Biosensors for environmental monitoring

Increasing environmental legislation which controls the release and the levels of certain chemicals in the environment has created a need for reliable monitoring of these substances in air, soil and especially water. Conventional analytical techniques, although highly precise, suffer from the disadvantages of high cost, the need for trained personnel and the fact that they are mostly laboratory bound. Biosensors because of their specificity, fast response times, low cost, portability, ease of use and a continuous real time signal, can present distinct advantages in certain cases. Their biological base makes them ideal for toxicological measurements which are suited for health and safety applications. Over the last 3-4 years there has been an increase in the number of publications concerning biosensors for environmental monitoring, especially in the field of pesticide measurements.This paper reviews some of the more important developments over the past 3-4 years.     

AIDS in Africa: distinguishing fact and fiction

The data widely purporting to show the existence and heterosexual transmission in Africa of a new syndrome caused by a retrovirus which induces immune deficiency are critically evaluated. It is concluded that both acquired immune deficiency (AID) and the symptoms and diseases which constitute the clinical syndrome (S) are of long standing in Africa, affect both sexes equally and are caused directly and indirectly by factors other than human immunodeficiency virus (HIV). Seropositivity to HIV in Africans usually represents no more than cross-reactivity caused by an abundance of antibodies induced by the numerous infectious and parasitic diseases which are endemic in Africa. The apparently high prevalence of AIDS and HIV seropositives is therefore not surprising and is not proof of heterosexual transmission of either HIV or AIDS.E. Papadopulos-Eleopulos is with the Department of Medical Physics, The Royal Perth Hospital, Perth 6000, Western Australia, Australia; V.F. Turner is with the Department of Emergency Medicine, The Royal Perth Hospital, Perth 6000, Western Australia, Australia, J.M. Papadimitriou is with the Department of Pathology, University of Western Australia, Perth, Western Australia. H. Bialy is with Bio/Technology, 65 Becker St, New York, NY 10012, USA.     

Cell-specific differences in membrane beta-glucosidase from normal and Gaucher cells.

Two isozymes of membrane-bound beta-glucosidase (beta-D-glucoside glucohydrolase, EC 3.2.1.21) with activity towards 4-methylumbelliferyl-beta-D-glucopyranoside have been identified in human cells. One of these isozymes was found to have a pH optimum of 5.0, a Km of 0.4 mM and to be rapidly inactivated at pH 4.0 ("acid-labile"). The second isozyme had a pH optimum of 4.5, a Km of 0.8 mM and was stable at pH 4.0 ("acid-stable"). Cultured long-term lymphoid lines and peripheral blood leukocytes contained both isozymes while cultured skin fibroblasts contained only the "acid-stable" form in detectable amounts. The specific activity of the "acid-stable" isozyme was severely reduced in cultured skin fibroblasts, cultured long-term lines and peripheral leukocytes from patients with Gaucher's disease. The specific activity of the "acid-labile" enzyme in the latter two cell types was apparently unaffected. The beta-glucosidase activity in all three cell types examined was predominantly particulate but the enzyme could be solubilized with low concentrations of Triton X-100. The solubilized enzyme required sodium taurocholate (0.2%) for maximum activity. Solubilized beta-glucosidase did not exhibit the cell-specific differences in pH optimum and Km shown by the membrane-bound enzyme.     

Persistent atrial paralysis: Case report with light microscopy and ultrastructural analyses

Persistent atrial paralysis in a patient with complete heart block and mild mitral insufficiency is presented. Left atrial specimens obtained during implantation of a permanent cardiac pulse generator showed evidence of hypertrophy and fibrosis; subcellular degenerative changes ranged from near normal to irreversible, thus suggesting that atrial paralysis may be due to the replacement of normal atrial muscle with nonfunctional fibrous tissue.     

Structure of bovine beta-lactoglobulin at 6A resolution.

Bovine β-lactoglobulin is a dimer with a molecular weight of 2 × 18,400. In solution it undergoes a pH-dependent transition at pH 7.0 between two alternative structures, named N and R. The structures of four different crystal forms have been determined by multiple isomorphous replacement with heavy-atoms. Two of them, lattices K and X, were crystallised at pH 6.5, corresponding to the N state in solution; and the other two, lattices Y and Z, were crystallised at pH 7.5, corresponding to the R state in solution. The figures of merit of the phase angles determined for these lattices were 0.76, 0.77, 0.80 and 0.80, respectively. The four structures that emerged are similar and show certain features suggestive of α-helices and pleated sheets, but the resolution is insufficient to trace the entire course of the polypeptide chain. No clear distinction can yet be made between the structures above or below pH 7.0, nor between the native molecule and the molecule from which the C-terminal leucine and histidine residues have been cleaved. Analyses at higher resolution are in progress.     

