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Biomedical subjects

M Kertesz

Publications and source records attributed to M Kertesz.

17 recordsLinked to original sources

Carbon nanotube actuators

Electromechanical actuators based on sheets of single-walled carbon nanotubes were shown to generate higher stresses than natural muscle and higher strains than high-modulus ferroelectrics. Like natural muscles, the macroscopic actuators are assemblies of billions of individual nanoscale actuators. The actuation mechanism (quantum chemical-based expansion due to electrochemical double-layer charging) does not require ion intercalation, which limits the life and rate of faradaic conducting polymer actuators. Unlike conventional ferroelectric actuators, low operating voltages of a few volts generate large actuator strains. Predictions based on measurements suggest that actuators using optimized nanotube sheets may eventually provide substantially higher work densities per cycle than any previously known technology.

Journal Article↗

Chloromethane Metabolism by Methylobacterium sp. Strain CM4

Methylobacterium sp. strain CM4 metabolized chloromethane quantitatively with a molar yield of 2.8 g of whole-cell protein/mol of C. This value was similar to that observed after growth with methanol (2.9 g of protein/mol of C) and about three times larger than the yield with formate (0.94 g of protein/mol of C). Chloromethane dehalogenation activity was inducible. MiniTn5 transposon insertion mutants with altered growth characteristics with chloromethane and other C1 compounds were isolated and characterized. Nine of these were unable to grow with chloromethane but were able to grow with methanol, methylamine, or formate. Seventy-three transposon mutants that were defective in the utilization of either methanol, methylamine, methanol plus methylamine, or formate could still grow with chloromethane. Based on the protein yield data and the properties of the transposon mutants, we propose a pathway for chloromethane metabolism that depends on methyltransferase and dehydrogenase activities.

Journal Article↗

Sample handling for proteome analysis.

The main factor limiting the sensitivity range for the identification of proteins isolated by two-dimensional (2-D) electrophoresis is sample handling: protein detection limits on the gel, losses during extraction and digestion, as well as interference of gel contaminants and detergents with the mass spectrometry (MS) detection increasing background noise. At the one hundred picomole level, losses are fairly negligible but when the amounts drop below 1 picomole (and subfemtomole peptide detection limits have been reported recently by MS), the losses become a critical point. In order to extend proteome analysis to include very low copy number proteins, methods must be developed to minimize losses and handling steps, maximize digestion and extraction yields, as well as to lower chemical noise. We present several methods that we have developed in our laboratory to: (i) increase the amount of material available in a sodium dodecyl sulfate (SDS)-free form which does not require staining, (ii) increase protein extraction and digestion yields and lower the contamination by autoproteolytic products, and (iii) allow direct modification of the peptide mixture to generate sequence tags.

Bacterial Proteins↗

Analysis of global responses by protein and peptide fingerprinting of proteins isolated by two-dimensional gel electrophoresis. Application to the sulfate-starvation response of Escherichia coli.

A set of 8 proteins (SSI, sulfate-starvation-induced proteins) was observed by comparative two-dimensional electrophoresis to be induced when Escherichia coli were grown using compounds other than sulfate or cysteine as the sole sulfur source. These proteins were isolated after two-dimensional gel electrophoresis, digested with trypsin and the masses of the resulting peptides determined by mass spectrometry. The list of peptide masses served as a protein fingerprint which was used to search the databases, allowing four of the SSI proteins (SSI2, 5, 7, 8) to be identified with a high degree of confidence. To identify the other SSI proteins, and to obtain sequence information for as many of the proteins as possible, automated on-line HPLC MS/MS (fragmentation analysis using coupled mass scanning devices) data collection was performed. The uninterpreted MS/MS spectra were used as peptide fingerprints to search the databases. Genes encoding two further proteins (SSI 1 and 3) were identified in the 8.5' region of the Escherichia coli genome. N-terminal sequencing of all of the proteins confirmed the results of protein and peptide fingerprinting and in addition showed that SSI 6 shows 50% similarity to the Bacillus subtilis orfM gene product. SSI 4 was not found in the databases by any of these methods. The methods described are of general use for the rapid analysis of complex cell responses. MS data accumulation takes about 5 min/protein for protein fingerprinting and 30 min for peptide fingerprinting and requires approximately 100 fmol of material. N-terminal sequencing however, takes about 5 h/protein and approximately 1 pmol to obtain a 10 amino acid sequence for a search.

