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Coagulation disorders and bleeding are the main problems among people with hemophilia. Although its common treatment is replacement therapy, the effects of herbs on bleeding treatment has been proven. In this study, the coagulation effect of the Terminalia bellirica (Gaertn.) Roxb. hydro-alcoholic extract was evaluated. 32 mice were randomly divided into four groups (n = 8) and received single doses of 2000, 1500, 1000 or 500 mg/kg/day. Blood samples were taken from the animals 14-16 days after treatment, and coagulation indices were examined including PT, aPTT, BT, CT, and platelets. The hydro-alcoholic extract of T. bellirica significantly reduces bleeding time in the BT test (10-fold) and increases platelet numbers (four-fold), showing the effect of this extract on initial homeostasis. A significant reduction of coagulation time in the CT test (five-fold) also indicates the effect of this plant extract on platelet aggregation. Moreover, the results showed the effectiveness of T. bellirica hydroalcoholic extract in the extrinsic coagulation pathway because all concentrations reduced the coagulation time in the PT test. Furthermore, given the ineffectiveness of the T. bellirica extract on the aPTT test, this extract probably had no effect on intrinsic coagulation factors. Considering the results, T. bellirica extract seems to have a coagulation effect on primary homeostasis and extrinsic pathway of secondary homeostasis. [ABSTRACT FROM AUTHOR]

Root and crown rot incited by an oomycete, Phytophthora melonis, causes significant yield losses in commercial pumpkin (Cucurbita pepo) production worldwide. Currently, resistant cultivars and knowledge of molecular mechanism of C. pepo against P. melonis are scarce. Here, we analysed the quantitative gene expression changes of 10 candidate gene markers (bHLH87, ERF014, HSF, MYB, PR-1, WRKY21, CPI, POD, PSK, SGT) in pumpkin roots and leaves at three time points (h post-inoculation, hpi) following inoculation with P. melonis in two resistant (Ghelyani and Tanbal), and two susceptible (Marmari and Khoreshti) varieties of pumpkin. Gene expression using quantitative real time PCR along a time course revealed the strongest transcriptomic response at 48 and 72 hpi in resistant genotypes, 1.1-2.7-fold in roots and leaves, respectively, with a high significant correlation (r = 0.857**-0.974**). We also found that CPI, PSK, SGT1 and POD act as a dual regulator that similarly modulate immunity not only against P. melonis, but also against other diseases such as early blight (Alternaria cucumerina), powdery mildew (Podosphaera xanthii), downy mildews (Pseudoperonospora cubensis), and pathogenic plant nematodes (Meloidogyne javanica). Furthermore, significantly higher activities of the ROS scavenging defence enzymes, catalase (1.6-fold increase) and peroxidase (6-fold increase) were observed in the roots of resistant cultivars at different hpi compared with non-inoculated controls. In addition, the biomass growth parameters including leaf and root length, stem and root diameter, root fresh weight and volume were significantly different among studied genotypes. Cumulatively, the transcriptome data provide novel insights into the response of pumpkins for improving pumpkin breeding to P. melonis. [ABSTRACT FROM AUTHOR]

The production of recombinant vitamin K dependent (VKD) proteins for therapeutic purposes is an important challenge in the pharmaceutical industry. These proteins are primarily synthesized as precursor molecules and contain pre–propeptide sequences. The propeptide is connected to γ‐carboxylase enzyme through the γ‐carboxylase recognition site for the direct γ‐carboxylation of VKD proteins that has a significant impact on their biological activity. Propeptides have different attitudes toward γ‐carboxylase and certain amino acids in propeptide sequences are responsible for the differences in γ‐carboxylase affinity. By aiming to replace amino acids in hFIX propeptide domain based on the prothrombin propeptide, pMT‐hFIX‐M14 expression cassette, containing cDNA of hFIX with substituted −14 residues (Asp to Ala) was made. After transfection of Drosophila S2 cells, expression of the active hFIX was analyzed by performing ELISA and coagulation test. A 1.4‐fold increase in the mutant recombinant hFIX expression level was observed in comparison with that of a native recombinant hFIX. The enhanced hFIX activity and specific activity of the hFIXD‐14A (2.2 and 1.6 times, respectively) were further confirmed by comparing coagulation activity levels of substituted and native hFIX. Enrichment for functional, fully γ‐carboxylated hFIX species via barium citrate adsorption demonstrated 2‐fold enhanced recovery in the S2‐expressing hFIXD‐14A relative to that expressed native hFIX. These results show that changing −14 residues leads to a decrease in the binding affinity to substrate, increase in γ‐carboxylation and activity of recombinant hFIX. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:515–520, 2018 [ABSTRACT FROM AUTHOR]

Insect expression systems have been used to achieve high expression of recombinant and complex proteins, but disability of insects in the synthesis of N-Glycan products similar to mammals has been a controversial conflict debate in recent years. Glycosylation products in insects contain high or low end of mannose units. The main reason for this inability is the low level of activity of a number of enzymes including β-N - (1 and 2) acetyl glucosamine transferase I and II, β- (1 and 4) galactosyl transferase, α-(2, 3) and α-(2, 6) sialyl transferase. In addittion, a hexoaminidase that remove N-acetyl glucosamine at the end of glycan products and prevents binding of galactose and Sialic acid to glycan products have been discovered in insects. So the insect cells can be engineered to produce glycan products similar with mammalians and remove blocking agents of synthesis of sialyl and galactose products. In this systematic review, the glycosylation pathways in mammals and insects and engineering of possible glycosylation pathways in S2 cells have been investigated. [ABSTRACT FROM AUTHOR]