bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of the fish Garra on the ecology of reservoirs and the occurrence of Microcystis blooms in semi‐arid tropical highlands: an experimental assessment using enclosures
Authors: DEJENIE TADESSE , ASMELASH TSEHAYE , ROUSSEAUX SARAH , GEBREGIORGIS TEKLIT , GEBREKIDAN ABREHA , TEFERI MEKONEN , NYSSEN JAN , DECKERS JOZEF , VAN DER GUCHT KATLEEN , VYVERMAN WIM , DE MEESTER LUC , DECLERCK STEVEN A. J.
Published: March 2009

1. Many man‐made reservoirs in the semi‐arid highlands of Northern Ethiopia (Tigray) are characterised by the occurrence of intensive blooms of cyanobacteria and a dominance of small riverine fishes belonging to the genus Garra.

2. We carried out enclosure experiments to test for the effect of these small fish on abiotic characteristics, phytoplankton biomass and zooplankton community structure in the pelagic of two reservoirs (Gereb Awso and Tsinkanet). Two experiments were carried out in each of the reservoirs, one at the end of the rainy season (highest water level) and one at the end of the dry season (lowest water level).

3. The presence of Garra in general increased the amount of suspended matter, nutrient concentrations (total nitrogen and total phosphorus), phytoplankton and Microcystis biomass (including the proportion of Microcystis in the phytoplankton community), and reduced water transparency. The positive effect of the presence of Garra on nutrient concentrations and phytoplankton productivity indicate that Garra has the potential to affect food web functioning indirectly through bottom‐up effects, by enhancing nutrient concentrations through sediment resuspension and excretion of nutrients. Indeed, population densities of the cladoceran zooplankton taxa Ceriodaphnia and Diaphanosoma also showed an overall increase in enclosures with Garra.

4. However, our data also provide some evidence for a potential of Garra to exert top‐down control on large bodied daphnids (Daphnia carinata, D. barbata), although such effect varied among experiments. The limited capability of Garra to control zooplankton communities mainly reflects the low efficiency of these small, riverine and benthos‐oriented fish in foraging on zooplankton and suggests the existence of an unoccupied niche for zooplanktivorous fish in the majority of the reservoirs.

5. Although the main effects of Garra on the pelagic food web seemed to be mediated by bottom‐up mechanisms, our results also indicate that one of the key variables, the relative abundance of Microcystis, was impacted by Daphnia‐mediated trophic cascade effects.
p97/valosin‐containing protein (VCP) is highly modulated by phosphorylation and acetylation
Authors: Mori‐Konya Chiho , Kato Naruyoshi , Maeda Ryota , Yasuda Kunihiko , Higashimae Naoki , Noguchi Masakatsu , Koike Masaaki , Kimura Yoko , Ohizumi Hiroshi , Hori Seiji , Kakizuka Akira
Published: March 2009

p97/valosin‐containing protein (VCP) is a member of the AAA family proteins, which plays various important roles in cells by using its ATPase activity. But mechanism of regulating its ATPase activity is mostly unknown. We report here that VCP is highly modified throughout the protein via acetylation and phosphorylation. In addition to six previously identified phosphorylation sites, we identified at least 14 serines, 14 threonines, 6 tyrosines and 22 lysines as potential modification sites. Interestingly, these sites included Lys251 and Lys524, which are very critical for the ATP binding in Walker A motif of D1 and D2 domains, respectively. It is notable that 16 sites are in the N‐terminal region and 16 sites are clustered in D2α domain (from Pro646 to Gly765). Indeed, amino acid substitution of Lys696 and Thr761 profoundly affect VCP ATPase activities. From these results, we propose that D2α domain acts as a VCP ATPase Regulatory domain or “VAR domain”. VCP modifications including those in this VAR domain may endorse adaptive and multiple functions to VCP in different cell conditions such as in the cell cycle and with abnormal protein accumulation.
Mortality of developing floodplain forests subjected to a drying climate and water extraction
Authors: HORNER GILLIS J. , BAKER PATRICK J. , MAC NALLY RALPH , CUNNINGHAM SHAUN C. , THOMSON JAMES R. , HAMILTON FIONA
Published: March 2009

