The possible roles of residues 79 and 80 of the Trp repressor from Escherichia coli K-12 in trp operator recognition

We constructed mutants of the Trp repressor from Escherichia coli K-12 with all possible single amino acid exchanges at positions 79 and 80 (residues 1 and 2 of the recognition helix). We tested these mutants in vivo by measuring the repression of synthesis of β-galactosidase with symmetric variants of α- and β-centered trp operators, which replace the lac operator in a synthetic lac system. The Trp repressor carrying a substitution of isoleucine 79 by lysine, showed a marked specificity change with respect to base pair 7 of the α-centered trp operator. Gel retardation experiments confirmed this result. Trp repressor mutant IR79 specifically recognizes a trp operator variant with substitutions in positions 7 and 8. Another mutant, with glycine in position 79, exhibited loss of contact at base pair 7. We speculate that the side chain of Ile79 interacts with the AT base pairs 7 and 8 of the α-centered trp operator, possibly with the methyl groups of thymines. Replacement of thymine in position 7 or 8 by uracil confirms the involvement of the methyl group of thymine 8 in repressor binding. Several Trp repressor mutants in position 80 (i.e. AI80, AL80, AM80 and AP80) broaden the specificity of the Trp repressor for α-centered trp operator variants with exchanges in positions 3, 4 and 5.     

An on-line advisory control system for the lactic acid fermentation process

A fuzzy expert system was developed for online diagnosing and controlling of bioprocesses. The system was constructed in object-oriented Smalltalk/V for diagnosing and controlling of bioprocesses. Lactic acid fermentation with an industrial strain ofLactobacillus casei was chosen as the model system. The performance of the fuzzy expert system and the knowledge base utilizing experts' knowledge and several facts obtained from the experiments were successfully validated with on-line fermentations. The fuzzy expert system could diagnose a fault on-line and give reasonable advice to the process operator. In order to achieve the diagnosing faculty, a database, a knowledge base, and both backward and forward chaining procedures were implemented employing the object-oriented programming environment. A defuzzifier was implemented in the system to achieve on-line control. In order to realize a decision-making system with a human operator and a fuzzy expert system, a new control strategy namedAdvice was also introduced. Several cultivations were carried out in order to collect knowledge on the effects concerned with inoculum properties to the process and to construct a database including standard time-course profiles. The performance of the fuzzy expert control system was successfully tested with on-line experiments.Visiting scientist from HUT at RIKEN     

Ethnography,storytelling and the fiction of Toshio Mori

Conclusion Anthropology has always been concerned with shocking the reader and with deconstructing the reader's view of the world by confrontation with that which is seemingly most alien. Nor is the call for self-reflexive research and writing new. Stanley Diamond's work offers one of the most eloquent visions of a self-reflexive anthropology as cultural critique.²⁶ In view of the overwhelming fascination in anthropology with post-modernism, it is useful to remind oneself that not only are the questions raised by post-modernists of long standing, but also that such questions can become an end in themselves, a way to avoid taking positions and to avoid responsibility.Fiction and ethnography should not be confused, nor should one be a substitute for the other. Ethnography has to travel on a fine line between positivist claims of representing some form of objective truth, an impressionistic pseudo-fiction or an ever more inward-turning contemplation of its own contradictions. To do ethnographic work is to accept the obligation to keep trying to elucidate while resisting the attraction of final, definitive interpretations and continually questioning the media of both research and writing that are being employed.     

Association of the Decline of Nashi Pears with an MLO

Mycoplasma-like organisms (MLOs) were constantly detected by the DAPI technique and by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified DNA in trees of Pyrus pyrifolia cvs Hosui and Kosui grafted on P. communis, seedlings rootstock with symptoms similar to the slow form of pear decline. These symptoms included upward curling of the leaves along the midrib. Leaves were abnormally thick and later turned reddish while major veins became swollen and brown. Trees with symptoms were usually 4–5 years old and were growing in the major pear areas of central Italy. The incidence of affected trees was particularly high in one orchard adjacent to a pear orchard strongly affected with the slow form of pear decline. In this case the distribution pattern of affected Nashi trees suggests that the causal agent was introduced from the adjacent pear orchard by an aerial vector. Although oriental pears are well-known hosts of the pear-decline agent when used as rootstocks of French cultivars, this is the first report of pear decline in P. pyrifolia varieties.     

