[Corneal neovascularization: review article]

Corneal neovascularization occurs through inordinate wound healing after infection, injury or surgery. Neovascularization is formation of new vascular structures in the locations which had not already vessels. The two overlapping mechanisms including vasculogenesis and angiogenesis are probably involved in neovascularization process, and the last mechanism is more involved in tumor growth and corneal and retinal disorders. In fact, corneal neovascularization is a visual threatening status that usually occurs along with inflammatory or infectious disorders of the eye surface. The studies of angiogenesis-related cancer showed that there is a balance between angiogenic factors [such as VEGF and FGF] and antiangiogenic molecules [such as angiostatin, endostatin and pigment epithelium-derived factor; EPDF] in cornea. Problems such as inflammation, infection, injury and lesions result in corneal neovascularization, which are due to stimulation of angiogenesis in this tissue. Corneal neovascularization may be influenced by matrix inetalloproteinase [MMPs] and other proteolytic enzymes. The application of new medical and surgical therapies such as angiostatic steroids, non-steroidal anti inflammatory drugs, argon laser photocoagulation and photodynamic therapy [PDT] in animal models had been efficient to some extent for inhibition of corneal neovascularization. In this study we reviewed neovascularization-dependent corneal disorders and molecular processes involved in this disorder, and also their potential therapies

[Bovine casein hydrolysis using green Ficus carica extract]

The problem of casein digestion is seen in some cases specially, among infants. In this study, it is attempted to consider the hydrolysis of cow's milk casein by means of Ficin enzyme in green Fig extract. Cow's milk casein intolerance occurs in a considerable number of infants and without early diagnosis and food replacement can lead to complications like malnutrition and growth retardation. Hydrolysis of cow's milk casein by chemical and enzymatical reactions is one of the main strategies for production of hydrolyzed milk for allergic infants. In this study there has been an effort to use green fig proteases for cow's milk hydrolysis as a new and cheap source. The aqueous extract was prepared from green fruit of F. carica using phosphate buffered saline. To evaluate the degree of hydrolysis, different concentrations of extract were added to casein and incubated for 1, 3 and 6 hours at various conditions including four buffering system including tris-HCl [pH 8.5] phosphate [pH 7], citrate [pH 5.5] and acetate [pH 4.5] buffers. Hydrolysis of casein was assessed by SDS-PAGE and band densitometry estimated using Scion lab soft ware. The results indicated that fig extract could hydrolyze pure casein completely at ratio 1/100 [enzyme to substrate] for one or three hours and at 1/500 hydrolyzed casein partially at 6 hours at phosphate buffer. Therefore, phosphate buffer system is more suitable than other buffer systems for casein hydrolysis. The results of this study showed that ficin can be used as a potential protease for the hydrolysis of milk casein which is used for nutritional targets

Purification of Kunitz soybean trypsin inhibitor using affinity chromathography

soybean trypsin inhibitor, a single chain protein with 181 amino acids and two disulfide bonds, has special tendency for pancreatic trypsin enzyme. This protein exists in soybean abundantly and showes high resistance to heating and chemicals. Various procedures such as combination of different chromatographic methods, immobilized metal affinity chromatography and preparative electrophoresis have been used to purify this protein. Pulverized soybean was defatted with methanol and the remainder was extracted after being solved in dionized water. Storage proteins in solution was eliminated by use of isollectric precipitation method. Trypsin inhibitor riched fraction was adsorbed on an affinity column with trypsin as the ligand. After washing unbonded proteins, trypsin inhibitor was eluted from the collumn by decreasing pH. Purity and activity of the inhibitor was assessed by SDS-PAGE and an spectrophotometric method, respectively. SDS-PAGE demonstrated a protein with a single band and molecular weight of 20 KDa with purity of more than 99 percent under reducing and non-reducing conditions. Activity of the purified inhibitor protein was 3600 TlU/mg. The results showed that overloading of the affinity collumn to gether with chromatography at pH of 6 to 8 increased the purity. The proposed method is simple and efficient for preparation of large amounts of pure trypsin inhibitor. In addition, this method does not need any further steps such as dialysis