River regulation and water extraction have altered the hydrology of rivers resulting in substantial changes to forest structure and the dieback of floodplain forests globally. Forest mortality, due to water extraction, is likely to be exacerbated by climate change‐induced droughts. In 1965, a plantation trial was established within a natural floodplain forest to examine the effect of planting density on timber production. We used data from this trial to investigate the effect of initial stand density on the structure and dynamics of Eucalyptus camaldulensis (Dehnh.) forests. Highest density stands (8000 trees ha−1) were dominated by many slender trees, mostly<10 cm in diameter, whereas the lowest density stands produced size distributions with a wider range of stem diameters and higher mean and maximum stem diameter. After 1996, the study area experienced a sharp decline in water availability due to a substantial lowering of the water table, reduced flooding frequency, a pronounced rainfall deficit and increased maximum temperatures. The drought coincided with a dramatic increase in mortality in the high‐density stands, yet remained little changed in low‐density treatments. Our results highlight the importance of initial stand density as a key determinant of the development of forest structure. Early thinning of high‐density stands is one component of a broader management approach to mitigate impacts of human‐induced drought and water extraction on developing floodplain forests.
Relative impacts of disturbance and temperature: persistent changes in microenvironment and vegetation in retrogressive thaw slumps
Authors: LANTZ TREVOR C. , KOKELJ STEVEN V. , GERGEL SARAH E. , HENRY GREG H. R.
Published: March 2009

In the Low Arctic, a warming climate is increasing rates of permafrost degradation and altering vegetation. Disturbance associated with warming permafrost can change microclimate and expose areas of ion‐rich mineral substrate for colonization by plants. Consequently, the response of vegetation to warming air temperatures may differ significantly from disturbed to undisturbed tundra. Across a latitudinal air temperature gradient, we tested the hypothesis that the microenvironment in thaw slumps would be warmer and more nutrient rich than undisturbed tundra, resulting in altered plant community composition and increased green alder (Alnus viridis subsp. fruticosa) growth and reproduction. Our results show increased nutrient availability, soil pH, snow pack, ground temperatures, and active layer thickness in disturbed terrain and suggest that these variables are important drivers of plant community structure. We also found increased productivity, catkin production, and seed viability of green alder at disturbed sites. Altered community composition and enhancement of alder growth and reproduction show that disturbances exert a strong influence on deciduous shrubs that make slumps potential seed sources for undisturbed tundra. Overall, these results indicate that accelerated disturbance regimes have the potential to magnify the effects of warming temperature on vegetation. Consequently, understanding the relative effects of temperature and disturbance on Arctic plant communities is critical to predicting feedbacks between northern ecosystems and global climate change.
Identification and function of Abdominal‐A in the silkworm, Bombyx mori
Authors: Pan M‐H. , Wang X‐Y. , Chai C‐L. , Zhang C‐D. , Lu C. , Xiang Z‐H.
Published: March 2009

Abdominal‐A (adb‐A) is a key gene in the development of insects. To understand its function in the silkworm, we cloned 1193 bp of the abd‐A gene of Bombyx mori (Bmabd‐A), including the complete coding sequence and part of the 3′ untranslated region sequence. Bmabd‐A has at least three mRNA splice variants with coding sequences of lengths 1032, 1044 and 1059 bp, encoding 343, 347 and 352 amino acids, respectively. Each splice variant of Bmabd‐A has three exons and differs only in second exon size. Bmabd‐A was expressed at low levels in unfertilized eggs, but increased gradually in fertilized eggs after laying 22 h. Bmabd‐A expression decreased in ant silkworms (newly hatched silkworms). After RNA interference for Bmabd‐A, the embryos had two mutant phenotypes, either completely or partially absent abdominal feet from the third to sixth abdominal segments, suggesting that Bmabd‐A is responsible for normal development of the third to sixth abdominal segments during embryonic development.
Transcriptome analysis of the salivary glands of the female tick Amblyomma americanum (Acari: Ixodidae)
Authors: Aljamali M. N. , Hern L. , Kupfer D. , Downard S. , So S. , Roe B. A. , Sauer J. R. , Essenberg R. C.
Published: March 2009

