Use of response surface methodology to optimise carotenogenesis in the microalga,Haematococcus pluvialis

The factors controlling biomass production and the synthesis of astaxanthin esters in the microalga Haematococcus pluvialis (CCAP 34/7) have been investigated using a statistical approach employing response surface methodology (RSM). The culture conditions required for optimal growth and carotenogenesis in this alga are very different. Of particular importance is the photon flux density: for growth the optimum is 50–60 μmol m⁻² s⁻¹ whereas the optimum for astaxanthin synthesis is much higher at ∼-1600 μmol m⁻² s⁻¹. The addition of low levels of NaCl to the medium also stimulates to a small extent synthesis of astaxanthin, but photon flux density remains the overriding factor. The optimal temperature for this strain is quite low at 14–15 °C. RSM has been shown to be a rapid and effective technique leading to the optimisation of algal culture conditions. This statistical approach can be applied readily to the majority of microalgae and their products.     

Deorphanization and Characterization of Human Olfactory Receptors in Heterologous Cells

Olfaction plays an indispensable role in human and animals in self and environmental recognition, as well as intra‐ and interspecific communication. Following the discovery of a family of olfactory receptors (ORs) by Buck and Axel in 1991, it has been established that the sense of smell begins with the molecular recognition of a chemical odorant by one or more ORs expressed in the olfactory sensory neurons. Therefore, characterization of the molecular interactions between odorant molecules and ORs is a key step in the elucidation of the general properties of the olfactory system and in the development of applications, i.e., design of new odorants, search for blockers, etc. The process putted in place at ChemCom to improve the expression of ORs at the cytoplasmic membrane of the HEK293 cell and assays enabling large‐scale deorphanization, and to characterize the interaction between chemical odorants and ORs is described. The family of human ORs includes ca. 400 putatively functional ORs which are GPCRs (G protein‐coupled receptors); to date over 100 human ORs have been deorphanized.     

Elastin-like polypeptide fusions enhance the accumulation of recombinant proteins in tobacco leaves

The production of recombinant proteins in plants is an active area of research and many different high-value proteins have now been produced in plants. Tobacco leaves have many advantages for recombinant protein production particularly since they allow field production without seeds, flowers or pollen and therefore provide for contained production. Despite these biosafety advantages recombinant protein accumulation in leaves still needs to be improved. Elastin-like polypeptides are repeats of the amino acids “VPGXG” that undergo a temperature dependant phase transition and have utility in the purification of recombinant proteins but can also enhance the accumulation of recombinant proteins they are fused to. We have used a 11.3 kDa elastin-like polypeptide as a fusion partner for three different target proteins, human interleukin-10, murine interleukin-4 and the native major ampullate spidroin protein 2 gene from the spider Nephila clavipes. In both transient analyses and stable transformants the concentrations of the fusion proteins were at least an order of magnitude higher for all of the fusion proteins when compared to the target protein alone. Therefore, fusions with a small ELP tag can be used to significantly enhance the accumulation of a range of different recombinant proteins in plant leaves. An erratum to this article can be found at     

Tritium labeling of thermolysin, elastase, and ribonuclease by exposure to tritium gas at low pressure

The bacterial metalloendoprotease thermolysin, bovine pancreatic ribonuclease, and porcine pancreatic elastase have been tritiated by exposure to subcurie amounts of tritium gas at pressures below 50 mTorr for periods of 1 to 6 h. Thermolysin, ribonuclease, and elastase have been purified to specific radioactivities of 15, 5, and 1 Ci/mol, respectively. Amino acid analyses of the tritiated enzymes revealed higher relative specific radioactivities for His, Pro, and Phe in all three proteins while Val and Ile were among the residues with the lowest relative specific radioactivities. The recovery of enzyme activity was always greater than 95% and the formation of tritiated decomposition products was not observed. This lowpressure gas exposure process requires less tritium gas and less time than the original method of Wilzbach to achieve equal or higher levels of tritium incorporation. In addition, the enzymes were completely active and did not show the presence of highly radioactive byproducts which have been observed in earlier studies of the Wilzbach labeling of proteins.     

Der Feldbegriff in seiner Anwendung auf das Problem der Zellteilung

Summary The authors show that in certain isolated tissues (cornea of frog, mouse cancer) the mitotic processes continue during at least one hour. They are very strongly stimulated by heat and also by mitogenetic radiation. In the case of the cancer cells new mitoses are promoted in considerable number. A detailed analysis of both energy factors leads to the conclusion that they effect a disturbance of the unstable constellations (structures) of the elementary particles in the cell-body. Thus a certain degree of disorganisation of its plasma seems to stimulate cell-division. This statement is, in the opinion of the authors a proof that cell-division is controlled by a field, the trajectories of the elementary particles of the cell-body being a function of their coordinates relative to the cell-axes.

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Résumé Les auteurs démontrent que dans certains tissus survivants (la cornée des yeux de grenouille et le cancer de souris) les mitoses continuent leur processus d'évolution durant au moins une heure. Par l'application de chaleur aussi bien que par l'irradiation mitogénétique on peut accélérer beaucoup le rhythme des mitoses en cours d'évolution et provoquer (dans les cas de cancer seulement) l'apparition d'un nombre considérable de nouvelles mitoses. L'analyse de l'action de ces deux facteurs d'énergie aboutit à la conclusion qu'il ne peut s'agir que d'un dérangement des constellations instables des éléments du corps cellulaire. On peut en déduire qu'un certain degré de désorganisation du plasme est favorable aux divisions cellulaires. En poursuivant et en développant cette conception, les auteurs arrivent à la conception du champ cellulaire de division, définissant les trajectoires des particules élémentaires comme fonction de ses coordonnés relativement aux axes des cellules.

     

A trypsin and chymotrypsin inhibitor from seeds of Bauhenia purpurea

A trypsin and chymotrypsin inhibitor was partially purified from Bauhenia purpurea seeds and separated from a second inhibitor by Ecteola cellulose chromatography. The factor inhibited bovine trypsin and chymotrypsin as well as pronase trypsin and elastase. It formed a complex with trypsin and with chymotrypsin, but a ternary complex could not be detected. Differences were detected in the effect on trypsin and on chymotrypsin, although one enzyme interfered with the inhibition of the other. The results obtained point to two active centers on the inhibitor for the trypsin and chymotrypsin inhibition such that the one cannot complex with the inhibitor after this inhibitor had complexed with the other.     

Electron-microscopic study of the rat masseter muscle following injection of lidocaine

The ultrastructure of rat masseter muscle was examined at 15 min, 1 and 6 h, and 1 and 2 days following a single injection of 2% lidocaine. Lesions developed within 15 min. The plasma membrane was disrupted and invaginated. The nuclei were pyknotic and the mitochondria appeared swollen. The myofibrils separated and became disoriented. By 1 and 6 h, these changes were severe. By 1 day, the macrophages appeared in damaged myofibers. The presence of a few presumptive myoblasts signaled the onset of regeneration. By 2 days, presumptive myoblasts formed within the basement membrane. The basal lamina proved most resistant to injury. Regeneration of masseter muscle following the damage produced by lidocaine appeared discontinuous in nature. The singly nucleated presumptive myoblasts seemed to arise within the lesions.