bluuebunny/crossref_metadata_2025 huggingface dataset:
Amino acid residues on the surface of soybean 4‐kDa peptide involved in the interaction with its binding protein
Authors: Hanada Kazuki , Nishiuchi Yuji , Hirano Hisashi
Published: June 2003

Soybean 4‐kDa peptide, a hormone‐like peptide, is a ligand for the 43‐kDa protein in legumes that functions as a protein kinase and controls cell proliferation and differentiation. As this peptide stimulates protein kinase activity, the interaction between the 4‐kDa peptide (leginsulin) and the 43‐kDa protein is considered important for signal transduction. However, the mechanism of interaction between the 4‐kDa peptide and the 43‐kDa protein is not clearly understood. We therefore investigated the binding mechanism between the 4‐kDa peptide and the 43‐kDa protein, by using gel‐filtration chromatography and dot‐blot immunoanalysis, and found that the 4‐kDa peptide bound to the dimer form of the 43‐kDa protein. Surface plasmon resonance analysis was then used to explore the interaction between the 4‐kDa peptide and the 43‐kDa protein. To identify the residues of the 4‐kDa peptide involved in the interaction with the 43‐kDa protein, alanine‐scanning mutagenesis of the 4‐kDa peptide was performed. The 4‐kDa peptide‐expression system in Escherichia coli, which has the ability to install disulfide bonds into the target protein in the cytoplasm, was employed to produce the 4‐kDa peptide and its variants. Using mass spectrometry, the expressed peptides were confirmed as the oxidized forms of the native peptide. Surface plasmon resonance analysis showed that the C‐terminal hydrophobic area of the 4‐kDa peptide plays an important role in binding to the 43‐kDa protein.
AtCYS1, a cystatin from Arabidopsis thaliana, suppresses hypersensitive cell death
Authors: Belenghi Beatrice , Acconcia Filippo , Trovato Maurizio , Perazzolli Michele , Bocedi Alessio , Polticelli Fabio , Ascenzi Paolo , Delledonne Massimo
Published: June 2003

In plants, cysteine protease inhibitors are involved in the regulation of protein turnover and play an important role in resistance against insects and pathogens. AtCYS1 from Arabidopsis thaliana encodes a protein of 102 amino acids that contains the conserved motif of cysteine protease inhibitors belonging to the cystatin superfamily (Gln‐Val‐Val‐Ala‐Gly). Recombinant A. thaliana cystatin‐1 (AtCYS1) was expressed in Escherichia coli and purified. AtCYS1 inhibits the catalytic activity of papain (Kd = 4.0 × 10−2 µm, at pH 7.0 and 25 °C), generally taken as a molecular model of cysteine proteases. The molecular bases for papain inhibition by AtCYS1 have been analysed taking into account the three‐dimensional structure of the papain–stefin B complex. AtCYS1 is constitutively expressed in roots and in developing siliques of A. thaliana. In leaves, AtCYS1 is strongly induced by wounding, by challenge with avirulent pathogens and by nitric oxide (NO). The overexpression of AtCYS1 blocks cell death activated by either avirulent pathogens or by oxidative and nitrosative stress in both A. thaliana suspension cultured cells and in transgenic tobacco plants. The suppression of the NO‐mediated cell death in plants overexpressing AtCYS1 provides the evidence that NO is not cytotoxic for the plant, indicating that NO functions as cell death trigger through the stimulation of an active process, in which cysteine proteases and theirs proteinaceous inhibitors appear to play a crucial role.
The molecular chaperone α‐crystallin incorporated into red cell ghosts protects membrane Na/K‐ATPase against glycation and oxidative stress
Authors: Derham Barry K. , Ellory J. Clive , Bron Anthony J. , Harding John J.
Published: June 2003

α‐Crystallin, a molecular chaperone and lens structural protein protects soluble enzymes against heat‐induced aggregation and inactivation by a variety of molecules. In this study we investigated the chaperone function of α‐crystallin in a more physiological system in which α‐crystallin was incorporated into red cell ‘ghosts’. Its ability to protect the intrinsic membrane protein Na/K‐ATPase from external stresses was studied. Red cell ghosts were created by lysing the red cells and removing cytoplasmic contents by size‐exclusion chromatography. The resulting ghost cells retain Na/K‐ATPase activity. α‐Crystallin was incorporated in the cells on resealing and the activity of Na/K‐ATPase assessed by ouabain‐sensitive 86Rb uptake. Incubation with fructose, hydrogen peroxide and methylglyoxal (compounds that have been implicated in diabetes and cataract formation) were used to test inactivation of the Na/K pump. Intracellular α‐crystallin protected against the decrease in ouabain sensitive 86Rb uptake, and therefore against inactivation induced by all external modifiers, in a dose‐dependent manner.
Interflavin electron transfer in human cytochrome P450 reductase is enhanced by coenzyme binding
Authors: Gutierrez Aldo , Munro Andrew W. , Grunau Alex , Wolf C. Roland , Scrutton Nigel S. , Roberts Gordon C. K.
Published: June 2003