Ticks infest a wide range of hosts while bypassing their immune, inflammatory and haemostatic responses during their extended feeding, which may last for more than two weeks. Here, we present a transcriptome analysis of 3868 expressed sequence tags (ESTs) from three cDNA libraries generated from the salivary glands of adult female Ambyomma americanum ticks at different stages of feeding. We applied a normalization step for one library, significantly decreasing the abundance of mitochondrial sequences amongst the 2292 sequences from the normalized library. Our ESTs include homologues that may modulate haemostatic, immune and inflammatory responses of the hosts. Other ESTs probably represent important components of the highly efficient secretory pathways for salivary proteins and concomitantly transmitted pathogens.
A venom metalloproteinase from the parasitic wasp Eulophus pennicornis is toxic towards its host, tomato moth (Lacanobia oleracae)
Authors: Price D. R. G. , Bell H. A. , Hinchliffe G. , Fitches E. , Weaver R. , Gatehouse J. A.
Published: March 2009

Three genes encoding clan MB metalloproteinases (EpMP1–3) were identified from venom glands of the ectoparasitic wasp Eulophus pennicornis. The derived amino acid sequences predict mature proteins of approximately 46 kDa, with a novel two‐domain structure comprising a C‐terminal reprolysin domain, and an N‐terminal domain of unknown function. EpMP3 expressed as a recombinant protein in Pichia pastoris had gelatinase activity, which was inhibited by EDTA. Injection of recombinant EpMP3 into fifth instar Lacanobia oleracea (host) larvae resulted in partial insect mortality associated with the moult to sixth instar, with surviving insects showing retarded development and growth. EpMP3 is expressed specifically in venom glands. These results suggest that EpMP3 is a functional component of Eulophus venom, which is able to manipulate host development.
Two hsp23 genes in the Mediterranean fruit fly, Ceratitis capitata: structural characterization, heat shock regulation and developmental expression
Authors: Kokolakis G. , Kritsidima M. , Tkachenko T. , Mintzas A. C.
Published: March 2009

In the present study, we characterized a 3320‐bp genomic DNA fragment encoding two medfly (Ceratitis capitata) homologues of the Drosophila melanogaster heat shock protein 23 (hsp23) gene, named Cchsp23‐αand ‐β. The two medfly hsp23 genes are transcribed in opposite directions and encode two almost identical proteins. Furthermore, the two genes exhibit a very high degree of similarity in their 5′ untranslated and proximal promoter regions. Phylogenetic analysis indicated that the CcHsp23 proteins are orthologous to Drosophila Hsp23 and Sarcophaga crassipalpis Hsp23. Structural analysis of the 5′ flanking regions of the Cchsp23 genes revealed the presence of several putative heat shock elements. Both CcHsp23 genes are induced by heat in a similar manner. In addition to heat‐induction, the Cchsp23 genes are expressed at several stages of normal development as well as in ovaries and testes. In general, the developmental expression patterns of the medfly genes are similar, suggesting that they are under similar regulatory mechanisms. However, the expression of the Cchsp23 genes differs significantly from the expression of the Drosophila hsp23 gene in certain embryonic and larval stages, suggesting differential regulation of the hsp23 genes in the two dipteran species. The expression of both Cchsp23 genes in adult flies is increased with age, especially in males.
Molecular characterization and expression of three heat shock protein70 genes from the carmine spider mite, Tetranychus cinnabarinus (Boisduval)
Authors: Li M. , Lu W. C. , Feng H. Z. , He L.
Published: March 2009