Alkanesulfonates↗

Evidence for two distinct phosphonate-degrading enzymes (C-P lyases) in Arthrobacter sp. GLP-1.

Arthrobacter sp. GLP-1 can utilize a wide range of organophosphonates as its sole source of phosphorus. The in-situ formation of sarcosine and methane from glyphosate and methanephosphonic acid respectively was studied. These two processes are differentially induced during phosphorus-deprivation. Methanephosphonic acid strongly inhibits glyphosate degradation (I50 10 microM), but glyphosate has very little effect on methane generation (I50 150 mM). The pattern of inhibition by other organophosphonates and organophosphonate analogues is also very different for the two systems. Degradation of glyphosate and methanephosphonic acid therefore represent distinct processes.

Arthrobacter↗

Mutagenicity and PAH content of airborn particulates and of fallen dusts from two Hungarian towns and emission samples from aluminum reduction and power plants.

Urban air particulates (suspended particles and settling dust), furthermore dust emitted by a Soderberg aluminum reduction plant and a coal burning power plant from an industrial town, Ajka (30,000 inhabitants) were analysed for PAH content (liquid chromatography) and mutagenicity (Salmonella microsome test). Air particulates from Papa--a town of similar size without considerable heavy industry--and corresponding plant emission from Inota, a third town in the study, served as controls. The dust content and the PAH concentration, as well as the mutagenic potency of the air in Ajka were higher than in Papa. Mutagenicity of the airborne particulates showed a clear seasonality with a winter maximum and a summer minimum in both towns. The mutagenic potency of air correlated well with the air BaP and total PAH content in Ajka, but not in Papa. The amounts of extractable organic material and mutagenic potency as calculated for unit quantity of airborne particulate matter was higher in the Papa samples. Similar differences between the two towns were observed in the case of fallen dust, too. On the basis of examination of emitted dust, it can be stated, that in the mutagenicity of urban air, the aluminum plant emission plays a considerably higher role than the power plant emission, which is the main component of air dust pollution in Ajka.

Air Pollutants↗

Biphasic changes in anterior pituitary Met-enkephalin concentration following reserpine treatment.

Met-enkephalin concentrations in the anterior pituitary gland were shown to decline dramatically within the first 24 h after reserpine treatment, with effects apparent as early as 6 h. This was followed by subsequent repletion and late augmentation of Met-enkephalin levels 3 weeks following reserpine. Treatment with the alpha-1-adrenergic agonist methoxamine had no effect, whereas the alpha-1-antagonist prazosin lowered Met-enkephalin concentrations. Treatment with the dopamine agonists bromocriptine or apomorphine had no effect, but haloperidol treatment increased anterior pituitary Met-enkephalin which was reversed by concomitant bromocriptine administration. We postulate that the changes in anterior pituitary Met-enkephalin following reserpine were related to alterations in the monoamine neurotransmitters. Adrenergic and dopaminergic mechanisms may have opposing roles in the maintenance of Met-enkephalin concentrations in the anterior pituitary gland.

Animals↗

[Met5]enkephalin concentrations in rat pituitary are maintained under opioid inhibition.

[Met5]Enkephalin concentrations in rat pituitary were examined following the administration of the opiate antagonist naloxone 5 mg/kg per day as single daily s.c. injections for 9 days. Compared to vehicle administration, there was a significant increase of [Met5]enkephalin concentrations in both pituitary lobes, ranging from 50-90% above control values. Similar treatment with the opioid agonist [D-Ala2,MePhe4,Met(o)5ol]enkephalin in doses up to 500 micrograms/kg per day did not result in any alteration of [Met5]enkephalin levels. These data suggest that opioid receptor mechanisms play a role in maintaining [Met5]enkephalin concentrations in both neurointermediate and anterior pituitary.

Animals↗

Dopamine receptors regulate Met-enkephalin content in pituitary.

Met-enkephalin-like immunoreactive material (ME-LI) in the neurointermediate lobe of rat pituitary was dramatically altered by the long-term administration of drugs altering dopamine receptor activity. Treatment with the dopamine receptor antagonist haloperidol resulted in significant increases of ME-LI and treatment with the dopamine receptor agonist bromocriptine resulted in significant decreases of ME-LI in the neurointermediate pituitary. The treatments had considerably less effect on ME-LI in the anterior lobe of the pituitary. These results suggest that dopamine receptor activation or inhibition influences the content of ME-LI in the rat pituitary gland.

