Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species.

BACKGROUND: More than a dozen of fungal immunomodulatory proteins (FIPs) have been identified to date, most of which are from Ganoderma species. However, little is known about the similarities and differences between different Ganoderma FIPs' bioactivities. In the current study, two FIP genes termed FIP-gap1 and FIP-gap2 from G. applanatum, along with LZ-8 and FIP-gsi, another two representative Ganoderma FIP genes from G. lucidum and G. sinense were functionally expressed in Pichia. Subsequently, bioactivities of four recombinant Ganoderma FIPs were demonstrated and compared. RESULTS: All the four Ganoderma FIP genes could be effectively expressed in P. pastoris GS115 at expression levels ranging from 197.5 to 264.3 mg L- 1 and simply purified by one step chromatography using HisTrap™ FF prepack columns. Amino acid sequence analysis showed that they all possessed the FIP conserved fragments. The homologies of different Ganoderma FIPs were from 72.6 to 86.4%. In vitro haemagglutination exhibited that FIP-gap1, FIP-gsi and LZ-8 could agglutinate human, sheep and mouse red blood cells but FIP-gap2 agglutinated none. Besides, the immunomodulation activities of these Ganoderma FIPs were as: rFIP-gap2 > rFIP-gap1 > rLZ-8 and rFIP-gsi in terms of proliferation stimulation and cytokine induction on murine splenocytes. Additionally, the cytotoxic activity of different FIPs was: rFIP-gap1 > rLZ-8 > rFIP-gsi > rFIP-gap2, examined by their inhibition of three human carcinomas A549, Hela and MCF-7. CONCLUSIONS: Taken together, four typical Ganoderma FIP genes could be functionally expressed in P. pastoris, which might supply as feasible efficient resources for further study and application. Both similarities and differences were indeed observed between Ganoderma FIPs in their amino acid sequences and bioactivities. Comprehensively, rFIP-gaps from G. applanatum proved to be more effective in immunomodulation and cytotoxic assays in vitro than rLZ-8 (G. lucidum) and rFIP-gsi (G. sinense).

[Estimation of water clarity in offshore marine areas based on modified semi-analysis spectra model].

The main objectives of the research described in the present paper are to develop a semi-analysis model of water clarity for case 2 waters without inputting the absorption and scattering coefficient, which are not easy to be obtained for offshore marine areas so far. Based on the Zsd (Secchi depth)inversion theory, a simple semi-analysis spectra model was established for offshore seawater clarity by analyzing the relationship between vertical diffuse attenuation coefficient K(d) (490) and the beam attenuation coefficient c(490) with remote sensing reflectance. This semi-analysis spectra model needed two band reflectance ratios on- ly, while tidal correction was produced for this model to improve the precision of the retrieving results. The semi-analysis spectra model was applied to ASD hyperspectral reflectance data measured in the Pearl River Estuary Ecological Zone (October 21, 23, 2012, November 2, 2012; N=20) and the Xuwen Coral Reef Protection Zone (January 13, 14, 2013, N=25) which covered different water body of tidal times and different pollution sources. The results indicated that the changing tendency of predicted values was consistent with the synchronous measurement values after comparing them. However, water clarity calculated by the ASD hyperspectral reflectance measured in spring tidal time, generated 0. 4 m deviation compared with in-situ water clarity, while water clarity calculated by the ASD hyperspectral reflectance measured in neap tidal time is close to the in-situ water clarity. So the tidal correction coefficient of 0.4 was further applied for the model. After modification, the coefficient of determination between the inversed and measured water clarity was 0. 663, the average absolute error was 0. 14 m and the average relative error was 19.5%. Research demonstrated that this semi-analysis inversion algorithm just needs two band reflectance ratio to complete the inversion of water clarity, which is simple and works relatively well for lower clarity (less than 2 meters) waters compared to He' (2004) and Doron' (2011) algorithms.

Use of reflectance ratios as a proxy for coastal water constituent monitoring in the pearl river estuary.

Spectra, salinity, total suspended solids (TSS, in mg/L) and colored dissolved organic matter (CDOM, ag(400) at 400 nm) sampled in stations in 44 different locations on December 18, 19 and 21, in 2006 were measured and analyzed. The studied field covered a large variety of optically different waters, the absorption coefficient of CDOM ([ag(400)] in m(-1)) varied between 0.488 and 1.41 m(-1), and the TSS concentrations (mg/L) varied between 7.0 and 241.1 mg/L. In order to detect salinity of the Pearl River Estuary, we analyzed the spectral properties of TSS and CDOM, and the relationships between field water reflectance spectra and water constituents' concentrations based on the synchronous in-situ and satellite hyper-spectral image analysis. A good correlation was discovered (the positive correlation by linear fit), between in-situ reflectance ratio R(680)/R(527) and TSS concentrations (R(2) = 0.65) for the salinity range of 1.74-22.12. However, the result also showed that the absorption coefficient of CDOM was not tightly correlated with reflectance. In addition, we also observed two significant relationships (R(2) > 0.77), one between TSS concentrations and surface salinity and the other between the absorption coefficient of CDOM and surface salinity. Finally, we develop a novel method to understand surface salinity distribution of estuarine waters from the calibrated EO-1 Hyperion reflectance data in the Pearl River Estuary, i.e. channels with high salinity and shoals with low salinity. The EO-1 Hyperion derived surface salinity and TSS concentrations were validated using in-situ data that were collected on December 21, 2006, synchronous with EO-1 Hyperion satellite imagery acquisition. The results showed that the semi-empirical relationships are capable of predicting salinity from EO-1 Hyperion imagery in the Pearl River Estuary (RMSE < 2‰).