The role of coenzyme binding in regulating interflavin electron transfer in human cytochrome P450 reductase (CPR) has been studied using temperature‐jump spectroscopy. Previous studies [Gutierrez, A., Paine, M., Wolf, C.R., Scrutton, N.S., & Roberts, G.C.K. Biochemistry (2002) 41, 4626–4637] have shown that the observed rate, 1/τ, of interflavin electron transfer (FADsq − FMNsq→FADox − FMNhq) in CPR reduced at the two‐electron level with NADPH is 55 ± 2 s−1, whereas with dithionite‐reduced enzyme the observed rate is 11 ± 0.5 s−1, suggesting that NADPH (or NADP+) binding has an important role in controlling the rate of internal electron transfer. In relaxation experiments performed with CPR reduced at the two‐electron level with NADH, the observed rate of internal electron transfer (1/τ = 18 ± 0.7 s−1) is intermediate in value between those seen with dithionite‐reduced and NADPH‐reduced enzyme, indicating that the presence of the 2′‐phosphate is important for enhancing internal electron transfer. To investigate this further, temperature jump experiments were performed with dithionite‐reduced enzyme in the presence of 2′,5′‐ADP and 2′‐AMP. These two ligands increase the observed rate of interflavin electron transfer in two‐electron reduced CPR from 1/τ = 11 s−1 to 35 ± 0.2 s−1 and 32 ± 0.6 s−1, respectively. Reduction of CPR at the two‐electron level by NADPH, NADH or dithionite generates the same spectral species, consistent with an electron distribution that is equivalent regardless of reductant at the initiation of the temperature jump. Spectroelectrochemical experiments establish that the redox potentials of the flavins of CPR are unchanged on binding 2′,5′‐ADP, supporting the view that enhanced rates of interdomain electron transfer have their origin in a conformational change produced by binding NADPH or its fragments. Addition of 2′,5′‐ADP either to the isolated FAD‐domain or to full‐length CPR (in their oxidized and reduced forms) leads to perturbation of the optical spectra of both the flavins, consistent with a conformational change that alters the environment of these redox cofactors. The binding of 2′,5′‐ADP eliminates the unusual dependence of the observed flavin reduction rate on NADPH concentration (i.e. enhanced at low coenzyme concentration) observed in stopped‐flow studies. The data are discussed in the context of previous kinetic studies and of the crystallographic structure of rat CPR.
31P NMR studies of energy metabolism in xanthosine‐5′‐monophosphate overproducing Corynebacterium ammoniagenes
Authors: Noguchi Yasushi , Shimba Nobuhisa , Kawahara Yoshio , Suzuki Ei‐ichiro , Sugimoto Shinichi
Published: June 2003

Corynebacterium ammoniagenes is an overproducer of xanthosine‐5′‐monophosphate (XMP) by consuming either glcose (glc) or glutamic acid (glu). Its energy metabolism was studied in vivo using 31P NMR spectroscopy coupled with a circulating fermentation system (CFS). CFS enabled us to validate directly the cellular dependency on carbon sources and changes in biomolecules produced according to alterations in the cellular energetic status. For the most efficient XMP production, the glutamic acid and glcose molar ratios (glu/glc) in the medium were adjusted to a molar ratio of 0.31. The 31P NMR illustrated the two distinct phases of the cellular energetic status due to the availability of the substrates from the medium. In the earlier phase, both glc and glu were utilized, resulting in average ATP and ADP concentrations in cells of 0.50 ± 0.17 µmol·g−1 of dry cell weight (DCW) and an undetermined level, respectively. The ADP concentration in the later phase increased to 2.15 ± 1.30 µmol·g−1 of DCW, while the ATP concentration decreased to an undetectable level in association with a remarkable decrease in XMP production. This decrease in the XMP‐producing ability was associated with an increase in production of the by‐product hypoxanthine. Because glu was found to be consumed completely during the earlier phase, glc was the only available substrate in the later phases. These findings by in vivo NMR indicate that changes in the carbon metabolism profoundly affect XMP production by C. ammoniagenes.
N‐Glycan structures of squid rhodopsin
Authors: Takahashi Noriko , Masuda Katsuyoshi , Hiraki Kenji , Yoshihara Kazuo , Huang Hung‐Hsiang , Khoo Kay‐Hooi , Kato Koichi
Published: June 2003

To determine the glycoforms of squid rhodopsin, N‐glycans were released by glycoamidase A digestion, reductively aminated with 2‐aminopyridine, and then subjected to 2D HPLC analysis [Takahashi, N., Nakagawa, H., Fujikawa, K., Kawamura, Y. & Tomiya, N. (1995) Anal. Biochem.226, 139–146]. The major glycans of squid rhodopsin were shown to possess the α1–3 and α1–6 difucosylated innermost GlcNAc residue found in glycoproteins produced by insects and helminths. By combined use of 2D HPLC, electrospray ionization‐mass spectrometry and permethylation and gas chromatography‐electron ionization mass spectrometry analyses, it was revealed that most (85%) of the N‐glycans exhibit the novel structure Manα1–6(Manα1–3)Manβ1–4GlcNAcβ1–4(Galβ1–4Fucα1–6)(Fucα1–3)GlcNAc.
Identification, cloning and characterization of two thioredoxin h isoforms, HvTrxh1 and HvTrxh2, from the barley seed proteome
Authors: Maeda Kenji , Finnie Christine , Østergaard Ole , Svensson Birte
Published: June 2003

