Tear protein interaction with hydrogel contact lenses

The design and synthesis of biomaterials covers a growing number of biomedical applications. The use of biomaterials in biological environment is associated with a number of problems, the most important of which is biocompatabUity. If the implanted biomaterial is not compatible with the environment, it will be rejected by the biological site. This may be manifested in many ways depending on the environment in which it is used. Adsorption of proteins takes place almost instantaneously when a biomaterial comes into contact with most biological fluids. The eye is a unique body site for the study of protein interactions with biomaterials, because of its ease of access and deceptive complexity of the tears. The use of contact lenses for either vision correction and cosmetic reasons or as a route for the controlled drug delivery, has significantly increased in recent years. It is relatively easy to introduce a contact lens Into the tear fluid and remove after a few minutes without surgery or trauma to the patient. A range of analytical techniques were used and developed to measure the proteins absorbed to some existing commercial contact lens materials and also to novel hydrogels synthesised within the research group. Analysis of the identity and quantity of proteins absorbed to biomaterials revealed the importance of many factors on the absorption process. The effect of biomaterial structure, protein nature in terms of size. shape and charge and pH of the environment on the absorption process were examined in order to determine the relative up-take of tear proteins. This study showed that both lysozyme and lactoferrin penetrate the lens matrix of ionic materials. Measurement of the mobility and activity of the protein deposited into the surface and within the matrix of ionic lens materials demonstrated that the mobility is pH dependent and, within the experimental errors, the biological activity of lysozyme remained unchanged after adsorption and desorption. The study on the effect of different monomers copolymerised with hydroxyethyl methacrylate (HEMA) on the protein up-take showed that monomers producing a positive charge on the copolymer can reduce the spoilation with lysozyme. The studies were extended to real cases in order to compare the patient dependent factors. The in-vivo studies showed that the spoilation is patient dependent as well as other factors. Studies on the extrinsic factors such as dye used in colour lenses showed that the addition of colourant affects protein absorption and, in one case, its effect is beneficial to the wearer as it reduces the quantity of the protein absorbed

A real-time biosensor system in vivo using luciferase as an intracellular reporter for the functional analysis of anti-cancer compounds based on Hsp70 inhibition

Introduction:Defective apoptosis (programmed cell death) is a major contributor to the process of cancer progression and metastasis. Over the last decade, new drug compounds were designed to enhance apoptosis in the tumor cells. Hsp70 chaperone is an anti-apoptotic protein and its inhibitors serve as suitable compounds in cancer treatment strategies. A number of different techniques such as x-ray crystallography are available for studying inhibitory effects of apoptosis-inducing compounds on Hsp70 activity. These techniques are expensive and time-consuming because of long sample preparation steps prior to analysis, and the functional analysis of new compounds synthesized cannot be examined in vivo. In order to overcome these problems, for the first time, a rapid, sensitive and inexpensive real-time biosensor system in vivo is introduced using intracellular reporter. In real-time biosensor system, because of the availability of a rapid and sensitive activity assay using biolominometer in vivo without cell lysis, luciferase has been considered as proper candidate. Methods and Results:Co-transformation of the cells was carried out with two expression vectors containing Hsp70 and firefly luciferase, and the assessment of new drug compounds on Hsp70 activity under the stress conditions was evaluated. Our result showed that Hsp70 plays a crucial role in suppressing inactivation of luciferase in vivo during heat-treatment. On the other hand, the activity of heat-inactivated luciferase was approximately regained in cells expressing Hsp70 after incubation at room temperature for 60 min. According to the results, the reactivation of thermally inactivated luciferase was inhibited in the cells by treating with Ver-155008 and PFT-μ compounds (as apoptosis-inducing compounds) with IC50 of 124 and 384 μM, respectively. Also, the sensitivity of this method for detecting Ver-155008 and PFT-μ compounds was as low as 8 and 23 μM, respectively, and it showed no response to doxorubicin anti-cancer drug which binds to DNA, and used as control. Conclusions:Real-time biosensor system using luciferase can be a class of in vivo biochemical tests used for simple, rapid and sensitive detection of apoptosis-inducing compounds based on Hsp70 inhibition to facilitate clinical and therapeutic studies in the years to come

Glutathione Peroxidase1 Gene Polymorphism (GPx1 Pro198Leu) in Association with Blighted Ovum

Objective: To evaluate salivary GPx-1 gene polymorphism in pregnant women suffering from blighted ovum. Method: In this case-control study, 34 blighted ovum patients and 34 healthy controls were studied. Genomic DNA was extracted from the saliva. The genotypes were determined by restriction fragment length polymorphism (RFLP-PCR) technique. Mad Calc (version 12.1) was used for statistical analysis. Result: The frequency of CC, CT, and TT genotypes of GPx-1 gene were 41%, 44% and 14%, respectively in blighted ovum patients and in healthy volunteers were 44%, 47%, and 8.82-9%, respectively. After statistical analysis, the study showed no significant association between this polymorphism and blighted ovum (with p = 0.63). Conclusion: These results indicated no significant association between GPx-1 (Pro198Leu) polymorphism and blighted ovum. However, further research is required to clarify the role of gene polymorphism in blighted ovum. [Indones J Obstet Gynecol 2016; 1: 15-18] Keywords: abortion, blighted ovum, glutathione peroxidase-1, GPx- 1, RFLP-PC

