Isolation, Purify and Characterization of Lectin from the Seeds of Artocarpus Species

Aim: To isolate, partially purify and characterize lectin from the seeds of Artocarpus species - using ammonium sulphate, gel-filtration and ion exchange chromatography. Methodology: ABO blood groups were screened for the assay of hemagglutinating property with seed lectins of both. heterophyllus and A. hirsutus. The seed lectins were extracted using suitable buffer system pH 7.0 and partially purified by gel filtration and ion exchange chromatographic techniques. Results and Discussion: Lectins were extracted from the defatted seed powders of A. heterophyllus and A. hirsutus. The extracts were used for optimization of assay buffer for hemagglutination activity and 50 mM Tris-HCl buffer, pH 8.8 containing 1 mM CaCl2 and 1 mM MnCl2 showed greater hemagglutinating property. Screening for optimum concentration of RBCs from different blood group (A, B, AB and O groups) indicated O group at 10 % concentration as ideal for the assay of hemagglutinating property with seed lectins of both A. heterophyllus and A. hirsutus. Conclusion: In this study, lectin from the seeds of Artocarpus heterophyllus and Artocarpus hirsutus were isolated and partially purified. The isolated lectins were characterized for their heamagglutination activity. Among the human blood types (A+, B+, AB+ and O+) used, all the blood groups showed agglutination while greater agglutination was observed with O+ blood group.

Development of an Orodispersible Film Containing Stabilized Influenza Vaccine

Most influenza vaccines are administered via injection, which is considered as user-unfriendly. Vaccination via oral cavity using an orodispersible film (ODF) might be a promising alternative. To maintain the antigenicity of the vaccine during preparation and subsequent storage of these ODFs, sugars such as trehalose and pullulan can be employed as stabilizing excipients for the antigens. In this study, first, β-galactosidase was used as a model antigen. Solutions containing β-galactosidase and sugar (trehalose or trehalose/pullulan blends) were pipetted onto plain ODFs and then either air- or vacuum-dried. Subsequently, sugar ratios yielding the highest β-galactosidase stability were used to prepare ODFs containing H5N1 whole inactivated influenza virus vaccine (WIV). The stability of the H5N1 hemagglutinin was assessed by measuring its hemagglutination activity. Overall, various compositions of trehalose and pullulan successfully stabilized β-galactosidase and WIV in ODFs. WIV incorporated in ODFs showed excellent stability even at challenging storage conditions (60 °C/0% relative humidity or 30 °C/56% relative humidity) for 4 weeks. Except for sugars, the polymeric component of ODFs, i.e., hypromellose, possibly improved stability of WIV as well. In conclusion, ODFs may be suitable for delivering of WIV to the oral cavity and can possibly serve as an alternative for injections.

SPECIFIC ASPECTS OF AVIBACTERIUM PARAGALLINARUM INACTIVATION WITH FORMALDEHYDE AND THIOMERSAL

The paper demonstrates results of testing different modes of Avibacterium paragallinarum inactivation with formaldehyde and thiomersal. The bacterium destruction by 0.20% and 0.10% formaldehyde proceeds at the constant rate thus indicating exponential dependence of the microorganism inactivation processes. This fact allows for calculation of the inactivation rate constant that amounts to 2.94 ± 0.37 h-1 for 0.10% formaldehyde and 5.86 ± 0.72 h-1 for 0.20% formaldehyde. Inactivation using formaldehyde at final concentration of 0.10% at 37 °С and continuous stirring (60 rpm) produces 7.0 dm3 of bacterin at concentration of 9.5 ± 0.2 lg microbial cells (mc)/cm3 in 4.3 ± 0.1 h. Thiomersal demonstrated bactericidal action against Avibacterium paragallinarum at concentration of 0.04% (1:2500) or higher. Herewith, inactivation process is specified by linearity and the inactivation rate constant amounts to 7.92±1.12 h-1. Under thiomersal sublethal concentration of 0.2% (1:5000) the survival curve is of irregular shape. However, the process of the microorganism death is not exponential, and under continuous decrease, the inactivation rate is going to zero thus making impossible the calculation of the inactivation rate constant. Inactivation mode involving use of 0.04% thiomersal at 37 °С allows production of 7.0 dm3 of bacterin at 9.5 ± 0.2 lg mc/cm3 concentration in 5.8 ± 0.1 h. Right after production, the hemagglutination activity of the thiomersal inactivated antigen was higher as compared to formaldehyde inactivated antigen (Р 0.05). The antigen produced using formaldehyde maintains high hemagglutination activity during storage that is critical for high quality vaccine production.

Purification and Characterization of a New Lectin from Loach Skin Mucus

Lectin from loach skin mucus plays an important role in pathogen defense. However, hardly can any paper relevant to the character of lectin from loach skin mucus be found in recent years. In this study, a kind of new lectin (LML), with a high hemagglutination activity of 166.23 × 103 HU/mg, was successfully isolated and purified from loach skin mucus. LML was a kind of glycoprotein with a molecular weight of 245 kDa. Also, the monosaccharide composition suggested that its carbohydrate chain was composed of rhamnose, arabinose, xylose, mannose, glucose, and galactose with a molar ratio of 2.02 : 11.66 : 2.06 : 1.00 : 14.09 : 6.00. Besides, LML depended on Ca2+ to induce hemagglutination and was strongly inhibited by D-lactose. The lectin exhibited powerful resistance to alkali and kept about 30% hemagglutination activity at pH 14.0, whereas its capacity of acid resistance was weak. The maximum hemagglutination activity of LML maintained at a temperature range from 20°C to 50°C. Moreover, the structure of LML was preliminarily studied, indicating it contained abundant glutamic acid, histidine, and serine, and its secondary structure contained α-helix (4.97%), β-sheet (27.55%), turns structure (49.78%), and unordered structure (17.70%).

Phytochemical Analysis of Muntingia calabura Extracts Possessing Anti-Microbial and Anti-Fouling Activities

The present study was carried out to determine the possible antifouling and anti microbial activity of plant extracts prepared from Muntingia calabura and phytochemical screening was demonstrated. The extracts from the following plant parts like stem, leaves, fruits, flower and stems were prepared in different aqueous solvents like ethanol, methanol, acetone, acetonitrile and water. The highest antimicrobial potentials were observed for the methanolic extracts against K. pneumonia, B. subtilis, B. megaterium and P. aeruginosa. Its efficacy was comparable to the standard drug, ampicillin. Significant amount of tannins, alkaloids, steroids and flavonoids were found. Interestingly, the present study showed anti fouling effect against pathogenic biofilm forming bacteria. These extracts also contain high hemagglutination activity. The present study provides evidence that solvent extract of M. calabura contains medicinally important bioactive compounds and this justifies the use of plant species as traditional medicine for treatment of various diseases.