Survival Rate and Enzyme Activities ofLactobacillus acidophilusFollowing Frozen Storage

The ability of two strains of Lactobacillus acidophilus, CRL 640 and CRL 800, to survive and retain their biological activities under frozen storage was determined. Freezing and thawing, as well as frozen storage, damaged the cell membrane, rendering the microorganisms sensitive to sodium chloride and bile salts. Both lactic acid production and proteolytic activity were depressed after 21 days at -20 degreesC, whereas beta-galactosidase activity per cell unit was increased. Cell injury was partially overcome after repair in a salt-rich medium. Copyright 1998 Academic Press.     

Dark states in monomeric red fluorescent proteins studied by fluorescence correlation and single molecule spectroscopy

Monomeric red fluorescent proteins (mRFPs) have become indispensable tools for studying protein dynamics, interactions and functions in the cellular environment. Their emission spectrum can be well separated from other fluorescent proteins, and their monomeric structure preserves the natural function of fusion proteins. However, previous photophysical studies of some RFPs have shown the presence of light-induced dark states that can complicate the interpretation of cellular experiments. In this article, we extend these studies to mRFP1, mCherry, and mStrawberry by means of fluorescence correlation spectroscopy and prove that this light-driven intensity flickering also occurs in these proteins. Furthermore, we show that the flickering in these proteins is pH-dependent. Single molecule spectroscopy revealed reversible transitions from a bright to a dark state in several timescales, even up to seconds. Time-resolved fluorescence spectroscopy showed multiexponential decays, consistent with a “loose” conformation. We offer a structural basis for the fluorescence flickering using known crystal structures and point out that the environment of Glu-215 is critical for the pH dependence of the flickering in RFPs. We apply dual-color fluorescence correlation spectroscopy inside live cells to prove that this flickering can seriously hamper cellular measurements if the timescales of the flickering and diffusion are not well separated.     

Comparison of the Respiratory Metabolism of Plantago lanceolata L. and Plantago major L.

Bryce, J. H. and ap Rees, T. 1985. Comparison of the respiratorymetabolism of Plantago lanceolata L. and Plantago major L.—J.exp. Bot. 36 1559–1565. The aim of this work was to discover if the respiratory metabolismof the roots of Plantago lanceolata L. differed from that ofthe roots of Plantago major L. Measurements of oxygen uptakeand dry weight of excised root systems during growth of seedlingsprovided evidence that the two species differed in the amountof respiration needed to support a given increase in dry weight.Excised root systems were given a 6-h pulse in [U-¹⁴C]sucrosefollowed by a 16.5-h chase in sucrose. The detailed distributionof ¹⁴C amongst the major components of the roots at the endof the pulse and the chase revealed no significant differencebetween the two species. Patterns of ¹⁴CO₂ production from [1-¹⁴C],[2-¹⁴C], [3,4-¹⁴C], and [6-¹⁴C]glucose of excised root systemsfrom plants of three ages were similar for the two species.It is suggested that there is no conclusive evidence for anysignificant inherent difference in the respiratory metabolismof the roots of the two species. Key words: ¹⁴C sugar metabolism, respiration, roots, Plantago     

Rapid recycling of triose phosphates in oak stem tissue

We report the carbon-13 and oxygen-18 isotope ratios in cellulose from the early and late wood of pedunculate oak (Quercus robur L.). The δ¹³ C value of the early wood correlates best with that of the late wood of the previous year. The δ¹⁸O value of the early wood correlates best with that of the late wood of the same year. We suggest that a biochemical explanation of these data is that there is a rapid cycle between hexose monophosphates and triose phosphates in oak stem tissue during cellulose synthesis. Evidence in support of this explanation is provided by the intramolecular distribution of ¹⁴C in labelled fructose extracted from cores of wood that had been supplied with [1?¹⁴C]- and [6-¹⁴C]glucose.     

Production and characterization of polyclonal and monoclonal antibodies against Aflatoxin B1 oxime-BSA in an enzyme-linked immunosorbent assay

From a single aflatoxin B₁ oxime — bovine serum albumin conjugate, polyclonal and monoclonal antibody preparations were produced. The four rabbit polyclonal antisera were specific for aflatoxin Bi in a microtitration plate enzyme — linked immunosorbent assay. The monoclonal antibodies showed a wide range of differing specificities, recognizing, for example, aflatoxins B₁, B₂, G₁ and G₂; B₁ and B₂; B₁ and G₁; and G₁ alone. No antibody preparations reacted with aflatoxin M₁. The significance of these results to the strategy of anti-aflatoxin antibody production for use in quantitative enzyme immunoassays is discussed.     

Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atrox

Background  

Bothrops atrox is responsible for the majority of snakebite accidents in the Brazilian Amazon region. Previous studies have demonstrated that the biological and pharmacological activities of B. atrox venom alter with the age of the animal. Here, we present a comparative proteome analysis of B. atrox venom collected from specimens of three different stages of maturation: juveniles, sub-adults and adults.