A novel approach to the synthesis of the purine anti-viral agent gancyclovir

651-654The aim of this research is to synthesize gancyclovir, a purine anti-viral drug in large scale via simple intermediates. Reaction of guanine 1 with acetic anhydride followed by the attachment of the 2-O-acetoxymethyl-1,3-di-O-benzyl glycerol 3 as a side chain, subsequently lead to the production of N2acetyl-9-(1,3-dibenzyloxy-2-propoxymethyl) guanine 6 and N2,acetyl-9-(1,3-dihydroxy-2-propoxy methyl)guanine 7 and finally synthesizing gancyclovir 8. Among many pathways to the synthesis of purine derivatives, our four step procedure results in good yield and is proved to be an economic way of large scale synthesis

A Novel Approach to the Synthesis of the Purine Antiviral Agent Gancyclovir.

651-654The aim of this research is to synthesize gancyclovir, a purine anti-viral drug in large scale via simple intermediates. Reaction of guanine 1 with acetic anhydride followed by the attachment of the 2-O-acetoxymethyl-1,3-di-O-benzyl glycerol 3 as a side chain, subsequently lead to the production of N2acetyl-9-(1,3-dibenzyloxy-2-propoxymethyl) guanine 6 and N2,acetyl-9-(1,3-dihydroxy-2-propoxy methyl)guanine 7 and finally synthesizing gancyclovir 8. Among many pathways to the synthesis of purine derivatives, our four step procedure results in good yield and is proved to be an economic way of large scale synthesis

Effect of Ramadan Fasting on Tear Proteins

Muslims abstain from eating, drinking and smoking from dawn to sunset during the holy month of Ramadan. Prolonged fasting is thought to be among risk factors for many diseases, e.g., cardiovascular, gastrointestinal and various infectious diseases. It could also play a part in several eye diseases, including dry eye syndrome, glaucoma, and cataract. Toxic and oxidative effects due to increased concentrations of some biochemicals as a result of reduction in tear volume thought to play an important role in damaging ocular tissue. Human tear is an important biological fluid similar to blood in many aspects. Tear film is composed of three basic layers i.e. lipid, aqueous and mucin. The tear film covering the ocular surface presents a mechanical and antimicrobial barrier, and endures an optical refractive surface. The aim of this study was to analyze and compare tear protein of volunteers during fasting. Using two reliable analytical methods, i.e. electrophoresis and high performance liquid chromatography (HPLC), we compared tear protein content of sixty volunteers (35 males and 25 females, 23–27 years old) during fasting in holly month of Ramadan (FAST: n=62) and one month before Ramadan (CTRL: n=60). The results showed that some identified tear proteins decreased during fasting. On the other hand, the activity of some enzymes such as lysozyme, lactoferrin and alpha amylase also decreased in fasting samples. Electrophoresis results showed that tear protein patterns in FAST (PP<0.005) than in CTRL

Thymoquinone synergistically potentiates temozolomide cytotoxicity through the inhibition of autophagy in U87MG cell line

Objective(s): Glioblastoma multiforme (GBM) is one of the most lethal forms of human cancer and temozolomide (TMZ) is currently part of the standard treatment for this disease. Combination therapy using natural substances can enhance the anti-cancer activity of TMZ. The purpose of this study was to evaluate the effect of TMZ in combination with thymoquinone (TQ) on human GBM cell line (U87MG). Materials and Methods: The cell line was treated with TMZ and/or TQ. Cell viability was assessed using trypan blue and MTT assay. The effect of TMZ and/or TQ on colony-forming ability of the cells was investigated. Apoptosis and autophagy were quantified by fluorescent dye staining. The expression level of two autophagy related genes (ATG) were assessed using RT-PCR. Furthermore, nitric oxide (NO) production was detected by Griess reaction. Results: After treatment with TMZ and/or TQ, the cell viability was reduced in a time- and dose-dependent manner, and the cell survival fraction (SF) was significantly decreased (P=0.000). Apoptosis index of U87MG cells was also significantly increased (P=0.000). Autophagy was significantly increased by TMZ (P=0.000) and decreased by TQ (P=0.018). Also TMZ and/or TQ significantly decreased NO production by U87MG cell (P=0.000). Conclusion: TQ enhanced the anti-cancer activity of TMZ by inhibition of autophagy at the transcriptional level and decreased the colony-forming ability and NO production of U87MG cell line