Latex of Ficus carica L. Induces Apoptosis Through Caspase and Bcl-2 Family in FaDu Human Hypopharynx Squamous Carcinoma Cells

Ficus carica L. (common fig), one of the first plants cultivated by humans, originated in the Mediterranean basin and currently grows worldwide, including southwest Asia and South Korea. It has been used as a traditional medicine for treatment of metabolic, cardiovascular, and respiratory diseases as well as hemorrhoids and skin infections. Its pharmacological properties have recently been studied in detail, but research on the anti-cancer effect of its latex has been only been studied on a limited basis on several cell lines, such prostate cancer, breast cancer, and leukemia. In this study, we investigated the anti-cancer activity of the latex of Ficus carica L.and its underlying mechanism in FaDu human hypopharynx squamous carcinoma cells. (See Ed. note above) We confirmed through SDS-PAGE analysis and gelatinolytic activity analysis that the latex of Ficus carica contains cysteine protease ficin. Our data showed that the latex inhibited cell growth in a dose-dependent manner. In addition, the latex treatment markedly induced apoptosis in FaDu cells as determined by FACS analysis, elevated expression level of cleaved caspase-9, -3 and PARP (poly (ADP-ribose) polymerase), and. increased the expression of Bax (pro-apoptotic factor) while decreasing the expression of Bcl-2 (anti-apoptotic factor). Taken together, these results suggested that latex containing the ficin inhibited cell growth and induced apoptosis by caspase and the Bcl-2 family signaling pathway in FaDu human hypopharynx squamous carcinoma cells. These findings point to the potential of latex of Ficus carica to provide a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.

[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

Quantittative analysis of amount and activity of ficin in Khorasan's fig tree latex retrieved from different organs of tree in different seasons

The fig tree latex has been identified as a rich resource of [ficin] an enzyme protein. This enzyme has a vast application in food substances, pharmaceutical and clinical diagnosis, Fig plant can be harvested well in Khorasan province. To determine optimum organ and season to retrieve latex for different purposes. In this study the measuring of the enzyme activity was based on its effect of casein substrate. Enzymatic activity was compared with st and ard papain solution activity. In this research calcium ion concentration as an enzyme activator was studied. The results show that the activity of the Khorasan native fig latex was 50-100 fold more than papain. The amount and the activity of ficin have been variant in different organs of the tree, and also varied with season. Calcium ion concentrations are also variable in different organs and are also season dependent. There is a relationship between calcium ion concentrations and specific activity of ficin. The optimum season for latex collection is fall and the best organ is the branch ends

Cold Agglutinin Disease due to Anti-Pr Cold Autoantibody in a Patient with Bone Marrow Transplantation / 대한수혈학회지

Cold agglutinin disease (CAD) is a most common autoimmune hemolytic anemia (AIHA) induced by cold antibody. CAD represents approximately 16-32% of AIHA cases and causative cold autoantibodies commonly show specificity against the I antigen. We report a case of cold agglutinin disease with anti-Pr cold autohemolysin. A 20 year old woman with a history of bone marrow transplantation was admitted with nausea, vomiting, and pallor. Direct antiglobulin tests were positive with IgG and C3d specific AHG reagents. Cold agglutinin titer was as high as 1:1024 at 4degrees C, 1:16 at room temperature, negative at 37degrees C. The agglutinin titer was diminished after treatment with protease, ficin and immunohematologic results of cold agglutinin was compatible with anti-Pr specificity. In unexpected antibody identification test, anti-M which showed reactivity at anti-human globulin phase was identified. Washed and prewarmed 16 units of A+, M antigen negative red blood cells were transfused. After two weeks, patient was improved with steroid therapy and experienced relief of fever and hemolysis, and she was discharged.

Production of CD44v6 Antibody Fragments and Comparision of Their Speciticities / 대한면역학회지

CD44v6 was known as tumor marker for tumor progression and metastasis in various kinds of carcinomas. The CD44v6 monoclonal antibody was produced by cell cultures or mouse ascite fluids using CD44v6 hybridoma cells, and its immunogloburin G (IgG) was purified by Protein A column. Using immobilized ficin and cysteine, the antibody fragment Fab was produced and purified by Protein A. Four CD44v6 scFv molecules were produced from the recombinant DNA and phage antibody technology and prurified by His-tag affinity chromatography. In order to inspect the function and specificity of each antibody molecule, western-blotting and ELISA against CD44v5-6 recombinant proteins and irnmunodetection in human ovarian carcinomas were estabilished. The results showed that immunodiagnosis did not distinguish the types of antibody fragments, but western-blotting and ELISA results did show some difference of their specificities and biological properties. These studies will contribute as a model study for the immunodiagnosis and therapy using the IgG, Fab and scFv of CD44v6 antibody to obtain the early detection of tumor progression and metastasis using immunoscintigraphy.