Three heat shock protein 70 (Hsp70) cDNAs were isolated from the carmine spider mite, Tetranychus cinnabarinus. They were tentatively named as TCHsp70‐1, TCHsp70‐2 and TCHsp70‐3. Structural analyses showed that all of the three TCHsp70 cDNAs held the full open reading frame (ORF). Putative protein sequences and a phylogenetic tree suggested that TCHsp70‐1 and TCHsp70‐3 were cytoplasm HSP70 and TCHsp70‐2 was endoplasmic reticulum HSP70. Comparison of deduced amino acid sequences of TCHsp70‐1 and TCHsp70‐3 showed 84.78% identity, TCHsp70‐1 and TCHsp70‐2 showed 57.33% identity, TCHsp70‐2 and TCHsp70‐3 showed 58.26% identity. Real‐time comparative quantitative PCR revealed that the relative expression of TCHsp70‐2 was lower than TCHsp70‐1 and TCHsp70‐3 at each temperature tested. TCHsp70‐1 and TCHsp70‐3 shared a similar expression pattern after cold and heat shock compared with their expression at normal temperature (26 °C), but the mRNA expression of TCHsp70‐1 was significantly higher and lower than that of TCHsp70‐3 at cold and heat shock temperatures (except for 34 °C), respectively. This result possibly indicated the expression patterns of TCHsp70 were affected by their location in different cellular compartments. The results also indicated that three TCHsp70s, especially TCHsp70‐1 and TCHsp70‐3, may play an important role in mediating tolerance to cold, thermal stress for Tetranychus cinnabarinus.
A shut‐down in expression of an insulin‐like peptide, ILP‐1, halts ovarian maturation during the overwintering diapause of the mosquito Culex pipiens
Authors: Sim C. , Denlinger D. L.
Published: March 2009

Short day‐length is used to programme adult diapause in the mosquito, Culex pipiens. The downstream endocrine event that halts ovarian maturation is a shut‐down in juvenile hormone (JH) production, and recent evidence suggests that the insulin signalling pathway may be a key upstream player in executing this developmental arrest. Genes encoding insulin‐like peptides‐1, ‐2 and ‐5 were identified in C. pipiens, and we report that transcript levels of insulin‐like peptides‐1 and ‐5 were significantly lower in diapausing females than in their nondiapausing counterparts. Genes encoding both insulin‐like peptides‐1 and ‐5 were suppressed using RNA interference in mosquitoes programmed for nondiapause, and ovarian maturation was monitored. Knocking down insulin‐like peptide‐1 with RNAi in nondiapausing mosquitoes resulted in a cessation of ovarian development akin to diapause, and this arrest in development could be reversed with an application of JH. Knocking down insulin‐like peptide‐5 did not alter ovarian development. These results are consistent with a role for insulin‐like peptide‐1 in the signalling pathway leading from the perception of short day‐lengths to the shut‐down in JH production that characterizes adult diapause in C. pipiens.
Transcriptional diversity and allelic variation in nicotinic acetylcholine receptor subunits of the red flour beetle, Tribolium castaneum
Authors: Rinkevich F. D. , Scott J. G.
Published: March 2009

Sequence analysis of 168 cDNA clones encoding 12 nicotinic acetylcholine receptor subunits, Tcasα1–Tcasα11 and Tcasβ1, from the red flour beetle, Tribolium castaneum, revealed extensive post‐transcriptional modification and multiple alleles. The greatest diversity was found for Tcasa6, where 18 unique transcripts, as a result of alternative and optional exon usage, were seen. A novel alternative exon 8d was found in one Tcasα6 transcript. Tcasα5 transcripts did not contain previously reported exons 8–10. Six subunits had transcripts that contained unspliced introns, which introduced premature stop codons. Intron 3′ splice site variants were seen at six intron boundaries across five subunits. A‐to‐I RNA editing was seen only in Tcasα6. Alleles were found for all subunit genes, except Tcasα1 and Tcasα10. Transcriptional and allelic diversity are discussed with respect to receptor function and potential interactions with insecticides.
Elucidation of structural changes in soy protein isolate upon heating by Raman spectroscopy
Authors: Herrero A. M. , Jiménez‐Colmenero F. , Carmona P.
Published: March 2009

