Characterization of polyhydroxyalkanoates (PHAs) biosynthesis by isolated Novosphingobium sp. THA_AIK7 using crude glycerol

Biodiesel-contaminated wastewater was used to screen for PHAs-producing bacteria by using crude glycerol as the sole carbon source. A gram-negative THA_AIK7 isolate was chosen as a potential PHAs producer. The 16S rRNA phylogeny indicated that THA_AIK7 isolate is a member of Novosphingobium genus which is supported by a bootstrap percentage of 100% with Novosphingobium capsulatum. The 1,487 bp of 16S rRNA gene sequence of THA_AIK7 isolate has been deposited in the GenBank database under the accession number HM031593. Polymer content of 45% cell dry weight was achieved in 72 h with maximum product yield coefficient of 0.29 g PHAs g?1 glycerol. Transmission electron micrograph results exhibited the PHAs granules accumulated inside the bacterial cell. PHAs polymer production in mineral salt media supplemented with 2% (w/v) of crude glycerol at initial pH 7 was extracted by the sodium hypochlorite method. Polymer film spectrographs from Nuclear magnetic resonance displayed a pattern of signal virtually identical to spectra of commercial PHB. Thermal analysis by Differential scanning calorimeter showed a melting temperature at 179°C. Molecular weight analysis by Gel permeation chromatography showed two main peaks of 133,000 and 700 g mol?1 with weight-average molecular weight value of 23,800 and number-average molecular weight value of 755. Endotoxinfree of PHAs polymer was preliminarily assessed by a negative result of the gel-clot formation, Pyrotell? Single test vial, at sensitivity of 0.25 EU ml?1. To our knowledge, this is the first reported test of endotoxin-free PHAs naturally produced from gram-negative bacteria which could be used for biomedical application.     

Importin β2 Mediates the Spatio-temporal Regulation of Anillin through a Noncanonical Nuclear Localization Signal

The compartmentalization of cell cycle regulators is a common mechanism to ensure the precise temporal control of key cell cycle events. For instance, many mitotic spindle assembly factors are known to be sequestered in the nucleus prior to mitotic onset. Similarly, the essential cytokinetic factor anillin, which functions at the cell membrane to promote the physical separation of daughter cells at the end of mitosis, is sequestered in the nucleus during interphase. To address the mechanism and role of anillin targeting to the nucleus in interphase, we identified the nuclear targeting motif. Here, we show that anillin is targeted to the nucleus by importin β2 in a Ran-dependent manner through an atypical basic patch PY nuclear localization signal motif. We show that although importin β2 binding does not regulate anillin''s function in mitosis, it is required to prevent the cytosolic accumulation of anillin, which disrupts cellular architecture during interphase. The nuclear sequestration of anillin during interphase serves to restrict anillin''s function at the cell membrane to mitosis and allows anillin to be rapidly available when the nuclear envelope breaks down to remodel the cellular architecture necessary for successful cell division.     

Cloning and characterization of cDNA encoding 3-methylcholanthrene inducible rat mRNA for UDP-glucuronosyltransferase

We have isolated cDNA clones of the mRNA for rat UDP-glucuronosyltransferase that catalyzes the glucuronidation of 4-nitrophenol, by using synthetic oligonucleotides as hybridization probes. The complete nucleotide sequence of the 1,927-base pairs cDNA insert has been determined. With untranslated sequences of 124 and 216 base pairs in the 5'- and 3'-terminal regions, respectively, the cDNA insert contained 1,587 base pairs that encode a complete primary structure of a putative precursor form of 4-nitrophenol UDP-glucuronosyltransferase with a calculated molecular weight of 60,114. The cDNA sequence also indicates the presence of 25 amino acids preceding the sequence determined by microsequence of the isolated protein. This extrapeptide, for the most part, consists of hydrophobic amino acids which are characteristic of the signal peptides as found for secretory proteins and most transmembrane proteins. Furthermore, the deduced amino acid sequence contains a putative halt transfer signal of a hydrophobic segment (residues 487-510), which is flanked on both sides by the peptide segments of highly charged amino acid residues (residues 463-486 and 511-529). These features are consistent with the properties of transmembrane proteins. Specific cDNA probes were used to analyze the induction of the enzyme in rat tissues by treatment with 3-methylcholanthrene. RNA blot analysis showed that 3-methylcholanthrene increased 10- to 15-fold the amount of hybridizable mRNA in liver. The livers and kidneys from 3-methylcholanthrene-treated rats were found to contain almost the same amount of hybridizable mRNA, although the basal level in the kidney was much higher than that of the liver, and the amounts in the lung were much lower than that of the liver and kidney.     

