Electrophysiological identification of antennal pH receptors in the ground beetle Pterostichus oblongopunctatus

Abstract. Electrophysiological responses of antennal taste bristles to 100 mm acetate and phosphate buffers were tested at pH 3–11 in the ground beetle Pterostichus oblongopunctatus (F.) (Coleoptera, Carabidae). Additionally, responses of these sensilla to 10 and 100 mm phosphate buffers were compared with each other. Generally, in response to these stimulating solutions, two sensory cells, classified as a salt cell (cation cell) and a pH cell, respectively, showed action potentials distinguished by differences in their amplitudes and polarity of spikes. The firing rate of the cation cell increased with increasing buffer concentration, and was influenced by buffer pH in a complicated way. The best stimulus for the second cell (pH cell) was pH of the stimulating buffer solution. As the pH of the stimulus solution increased, higher rates of firing were produced by the pH cell. For example, the number of action potentials elicited by 100 mm phosphate buffer at pH 11.1 was approximately 16-fold higher compared with that at pH 8.1, and firing rates during the first second of the response were 27.9 and 1.7 imp/s, respectively. The pH cell did not fire or fired at very low frequency (first second response below 5 imp/s) at pH 3–6. This level of acidity probably represents the pH preferences of this ground beetle in its forest habitat and hibernating sites. By contrast to the cation cell, the pH cell responded to increases in buffer concentration by decreasing its firing rate.     

Identification of a novel, multifunctional β-defensin (human β-defensin 3) with specific antimicrobial activity

Previous studies have shown the implication of beta-defensins in host defense of the human body. The human beta-defensins 1 and 2 (hBD-1, hBD-2) have been isolated by biochemical methods. Here we report the identification of a third human beta-defensin, called human beta-defensin 3 (hBD-3; cDNA sequence, Genbank accession no. AF295370), based on bioinformatics and functional genomic analysis. Expression of hBD-3 is detected throughout epithelia of many organs and in non-epithelial tissues. In contrast to hBD-2, which is upregulated by microorganisms or tumor necrosis factor-alpha (TNF-alpha), hBD-3 expression is increased particularly after stimulation by interferon-gamma. Synthetic hBD-3 exhibits a strong antimicrobial activity against gram-negative and gram-positive bacteria and fungi, including Burkholderia cepacia. In addition, hBD-3 activates monocytes and elicits ion channel activity in biomembranes, specifically in oocytes of Xenopus laevis. This paper also shows that screening of genomic sequences is a valuable tool with which to identify novel regulatory peptides. Human beta-defensins represent a family of antimicrobial peptides differentially expressed in most tissues, regulated by specific mechanisms, and exerting physiological functions not only related to direct host defense.     

REPuter: the manifold applications of repeat analysis on a genomic scale

The repetitive structure of genomic DNA holds many secrets to be discovered. A systematic study of repetitive DNA on a genomic or inter-genomic scale requires extensive algorithmic support. The REPuter program described herein was designed to serve as a fundamental tool in such studies. Efficient and complete detection of various types of repeats is provided together with an evaluation of significance and interactive visualization. This article circumscribes the wide scope of repeat analysis using applications in five different areas of sequence analysis: checking fragment assemblies, searching for low copy repeats, finding unique sequences, comparing gene structures and mapping of cDNA/EST sequences.     

Linking a farm model and a location optimization model for evaluating energetic and material straw valorization pathways—A case study in Baden‐Wuerttemberg

Diminishing fossil carbon resources, global warming, and increasing material and energy needs urge for the rapid development of a bioeconomy. Biomass feedstock from agro‐industrial value chains provides opportunities for energy and material production, potentially leading to competition with traditional food and feed production. Simulation and optimization models can support the evaluation of biomass value chains and identify bioeconomy development paths, potentials, opportunities, and risks. This study presents the linkage of a farm model (EFEM) and a techno‐economic location optimization model (BIOLOCATE) for evaluating the straw‐to‐energy and the innovative straw‐to‐chemical value chains in the German federal state of Baden‐Wuerttemberg taking into account the spatially distributed and price‐sensitive nature of straw supply. The general results reveal the basic trade‐off between economies of scale of the energy production plants and the biorefineries on the one hand and the feedstock supply costs on the other hand. The results of the farm model highlight the competition for land between traditional agricultural biomass utilization such as food and feed and innovative biomass‐to‐energy and biomass‐to‐chemical value chains. Additionally, farm‐modeling scenarios illustrate the effect of farm specialization and regional differences on straw supply for biomass value chains as well as the effect of high straw prices on crop choices. The technological modeling results show that straw combustion could cover approximately 2% of Baden‐Wuerttemberg's gross electricity consumption and approximately 35% of the district heating consumption. The lignocellulose biorefinery location and size are affected by the price sensitivity of the straw supply and are only profitable for high output prices of organosolv lignin. The location optimization results illustrate that economic and political framework conditions affect the regional distribution of biomass straw conversion plants, thus favoring decentralized value chain structures in contrast to technological economies of scale.     