Changes in protein secondary and tertiary structure of soy protein isolate (SPI) upon heating process were investigated by Raman spectroscopy. SPI refrigerated and heated at 70, 100 and 200 °C was analysed. Non‐significant differences in secondary structure (α‐helix, β‐sheet, unordered and turn) were observed in SPI upon heating at these temperatures. However, comparison of the Raman spectra of SPI refrigerated and heated at 70, 100 and 200 °C reveals changes in hydrophobic group environments. An intensity decreasing trend of the 1340 cm−1band attributable to tryptophan vibrations was observed, which involves an increase in solvent exposure of tryptophan residues. The spectral results also showed intensity increasing trends for the 1450 and 2935 cm−1bands of SPI upon heating, which can be ascribed to breaking of hydrophobic contacts and subsequent solvent exposure of the corresponding hydrophobic aliphatic groups. Finally, it was found tertiary structural transitions towards greater proportions of buried tyrosine residues upon heating of SPI.
Assessment of the microbiological conditions of red‐meat carcasses from bacterial counts recovered by sampling via excision or swabbing with cotton wool
Authors: Martínez Bernardo , Celda María Felicidad , Millán María Encarnación , Espacio Aurora , Cano Monserrat , López‐Mendoza María Carmen
Published: March 2009

Samples from 240 carcasses were collected from four animal species (porcine, ovine, bovine and equine). Two samples were taken from each carcass, one using the excision method (EX) and the other the wet–dry swabbing method (SW). Eight areas from each carcass were sampled. Most of the samples obtained by SW revealed total aerobic viable counts (TVC) levels of between 3.1 and 4.0 log CFU cm−2, while most of the values corresponding to excision were located between 4.1 and 5.0 log CFU cm−2. Moreover, Enterobacteriaceae (EC) counts were only detected above 3.0 log CFU cm−2 in 0.85% of the carcasses when the samples were collected by swabbing, while the excision method revealed that 13.75% of the carcasses presented EC greater than 3.0 log CFU cm−2. TVC and EC by EX revealed statistically significant differences compared to SW, while no significant linear relationship was found between carcass surface bacterial counts obtained by SW and EX.
Canarium zeylanicum seed oil: an edible oil with beneficial qualities
Authors: Seneviratne Kapila N. , Kotuwegedara Roshan T.
Published: March 2009

The quality parameters, fatty acid composition, phenolic compounds and antioxidant properties of the seed oil of Canarium zeylanicum were determined. The results indicate that the seed oil of Canarium zeylanicum is an excellent source of essential fatty acids which contains C18:2 (49.35 ± 0.35%) and C18:3 (19.09 ± 0.01%). The total phenol contents of the seed oil and the seed hull of Canarium zeylanicum were 66 ± 6 and 2014 ± 14 mg kg−1respectively. The identified phenolic acids in the methanolic extracts of seed oil include 3,4‐dihydroxybenzoic acid, p‐hydroxybenzoic acid, vanillic acid and ellagic acid. Diphenyl‐2‐picrylhydrasyl radical scavenging activity, inhibition of deoxyribose degradation and the reducing power of the methanolic extracts of seed oil and seed hull of Canarium zeylanicum are comparable with those of butylated hydroxytoluene solutions.
LC‐MS method development to evaluate major triterpenes in skins and cuticular waxes of grape berries
Authors: Orbán Norbert , Kozák Imre Olivér , Drávucz Mónika , Kiss Attila
Published: March 2009

Predominant triterpenes (free oleanolic acid, β‐sitosterol and β‐sitosterol‐3‐O‐β‐d‐glucoside) in skin and cuticular wax of grape berries have been analysed by liquid chromatography–mass spectrometry (LC‐APCI‐MS). The validity of the developed method was established by determining linearity, recovery, precision, accuracy, limit of detection and quantification. Detection limits were in the range of 0.12–0.25 mg L−1, and linearity values ranged up to 105 mg L−1. The repeatability of the method was good. High variability was found among the measured grape varieties based on triterpene content, quantitites of oleanolic acid of cuticular wax ranged from 31.53 mg kg−1 to 162.01 mg kg−1 of the twelve analysed samples. The highest sitosterol content was measured in the sample ‘Othello’ (73.12 mg kg−1), while maximum sitosterol glucoside content was also found in ‘Othello’ variety (13.68 mg kg−1). The results showed that the study on triterpenes could be an informative tool to characterise grape varieties.
Functional properties of isolated porcine blood proteins
Authors: Álvarez Carlos , Bances Margarita , Rendueles Manuel , Díaz Mario
Published: March 2009