Transcription of a zebrafish gene of the hairy-Enhancer of split family delineates the midbrain anlage in the neural plate

her5 encodes a basic helix-loop-helix (bHLH) protein with all features characteristic of the Drosophila hairy-E(spl) family. her5 is expressed in a band of cells within the neural anlage from about 90% epiboly on to at least 36 h postfertilization (hpf). After completion of brain morphogenesis, her5-expressing cells are located in the caudal region of the midbrain, at the boundary with the rhombencephalon. Labelling of cells within the her5 expression domain in the neural plate by injection of fluorescein-dextran allows their labelled progeny to be localized in the 36-hpf-old embryo using an anti-fluorescein antibody. This shows that the her5 expression domain corresponds to the midbrain primordium, including both the tectum and the tegmentum, in the neural plate. A possible function for her5 in regionalization of the brain and/or control of the midbrain-hindbrain boundary is discussed.     

Improved correlation between soil exchangeable K and plant K contents on a tissue water basis

In acid volcanic soils, plant roots are thought to be injured by acidity (low pH) and/or solubilized aluminium (Al) ions. An attempt was made to separate the effects of low pH from those of Al on the elongation and viability of alfalfa (Medicago sativa L.) radicles in water culture. Root elongation was irreversively curtailed by 20 hours treatment at pH 4.0 without Al or 20 mmol m^-3 Al at pH 5.0. Viability of surface cells of root tips was detected as a degrading activity of fluorescein diacetate (FDA) by cellular esterases and subsequent accumulation of derived fluorescein within cells. Large numbers of the surface cells lost their viability after four hours exposure at the low pH. In contrast, surface cells maintained both FDA degrading activity and ability to accumulate fluorescein 20 h after initial exposure to the Al solution (20 mmol Al m^-3, pH 5.0). These results suggest that there are some significant differences in the mechanisms of phytotoxicity to alfalfa root between the two stress factors.     

Biochemical and serological evidence for an RNase E-like activity in halophilic Archaea.

Endoribonuclease RNase E appears to control the rate-limiting step that mediates the degradation of many mRNA species in bacteria. In this work, an RNase E-like activity in Archaea is described. An endoribonucleolytic activity from the extreme halophile Haloarcula marismortui showed the same RNA substrate specificity as the Escherichia coli RNase E and cross-reacted with a monoclonal antibody raised against E. coli RNase E. The archaeal RNase E activity was partially purified from the extreme halophilic cells and shown, contrary to the E. coli enzyme, to require a high salt concentration for cleavage specificity and stability. These data indicate that a halophilic RNA processing enzyme can specifically recognize and cleave mRNA from E. coli in an extremely salty environment (3 M KCI). Having recently been shown in mammalian cells (A. Wennborg, B. Sohlberg, D. Angerer, G. Klein, and A. von Gabain, Proc. Natl. Acad. Sci. USA 92:7322-7326, 1995), RNase E-like activity has now been identified in all three evolutionary domains: Archaea, Bacteria, and Eukarya. This strongly suggests that mRNA decay mechanisms are highly conserved despite quite different environmental conditions.     

Economic history of ostrich fern,Matteuccia struthiopteris,the edible fiddlehead

Although the ostrich fern, Matteuccia struthiopteris,is distributed throughout much of the Northern Hemisphere, its consumption as a spring vegetable has been largely restricted to the tables of the Maritime Provinces in Canada and neighboring Maine in the United States. However, in little more than a decade the demand has expanded beyond these traditional boundaries, due to the present availability of the frozen product. The heart of the industry is undoubtedly New Brunswick. The harvest of 200 t/yr is approximately 4 times that of Maine, and from an historical vantage the traditional use as a spring vegetable is with the New Brunswickers as well. In light of the present high demand for the product and the reports of localized overpicking, a review of the economic history, resource management and attempts at cultivation is presented.