Two soluble glycosyltransferases glycosylate less efficiently in vivo than their membrane bound counterparts

Many Golgi glycosyltransferases are type II membrane proteins which are cleaved to produce soluble forms that are released from cells. Cho and Cummings recently reported that a soluble form of alpha1, 3- galactosyltransferase was comparable to its membrane bound counterpart in its ability to galactosylate newly synthesized glycoproteins (Cho,S.K. and Cummings,R.D. (1997) J. Biol. Chem., 272, 13622-13628). To test the generality of their findings, we compared the activities of the full length and soluble forms of two such glycosyltransferases, ss1,4 N-Acetylgalactosaminyltransferase (GM2/GD2/ GA2 synthase; GalNAcT) and beta galactoside alpha2,6 sialyltransferase (alpha2,6-ST; ST6Gal I), for production of their glycoconjugate products in vivo . Unlike the full length form of GalNAcT which produced ganglioside GM2 in transfected cells, soluble GalNAcT did not produce detectable GM2 in vivo even though it possessed in vitro GalNAcT activity comparable to that of full length GalNAcT. When compared with cells expressing full length alpha2,6-ST, cells expressing a soluble form of alpha2,6-ST contained 3-fold higher alpha2,6-ST mRNA levels and secreted 7-fold greater alpha2,6-ST activity as measured in vitro , but in striking contrast contained 2- to 4-fold less of the alpha2,6-linked sialic acid moiety in cellular glycoproteins in vivo . In summary these results suggest that unlike alpha1,3-galactosyltransferase the soluble forms of these two glycosyltransferases are less efficient at glycosylation of membrane proteins and lipids in vivo than their membrane bound counterparts.      

Study of response of Swiss Webster mice to light subunit of mushroom tyrosinase

The light subunit of mushroom, Agaricus bisporus, tyrosinase (LSMT), has been identified as an extrinsic component of the enzyme. Its function is unknown, but it can cross an epithelial cell layer, which suggests that it can be absorbed by the intestine. A similar capability has been demonstrated for the HA-33 component of the progenitor toxin from Clostridium botulinum, which is the closest structural homolog of LSMT. Unlike HA-33, LSMT appears to be non-immunogenic as shown by preliminary tests in Swiss Webster mice. We investigated the immunogenicity and histopathology of LSMT in mice to determine its safety in vivo. LSMT did not evoke generation of antibodies after prolonged periods of intraperitoneal administration. Histopathological observations confirmed the absence of responses in organs after twelve weekly administrations of LSMT. We found that LSMT is not toxic and is less immunogenic than the C. botulinum HA-33 protein, which supports further research and development for pharmaceutical application.     

Transcriptomic response to differentiation induction

Background  

Microarrays used for gene expression studies yield large amounts of data. The processing of such data typically leads to lists of differentially-regulated genes. A common terminal data analysis step is to map pathways of potentially interrelated genes.     

Extracellular polymeric substances of the marine fouling diatom amphora rostrata Wm.Sm

Amphora rostrata was grown under continuous illumination at 27°C in batch cultures using f/2 medium. Cell biomass (measured as chllorophyll a and cell counts) reached a maximum on day 7. Thereafter, cell biomass as chl a showed a small decrease. Planktonic('free') and biofilm extracellular polymeric substances (EPS) from the adherent cells of A. rostrata were studied. Both types of EPS were produced during the logarithmic phase of growth. However, production was higher during the stationary growth phase. Enhanced EPS production was associated with nutrient deficient conditions. Planktonic and biofilm EPS were purified by gel filtration using Sephadex G‐200 and ion exchange chromatography using DEAE‐cellulose. Both polymers showed the presence of a single peak. Capillary gas Chromatographie analysis of both planktonic and biofilm EPS showed that fucose (36.7%) and galactose (27.6%) were the most abundant monosaccharides, with small quantities of rhamnose, xylose, arabinose, mannose and glucose. Other chemical analysis showed the presence of sulphate, uronic acids, hexoamines, pyruvate and proteins in both the planktonic and bio‐film EPS. Uronic acid, pyruvate and sulphate together were found to contribute ～50 to 60% (W/W) to the EPS of A. rostrata. Such a high content of non‐sugar components indicates their importance to the diatom in metal binding, desiccation prevention and flexibility.     

Morphometric and phytochemical characterization of chaura fruits (Gaultheria pumila): a native Chilean berry with commercial potential

Background

For the first time, a morphometric characterization of chaura (Gaultheria pumila) fruits has been conducted between natural populations growing in the Villarrica National Park, Araucania Region, Chile. Chaura is a native Ericaceae from Chile that produces aromatic and tasty fruits which could be of agricultural interest.

Results

To influence the decision for a further domestication of G. pumila, both the fruit sizes (indicator of productivity) and the nutritional properties of the fruits have been determined from different subpopulations. Samples were a total of 74 plants and 15 fruits per plant which were randomly harvested following its natural distribution around the Villarrica volcano. Altogether, fresh weight, shape, color, diameter in the pole and the equatorial dimensions were determined as phenotypic traits of the G. pumila fruits. Meanwhile the total soluble solids, anthocyanin and pectin contents were calculated as nutritional traits of the Chaura fruits. Results showed a high phenotypic diversity between the sampled population with three main fruit shapes and three predominant colors. The round shapes were the most abundant, whereas a significant correlation was found among fruit size with weight and color. The highest fresh weight (597.3 mg), pole diameter (7.1 mm) and equatorial diameter (6.5 mm) were estimated in the pink color fruits.

Conclusions

The total amount of anthocyanin was higher in red fruits, while the maximum pectin content was obtained in the round white fruits. Overall results must pave the way for a further domestication and introduction of the Chaura species in the agro-productive system in Chile.     

Aspects and altitudes modify the requirement of disturbance in oak (Quercus leucotrichophora A. Camus) belt of Garhwal Himalaya

Present study measures the impact of forest disturbance on population structure and regeneration status of a Himalayan banj oak (Qsuercus leucotrichophora A. Camus) forest at different aspects and altitudes. The whole study was carried out by placing 300 systematically selected sample plots in banj oak forest. The study revealed that moderately disturbed forest patches were present in all elevation ranges and both north and south facing aspects whereas most of the highly disturbed patches were situated near middle and lower stretches of forests or close to habitations. Density of primary diameter class (5–15 cm) was recorded highest in moderately disturbed zone in upper elevation ranges and north facing aspect and ‘fair’ category of regeneration was most frequent in all elevation ranges and aspects. The paper concludes a positive effect of mid-level disturbance on plant community for better regeneration and study recommends a minimum resource extraction and silvicultural practices in banj-oak belt of Himalaya for a minimum canopy opening which not only be able to provide biomass to local communities for their daily needs but also would be able to maintain and improve forest health.