The isolated proteins contained in the blood from slaughterhouses could be recovered and used to improve the functional and nutritional properties of food products. In this work some functional properties, namely solubility, emulsifying capacity, gelling and foaming capacity have been tested for different protein fractions of porcine blood. The blood proteins studied were globins (α, β and γ), albumin and fibrinogen and the whole plasma from the plasmatic fraction, and haemoglobin and globin obtained from the haemoglobin by a chemical method from the cellular fraction. The effect of pH and protein concentration on functional properties was determined. Results indicate that blood proteins present good functional properties specially solubility, gellation and emulsifying capacities that could make blood protein useful as food additives in the formulation of food products.
Losses of vitamin C from fresh strawberries in a commercial supply chain
Authors: Russell L. F. , LeBlanc D. I. , McRae K. B. , Ryan D. A. J.
Published: March 2009

Fresh strawberries were evaluated through a commercial supply chain, from harvest to immediately prior to retail display. These samples were compared with controls that were refrigerated immediately after harvest and kept at a constant temperature for the same period of time. Temperature data were collected throughout all stages of the supply chain. Moisture and vitamin C (ascorbic acid) contents were determined at selected points of the supply chain using recognised methodology. Significant differences were found in the vitamin C concentrations of fresh strawberries at various stages of the supply chain but not in their moisture content. The ascorbic acid concentration decreased with time spent in the supply chain in all trials but the absolute vitamin C content differed from trial to trial. This result indicates that both pre‐ and post‐harvest factors influence the vitamin C content of fresh strawberries available to consumers on the retail market.
Quality of applesauces processed by pulsed electric fields and HTST pasteurisation†
Authors: Jin Z. T. , Zhang H. Q. , Li S. Q. , Kim M. , Dunne C. P. , Yang T. , Wright A. O. , Venter‐Gains J.
Published: March 2009

A pilot plant scale continuous flow pulsed electric field (PEF) and high temperature short time (HTST) processing system was integrated with an aseptic packaging machine. Fuji applesauce and blueberry applesauce were processed with PEF followed by HTST pasteurisation (PEF + HTST). PEF + HTST processed Fuji applesauce from fresh Fuji apples demonstrated high and stable sensory scores during 9 months storage at 27 °C, and had comparative sensory quality with Meal Read‐to‐Eat (MRE) and commercial applesauce products stored at 4 °C. PEF + HTST processed blueberry applesauce from pre‐pasteurised materials had lower sensory scores than PEF + HTST processed Fuji applesauce and was significantly less stable during the storage at 27 °C. PEF + HTST processed applesauces had aerobic count and mould and yeast count of <10 cfu mL−1 during storage. Electrical conductivity, pH and °Brix, were not significantly changed throughout storage time (P > 0.05).
Partial purification and characterisation of banana [Musa (AAA Group) ‘Gros Michel’] polyphenol oxidase
Authors: Chaisakdanugull Chitsuda , Theerakulkait Chockchai
Published: March 2009

