Electrophysiological identification of cold receptors on the antennae of the ground beetle Pterostichus aethiops

Abstract.  In single-sensillum extracellular electrophysiological recordings, terminal campaniform sensilla at the tip of antennae of the ground beetle Pterostichus aethiops (Pz., 1797) show action potentials of three sensory cells, A-, B- and C-cells, distinguished by differences in their spike amplitudes. Only the A-cell, with the largest spike amplitude, is highly sensitive to temperature fluctuations, showing remarkable changes in its firing rate induced by changes in temperature of 0.1 °C. The firing rate of A-cells at 23 °C varies from 15–52 Hz among different beetles. Mean impulse frequency of A-cells is found to be a function of steady temperature, the firing rate decreasing with temperature increase. A-cells respond to a rapid temperature drop with a strong phasic-tonic reaction; larger decreases in temperature evoke higher peak frequency values. Maximum peak frequencies, varying from 380–630 Hz in different beetles, are induced by temperature decreases of 3–10 °C, whereas temperature rise strongly inhibits impulse activity of the A-cell. The first manifestation of rapid warming in the nerve impulse sequence is a very long interspike period, followed by diminished activity. Both the length of the long interspike period and the rate of following impulse activity are functions of temperature change; hence, A-cells respond to temperature changes as typical cold receptors, similar to coeloconic and short hair-like sensilla in other insects.     

Effect of amino acid substitutions on the candidacidal activity of LFampin 265-284

LFampin 265-284, derived from bovine lactoferrin, has broad-spectrum antimicrobial activity against the yeast Candida albicans and several Gram-positive and Gram-negative bacteria. A glycine substitution scan was used to identify residues that are important for its candidacidal activity. Each single substitution of a positively charged residue led to considerable reduction in candidacidal activity, for each residue to a different extent. Substitution within the helix-facilitating N-terminal sequence DLIW had less severe effect; substitution of Ile and Trp led to a somewhat reduced potency. No substantial effects were found on the propensity to adopt a helical structure or to bind to C. albicans cells.     

Biochemical markers and genetic research of ADHD

ADHD (attention hyperactivity disorder) is a polygenetic disorder with various candidate genes. At this time, more than thirty dopaminergic, noradrenergic, serotonergic and GABA-ergic genes are known. The research of only some candidate genes (DRD4, DAT, DRD5, DBH, 5HTT, HTR1B and SNAP25) brought relatively consistent results confirming the heredity of ADHD syndromes. The results of research of other genes (DRD2, DRD3, MAO, ADR2A, GABA A3, GABA B3) are not clear yet. This paper summarizes the most important genetic data in correlations with biochemical periphery parameters (especially for DBH, HVA, MHPG, serotonin). Hypothetically, certain subgroups of ADHD may be identified by correlation of biochemical characteristics and some candidate genes. The paper discusses some implications for future research. Review.     

Environmental signals from leaves--a physiognomic analysis of European vegetation

Leaf physiognomic traits vary predictably along climatic and environmental gradients. The relationships between leaf physiognomy and climate have been investigated on different continents, but so far an investigation based on European vegetation has been missing. A grid data set (0.5 degrees x 0.5 degrees latitude/longitude) has been compiled in order to determine spatial patterns of leaf physiognomy across Europe. Based on distribution maps of native European hardwoods, synthetic chorologic flora lists were compiled for all grid cells. Every synthetic chorologic flora was characterised by 25 leaf physiognomic traits and correlated with 16 climatic parameters. Clear spatial patterns of leaf physiognomy have been observed, which are statistically significant related to certain, temperature-related climate parameters. Transfer functions for several climatic parameters have been established, based on the observed relationships. The study provides evidence that synthetically generated floras represent a powerful tool for analysing spatial patterns of leaf physiognomy and their relationships to climate. The transfer functions from the European data set indicate slightly different relationships of leaf physiognomy and environment compared with results obtained from other continents.     

General Description of Partly Meromictic Hypertrophic Lake Verevi,its Ecological Status,Changes during the Past Eight Decades,and Restoration Problems