Two thioredoxin h isoforms, HvTrxh1 and HvTrxh2, were identified in two and one spots, respectively, in a proteome analysis of barley (Hordeum vulgare) seeds based on 2D gel electrophoresis and MS. HvTrxh1 was observed in 2D gel patterns of endosperm, aleurone layer and embryo of mature barley seeds, and HvTrxh2 was present mainly in the embryo. During germination, HvTrxh2 decreased in abundance and HvTrxh1 decreased in the aleurone layer and endosperm but remained at high levels in the embryo. On the basis of MS identification of the two isoforms, expressed sequence tag sequences were identified, and cDNAs encoding HvTrxh1 and HvTrxh2 were cloned by RT‐PCR. The sequences were 51% identical, but showed higer similarity to thioredoxin h isoforms from other cereals, e.g. rice Trxh (74% identical with HvTrxh1) and wheat TrxTa (90% identical with HvTrxh2). Recombinant HvTrxh1, HvTrxh2 and TrxTa were produced in Escherichia coli and purified using a three‐step procedure. The activity of the purified recombinant thioredoxin h isoforms was demonstrated using insulin and barley α‐amylase/subtilisin inhibitor as substrates. HvTrxh1 and HvTrxh2 were also efficiently reduced by Arabidopsis thaliana NADP‐dependent thioredoxin reductase (NTR). The biochemical properties of HvTrxh2 and TrxTa were similar, whereas HvTrxh1 had higher insulin‐reducing activity and was a better substrate for Arabidopsis NTR than HvTrxh2, with a Km of 13 µm compared with 44 µm for HvTrxh2. Thus, barley seeds contain two distinct thioredoxin h isoforms which differ in temporal and spatial distribution and kinetic properties, suggesting that they may have different physiological roles.
Dynamic model of Escherichia coli tryptophan operon shows an optimal structural design
Authors: Bhartiya Sharad , Rawool Subodh , Venkatesh K. V.
Published: June 2003

A mathematical model has been developed to study the effect of external tryptophan on the trp operon. The model accounts for the effect of feedback repression by tryptophan through the Hill equation. We demonstrate that the trp operon maintains an intracellular steady‐state concentration in a fivefold range irrespective of extracellular conditions. Dynamic behavior of the trp operon corresponding to varying levels of extracellular tryptophan illustrates the adaptive nature of regulation. Depending on the external tryptophan level in the medium, the transient response ranges from a rapid and underdamped to a sluggish and highly overdamped response. To test model fidelity, simulation results are compared with experimental data available in the literature. We further demonstrate the significance of the biological structure of the operon on the overall performance. Our analysis suggests that the tryptophan operon has evolved to a truly optimal design.
Expression, localization and potential physiological significance of alcohol dehydrogenase in the gastrointestinal tract
Authors: Vaglenova Julia , Martínez Susana E. , Porté Sergio , Duester Gregg , Farrés Jaume , Parés Xavier
Published: June 2003

ADH1 and ADH4 are the major alcohol dehydrogenases (ADH) in ethanol and retinol oxidation. ADH activity and protein expression were investigated in rat gastrointestinal tissue homogenates by enzymatic and Western blot analyses. In addition, sections of adult rat gastrointestinal tract were examined by in situ hybridization and immunohistochemistry. ADH1 and ADH4 were detected along the whole tract, changing their localization and relative content as a function of the area studied. While ADH4 was more abundant in the upper (esophagus and stomach) and lower (colorectal) regions, ADH1 was predominant in the intestine but also present in stomach. Both enzymes were detected in mucosa but, in general, ADH4 was found in outer cell layers, lining the lumen, while ADH1 was detected in the inner cell layers. Of interest were the sharp discontinuities in the expression found in the pyloric region (ADH1) and the gastroduodenal junction (ADH4), reflecting functional changes. The precise localization of ADH in the gut reveals the cell types where active alcohol oxidation occurs during ethanol ingestion, providing a molecular basis for the gastrointestinal alcohol pathology. Localization of ADH, acting as retinol dehydrogenase/retinal reductase, also indicates sites of active retinoid metabolism in the gut, essential for mucosa function and vitamin A absorption.
Structural elucidation of polysaccharide part of glycoconjugate from Treponema medium ATCC 700293
Authors: Hashimoto Masahito , Asai Yasuyuki , Jinno Takayoshi , Adachi Seiji , Kusumoto Shoichi , Ogawa Tomohiko
Published: June 2003

Glycoconjugates are distributed on the cell surfaces of some small‐sized treponemes and have been reported to be completely different from lipopolysaccharides. We separated a glycoconjugate fraction from Treponema medium ATCC 700293, a medium‐sized oral spirochete, to assess its immunobiological activities and elucidate the chemical structure of its polysaccharide part using phenol/water extraction, hydrophobic chromatography, and gel filtration. The glycoconjugate showed negligible or weak endotoxic and immunobiological properties. The chemical structure of the polysaccharide part was shown by two‐dimensional NMR and MALDI‐TOF‐MS to be a tetrasaccharide backbone with two amino acids:

[→4)β‐d‐GlcpNAc3NAcA(1→4)β‐d‐ManpNAc3NAOrn(1→3)β‐d‐GlcpNAc(1→3)α‐D‐Fucp4NAsp(1→]

where GlcNAc3NAcA is 2,3‐diacetamido‐2,3‐dideoxyglucuronic acid, ManNAc3NAOrn is Nδ‐(2‐acetamido‐3‐amino‐2,3‐dideoxymannuronyl)ornithine, and Fuc4NAsp is 4‐(α‐aspartyl)amino‐4,6‐dideoxygalactose.
Calcium and polyamine regulated calcium‐sensing receptors in cardiac tissues
Authors: Wang Rui , Xu Changqing , Zhao Weimin , Zhang Jing , Cao Kun , Yang Baofeng , Wu Lingyun
Published: June 2003