Animals↗

Probing protein function using a combination of gene knockout and proteome analysis by mass spectrometry.

Recently the determination of the genome sequences of three procaryotes (Haemophilus influenzae, Methanococcus jannaschii and Mycoplasma genitalium) as well as the first eucaryotic genome (Saccharomyces cerevisiae) were completed. Between 40-60% of the genes were found to code for proteins to which no function could be assigned. We describe an approach which combines proteome analysis (mapping of expressed proteins isolated by two-dimensional polyacrylamide gel electrophoresis to the genome) with genetic manipulations to study the complex pattern of protein regulation occurring in Escherichia coli in response to sulfate starvation. We have previously described the upregulation of eight spots on two-dimensional (2-D) gels in response to sulfate starvation and the assignment of six of these to entries in the E. coli genome sequence (Quadroni et al., Eur. J. Biochem. 1996, 239, 773-781). Here we describe the identification of the remaining two proteins which are encoded in a sulfate-controlled operon in the 21.5' region of the E. coli genome. Upregulated protein spots were cut from multiple 2-D gels collected and run on a modified funnel gel to concentrate the proteins and remove the sodium dodecyl sulfate before digestion. The peptide masses obtained from the digests were used to search the SwissProt database or a six-frame translation of the EMBL DNA database using a peptide mass fingerprinting algorithm. A digest can be reanalyzed after deuterium exchange to obtain a second, orthogonal data set to increase the confidence level of protein identification. The digests of the remaining unidentified proteins were used for peptide fragment generation using either post-source decay in a matrix-assisted laser desorption ionization (MALDI) time-of-flight mass spectrometer or collision-induced dissociation (CID) coupled mass spectrometry (MS/MS) with triple stage quadrupole or ion trap mass spectrometers. The spectra were used as peptide fragment fingerprints to search the SwissProt and EMBL databases.

Amino Acid Sequence↗

Met-enkephalin-like immunoreactivity in neurointermediate pituitary is decreased by DA receptor stimulation.

The effect of dopamine receptor stimulation by administration of the dopamine analogue bromocriptine on Met-enkephalin-LI was examined in rat hypothalamus, and neurointermediate and anterior lobes of pituitary. Bromocriptine treatment resulted in a dramatic decline of Met-enkephalin-LI in neurointermediate pituitary which was significant by 3 days of treatment. Maximal reduction of Met-enkephalin-LI ranged between 60-70% of pretreatment values and was maintained as long as bromocriptine was administered (4 weeks), with no evidence of desensitization or "escape." The effects of bromocriptine on neurointermediate lobe were of long duration and persisted for at least 4 days after discontinuation of treatment. No significant effects of bromocriptine were detected on Met-enkephalin-LI in hypothalamus or anterior pituitary. Whether these differences represent truly different regional regulation of Met-enkephalin-LI or whether the changes are more sensitively reflected in an area such as neurointermediate lobe that largely consists of nerve terminals, remains to be shown.

Animals↗

Met-enkephalin concentrations in striatum respond reciprocally to alterations in dopamine neurotransmission.

The striatum is richly innervated by both enkephalinergic and dopaminergic neurons, providing an anatomic framework from which intimate functional interrelationships between these neuronal systems may be postulated. Accordingly, many functional processes within dopamine neurons have been shown to be modulated by opioid peptides. In the present study we confirm predictable reciprocal effects in enkephalin neurons, brought about by modification of dopamine neurotransmission. Dopamine receptor blockade reliably increased striatal Met-enkephalin concentrations by about 50%, whereas chronic treatment with a potent long-acting dopamine receptor agonist was necessary to demonstrate a small 10-20% decrease in Met-enkephalin concentrations. Depletion of presynaptic dopamine also resulted in a marked 50-60% augmentation of Met-enkephalin levels, that could be prevented by concomitant treatment with a dopamine analogue. Increasing dopamine turnover and release by a mu-opioid agonist decreased Met-enkephalin concentrations, as might have been predicted. Thus we have shown a marked dopaminergic influence that maintains striatal Met-enkephalin concentrations by near maximal tonic inhibitory effects.

Animals↗