Polyphenol oxidase (PPO) from pulp of banana [Musa (AAA Group) ‘Gros Michel’] was extracted and precipitated with 80% saturated ammonium sulphate followed by conventional column chromatography on Sephacryl S‐200 HR and fast protein liquid chromatography on Mono Q column. The lyophilised PPO obtained from Sephacryl S‐200 HR column was used for characterisation and inhibition studies. The partially purified PPO obtained from the Mono Q column exhibited at least three isoenzymes. The banana PPO had optimum pH for activity at 7 and it was stable around the same pH. Only 48% of initial enzyme activity was lost after heating at 70 °C for 30 min. The enzyme was completely inhibited by 2 mm sodium metabisulphite, 2 mm l‐cysteine, 4 mm ascorbic acid, and 100 mm 4‐hexylresorcinol. The Km and Vmax of banana PPO for dopamine were 2.08 mm and 0.124 mm min−1 respectively.
Determination of the calorific value of gum arabic in rats using modified versions of the growth bioassay and the digestibility assay
Authors: Smith Chris J. , Menzies Alan R. , Williams Peter A. , Phillips Glyn O.
Published: March 2009

The calorific values of gum arabic (GA), cellulose (Solka Floc) and sucrose were measured in a rat model using modifications to the growth assay and energy balance procedures simultaneously. Although the two procedures gave consistent results for cellulose (Solka Floc) and sucrose, i.e. 0 and >3.5 kcal g−1, respectively, contradictory results were obtained for GA. The growth assay procedure indicated that GA had zero calorific value while the energy balance procedure indicated a value of 3.1 kcal g−1. Similar results have been reported by other researchers using the two different techniques separately. Results of this type have led to confusion as to the actual calorific value of GA. The anomalous results for GA may be explained in terms of losses resulting from processes which provide no calories for the rat, for example bacterial fermentations in the caecum. Other sources of loss may also have a role. We have proposed that the calorific value of zero as determined using the growth assay is the correct value in the context of the nutritive value of GA.
Volatile compounds from the fruiting bodies of three Hygrophorus mushroom species from Northern Greece
Authors: Ouzouni Paraskevi K. , Koller Wolf‐Dietrich , Badeka Anastasia V. , Riganakos Kyriakos A.
Published: March 2009

The volatile composition of the wild edible mushroom species Hygrophorus chrysodon, Hygrophorus eburneus and Hygrophorus russocoriaceus was analysed by headspace‐solid phase microextraction combined with gas chromatography–mass spectrometry. Forty‐five volatile compounds were detected, including alcohols, aldehydes, ketones, hydrocarbons, esters and terpenes. The most abundant compounds in the three species were 3‐methyl butanal, hexanal, p‐cymene, 3‐octanone, 1‐octen‐3‐one, 3‐octanol, 1‐octen‐3‐ol and benzoic acid methyl ester. The concentration of eight‐carbon (C8) compounds relative to the total volatiles varied widely ranging from 24.14% to 61.41% among the three species. Finally, the presence of sesquiterpenes only in Hygrophorus russocoriaceus, confirms the characteristic distinctive cedar odour of this species.
How does the pitch and pattern of a signal affect auditory arousal thresholds?
Authors: BRUCK DOROTHY , BALL MICHELLE , THOMAS IAN , ROUILLARD VINCENT
Published: March 2009

How arousal thresholds vary with different sounds is a critical issue for emergency awakenings, especially as sleepers are dying in fires despite having a working smoke alarm. Previous research shows that the current high‐pitched (3000+ Hz) smoke alarm signal is significantly less effective than an alternative signal, the 520 Hz square wave, in all populations tested. However, as the number of sounds tested has been small further research is needed. Here we measured auditory arousal thresholds (AATs) across signals with a range of characteristics to determine the most effective waking signal. Thirty‐nine young adults participated over three nights. In Part A, nine signals were presented in stage 4 sleep with ascending decibel levels. Signals were short beeps in the low‐ to mid‐frequency range with different spectral complexities: square waves, pure tones, whoops and white noise. Part B manipulated temporal patterns, inserting silences of 0, 10 and 21 s after each 12 s of beeps. It was found that the low‐frequency (400 and 520 Hz) square waves yielded significantly lower AATs than the alternatives. A trend was found across the three temporal manipulations, with a 10 s intervening silence showing some advantage. These findings support earlier research indicating that the best sound for awakening from deep sleep is a low‐frequency square wave. It is argued that the signal with the lowest response threshold when awake may be the same as the most arousing signal when asleep, especially where the sleeper processes the signal as meaningful.
Sleep and time course of consolidation of visual discrimination skills in patients with narcolepsy–cataplexy
Authors: CIPOLLI CARLO , CAMPANA GIANLUCA , CAMPI CLAUDIO , MATTAROZZI KATIA , MAZZETTI MICHELA , TUOZZI GIOVANNI , VANDI STEFANO , VIGNATELLI LUCA , PLAZZI GIUSEPPE
Published: March 2009