Activation of a calcium‐sensing receptor (Ca‐SR) leads to increased intracellular calcium concentration and altered cellular activities. The expression of Ca‐SR has been identified in both nonexcitable and excitable cells, including neurons and smooth muscle cells. Whether Ca‐SR was expressed and functioning in cardiac myocytes remained unclear. In the present study, the transcripts of Ca‐SR were identified in rat heart tissues using RT‐PCR that was further confirmed by sequence analysis. Ca‐SR proteins were detected in rat ventricular and atrial tissues as well as in isolated cardiac myocytes. Anti‐(Ca‐SR) Ig did not detect any specific bands after preadsorption with standard Ca‐SR antigens. An immunohistochemistry study revealed the presence of Ca‐SR in rat cardiac as well as other tissues. An increase in extracellular calcium or gadolinium induced a concentration‐dependent sustained increase in [Ca2+]i in isolated ventricular myocytes from adult rats. Spermine (1–10 mm) also increased [Ca2+]i. Pre‐treatment of cardiac myocytes with thapsigargin or U73122 abolished the extracellular calcium, gadolinium or spermine‐induced increase in [Ca2+]i. The blockade of Na+/Ca2+ exchanger or voltage‐dependent calcium channels did not alter the extracellular calcium‐induced increase in [Ca2+]i. Finally, extracellular calcium, gadolinium and spermine all increased intracellular inositol 1,4,5‐triphosphate (IP3) levels. Our results demonstrated that Ca‐SR was expressed in cardiac tissue and cardiomyocytes and its function was regulated by extracellular calcium and spermine.
Are UV‐induced nonculturable Escherichia coli K‐12 cells alive or dead?
Authors: Villarino Andrea , Rager Marie‐Noëlle , Grimont Patrick A. D. , Bouvet Odile M. M.
Published: June 2003

Cells that have lost the ability to grow in culture could be defined operationally as either alive or dead depending on the method used to determine cell viability. As a consequence, the interpretation of the state of ‘nonculturable’ cells is often ambiguous. Escherichia coli K12 cells inactivated by UV‐irradiation with a low (UV1) and a high (UV2) dose were used as a model of nonculturable cells. Cells inactivated by the UV1 dose lost ‘culturability’ but they were not lysed and maintained the capacity to respond to nutrient addition by protein synthesis and cell wall synthesis. The cells also retained both a high level of glucose transport and the capacity for metabolizing glucose. Moreover, during glucose incorporation, UV1‐treated cells showed the capacity to respond to aeration conditions modifying their metabolic flux through the Embden–Meyerhof and pentose‐phosphate pathways. However, nonculturable cells obtained by irradiation with the high UV2 dose showed several levels of metabolic imbalance and retained only residual metabolic activities. Nonculturable cells obtained by irradiation with UV1 and UV2 doses were diagnosed as active and inactive (dying) cells, respectively.
Infection of Tomicus piniperda (Col., Scolytidae) with Canningia tomici sp. n. (Microsporidia, Unikaryonidae)
Authors: Kohlmayr B. , Weiser J. , Wegensteiner R. , Händel U. , Zizka Z.
Published: June 2003

Canningia tomici sp. n. (Microsporidia, Unikaryonidae) infects the midgut epithelium, the gut muscules, Malpighian tubules, connective tissues, adipose tissues and the gonads of the pine shoot beetle, Tomicus piniperda (L.) (Coleoptera, Scolytidae). The infection is present in populations of Tomicus piniperda in Europe and in the United States. Uninucleate oval single spores occur in two sizes: 2.8 ± 0.4 × 1.4 ± 0.4 μm and 3.8 ± 0.3 × 2.0 ± 0.2 μm. The polar filament of this microsporidium is fixed subapically in a flat anchoring disc. The thick posterior lamellae of the binary polaroplast are asymmetric due to the lateral fixation of the polar filament.
Prey consumption during development as well as longevity and reproduction of Typhlodromus pyri Scheuten (Acari, Phytoseiidae) at higher temperatures in the laboratory
Authors: Sengonca C. , Khan I. A. , Blaeser P.
Published: June 2003

The predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae) has been reported as an important predator of the European red mite, Panonychus ulmi (Koch) (Acari, Tetranychidae) in apple culture and vineyards at below 25 °C. However, sufficient biological data was lacking on its efficiency at temperatures above 25 °C. Therefore, the purpose of the present laboratory work was to obtain experimental data on prey consumption during development as well as longevity and reproduction of T. pyri on apple leaf discs and in Plexiglas cells at constant temperatures of 25 ± 2 °C and 30 ± 2 °C with P. ulmi as prey.

The results showed that mean daily and total prey consumption by both the nymphs and adults of T. pyri decreased significantly on both the arenas as the temperature was increased from 25 °C to 30 °C, whereby adult prey consumption, both mean daily and total, was higher than that of nymphs. Prey consumption by both the nymphs and adults was significantly higher in the Plexiglas cells than on the leaf discs at both temperatures. Mean total prey consumption during nymphal development was 16.1 (♀♀) and 12.8 (♂♂) at 25 °C compared to 7.0 (♀♀) and 5.8 (♂♂) preys at 30 °C on the apple leaf discs and 46.0 (♀♀) and 38.5 (♂♂) at 25 °C compared to 25.2 (♀♀) and 20.3 (♂♂) preys at 30 °C in the Plexiglas cells. Mean duration of nymphal development was similar for the two sexes at the same temperature, but it was longer at 25 °C than at 30 °C. It was 6.0 and 4.0 days on the apple leaf discs while 7.0 and 6.0 days in the Plexiglas cells at 25 °C and 30 °C, respectively. Mean daily and total prey consumption by both male and female adults also decreased with the increasing temperature, whereby the females consumed more than double the mean total number of prey than the males on both the arenas of observation and at both temperatures: 355.4 versus 149.7 preys at 25 °C and 192.2 versus 85.6 preys at 30 °C on the leaf discs and 826.8 versus 374.5 preys at 25 °C and 488.9 versus 187.9 preys at 30 °C in the Plexiglas cells. Longevity of the females was longer than males on both arenas and at both temperatures and it was longer at 25 °C than at 30 °C. Mean total longevity on the apple leaf discs was 68.3 (♀♀) and 50.8 (♂♂) days at 25 °C compared to 52.5 (♀♀) and 36.8 (♂♂) days at 30 °C, while in the Plexiglas cells it was 91.0 (♀♀) and 65.8 (♂♂) days at 25 °C compared to 75.3 (♀♀) and 48.5 (♂♂) days at 30 °C. Reproduction in females also decreased significantly with increasing temperature. It decreased from 62.0 to 39.0 eggs/female on the leaf discs and 75.0 to 47.1 eggs/female in the Plexiglas cells. The females laid significantly higher numbers of eggs at both temperatures in the Plexiglas cells than on the leaf discs. Oviposition period in females was 30 days at 25 °C on both the arenas, while at 30 °C it was 26 days on the apple leaf discs and 27 days in the Plexiglas cells.
The impact of thiamethoxam on bumble bee broods (Bombus terrestris L.) following drip application in covered tomato crops
Authors: Sechser B. , Freuler J.
Published: June 2003

