Evaluating pre-treatment methods in the isolation and purification of monoclonal IgG antibody

In this study, the effects of different sample preparation techniques on the separation of monoclonal antibody IgG I were inve st igated experimentally. Monoclonal IgG I was obtained from hybridoma cell line TB /C3 transfected with bcl-2 carrier plasmid. which was grown in serum-free medium. Three different pre-treatment techniques prior to Protein G affinity chromat ography have been used in order to concentrate and partial purify the monoclonal antibody . The pre-treatments researched in this paper are precipitation of the antibody by ammonium sulfate, dilution of the antibody in the binding buffer of affinity chromatography and ultra filtration through an Amicon Ultra- 15 filter with molecular weight cut-off at 100 kDa. Purification through direct application of the antibody onto the Protein G affinity column without pre-treatments was used as a control method. The results indicate that the ultra filtration through an Amicon filter was an effective method for both concentration and partial purification of the antibodies in laboratory scale

A proanthocyanidin from cinnamomum zeylanicum stimulates phosphorylation of insulin receptor in 3t3-l1 adipocytes

Non-insulin-dependent diabetes mellitus is due to in large part to insulin resistance, a state where the target cells are no longer responding to ordinary levels of circulating insulin. A drug that promotes the initiation of insulin receptor (IR) signaling by enhancing IR phosphorylation should be useful in the treatment of Type 2 diabetes. To investigate a compound that is able to phosphorylate IR, a proanthocyanidin, cinnamtannin B1, has been isolated from the stem bark of Cinnamomum zeylanicum Blume (Lauraceae). The structure of the compound was established by spectroscopic methods. Cinnamtannin B1 (0.11 mM) activates the phosphorylation of insulin receptor â-subunit on 3T3-L1 adipocytes. Like insulin, cinnamtannin B1-stimulated phosphorylation of insulin receptor was inhibited by wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PI3K) and cytochalasin B, an inhibitor of the glucose transporter (GLUT4); otherwise the phosphorylation was enhanced by vanadate, a tyrosine phosphatase inhibitor. These results suggest that the activity of cinnamtannin B1 is specially mediated by phosphorylation of IR through activation of the PI3K

Effect of various processing parameters on the quality of papaya fruit tea

Exotic fruits from which drinks are ma de in most tropical countries are so abundant and such in a great variety, that it would probably be impossible for people to sample all of them, but they worth a try. Some of the great varieties of fruits used in drinks are bananas, guavas, papayas, oranges, pineapples, grapes, mangoes, watermelon, coconuts, longans, rambutans, strawberries, lime and tamarind

Generation of New Hybridoma UTM-Ha Secreting Monoclonal Antibody Specific to Acanthamoeba species Isolated from Corneal Infection

Acanthamoeba Keratitis is an important corneal infection that caused impaired vision. The Universiti Malaysia Terengganu (UMT) researcher found new species of Acanthamoeba that was being isolated from eye infection in Hospital Kuala Lumpur, which have the characteristic between Acanthamoeba polyphaga and A. castellanii named as HKL-Acantha. This study was aimed to generate specific monoclonal antibody against HKL-Acantha that eventually could be used in diagnosing eye infection among Malaysian. Two Balb/c mice were immunized with sonicated HKL-Acantha through intraperitoneal injection, and anti HKL-acantha IgG, IgM, IgA were measured using ELISA test. Sera from infected mice showed detectable anti-HKL-acantha IgM. The spleen from animal with high antibody titer around (1:81000) was used as a partner in fusion with SP2/0-Ag14 myeloma cells to produce new hybridoma cells, which were then selected and cloned using the selection medium. Two positive hybridoma clones secreting IgM antibodies were obtained and named as hybridoma UTM-Ha1 and UTM-Ha2. Both clones were secreting monoclonal IgM antibody specific to HKL-Acantha. This study suggests the potential of both hybridomas UTM-Ha1 and UTM-Ha2 to generate specific monoclonal IgM against HKL-acantha. This newly generated monoclonal IgM could be used to diagnose the presence of HKL- acantha in patients with eye infection in future

