl cyclic adenosine 3':5'-monophosphate (dibutyryl cyclic AMP) and 1 mM theophylline. The culture fluid was replenished daily. Stimulated cultures secreted many times more chorionic gonadotropin and estrogens than did control cultures in the absence of increased cellular proliferation. Scanning electron microscopy revealed remarkable surface changes of stimulated cells. Control cells (not stimulated) were smooth or provided with varying numbers of microvilli (Fig. 1). The latter, usually, were short and thin. The surface features of stimulated cells were considerably different. There was marked increase of microvilli which appeared elongated and thick. Many cells were covered with confluent polypoid projections (Fig. 2). Transmission electron microscopy demonstrated marked activity of cytoplasmic organelles. Mitochondria were increased in number and size; some giant forms with numerous cristae were observed.

A in Vivo Assay for Standardizing Heparin Preparations

1979 ◽  
Vol 41 (03) ◽  
pp. 576-582
Author(s):  
A R Pomeroy
Keyword(s):  
In Vitro Assays ◽  
British Pharmacopoeia ◽  
In Vitro Method ◽  
Standard Preparation ◽  
Reliable Estimation ◽  
Assay Procedure ◽  
Accuracy And Precision ◽  
Heparin Preparation

SummaryThe limitations of currently used in vitro assays of heparin have demonstrated the need for an in vivo method suitable for routine use.The in vivo method which is described in this paper uses, for each heparin preparation, four groups of five mice which are injected intravenously with heparin according to a “2 and 2 dose assay” procedure. The method is relatively rapid, requiring 3 to 4 hours to test five heparin preparations against a standard preparation of heparin. Levels of accuracy and precision acceptable for the requirements of the British Pharmacopoeia are obtained by combining the results of 3 to 4 assays of a heparin preparation.The similarity of results obtained the in vivo method and the in vitro method of the British Pharmacopoeia for heparin preparations of lung and mucosal origin validates this in vivo method and, conversely, demonstrates that the in vitro method of the British Pharmacopoeia gives a reliable estimation of the in vivo activity of heparin.

Potentially Thrombogenic Materials in Factor IX Concentrates

1975 ◽  
Vol 33 (03) ◽  
pp. 617-631 ◽  
Author(s):  
H. S Kingdon ◽  
R. L Lundblad ◽  
J. J Veltkamp ◽  
D. L Aronson
Keyword(s):  
Human Plasma ◽  
Antithrombin Iii ◽  
Factor Ix ◽  
In Vitro Assays ◽  
Substantial Agreement ◽  
Coefficient Of Correlation

SummaryFactor IX concentrates manufactured from human plasma and intended for therapeutic infusion in man have been suspected for some time of being potentially thrombogenic. In the current studies, assays were carried out in vitro and in vivo for potentially thrombogenic materials. It was possible to rank the various materials tested according to the amount of thrombogenic material detected. For concentrates not containing heparin, there was substantial agreement between the in vivo and in vitro assays, with a coefficient of correlation of 0.77. There was no correlation between the assays for thrombogenicity and the antithrombin III content. We conclude that many presently available concentrates of Factor IX contain substantial amounts of potentially thrombogenic enzymes, and that this fact must be considered in arriving at the decision whether or not to use them therapeutically.

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