The side effects of the neonicotinyl compound thiamethoxam on the brood of bumble bees (Bombus terrestris L.) were investigated on tomatoes in plastic tunnels and glasshouses.

Preceeding trials had revealed a strong contact and stomach activity of the compound by foliar application. A decisive improvement was obtained by replacing the foliar by drip irrigation of thiamethoxam at rates between 150 and 161 g ai/ha. The mortality figures of all bumble bee stages dropped to the level in the untreated control. No dead bumble bees were found in the thiamethoxam treated tunnels/glasshouses on the ground.

Thiamethoxam could be rated as being harmless to bumble bees with a single application through the irrigation system. No spilling of this compound during the application should occur to avoid intoxication of adult bumble bees by oral uptake or contact contamination during the cleaning process. If multiple applications via the irrigation system or under any hydroponic growing conditions is considered, further sequential testing of this mode of use is recommended.
Patterns of bark beetle occurrence in Norway spruce stands of national parks in Tatra Mts. in Poland and Slovakia
Authors: Grodzki W. , Jakuš R. , Gazda M.
Published: June 2003

This paper presents the results of a research project concerning bark beetle populations in Norway spruce stands managed under a nature protection regime in national parks in Tatra Mountains in Poland and Slovakia. The preliminary data on bark beetle occurrence and related selected stand characteristics are presented. The dynamics of a bark beetle outbreak that occurred from 1994–97 in the stands localized on both sides of the state border and in various nature protection regimes is analyzed retrospectively. The data concerning the tree/stand characteristics, i.e. DBH and increment of trees are discussed in relation to its potential role in the mechanism for the spread of bark beetle populations. The patterns of the spatial distribution of trees attacked during the research period (1999–2001) are described.
Virus vector aphid activity and seed potato tuber virus infection in Hungary
Authors: Basky Z.
Published: June 2003

An aphid and virus survey was conducted from 1993 and 1999 on the eastern lowlands in Hungary. Aphid flight was monitored with two yellow pan traps placed on the edge and in the middle of seed potato fields. From 1993 and 1997, aphid flight was monitored with a Rothamsted type suction trap. The aphid and virus survey was continued in four different regions of Hungary in 2000. Aphid flight activity recorded by the suction trap and the yellow pan traps was different. Cumulative vector intensity based on yellow pan trap catches revealed a greater variation between years than among the different locations. In 4 years out of 8, aphid flight was less intensive and quality seed potato was harvested regardless of the production site. Cumulative vector intensity is a good means to forecast the time of haulm destruction in seed potatoes.
Interactions amongst Mycorrhiza, Azotobacter chroococcum and Root Characteristics of Wheat Varieties
Authors: Behl R. K. , Sharma H. , Kumar V. , Narula N.
Published: June 2003

A field experiment was conducted in a randomized block design with three replications over 2 years to evaluate the effect of wheat cultivar and dual inoculation of Azotobacter chroococcum (Azc) and arbuscular mycorrhiza fungi (AMF, Glomus fasciculatum) on root characters and AMF infection in three crosses of wheat. The experimental material comprised four wheat parents, WH‐147, WH‐157, WH‐542 and PBW‐175, and three F1 crosses, WH‐147 ×WH‐157, WH‐147 × WH‐542 and WH‐147 × PBW‐175. Comparison of treatment averages, i.e. control (mineral nutrients 60 kg N + 30 kg P2O5 + 12.5 kg ZnSO4 ha−1, as in other two treatments), AMF and AMF + Azc, revealed that inoculation of Azc led to an increase in AMF infection in roots. Maximum root biomass was obtained in F1 hybrids WH‐147 × WH‐157 in the AMF treatment and in WH‐147 × PBW‐175 receiving AMF + Azc. Total root length and AMF infection of roots was maximum in WH‐147 × PBW‐175 for all the treatments during both years. A positive association between AMF infection in roots and Azotobacter survival in the rhizosphere was apparent. Similarly, maximum A. chroococcum counts were observed 80 and 120 days after sowing in the AMF + Azc treatment in cross WH147 × PBW175.
Chilling Tolerance of Maize Seedlings in the Field during Cold Periods in Spring is Related to Chilling‐Induced Increase in Abscisic Acid Level
Authors: Janowiak F. , Luck E. , Dörffling K.
Published: June 2003

