Expression and characterization of HlyX hemolysin from Leptospira interrogans serovar Copenhageni: potentiation of hemolytic activity by LipL32

The HlyX, a putative hemolysin identified from the Leptospira genomes, was cloned, expressed in Escherichia coli, purified, and its hemolytic activity was confirmed. Mouse polyclonal antiserum against the recombinant HlyX recognized HlyX-related antigens in a panel of Leptospira species extracts and it was also able to abolish the hemolytic activity of HlyX. A mixture of HlyX and LipL32, a known hemolysin from Leptospira, induced hemolysis in a synergistic way that was fully inhibited by antiserum against either protein. Moreover, sera from patients with leptospirosis also recognized the recombinant HlyX, showing that it is presented to the host immune system during Leptospira infection.     

Accumulation of furanocoumarins in Ruta graveolens L. shoot culture

Ruta graveolens L. shoots cultured in stationary liquid phase produced furanocoumarins: psoralen, bergapten, xanthotoxin, isopimpinellin and imperatorin at the amount totalling almost 1 g/100 g dry wt of the shoots. The dominating metabolites were therapeutically important compounds: xanthotoxin – 0.33 g/100 g dry wt and bergapten – 0.32 g/100 g dry wt. Maximum contents of the majority of the compounds were observed on 28th day of culture.     

A comparison of the GHG emissions caused by manufacturing tissue paper from virgin pulp or recycled waste paper

Purpose

The aim of this work is to compare greenhouse gas (GHG) emissions from producing tissue paper from virgin pulp (VP) or recycled waste paper (RWP). In doing so, the study aims to inform decision makers at both company and national levels which are the main causes of emissions and to suggest the actions required to reduce pollution.

Methods

An attributional life cycle assessment (LCA) was performed in order to estimate and compare the GHG emissions of the two processes. LCA allows us to assess how the choice of raw material for VP and RWP processes influences total GHG emissions of tissue paper production, what are the main drivers behind these emissions and how do the direct materials; energy requirements and transportation contribute to the generation of emissions. The cradle-to-gate approach is carried out.

Results and discussion

The results show that demands for both thermal energy and electricity are higher for the RWP than for the VP if only the manufacturing stages are considered. However, a different picture emerges when the analysis looks at the entire life cycle of the production. GHG from the VP are about 30 % higher than the RWP, over the life cycle emitting 568 kg CO₂ eq more per kilogram of tissue paper. GHG emissions from the wood pulping alone were 559 g CO₂ eq per kilogram of tissue paper, three times higher than waste paper collection and transportation.

Conclusions

In terms of GHG emissions from cradle to gate, the recycled process less intensive than the virgin one for two reasons. First, as shown in the results the total GHG emissions from RWP are lower than those from VP due to relatively lower energy and material requirements. Second is the non-recyclability nature of tissue paper. Because the tissue paper is the last use of fibre, using RWP as an input would be preferable over using VP. The environmental profile of the tissue products both from RWP and VP can be improved if the following conditions are considered by the company. First, the company should consider implementing a cogeneration unit to simultaneously generate both useful heat and electricity. Second, it may consider changing the VP mix, in order to avoid the emissions associated with long distance transpiration effort. Third, there is the option of using sludge as fuel, which would reduce the total fossil fuel requirement.     

<Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> Vaginitis: Microbiology and In Vitro Antifungal Susceptibility

Genitourinary infections by Saccharomyces cerevisiae are rare. Here, we describe eight S. cerevisiae vulvovaginitis episodes where molecular (Affirm VPIII) and conventional microbiological methods (culture and carbohydrate assimilation) have proven to be inadequate for diagnostic purposes. DNA sequencing allowed the correct identification of the pathogen. All isolates were susceptible to most antifungal agents, with two of them also found to be susceptible-dose-dependent to itraconazole.     

