Pyrrolizidine Alkaloids-Free Extract from the Cell Culture of Lithospermum officinale with High Antioxidant Capacity

Background: We examined anti-Toxoplasma effect of hydroalcoholic extract of Terminalia chebula Retz (T. chebula) in cell culture and murine model. Methods: The study was conducted in Shahrekord University of Medical Sciences, Iran in 2017. Half maximal effective (concentration (EC50) of T. chebula extract and pyrimethamine was determined in infected Hela cells by using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) method. In the animal model, BALB/c mice were injected with tachyzoites (104) of T. RH strain intraperitoneally. 24h after the injection, the test groups were orally treated with 100, 200, 400 and 800 mg/kg of T. chebula extract for 7 days. The survival rate of the mice was determined and blood samples were collected to determine the amount of serum Malondialdehyde (MDA) and antioxidant capacity. Then peritoneal fluid of the mice was collected to count the number of tachyzoites and after necropsy, the pathologic changes, including the weight of liver, spleen and kidneys were investigated. The analysis of data was accomplished using SPSS. Results: EC50 values were 94.7μg/mL and 290.50μg/mL for T. chebula and pyrimethamine respectively. In the animal model, the extract of T. chebula in concentration of 100 mg/kg showed the same anti-Toxoplasma effect as pyrimethamine. This concentration of the extract decreased number of intraperitoneal tachyzoites and increased the survival rate of the mice. This extract reduced the levels of serum MDA and tissue inflammation and increased serum antioxidant capacity. Conclusion: Regarding the positive effect of extract, after more clinical trials in the animal model and standardization of the extract, it can be used as an alternative or complementary therapy for toxoplasmosis.

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A T.E.M. study of mouse mammary epithelial cell secretory differentiation cultured on collagen and Matrigel

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100085241 ◽

1991 ◽

Vol 49 ◽

pp. 188-189

Author(s):

W.N. Bentham ◽

V. Rocha

Keyword(s):

Cell Culture ◽

Mammary Epithelial ◽

Secretory Process ◽

Cell Culture System ◽

Protein Maturation ◽

Cell Culture Techniques ◽

Culture Techniques ◽

Mouse Mammary Epithelial Cell ◽

Secretory Differentiation

It has been an interest of our lab to develop a mammary epethelial cell culture system that faithfully duplicates the in vivo condition of the lactating gland. Since the introduction of collagen as a matrix on which cells are cultivated other E.C.M. type matrices have been made available and are used in many cell culture techniques. We have previously demonstrated that cells cultured on collagen and Matrigel do not differentiate as they do in vivo. It seems that these cultures often produce cells that show a disruption in the secretory process. The appearance of large ribosomal studded vesicles, that specifically label with antibody to casein, suggest an interruption of both protein maturation and secretion at the E.R. to golgi transition. In this report we have examined cultures on collagen and Matrigel at relative high and low seeding densities and compared them to cells from the in vivo condition.

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Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111240 ◽

1984 ◽

Vol 42 ◽

pp. 226-227 ◽

Cited By ~ 1

Author(s):

K. Pegg-Feige ◽

F. W. Doane

Keyword(s):

Electron Microscopy ◽

Cell Culture ◽

Immunoelectron Microscopy ◽

Detection Method ◽

Solid Phase ◽

Protein A ◽

Plant Viruses ◽

Rapid Diagnosis ◽

Virus Diagnosis ◽

Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

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Automated cell counting of astrocytes on patterned substrates containing aliphatic and charged properties

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010014124x ◽

1995 ◽

Vol 53 ◽

pp. 974-975

Author(s):

W. Shain ◽

H. Ancin ◽

H.C. Craighead ◽

M. Isaacson ◽

L. Kam ◽

...

Keyword(s):

Cell Culture ◽

Cell Attachment ◽

Culture Method ◽

Cell Counting ◽

Data Sets ◽

Nuclear Staining ◽

Double Positive ◽

A Cell ◽

Wafer Test ◽

Cell Densities

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.

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