Recent studies on chilling tolerance mechanisms in maize revealed a significant positive correlation between genotypic chilling tolerance and chilling‐induced accumulation of the stress hormone abscisic acid (ABA) under controlled growth chamber conditions. Based on this and other results, the hypothesis was developed that chilling tolerance in maize is related to the ability to accumulate large amounts of ABA rapidly, as a protection against chilling injury. The objective of the present study was to test this hypothesis under field conditions during natural cold weather periods in spring, which often cause severe chilling injury in maize fields. In two experiments at two locations in Europe with contrasting climates, eight maize genotypes with different genotypic chilling tolerance were cultivated in spring in the field according to agronomic practice for maize. Before and at the end of cold weather periods, ABA levels and water contents were determined in the third leaves. It was found that chilling‐tolerant genotypes accumulated higher amounts of ABA during the chilling period than chilling‐sensitive genotypes. A significant positive correlation between chilling tolerance and the levels of ABA in the leaves was found. These results support the above‐mentioned hypothesis. In contrast to earlier studies performed under growth chamber conditions, the water content in chilled leaves was mostly higher than in non‐chilled leaves. The increase in ABA is therefore not attributable to chilling‐induced water deficit, but probably to the low temperature itself.
Considerations for anaesthesia in children with haematological malignancy undergoing short procedures
Authors: Culshaw Valerie , Yule Murray , Lawson Ros
Published: June 2003

As a result of increased use of risk‐directed treatment regimes, there is a regular requirement for short‐lasting but painful procedures to be performed on children to aid in diagnosis or treatment. The aim of any anaesthetic technique is to provide analgesia and amnesia with minimal side‐effects and early return to former activity levels. We review the implications of haematological malignancy in children with regard to anaesthesia and the consequences arising from both the disease and ensuing treatment. We outline some of the current anaesthetic techniques in use and review the advantages and disadvantages of each.
Life threatening unilateral pulmonary overinflation might be more successfully treated by contralateral selective intubation than by emergency pneumonectomy
Authors: Holzki Josef , Kellner Max
Published: June 2003

During a period of 3 years, three infants were admitted to our hospital for unilateral emergency pneumonectomy due to life threatening overinflation of one lung, preventing adequate ventilation of the unaffected contralateral side. All three patients were able to be stabilized by unilateral selective bronchial intubation of the unaffected lung after bronchoscopy, ruling out a flap valve mechanism. No emergency pneumonectomies were required. In one patient, lung function of the overinflated side (three lobes) recovered fully and, in the two remaining patients, one lobe of the overinflated side recovered. The overinflated lobes were removed later by elective surgery, thus not exposing the children to a potentially dangerous emergency operation. These results are in contrast with reports in the literature. Emergency pneumonectomy in neonates and infants due to overinflation of one lung may be avoided by selective unilateral intubation of the main stem bronchus of the compressed lung.
Postobstructive pulmonary oedema during anaesthesia in children with mucopolysaccharidoses
Authors: Walker R.W.M. , Colovic V. , Robinson D.N. , Dearlove O.R.
Published: June 2003

We present case reports of five patients with severe forms of mucopolysaccharidoses who developed postobstructive pulmonary oedema during anaesthesia. The difficulties of anaesthesia in these patients and the particular predisposition that these patients exhibit for the development of postobstructive pulmonary oedema is discussed.
Double trouble: prolapsing epiglottis and unexpected dual pathology in an infant
Authors: De Beer David , Chambers Neil
Published: June 2003

A 3‐week‐old full‐term female neonate was admitted with a 4‐day history of episodic stridor, desaturations and difficult feeding. Initial assessment using fluoroscopy suggested distal tracheomalacia. Inhalational induction for examination under anaesthesia of the upper airway at 4 weeks of age caused almost complete airway obstruction due to severe anterior, or epiglottic, laryngomalacia. This airway obstruction was unresponsive to continuous positive airway pressure, the use of an oropharyngeal airway and hand ventilation and required urgent tracheal intubation using suxamethonium. Epiglottopexy, a relatively unknown procedure, was performed uneventfully 2 days later, with complete relief of the respiratory compromise. However, the infant remained desaturated postoperatively. A ventilation perfusion scan subsequently revealed multiple pulmonary arteriovenous malformations, unsuitable for embolization and requiring nocturnal home oxygen therapy. Review at 3 months of age found a thriving infant with no airway obstruction and good epiglottic positioning on examination under anaesthesia. Although the patient's oxygen requirements had diminished, the long‐term outcome remains uncertain.
Anaesthesia for an adolescent with mitochondrial encephalomyopathy–lactic acidosis–stroke‐like episodes syndrome
Authors: Bolton P. , Peutrell J. , Zuberi S. , Robinson P.
Published: June 2003

Anaesthesia for patients with mitochondrial disorders occurs with regularity, either for a diagnostic or therapeutic procedure associated with the syndrome or for unassociated surgery. We report the case of a 17‐year‐old boy with mitochondrial encephalomyopathy–lactic acidosis–stroke‐like episodes (MELAS) syndrome who underwent a laparotomy for fundoplication. The potential hazards of anaesthesia for this patient, and the steps taken to avoid them, are discussed.
Anaesthetic management of a child with Pallister–Killian syndrome
Authors: Iacobucci T. , Galeone M. , De Francisci G.
Published: June 2003

Pallister–Killian syndrome is characterized by tetrasomy of the short arm of chromosome 12p, which produces mental retardation of varying degrees and dysmorphic characteristics. We describe anaesthesia in a 2‐year‐old child affected by this syndrome who underwent surgery for orchidopexy. Anaesthetic consisted of an inhalation mixture of O2, N2O and sevoflurane, together with an inguinal block with ropivacaine and administration of alfentanil plus ketorolac. Tracheal intubation was uneventful. No complications of any type were observed.
Does ketamine or magnesium affect posttonsillectomy pain in children?
Authors: O'Flaherty Jennifer E. , Lin Charles X.
Published: June 2003

Background: Many previous studies have suggested a role for the N‐methyl‐D‐aspartate (NMDA) receptor antagonists ketamine and magnesium in decreasing postoperative pain and analgesic requirements in adults, but none has investigated these medications in children.

