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Radiation Carcinogenesis: Tissue Culture Model
Published in Kedar N. Prasad, Handbook of RADIOBIOLOGY, 2020
Protease inhibitors dramatically suppress the radiation-induced transformation of cells in culture.10 A protease inhibitor, such as antipain, by itself does not induce transformations; however, these inhibitors markedly reduce the frequency of transformation. They completely reduce the transformation induced by 400 rads + TPA. A soybean trypsin inhibitor has produced similar results (Figure 17.5).19 Other protease inhibitors from the potato also reduced the radiation-induced transformation frequency by a factor of 2- to 3-fold.20
Inherited Differences in Alpha1-Antitrypsin
Published in Stephen D. Litwin, Genetic Determinants of Pulmonary Disease, 2020
The trypsin-inhibiting activity of serum as reported by several laboratories indicates that between 0.9 and 1.5 mg trypsin is inhibited by 1 ml of serum. These values were determined with commercially available bovine trypsin without correcting for the differences in activity of the trypsin preparations used. As has been pointed out by Laskowski [31], commercial trypsin preparations are often only 50-60% active. Differences in activity between preparations of trypsin can usually explain discrepancies in values that have occasionally been reported. Attempts have been made to standardize trypsin with soybean trypsin inhibitor [32]. However, this has not been satisfactory because inhibitor preparations are frequently also not fully active and therefore cannot be used to quantitate the amount of enzymatically active trypsin in a given preparation.
Interaction of Amebas with Cells
Published in Roberto R. Kretschmer, Amebiasis: Infection and Disease by Entamoeba histolytica, 2020
Much interest has focused on the effects of extracts of amebas on target cells as a model for the investigation of the histolytic activities of the parasite. Homogenates of trophozoites added to monolayers of tissue cultured cells caused the cells to round up and be released from the culture dish. Cells recovered from suspension could be replated and gave rise to viable monolayer cultures as before indicating that the effect was cytotoxic but not cytopathic and resembled that caused by trypsin treatment.53 Like trypsin too, the effect was inhibited by serum although not by classical trypsin inhibitors. A number of groups have partially purified what may be a family of toxin-like molecules with proteinase activity which are thought to be candidates for the histolytic activity of the parasite.54-60 These included a 25 to 45 kDa species57,58 and a group of four proteins with molecular weights of 25, 58, 65, and 68 kDa.56,61 These last four were present in much higher amounts in the more virulent strains of E. histolytica.
The effects of minocycline on proliferation, differentiation and migration of neural stem/progenitor cells
Published in International Journal of Neuroscience, 2020
Fatemeh Shamsi, Zahra Zeraatpisheh, Hamed Alipour, Abbas Nazari, Hadi Aligholi
The ganglionic eminence of 13.5 day old embryo mice was harvested using a stereomicroscope. After mechanical cutting, the specimen was dissociated by adding 0.05% Trypsin/EDTA (Invitrogen, USA) for 5 min at 37°C. Then, trypsin was inhibited by soybean trypsin inhibitor (Sigma, USA). Next, the cells were plated in Dulbecco’s modified Eagle’s medium/F12 (Invitrogen, USA) containing 1% N2 supplement (Invitrogen, USA), 2% B27 supplement (Invitrogen, USA), 1%penicillin/streptomycin, 1% glutamax (Invitrogen, USA), 20 ng/ml epidermal growth factor (EGF; Miltenybiotech, Germany) and incubated at 37°C and 5% CO2. Cells proliferated during the primary culture as free-floating clusters (neurospheres). The mature spheres were passaged every 7 days and replated into fresh growth medium. The cells obtained from the second passage were used for the rest of the study.
Celastrol enhances Atoh1 expression in inner ear stem cells and promotes their differentiation into functional auditory neuronal-like cells
Published in Organogenesis, 2018
Zhao Han, Yu-yan Gu, Ning Cong, Rui Ma, Fang-lu Chi
The BALB/C6 mice, aged 4–7 weeks, were purchased from the Shanghai Laboratory Animal Center (SLAC) and maintained in the pathogen-free environment. All mice experiments were conducted in strict accordance with protocol approved by the institutional committee of animal care and use. The sphere-forming and self-renewing inner ear stem cells were isolated from the utricular sensory epithelium of adult BALB/C6 mice following previously established method.21 Briefly, Mice were first euthanized by cervical dislocation, and the utricular maculae were carefully dissected from mice and rinsed with ice-cold PBS buffer. The sensory sheets were isolated by gentle treatment with thermolysin in DMEM/high glucose-F12 mixture (1:1) and followed by trypsin digestion in PBS. The excessive enzymatic activity was quenched by soybean trypsin inhibitor (Worthington). The cells were dispersed by repeated pipetting and resuspended in mixed culture medium (DMEM/high glucose 1:1 F12 with complete supplementation, Gibco, Grand Island, NY, USA). The single-cell suspension was prepared by passing through the 70 μm cell strainer (BD, Franklin Lakes, NJ, USA) and subjected to continuous culture in humidified incubator supplemented with 5% CO2 at 37°C.
Antibacterial action mechanisms and mode of trypsin inhibitors: a systematic review
Published in Journal of Enzyme Inhibition and Medicinal Chemistry, 2022
Amanda Maria de Souza Nascimento, Victor Hugo de Oliveira Segundo, Ana Júlia Felipe Camelo Aguiar, Grasiela Piuvezam, Thaís Souza Passos, Karla Suzanne Florentino da Silva Florentino da Silva Chaves Damasceno, Ana Heloneida de Araújo Morais
In this protocol, the scope of the review was registered, which included in vitro studies (bacterial culture), in vivo (rats and mice), and using cells, in which trypsin inhibitor-type peptides were administered to discover the mechanisms involved in the antibacterial action of these inhibitors. Primary endpoints were considered a mechanism of action, and secondary endpoints were a mode of action antibacterial of trypsin inhibitor-type peptides or proteins. It is noteworthy that antimicrobials can be classified per the mechanism of action, or even as to the mode of action or effect on microorganisms [classification based on pharmacodynamic parameters such as minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)]21.