Cinnamtannin B1: a small compound having antidiabetes properties

Type 2 diabetes mellitus is a chronic metabolic disease that results from defects in insulin secretion and insulin receptor kinase. Investigation of novel small active molecule that can potentiate insulin action or having a similar action as insulin is important in the treatment of diabetes. World ethnobotanical information on medicinal plants reports almost 800 plants used in the treatment of diabetes mellitus. However, only a small number of them have been studied thoroughly. Recent study conducted on Cinnamomum reported that it has a great activity in activating insulin receptor kinase and inhibiting insulin receptor phosphatase leading to increased insulin sensitivity and function as a mimetic for insulin. Our study was designed to investigate insulin-mimetic activity of cinnamtannin B1 isolated from Cinnamomum zeylanicum on adipocyte cells. The insulin-mimetic activity of cinnamtannin B1 was evaluated by monitoring preadipocytes differentiation, glucose uptake and phosphorylation of insulin receptor -subunit in 3T3-L1 adipocytes. To determine whether cinnamtannin B1 able to promote differentiation of preadipocytes, we cultured 3T3-L1 preadipocytes in the presence of cinnamtannin B1, or combination of cinnamtannin B1 and insulin, and then cell proliferation was measured at several points during the course of growth. Investigation of role of cinnamtannin B1 on tyrosine posphorylation of insulin receptor of 3T3-L1 cells was done by immunoprecipitation of cells lysate with anti-insulin receptor -subunit antibody and the immunocomplex samples were subjected to SDS-PAGE, transferred to nitrocellulose membranes, and immunoblotted with monoclonal anti- phosphotyrosine antibody. Evaluation of glucose uptake by adipocyte cells after treatment with cinnamtannin B1 was carried out by analyzing of radioactive glucose uptake with liquid scintillation counter. Based on these experiments, it was found that, a mixture cinnamtannin B1 with differentiation cocktail was able to induce differentiation of preadipocytes cells. Cinnamtannin B1 was found to active to stimulate phosphorylation of insulin receptor -subunit by positively exhibited phosphorylation at 170-kDa. The mixture of cinnamtannin B1 was also able to stimulate glucose uptake from a basal value. The results demonstrated that activity of cinnamtannin B1 on adipocyte cells was found to mimicking insulin action. It acted directly on insulin receptor -subunit by activation of PI3-kinase that stimulates glucose transporter-4 (GLUT-4) translocation. Stimulation of GLUT4 translocation therefore stimulates glucose uptake lead to glucose disposal process in adipocytes. Based on the work that has been carried out, it was suggested that cinnamtannin B1 could be one of the potential lead drug compound in the treatment of type 2 diabetes

Development of DiabecineTM tablet and confirmation of its physical properties and pharmaceutical safety analysis

Herbal medicine is usually made using dry powdered herbs in the form of capsule. Capsule form herbal supplement suffers lower shelf life as compared to compact herbal powder in tablet form. In this study, Diabecine™, a blend of herbal medicine traditionally used as herbal supplement for diabetic patients was selected and transformed into a compressed tablet. Direct compression method and minimal usage of excipients were the aims of this study. By using direct compression, the blend of 40% fine powder herbs and 60% of excipients performed the best and fulfill the pharmaceutical standard. The safety data of microbial and heavy metal testing obtained met the safety requirements for herbal supplement category under the National Pharmaceutical Control Bureau of Malaysia. In conclusion, the tablet formulation of DiabecineTM is suitable to be manufactured by using direct compression method. This research implicates the possibility of producing tablets with high dose of herbal powder by direct compression method