Methods: This randomized, double‐blind, placebo‐controlled study evaluated the effects of ketamine and magnesium in children undergoing tonsillectomy. Eighty patients, aged 3–12 years, were randomly assigned to four groups. Patients received either ketamine 0.15 mg·kg−1, magnesium sulphate 30 mg·kg−1, ketamine 0.15 mg·kg−1 plus magnesium sulphate 30 mg·kg−1, or placebo intravenously 5 min prior to the start of surgery. Intraoperative analgesia was standardized, and included fentanyl and dexamethasone.

Results: There were no differences among the groups with respect to pain assessment postoperatively. Compared with placebo, the treatment groups did not require less fentanyl in the postanaesthesia recovery room or consume less codeine in the first 24‐h postoperatively. There was no evidence of synergism between ketamine and magnesium. There were no differences among the groups in the incidence of nausea, vomiting, sedation, bleeding, or dreaming postoperatively.

Conclusion: This study did not demonstrate a decrease in pain or analgesic consumption in children undergoing tonsillectomy when pretreated with a small dose of ketamine and/or magnesium.
Differential lung ventilation in an infant using LMATM and a long tracheal tube
Authors: Arai Toshimi , Yamashita Masao
Published: June 2003

This is a case report of differential lung ventilation in an infant using a conventional laryngeal mask airway (LMATM) and a commercially available longer paediatric tracheal tube. This 2‐month‐old infant had her left‐sided congenital diaphragmatic hernia repaired on the sixth day of her life, and had been mechanically ventilated. The right middle lobe became grossly emphysematous and herniated into the left side of the chest, and the right lower lobe became totally atelectatic. A right middle lobe lobectomy was planned. To avoid hyperinflating the right middle lobe, and to expand the right lower lobe without providing a high airway pressure on the left lung, the dependent lung (i.e. left lung) was ventilated with an LMA, and the right lower lobe was expanded with a long tracheal tube inserted through the LMA via the swivel connector. This combination of a conventional LMA and a commercially available longer paediatric tracheal tube could be another way of providing differential ventilation in infants.
Comparison of patient‐controlled analgesia with and without a background infusion after appendicectomy in children
Authors: Yildiz Karamehmet , Tercan Elvan , Dogru Kudret , Ozkan Ugur , Boyaci Adem
Published: June 2003

Background: There have been many studies using patient‐controlled analgesia (PCA) and opioids for postoperative analgesia in children. In this study, we investigated the efficacy, usefulness and analgesic consumption of two different PCA programmes [bolus dose alone (BD) or bolus dose with background infusion (BD + BI)] to evaluate postoperative analgesia for children after emergency appendicectomy.

Methods: Forty children, aged between 6 and 15 years and ASA class I or II, undergoing emergency appendicectomy were randomly allocated into two groups. The children were given a loading dose of pethidine 0.3 mg·kg−1 and 150 μg·kg−1 bolus intravenously in group BD (n = 20) and pethidine 0.3 mg·kg−1 loading dose, 75 μg·kg−1 bolus and 15 μg·kg−1·h−1 background infusion in group BD + BI (n = 20). The lockout interval was 20 min in both groups.

Results: There were no significant differences in pain, sedation and nausea scores during the 24‐h postoperative period between the groups (P > 0.05). Pethidine consumption was significantly lower in group BD + BI than that in group BD for the first 24‐h period (P < 0.05).

Conclusions: We demonstrated that both these PCA programmes were effective and reliable for postoperative pain relief in children. We believe that giving information about PCA to the children and their parents is useful during the preoperative period. However, the background infusion with lower bolus dose in PCA did not increase pethidine consumption.
Comparison of 0.2% ropivacaine and 0.25% bupivacaine for axillary brachial plexus blocks in paediatric hand surgery
Authors: Thornton K.L. , Sacks M.D. , Hall R. , Bingham R.
Published: June 2003

Background:  The purpose of this study was to compare the use of ropivacaine 0.2% with bupivacaine 0.25% for axillary brachial plexus block in children undergoing hand surgery.

Methods:  In a double‐blind, randomized study, 35 children undergoing hand surgery received axillary brachial plexus blocks with 0.5 ml·kg−1 of either 0.2% ropivacaine or 0.25% bupivacaine. Pain scores were noted at 0, 3, 6, 12 and 24 h after surgery. The time to first dose of codeine phosphate and the total doses of all analgesics given were recorded.

Results:  There was no significant difference between the two groups in pain scores, the time to first dose of codeine phosphate or in analgesic requirements in the first 24 h.

Conclusions:  Ropivacaine 0.2% is as effective as bupivacaine 0.25% for axillary brachial plexus blocks in children undergoing hand surgery.
The dose of caudal epidural analgesia and duration of postoperative analgesia
Authors: Schrock Charles R. , Jones Matthew Barry
Published: June 2003

Background: Single dose caudal epidural is commonly utilized for postoperative analgesia in children. Previous studies have determined the optimal concentration of local anaesthetic, and the minimal volume to produce a desired dermatomal distribution. However, none has sought the optimal volume to administer. The specific aim of this study was to determine whether the volume of caudal epidural local anaesthetic influenced the duration of postoperative analgesia.