Enhanced Terahertz Transmission Through Bullseye Plasmonics Lenses Fabricated Using Micromilling Techniques

Imaging applications at terahertz frequencies are, in general, limited to relatively low spatial resolution due to the effects of diffraction. By using a subwavelength aperture in the near-field, however, it is possible to achieve subwavelength resolution, although low transmission through the aperture limits the sensitivity of this approach. Plasmonic lenses in the form of bullseye structures, which consist of a circular subwavelength aperture surrounded by concentric periodic corrugations, have demonstrated enhanced transmission, thereby increasing the utility of near-field imaging configurations. In this paper, the design, fabrication, and experimental performance of plasmonic lenses optimized for 300 GHz are discussed. While nanofabrication techniques are required for optical applications, microfabrication techniques are sufficient for terahertz applications. The process flow for fabricating a double-sided bullseye structure using a precision micromilling technique is described. Transmission and beam profile measurements using a customized terahertz testbed are presented.

     

Functional analysis of a dominant mutation of human connexin26 associated with nonsyndromic deafness

Cx26 has been implicated in dominant (DFNA3) and recessive (DFNB1) forms of nonsyndromic sensorineural deafness. While most homozygous DFNB1 Cx26 mutations result in a simple loss of channel activity, it is less clear how heterozygous mutations in Cx26 linked to DFNA3 cause hearing loss. We have tested the ability of one dominant mutation (W44C) to interfere with wild-type human Cx26 (HCx26wt). HCx26wt induced robust electrical conductance between paired oocytes, and facilitated dye transfer between transfected HeLa cells. In contrast, oocyte pairs injected with only W44C were not electrically coupled above background levels, and W44C failed to dye couple transfected HeLa cells. Moreover, W44C dramatically inhibited intercellular conductance of HCx26wt when co-expressed in an equal ratio, and the low levels of residual conductance displayed altered gating properties. A nonfunctional recessive mutation (W77R) did not inhibit the ability of HCx26wt to form functional channels when co-injected in the same oocyte pairs, nor did it alter HCx26wt gating. These results provide evidence for a functional dominant negative effect of the W44C mutant on HCx26wt and explain how heterozygous Cx26 mutations could contribute to autosomal dominant deafness, by resulting in a net loss, and/or alteration, of Cx26 function.     

Asymmetric division and cosegregation of template DNA strands in adult muscle satellite cells

Satellite cells assure postnatal skeletal muscle growth and repair. Despite extensive studies, their stem cell character remains largely undefined. Using pulse-chase labelling with BrdU to mark the putative stem cell niche, we identify a subpopulation of label-retaining satellite cells during growth and after injury. Strikingly, some of these cells display selective template-DNA strand segregation during mitosis in the muscle fibre in vivo, as well as in culture independent of their niche, indicating that genomic DNA strands are nonequivalent. Furthermore, we demonstrate that the asymmetric cell-fate determinant Numb segregates selectively to one daughter cell during mitosis and before differentiation, suggesting that Numb is associated with self-renewal. Finally, we show that template DNA cosegregates with Numb in label-retaining cells that express the self-renewal marker Pax7. The cosegregation of 'immortal' template DNA strands and their link with the asymmetry apparatus has important implications for stem cell biology and cancer.     

Loss-of-function and residual channel activity of connexin26 mutations associated with non-syndromic deafness

Connexins are the protein subunits of gap junction channels that allow a direct signaling pathway between networks of cells. The specific role of connexin channels in the homeostasis of different organs has been validated by the association of mutations in several human connexins with a variety of genetic diseases. Several connexins are present in the mammalian cochlea and at least four of them have been proposed as genes causing sensorineural hearing loss. We have started our functional analysis by selecting nine mutations in Cx26 that are associated with non-syndromic recessive deafness (DFNB1). We have observed that both human Cx26 wild-type (HCx26wt) and the F83L polymorphism, found in unaffected controls, generated electrical conductance between paired Xenopus oocytes, which was several orders of magnitude greater than that measured in water-injected controls. In contrast, most recessive Cx26 mutations (identified in DFNB1 patients) resulted in a simple loss of channel activity. In addition, the V37I mutation, originally identified as a polymorphism in heterozygous unaffected individuals, was devoid of function and thus may be pathologically significant. Unexpectedly, we have found that the recessive mutation V84L retained functional activity in both paired Xenopus oocytes and transfected HeLa cells. Furthermore, both the magnitude of macroscopic junctional conductance and its voltage-gating properties were indistinguishable from those of HCx26wt. The identification of functional differences of disease causing mutations may lead to define which permeation or gating properties of Cx26 are necessary for normal auditory function in humans and will be instrumental in identifying the molecular steps leading to DFNB1.