Cinnamtannin B1: a small compound having antidiabetes properties

Type 2 diabetes mellitus is a chronic metabolic disease that results from defects in insulin secretion and insulin receptor kinase. Investigation of novel small active molecule that can potentiate insulin action or having a similar action as insulin is important in the treatment of diabetes. World ethnobotanical information on medicinal plants reports almost 800 plants used in the treatment of diabetes mellitus. However, only a small number of them have been studied thoroughly. Recent study conducted on Cinnamomum reported that it has a great activity in activating insulin receptor kinase and inhibiting insulin receptor phosphatase leading to increased insulin sensitivity and function as a mimetic for insulin. Our study was designed to investigate insulin-mimetic activity of cinnamtannin B1 isolated from Cinnamomum zeylanicum on adipocyte cells. The insulin-mimetic activity of cinnamtannin B1 was evaluated by monitoring preadipocytes differentiation, glucose uptake and phosphorylation of insulin receptor β-subunit in 3T3-L1 adipocytes. To determine whether cinnamtannin B1 able to promote differentiation of preadipocytes, we cultured 3T3-L1 preadipocytes in the presence of cinnamtannin B1, or combination of cinnamtannin B1 and insulin, and then cell proliferation was measured at several points during the course of growth. Investigation of role of cinnamtannin B1 on tyrosine posphorylation of insulin receptor of 3T3-L1 cells was done by immunoprecipitation of cells lysate with anti-insulin receptor β-subunit antibody and the immunocomplex samples were subjected to SDS-PAGE, transferred to nitrocelluose membranes, and immunoblotted with monoclonal anti-phosphotyrosine antibody. Evaluation of glucose uptake by adipocyte cells after treatment with cinnamtannin B1 was carried out by analyzing of radioactive glucose uptake with liquid scintillation counter. Based on these experiments, it was found that, a mixture cinnamtannin B1 with differentiation cocktail was able to induce differentiation of preadipocytes cells. Cinnamtannin B1 was found to active to stimulate phosphorylation of insulin receptor β-subunit by positively exhibited phosphorylation at 170-kDa. The mixture of cinnamtannin B1 was also able to stimulate glucose uptake from a basal value. The results demonstrated that activity of cinnamtannin B1 on adipocyte cells was found to mimicking insulin action. It acted directly on insulin receptor β-subunit by activation of PI3-kinase that stimulates glucose transporter-4 (GLUT-4) translocation. Stimulation of GLUT4 translocation therefore stimulates glucose uptake lead to glucose disposal process in adipocytes. Based on the work that has been carried out, it was suggested that cinnamtannin B1 could be one of the Annals of Medicine & Healthcare Research Page 452 potential lead drug compound in the treatment of type 2 diabetes

Human papilloma virus E6/E7 messenger RNA as a biomarker for detecting the risk evaluation of cervical cancer progression: overview of recent clinical trials

Cervical cancer is one of the major women health problems, which is considered to be responsible for significant percentage of cancer related deaths in low and middle income countries. While Human Papilloma virus (HPV) DNA testing has an established role in cervical cancer prevention, there is a need to use other biomarkers with higher specificity and prognostic value to recognize patients at risk of progressive illness. There are evidences suggest that, direct detection of HPV messenger RNA transcripts may establish a more specific method for defining clinically important infection than viral DNA detection. Our objective was to provide an overview of the literature on specificity of HPV mRNA testing compared to DNA testing for detecting the risk evaluation of cervical intraepithelial neoplasia and invasive cervical cancer. We focused on recent clinical studies that support the application of HPV E6/E7 mRNA as a marker in advanced cervical cancer screening program. We provide overview of sample size, recruitment setting, age, HPV mRNA and HPV DNA assays for researches included in our review. The pooled review of clinical studies provided evidence that HPV mRNA might be a relevant diagnostic biomarker but additional studies need to be developed in order to make strong conclusio

Design, fabricate and testing of stationary magnetic stirrer bar for uniform mixing of cell culture

Magnetic stirrer bar is widely used in bio-chemical field for mixing of cell cultures. Increases in rotation speed often caused the magnetic stirrer bar to be spinoff from its position and leads to death of the cell cultures. Design of the magnetic stirrer bar accessory, three point arms is the main focus of the research. The attachment of this additional accessory has eliminated the spin-off effect. The hydrodynamic behavior of this design has been simulated using GAMBIT and FLUENT simulation software. The simulated fluid movement has shown to have a uniform mixing and break the vortex caused by normal magnetic bar. The accessory has been fabricated and being tested for real mixing of cell lives

Development of Diabecine™ tablet and confirmation of its physical properties and pharmaceutical safety analysis

Herbal medicine is usually made using dry powdered herbs in the form of capsule. Capsule form herbal supplement suffers lower shelf life as compared to compact herbal powder in tablet form. In this study, Diabecine™, a blend of herbal medicine traditionally used as herbal supplement for diabetic patients was selected and transformed into a compressed tablet. Direct compression method and minimal usage of excipients were the aims of this study. By using direct compression, the blend of 40% fine powder herbs and 60% of excipients performed the best and fulfill the pharmaceutical standard. The safety data of microbial and heavy metal testing obtained met the safety requirements for herbal supplement category under the National Pharmaceutical Control Bureau of Malaysia. In conclusion, the tablet formulation of Diabecine™ is suitable to be manufactured by using direct compression method. This research implicates the possibility of producing tablets with high dose of herbal powder by direct compression method