Long-term storage of industrial microbial strains

Morphofunctional characteristics of industrial microorganisms belonging to different genera, species, and strains were investigated after 15 to 20 years of storage in liquid nitrogen. The taxonomic position of microorganisms, the cell physiological state prior to storage, and the cryoconservation regime were found to affect microbial cryoresistance. Protective media, density of cell suspensions, freezing rate, and heating temperature are the parameters important for development of efficient technologies for cryoconservation of industrial microorganisms at ?196°C.     

Studies of plasma dynamics in megampere X-pinches

Results are presented from experimental studies of the dynamics and parameters of the plasma generated by megampere (up to 2.3 MA) currents flowing through X-pinches formed from crossed wires made of various materials. The experiments were performed with different numbers of wires and different wire diameters. The X-ray yield in neon-like molybdenum lines (in the 2.5- to 3-keV photon energy range) from a <20 μm hot spot was larger than 10 J, and the soft X-ray power emitted from this spot reached 120 GW. The hot-spot lifetime estimated by comparing the measured and calculated emission intensities and line profiles of helium-like Fe and Cr ions was ～10 ps. Hard X-ray emission in the photon energy range ≥800 keV was also detected.     

Influence of rabbit cold adaptation on the depressor muscarinic cholinergic reaction of the arterial pressure of the hind limb vessels and the small intestine in situ and the systemic arterial pressure

Modification of the main parameters of cholinergic reactions in rabbits, i.e., of the specific sensitivity to agonists (EC ₅₀), maximal reaction values (P _m ) of arterial pressure in the hind limb vessels and small intestine in situ, and systemic arterial pressure after adaptation to cold for 1, 5, 10, and 30 days has been investigated. The depressor reaction to acetylcholine (muscarinic cholinergic receptors agonist) was established to correspond to the model p = (P _m A ⁿ )/(EC ₅₀ ⁿ + A ⁿ ) with n =1. In the control EC ₅₀ was equal to 0.85, 1.01, and 1.21 nmol/kg, while P _m equaled 100, 32.6, and 61.2 mm Hg for the hind limb vessels, small intestine in situ, and systemic arterial pressure, respectively.     

Oleic acid Enhances Dengue Virus But Not Dengue Virus-Like Particle Production from Mammalian Cells

Despite the recent introduction of a commercial vaccine, the mosquito-transmitted dengue virus is still a worldwide public health problem. Based on the live attenuated vaccine strategy, the commercial vaccine has a less than optimal protective profile. Virus-like particles (VLPs) offer an attractive alternate vaccination strategy due to the effectively native presentation of epitopes in the absence of any infectious genetic material. However, the production of amounts of VLP in a platform that can support commercial development remains a major obstacle. This study generated two DENV 2 VLPs [codon-optimized and chimeric DENV/Japanese encephalitis virus (JEV)] and directly compared yields of these constructs by western blotting and dot blot hybridization. The effect of oleic acid supplementation, a process known to increase DENV production in natural infection, was also investigated. Results showed that the chimeric construct gave a two- to threefold higher yield than the codon-optimized construct and that while oleic acid increased DENV virion production in natural infection, it inhibited VLP production. These results suggest that further optimization of DENV VLP expression is possible, but it will require more understanding of how native DENV infection remodels the host cell machinery.     

Ambiguity in a trans-disciplinary stakeholder assessment of neglected and underutilized species in China, Cambodia, Thailand and Vietnam

Neglected and Underutilized Crop Species (NUS) are grown primarily by traditional farmers in developing countries. They were once more widely grown but are today falling into disuse for a variety of agronomic, genetic, economic and cultural factors. While these crops continue to be maintained by socio-cultural preferences and use practices, most of them remain inadequately characterized and neglected by research and conservation. The aim of our work was to improve the agricultural portfolio by identifying promising neglected and underutilised species in China, Cambodia, Northeastern Thailand and Northern Vietnam. We began with preparing a “Masterlist” containing 260 species, based on farmer interviews and literature review. After an initial pre-selection we identified 17 NUS for China, 13 for Cambodia, 12 for Thailand, and 22 for Vietnam as of highest priority. These NUS then underwent a multi-criteria and trans-disciplinary assessment involving 511 stakeholders such as scientists, farmers, NGOs and policy makers. Based on the assessment we identified the most promising NUS for each country. We also identified some limitations for the promotion of so-called priority NUS regarding to some substancial disagreement between the involved stakeholders. In China and Cambodia, for example, farmers judged the potential of NUS significantly lower than other stakeholders, and in Vietnam scientists and policy makers had substantially different opinions on NUS compared to farmers. Addressing these ambiguous views will be highly relevant to the develoment of an improved agricultural portfolio.