Methods: Fifty‐four children aged 1–6 years and ASAPS I‐II scheduled for elective inguinal herniorraphy were enrolled in this randomized and blinded clinical trial. They received a standardized general anaesthetic with one of three possible doses of caudal epidural analgesic: 0.7, 1.0, or 1.3 ml·kg−1 of 0.175% bupivacaine with 1 : 200 000 epinephrine. The patients were assessed by blinded observers during in‐hospital recovery and by parents at home.

Results: The principal outcome measure of time until first postoperative analgesic requirement was similar between the groups (4.2, 3.6, and 4.8 h respectively). Other effects which might be altered by epidural analgesia, including time until first void, ambulation, and discharge readiness did not differ between groups.

Conclusions: Increasing local anaesthetic dose and volume do not increase the duration of postoperative analgesia of caudal epidural in children undergoing inguinal herniorraphy.
Comparison between instillation of bupivacaine versus caudal analgesia for postoperative analgesia following inguinal herniotomy in children
Authors: Machotta Andreas , Risse Anna , Bercker Sven , Streich Rita , Pappert Dirk
Published: June 2003

Background: In this study we compare the postoperative pain relief for inguinal herniotomy in children provided by instillation of bupivacaine into the wound with that provided by a caudal block.

Methods: Fifty‐eight children aged 0–5 years having elective unilateral hernia repair were studied in this prospective, randomized, single‐blind study. Anaesthesia was induced and maintained with oxygen, nitrous oxide, sevoflurane and propofol. Patients were randomly assigned to receive caudal analgesia with 1.0 ml·kg−1 body weight (BW) bupivacaine 0.25% or wound instillation with 0.2 ml·kg−1 BW bupivacaine 0.5% at the end of surgery. Pain was assessed over 24 h using a modified 10‐point objective pain scale. During the first postoperative hour in the postanaesthesia care unit (PACU), intravenous (i.v.) piritramide (0.05 mg·kg−1) was administered to any child scoring 5 or more points on the pain scale. On the ward, rectal acetaminophen was administered by a staff nurse when considered necessary.

Results: Thirty children in the caudal group and 28 children in the wound instillation group were studied. There were no statistically significant differences between the groups regarding need for i.v. opioids, discharge time from the PACU and administration of acetaminophen. No statistically significant differences in postoperative pain score were observed in 16 of a total of 17 postoperative observations. No complications and no adverse effects were observed.

Conclusion: Instillation of bupivacaine into a wound provides postoperative pain relief following hernia repair, which is as effective as that provided by a postoperative caudal block.
Double‐blind randomized placebo‐controlled trial of the effect of ketamine on postoperative morphine consumption in children following appendicectomy
Authors: Dix P. , Martindale S. , Stoddart P.A.
Published: June 2003

Background: Ketamine has an opioid sparing effect following surgery in adults. This study investigated whether a similar effect is seen following appendicectomy in paediatric patients.

Methods: Seventy‐five ASA 1 or 2 children aged 7–16 years were recruited, and randomly allocated to one of three groups. Following a standard anaesthetic for appendicectomy, all were prescribed patient controlled analgesia (PCA) morphine with paracetamol and nonsteroidal anti‐inflammatory drugs (NSAIDS) as required for postoperative analgesia. In addition the control group received a saline infusion postoperatively, the ketamine bolus group received 500 μg·kg−1 intravenous (i.v.) ketamine preincision and a saline infusion postoperatively, and the ketamine infusion group received 500 μg·kg−1 i.v. ketamine preincision and a ketamine infusion at 4 μg·kg−1 min−1 postoperatively. Morphine consumption, rescue analgesia requirement and side‐effects were recorded postoperatively.

Results: There was no difference in morphine consumption between the groups. The ketamine infusion group required more doses of rescue analgesia and reported more side‐effects than the control group. Five patients, all in the ketamine infusion group, reported hallucinations.

Conclusions: In this paediatric population intravenous ketamine did not have a morphine sparing effect. The increased incidence of side‐effects, especially hallucinations, reported by patients given a ketamine infusion may limit the further use of postoperative ketamine in children.
Erythropoietin therapy and acute preoperative normovolaemic haemodilution in infants undergoing craniosynostosis surgery
Authors: Meneghini Luisa , Zadra Nicola , Aneloni Vittorio , Metrangolo Salvatore , Faggin Roberto , Giusti Franca
Published: June 2003

Background: A retrospective study was performed to evaluate whether pretreatment with erythropoietin and iron combined with acute preoperative normovolaemic haemodilution (APNH) could decrease homologous blood transfusion in craniosynostosis (CS) surgery. A treated group was compared with a historical group of infants who underwent surgery with no pretreatment.

Methods: The charts of 25 healthy infants who underwent CS surgery were reviewed. Nine of them underwent surgery with no treatment beforehand. Sixteen infants were given erythropoietin at a dosage of 300 U·kg −1 two times per week and iron (elemental iron 10 mg·kg−1·day−1) for 3 weeks before surgery. On the day of surgery APNH was performed after induction of general anaesthesia; a precalculated amount of autologous blood was withdrawn and replaced by hydroxyethyl starch 6%.

Results: Eleven of the 16 infants of the study group received only autologous blood. Five of 16 received homologous blood transfusion vs seven of nine infants in the control group.

Conclusions: APNH combined with erythropoietin was effective in reducing homologous blood requirements during CS surgery. Further studies are necessary on a larger scale to assess the role of this technique in avoiding homologous blood transfusion and to evaluate how infants can benefit from this combined approach.