Apoptosis is not required for acantholysis in pemphigus vulgaris

The autoimmune blistering skin disease pemphigus vulgaris (PV) is caused primarily by autoantibodies against desmosomal cadherins. It was reported that apoptosis can be detected in pemphigus skin lesions and that apoptosis can be induced by PV-IgG in cultured keratinocytes. However, the role of apoptosis in PV pathogenesis is unclear at present. In this study, we provide evidence that apoptosis is not required for acantholysis in PV. In skin lesions from two PV patients, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) positivity, but not cleaved caspase-3, was detected in single keratinocytes in some lesions but was completely absent in other lesions from the same patients. In cultures of human keratinocytes (HaCaT and normal human epidermal keratinocytes), PV-IgG from three different PV patients caused acantholysis, fragmented staining of Dsg 3 staining, and cytokeratin retraction in the absence of nuclear fragmentation, TUNEL positivity, and caspase-3 cleavage and hence in the absence of detectable apoptosis. To further rule out the contribution of apoptotic mechanisms, we used two different approaches that are effective to block apoptosis induced by various stimuli. Inhibition of caspases by z-VAD-fmk as well as overexpression of Fas-associated death domain-like interleukin-1beta-converting enzyme (FLICE)-like inhibitory proteins FLIP(L) and FLIP(S) to inhibit receptor-mediated apoptosis did not block PV-IgG-induced effects, indicating that apoptosis was not required. Taken together, we conclude that apoptosis is not a prerequisite for skin blistering in PV but may occur secondary to acantholysis.     

Histatin 5-derived peptide with improved fungicidal properties enhances human immunodeficiency virus type 1 replication by promoting viral entry

Antimicrobial peptides are found in a number of body compartments and are secreted at mucosal surfaces, where they form part of the innate immune system. Many of these small peptides have a broad spectrum of inhibitory activity against bacteria, fungi, parasites, and viruses. Generally, the peptide's mode of action is binding and disruption of membranes due to its amphipathic properties. Histatin 5 is a salivary peptide that inhibits Candida albicans, an opportunistic fungus that causes oropharyngeal candidiasis in a majority of human immunodeficiency virus type 1 (HIV-1)-infected patients progressing towards AIDS. Previously, we increased the fungicidal properties of histatin 5 by replacing amino acids in the active domain of histatin 5 (Dh-5) (A. L. Ruissen, J. Groenink, E. J. Helmerhorst, E. Walgreen-Weterings, W. van't Hof, E. C. Veerman, and A. V. Nieuw Amerongen, Biochem. J. 356:361-368, 2001). In the current study, we tested the anti-HIV-1 activity of Dh-5 and its derivatives. Although Dh-5 inhibited HIV-1 replication, none of the peptide variants were more effective in this respect. In contrast, one of the derivatives, Dhvar2, significantly increased HIV-1 replication by promoting the envelope-mediated cell entry process. Most likely, Dhvar2 affects membranes, thereby facilitating fusion of viral and cellular membranes. This study shows that modification of antimicrobial peptides in order to improve their activity against a pathogen may have unpredictable and unwanted side effects on other pathogens.     

Chemical modification of the active site of ferredoxin-NADP reductase and conformation of the binary ferredoxin/ferredoxin-NADP reductase complex in solution

Ferredoxin-NADP⁺ reductase (EC 1.18.1.2) was chemically modified by the triplet probe eosin isothiocyanate (eosin-NES). Incorporation of 1 mol eosin-NCS/mol ferredoxin-NADP⁺ reductase completely inhibited binding of NADP⁺/NADPH to the enzyme. Binding of eosin without the reactive group to the enzyme was shown to be reversible but to compete with NADP⁺/NADPH with a K_i of approx. 5 μM. The binding site of eosin-NCS has been located in the primary sequence ferredoxin-NADP⁺ reductase. After specific cleavage of arginine with trypsin a single labelled peptide was obtained and identified as the fragment from residue 179–228 in the primary sequence. Binding of eosin-NCS occurred in either of two predicted helices (residues 179–189 or 212–228) which are both part of an /β structure characteristic for nucleotide binding folds. The rotational diffusion in solution of the eosin-labelled ferredoxin-NADP⁺ reductase and its complex with ferredoxin was measured with laser flash spectroscopy under photoselection. From the measured rotational correlation times and the known structure of ferredoxin-NADP⁺ reductase at 3.7 Å resolution, we propose that ferredoxin is bound to ferredoxin-NADP⁺ reductase between the two domains of the flavoprotein. The two ferredoxin-NADP⁺ reductase domains and ferredoxin form a triangle which results in a highly integrated binary complex.