The present study describes generally the ecosystem of Lake Verevi while more detailed approaches are presented in the same issue. The main task of the article is to estimate long-term changes and find the best method for the restoration of good ecological status. Lake Verevi (surface 12.6 ha, mean depth 3.6 m, maximum depth 11 m, drainage area 1.1 km², water exchange 0.63-times per year) is a hypertrophic hard-water lake located in town Elva (6400 inhabitants). Long-term complex limnological investigations have taken place since 1929. The lake has been contaminated by irregular discharge of urban wastewaters from oxidation ponds since 1978, flood from streets, and infiltrated waters from the surrounding farms. The so-called spring meromixis occurred due to extremely warm springs in recent years. The index value of buffer capacity of Lake Verevi calculated from natural conditions is on the medium level. Water properties were analysed according to the requirements of the EU Water Framework Directive. According to the classification, water quality as a long-term average of surface layers is moderate-good, but the water quality of bottom layers is bad. Values in deeper layers usually exceed 20–30 times the calculated reference values by Vighi and Chiaudani’s model. Naturally, at the beginning of the 20th century the limnological type of the lake was moderately eutrophic. During the 1980s and 1990s the ecosystem was out of balance by abiotic characteristics as well as by plankton indicators. Rapid fluctuations of species composition and abundance can be found in recent years. Seasonal variations are considerable and species composition differs remarkably also in the water column. The dominating macrophyte species vary from year to year. Since the annual amount of precipitation from the atmosphere onto the lake surface is several times higher, the impact of swimmers could be considered irrelevant. Some restoration methods were discussed. The first step, stopping external pollution, was completed by damming the inlet. Drainage (siphoning) of the hypolimnetic water is discussed. Secondary pollution occurs because Fe:P values are below the threshold. The authors propose to use phosphorus precipitation and hypolimnetic aeration instead of siphoning.     

Lactoferrampin, an antimicrobial peptide of bovine lactoferrin, exerts its candidacidal activity by a cluster of positively charged residues at the C-terminus in combination with a helix-facilitating N-terminal part

The antimicrobial activity of bovine lactoferrin (bLF) is attributed to lactoferricin, which is situated in the N1-domain of bLF. Recently, another antimicrobial domain consisting of residues 268-284, designated lactoferrampin (LFampin), has been identified in the N1-domain of bLF, which exhibited antimicrobial activity against Candida albicans and several bacteria. In the present study, the candidacidal activity of a series of peptides spanning this antimicrobial domain was investigated in relation to the charge and the capacity to form a helical conformation in hydrophobic environments. C-Terminal truncation of LFampin resulted in a drastic decrease in candidacidal activity. Positively charged residues clustered at the C-terminal side of the LFampin domain appeared to be crucial for the candidacidal activity. The ability to adopt helical conformations did not change when LFampin was truncated at the C-terminal side. N-Terminally truncated LFampin peptides, truncated up to the sequence 270-284, were more reluctant to adopt a helical conformation. Therefore, we conclude that the C-terminal part of LFampin 265-284, which is the most active peptide, is crucial for its candidacidal activity, due to the presence of clustered positive charges, and that the N-terminal part is essential for activity as it facilitates helix formation.     

Electrophysiological identification of hygroreceptor neurons from the antennal dome-shaped sensilla in the ground beetle Pterostichus oblongopunctatus

This study gives the first electrophysiological evidence of hygroreceptors in carabids. Extracellular recordings from the antennal dome-shaped sensilla of the carabid beetle Pterostichus oblolongopunctatus (Coleoptera, Carabidae) clearly show the presence of moist and dry neuron antagonistically responding to humidity changes. The cold neuron of the same sensillum did not respond to changes in humidity. For the first time, we demonstrate that the binary system of two antagonistic hygroreceptor neurons discriminates differences between steady-state humidity levels more sensitively than either neuron separately. Another advantage of the binary system is that it guarantees immediate and strong phasic-tonic response to rapid humidity changes in either direction. In the hygrosensing system of carabids, this would allow detection of subtle step-changes in humidity with greater sensitivity than differences in steady-state values of humidity. Thus, construction of the hygrosensing system with opposing receptor neurons may allow insects to detect environmental humidity differences critical for their habitat and microhabitat selection, and survival with great precision.     

A one-enzyme strategy to release an antimicrobial peptide from the LFampin-domain of bovine lactoferrin

Antimicrobial peptides have been found throughout living nature, yet antimicrobial sequences may still lie hidden within a wide variety of proteins. A rational strategy was developed to select interesting domains, based on the presumed common features of antimicrobial peptides, and to release these from accessible and safe proteins. In silico proteolysis simulations of bovine lactoferrin (bLF) with selected endoproteinases predicted the liberation of peptides that encompasses a cationic amphipathic alpha-helix. Three predicted peptides were synthesized and tested for their biological activity, demonstrating that one single enzyme was sufficient to obtain an antimicrobial peptide. The proof of principle demonstrated that a 32-mer fragment isolated from the endoproteinase AspN digestion of bLF possessed strong antimicrobial activity. Moreover, desalted crude digest had improved activity over native bLF. Hence, selective digestion of bLF increases its antimicrobial activity by release of antimicrobial stretches.