Bioenergetics of the range caterpillar,Hemileuca oliviae (Ckll.)

Summary The energy budget of the New Mexico range caterpillar, Hemileuca oliviae (Ckll.), was estimated on caterpillars reared on an artificial diet under constant conditions. There was a 54% average age-specific decrease in assimilation efficiency in the larval stages. Net production efficiency was relatively constant at 52%. Of the 26.77 cal/mg produced by the feeding larvae, 0.53 cal/mg were expended by the pupae and 0.59 cal/mg were expended by the adults. Since maintenance costs of the non-feeding stages accounted for only four percent of the larval production, that production was adequate for adult development and reproduction.     

Effects of radiation,temperature and humidity on photosynthesis,transpiration and water use efficiency of oilseed rape (Brassica campestris L.)

The net photosynthetic rate (F), transpiration rate (Q) and water use efficiency (F/Q) of oilseed rape (Brassica campestris L. cv. Span) was studied under a range of atmospheric conditions by gas exchange techniques. The plants were at the full bloom/pod initiation stage of development at the time of measurement. The environmental conditions consisted of various levels of photosynthetically active radiation (100 to 2800 (μmol m^?2 s^?1 PAK: 400–700 nm), air temperature (10 to 42°C) and vapour pressure deficit (0.7 to 2.1 kPa VPD). The peak values ofF were recorded at 1600 μmol m^?2 s^?1 PAR, 20°C air temperature and 1.2 kPa VPD of air in the chamber. Q increased with increasing PAR, air temperature and VPD. However, theF/Q remained high and almost constant from 600 to 1600 μmol m^?2 s^?1 PAR, but declined at the low and high photon flux densities.F/Q decreased progressively with increase in air temperature and VPD of air in the chamber.     

Separation of urine proteins on the anion-exchange resin mono QTM

Proteins excreted in urine due to renal failure were separated on Mono Q^TM, a new strong anion exchanger designed for fast high-resolution protein separations. The separation procedure was divided into two steps. The first step involved removal of low-molecular- weight substances by rapid desalting on a Sephadex G-25 Superfine column. In the second step, the total protein fraction (3–6 ml) was loaded onto the Mono Q column with the aid of a superloop. The proteins were adsorbed onto the top of the ion-exchanger column and gradually displaced by a combined pH and salt gradient in 40 min. The choice of ion exchanger and initial operating conditions were based on data obtained from electrophoretic titration curve experiments. Identification of separated proteins was achieved by fused rocket electrophoresis and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, respectively.     

Characterization of human loricrin. Structure and function of a new class of epidermal cell envelope proteins

We have isolated and characterized a full-length cDNA clone encoding human loricrin. Curiously, this protein displays major differences from the recently described mouse loricrin (Mehrel, T., Hohl, D., Nakazawa, H., Rothnagel, J.A., Longley, M.A., Bundman, D., Cheng, C.K., Lichti, U., Bisher, M.E., Steven, A. C., Steinert, P.M., Yuspa, S.H., and Roop, D.R. (1990) Cell 61, 1103-1112). Although both proteins are glycine-serine-cysteine-rich, the sequences have not been conserved. However, analysis of the sequences reveals a common motif of quasi-peptide repeats of an aliphatic or aromatic amino acid residue followed by several glycine and/or serine and cysteine residues. These sequences are interspersed and flanked by short glutamine- or glutamine/lysine-rich peptides. Thus loricrins consist of a family of cell envelope proteins of highly variable sequences that nevertheless retain common structural elements. We show that unlike all other putative protein components of the cell envelope, loricrins are highly insoluble, due at least in part to cross-linking by disulfide bonds. Furthermore, we have isolated four peptides from purified human cell envelopes that contain recognizable loricrin sequences and which are cross-linked by the N epsilon-(gamma-glutamyl)lysine isodipeptide bond. The presence of such bonds thus affords an explanation for the extraordinary insolubility of loricrin by cross-linking to the cell envelope and can also explain the low steady-state levels of monomeric loricrin in cytoskeletal extracts of epidermis. This study represents the first report of this isodipeptide cross-link in a protein component of the cornified cell envelope. We propose a model for the structure of loricrin in which (i) the unusual glycine-serine-rich sequences adopt a flexible loop conformation, indexed on the recurrent aliphatic residues; (ii) inter- or intramolecular isodipeptide and disulfide cross-links induce or stabilize folding of loricrin so as to form a more compact rosette-like structure; and (iii) the presence of the flexible glycine-rich loops necessarily will impact a flexible character to the cell envelope and entire epithelium.