Evaluation of grass pollen quality control for allergy test kit production using pollen morphological characters

Grass pollen is one of the leading causes of allergies in throughout the world. The symptom level depends on the personal immunity. Pollen allergy test kits are suitable for detecting allergic symptoms, which they are produced from pure pollen of specific species. The production of pollen allergy testing kits requires accurate botanical standards and identification for the process of quality control of raw materials basing on Thai Food and drug ministration (Thai FDA). To produce allergy test kit, we focused on the pollen of these 6 common grass species and also use pollen morphological characters for detection and identification. Therefore, this study focused on the method for qualification control and standardization for prototype product of grass pollen allergy testing kit. Six common grass species as Megathyrsus maximus (Jacq.) B.K.Simon & S.W.L. Jacobs, Oryza sativa L., Sorghum bicolor (L.) Moench, Urochloa eminii (Mez) Davidse, Urochloa mutica (Forssk.) T.Q.Nguyen and Zea mays L.). The criteria were: 1) common in Thailand and Southeast Asia; 2) high density in the atmosphere; and 3) severity of the allergic reaction. The modified acetolysis were applied for samples preparation. The pollen morphological (polar and equatorial axes, exine, porus, annulus and ornamentation) were observed and measurement under light and scanning electron microscope. The character matrix was analyzed based on a regression tree and Tukey’s HSD test to assess support for the different pollen morphological characters. The distinguished character is ornamentation. The results were successfully described the pollen species and showed that the pollen morphologies were useful for contaminated detection. In addition, this method was suitable applied for qualification control of the raw materials for grass pollen allergy testing kits production

The albumin/starch scaffold and its biocompatibility with living cells

Abstract Combinations of protein and carbohydrate scaffold, bovine serum albumin (BSA) and cassava starch, have been used as materials to fabricate 3D scaffolds using an in-house developed freeze-drying method. Using this method, the BSA/starch generated a stable foam-type scaffold while the BSA alone could not maintain a rigid structure. The albumin/starch scaffolds had a porous structure and indicated the secondary structures of protein in the Fourier transform infrared spectroscopy (FTIR) study. Small and wide-angle X-ray scattering analysis (SAXS and WAXS) data indicated that the BSA molecules were uniformly dispersed within the starch framework and thereby characterized the size of the scaffold. The alcohol treatment changed the protein structure within the scaffold from an α-helix into a random coil. Interestingly, the BSA/starch scaffold with methanol treatment had better mechanical properties than that with the ethanol treatment. Cytotoxicity analyzed using colorimetric assay of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) with HepG2 cells showed that the cell viability for the BSA/starch scaffold under all tested conditions was greater than 90 %. Thus, all obtained materials (based on the BSA/starch scaffolds) could be used as a matrix for culturing living cells. All these results indicated that the BSA/starch scaffolds could be used for tissue formation in tissue bioengineering

Nutritional Profiles, Phytochemical Analysis, Antioxidant Activity and DNA Damage Protection of Makapuno Derived from Thai Aromatic Coconut

Makapuno is a natural mutant coconut cultivar with jelly-like endosperm. Here, we investigated the nutritional compositions, active ingredients, and antioxidant activities of Makapuno meat and water. The contents of macronutrients, sugars, vitamins, amino acids, and fatty acids were reported. We found that Makapuno meat has higher dietary fiber with lower protein and fat content compared to normal coconut meat. Medium-chain fatty acids were the major fat component of Makapuno meat and water. Phytochemical analysis revealed that while flavonoid content was lower, the total phenolic, alkaloid, and tannin contents of Makapuno meat were comparable with those of mature coconut. However, Makapuno water contained higher alkaloid content when compared to mature and young coconuts. The antioxidant activities, as examined by DPPH, FRAP, and ABTS assays, showed that Makapuno meat and water had antioxidant activities, and Makapuno water exhibited protective activity against DNA damage. Hence, this research provides the nutraceutical importance of Makapuno, which could be used in the food industry

Evaluating the effect of pore size for 3d-printed bone scaffolds

The present study investigated the influence of pore size of strut-based Diamond and surface-based Gyroid structures for their suitability as medical implants. Samples were made additively from laser powder bed fusion process with a relative density of 0.3 and pore sizes ranging from 300 to 1300 μm. They were subsequently examined for their manufacturability and mechanical properties. In addition, non-Newtonian computational fluid dynamics and discrete phase models were conducted to assess pressure drop and cell seeding efficiency. The results showed that both Diamond and Gyroid had higher as-built densities with smaller pore sizes. However, Gyroid demonstrated better manufacturability as its relative density was closer to the as-designed one. In addition, based on mechanical testing, the elastic modulus was largely unaffected by pore size, but post-yielding behaviors differed, especially in Diamond. High mechanical sensitivity in Diamond could be explained partly by Finite Element simulations, which revealed stress localization in Diamond and more uniform stress distribution in Gyroid. Furthermore, we defined the product of the normalized specific surface, normalized pressure drop, and cell seeding efficiency as the indicator of an optimal pore size, in which this factor identified an optimal pore size of approximately 500 μm for both Diamond and Gyroid. Besides, based on such criterion, Gyroid exhibited greater applicability as bone scaffolds. In summary, this study provides comprehensive assessment of the effect of pore size and demonstrates the efficient estimation of an in-silico framework for evaluating lattice structures as medical implants, which could be applied to other lattice architectures

Generation of a Single-Chain Variable Fragment Antibody against Feline Immunoglobulin G for Biosensor Applications

For many decades, feline infectious disease has been among the most common health problems and a leading cause of death in cats. These diseases include toxoplasmosis, feline leukemia virus (FeLV), and particularly feline immunodeficiency virus (FIV) disease. Early diagnosis is essential to increase the chance of successful treatment. Generally, measurement of the IgG level is considered to be indicative of an individual’s immune status for a particular pathogen. The antibodies specific to feline IgG are crucial components for the development of a detection kit. In this study, feline IgG-bound scFv was selected using phage display technology. Three rounds of biopanning were conducted against purified feline IgG. Through an indirect enzyme-linked immunosorbent assay (ELISA), two scFv clones demonstrating the best binding ability to feline IgG were chosen for biochemical characterization. In addition, the selected scFv (N14) was expressed and purified in a bacterial system. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the size of the purified N14 was 29 kDa. A sandwich ELISA was used to evaluate the binding capacity of the purified scFv to feline IgG. As expected, the purified N14 had the capacity to bind feline IgG. Furthermore, N14 was modified to create a scFv-alkaline phosphatase (scFv-AP) fusion platform. The surface plasmon resonance (SPR) results revealed that N14-AP bound to feline IgG with an affinity binding value of 0.3 ± 0.496 μM. Additionally, the direct ELISA demonstrated the binding capacity of N14-AP to feline IgG in both cell lysate and purified protein. Moreover, N14-AP could be applied to detect feline IgG based on electrosensing with a detection limit of 10.42 nM. Overall, this study successfully selected a feline IgG-bound scFv and developed a scFv-AP platform that could be further engineered and applied in a feline infectious disease detection kit