One of the adhesive cells culturing difficulties, is the damage they bear when dislodgement from culture flask. These injuries often cause a lot of cells destruction and mortality. In this article two methods of dislodgment of adhesive cells from culture Basks have been briefly explained. The first method involves 0.0%-0.07%. TRYPSIN solution containing 1 m mol of EDTA and the second one contains 5-10m mol EDTA solution having albumin 1mg/ml.In albumin method the less EDTA solution is concentrated, the more time is needed to collect the cells. In TRYPSINizing method, for the cells of which adhesive power is higher, or if under specific care cell adhesiveness rises, higher TRYPSIN percentage should be used.

Herein, the protein corona formation on the spherical metal nanoparticles is studied to investigate the possible effects of silver nanoparticles (AgNPs) on the protein activity and conformation. The digestion capability of TRYPSIN was monitored on the human serum albumin (HSA) at standard enzymatic hydrolysis conditions in the absence and presence of different concentrations of AgNPs. So, the ratio of enzyme: HSA, the duration and the temperature of nanoparticle treatment were evaluated. The activity of treated TRYPSIN molecules, in the form of hard (HC) and soft corona (SC) was studied using sodium dodecyl sulfate poly acrylamide gel electrophoresis (SDS-PAGE), and nano liquid chromatography electrospray (ion trap) mass spectrometry (nano LC-ESI/MS, LC-ESI/MS). In addition, the characteristics of silver nanoparticles and the formation of HSA corona on the nanoparticle surface were monitored using ultra-violet/visible spectroscopy (UV-Vis), Dynamic light scattering (DLS) and fluorescence spectrophotometry. The results demonstrated that not only the corona formation but also the AgNPs/TRYPSIN interaction decreases the hydrolysis potency of TRYPSIN. Furthermore, interaction of AgNPs/HSA could influence both nanoparticles and HSA molecule features, companied with fluorescence study, that the HSA secondary structure. Also, LC-ESI/MS data revealed the most affected HSA triptics have α-helix structure.

Curcumin, a polyphenol yellow orange pigment present in Indian spice “turmeric”, is a member of the ginger family (Zingiberaceae). The extract of turmeric was obtained by using soxhlet apparatus at 45-55 oC, using methanol as solvent and then curcumin was isolated from methanolic extract by column chromatography. In the present study, a series of four new derivatives of curcumin (3a-d) were synthesized by reacting curcumin (1) with different alkyl halides (2a-d) using DMF and lithium hydride. The structures of natural curcumin and its synthetic derivatives were confirmed with the help of their 1H-NMR spectra and all these were screened against acetylcholinesterase, butyrylcholinesterase, lipoxygenase and TRYPSIN enzymes. Curcumin and its derivatives exhibited moderate inhibitory potential against all aforesaid enzymes.

In recent years, increased interest is seen among scientist and pharmaceutical industries to replace the synthetic antibiotics with antimicrobial agents that are perceived as “natural”. Currently, peptides with known sequences have been identified which exhibit antimicrobial effects. These peptides are produced in vitro and in vivo by proteolysis of naturally occurring parent proteins. Ovotransferrin (OT) is an egg white protein with iron binding properties. The purpose of this research was to isolate, purify and characterize novel antimicrobial peptides from ficin-and TRYPSIN-treated OT. OT was purified from chicken egg white by ammonium sulfate precipitation followed by DEAE- Sephadex ion exchange chromatography and subjected to treatment with different concentrations of ficin and TRYPSIN. The produced peptides were separated by Sephadex G-50 gel filtration chromatography. The antimicrobial effects of selected peptides against Staphylococcus aureus (G+) and Salmonella typhimurium (G-) was evaluated. Results showed that purified peptides decreased the number of S. aureus and S. typhimurium by 97% and 28% respectively. Taken together, the results of this study suggest that it is possible to substitute the antimicrobial agents of chemical origin with the peptides which are obtained from OT.

Background: Despite the extensive information available in the literature, cell surface marker signature of human Amniotic Epithelial Cells (hAECs) remains controversial. The aim of the present study was to characterize immunopheno-typic features, proliferative capacity and immunogenicity of hAECs. We also tested whether expression of some cell surface markers is influenced by the type of TRYPSIN used for tissue digestion.Methods: Single cell suspensions of amniotic membranes from four human placentas were isolated by enzymatic digestion and expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, HLA-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. The differential impact of four TRYPSIN types on the yield and expression of CD105 and HLA-I was also determined. The proliferative capacity of cultured hAECs was assessed and compared in the presence and absence of Epidermal Growth Factor (EGF). To test their immunogenicity, hAECs were injected into Balb/c mice and the reactivity of hyperimmunized sera was examined by immunofluorescence staining.Results: Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73 and the embryonic marker, SSEA-4. A large proportion of the cells also expressed STRO-1 and OCT-4. The purified cells also expressed HLA-G and HLA-I. A very small proportion of hAECs expressed CD34, CD38, CD44, CD133 and HLA-DR. The type of TRYPSIN used for enzymatic digestion affected both the percentage and expression of HLA-I and CD105. hAECs revealed substantial proliferative capacity only when cultured in the medium supplemented with EGF. These cells were shown to be capable of inducing high amounts of anti-donor antibodies.Conclusion: Here we provided evidence that hAECs are immunogenic cells with high level of HLA-I expression. Furthermore, this work highlighted the impact of isolation procedure on the immunophenotype of hAEC.