The level of procedural skills improves in normal individuals when the acquisition is followed by a period of sleep rather than wake. If sleep plays an important role in the consolidation process the advantage it provides should be reduced or delayed when its organization is altered, as in patients with chronic sleep disorders. To test this prediction in patients with narcolepsy–cataplexy (NC), who usually have a more fragmented organization of sleep than normals, we compared the initial, intermediate and delayed level of consolidation of visual skills. Twenty‐two drug‐naive NC patients and 22 individually‐matched controls underwent training at a texture discrimination task (TDT) and were re‐tested on the next morning (after a night spent in laboratory with polysomnography) and after another six nights (spent at home). TDT performance was worse in patients than controls at training and at both retrieval sessions and the time course of consolidation was different in NC patients (who improved mainly from next‐day to 7th‐day retrieval session) compared with controls. Moreover, the less‐improving patients at next‐day retrieval had a wider disorganization of sleep, probably because of an episode of rapid eye movement (REM) sleep at sleep onset REM, on post‐training night more frequently than more‐improving patients. These findings suggest that the time course of the consolidation process of procedural skills may be widely influenced by the characteristics of sleep organization (varying night‐by‐night much more in NC patients than controls) during post‐training night.
Bioavailability, distribution and depletion of monensin in chickens
Authors: HENRI J. , BUREL C. , SANDERS P. , LAURENTIE M.
Published: March 2009

The pharmacokinetics of monensin including apparent volume of distribution, total body clearance, systemic bioavailability, partition coefficients and tissue residues were determined in chickens. The drug was given by intravenous injection in the left wing vein at the dose of 0.46 mg/kg and by intracrop administration at the dose of 4 mg/kg according to a destructive sampling. The pharmacokinetic variables were compared after noncompartmental, naïve averaged, naïve pooled and nonlinear mixed‐effects modelling analyses. Partition coefficients and tissue residues were determined after a treatment with feed additives (125 mg/kg of feed) of 33 days. The clearance, volume of distribution and bioavailabilty were approximately 2.2 L/h/kg, approximately 9 L/kg and approximately 30% respectively except with nonlinear mixed effects models that presented values of 1.77 L/h/kg, 14.05 L/kg and 11.36% respectively. Tissue/plasma partition coefficients were estimated to 0.83, 3.39 and 0.51 for liver, fat and thigh muscle respectively. Monensin residues after treatment were not detected 6 h after withdrawal except for fat where monensin was still quantifiable 12 h after. Pharmacokinetic variables seem to be inaccurate when assessed with non linear mixed‐effects modelling associated to destructive sampling in chickens. Values varied slightly with noncompartmental, naïve averaged and naïve pooled analyses. The absorption, elimination and partition parameters will be incorporated into a physiologically based pharmacokinetic model and the depletion study will be used to test the ability of this model to describe monensin residues in edible tissues under different dosage regimens.
Absence of the interstitial cell of Cajal network in mitochondrial neurogastrointestinal encephalomyopathy
Authors: Zimmer V. , Feiden W. , Becker G. , Zimmer A. , Reith W. , Raedle J. , Lammert F. , Zeuzem S. , Hirano M. , Menges M.
Published: March 2009

Abstract  Mitochondrial neurogastrointestinal enceph‐alomyopathy (MNGIE) is a rare autosomal‐recessive multisystemic disorder with predominant gastrointestinal involvement, presenting with variable degrees of gut dysmotility up to frank chronic intestinal pseudoobstruction. Despite major advances in understanding its basic molecular pathogenesis in recent years, the distinct mechanisms and pathoanatomical substrate underlying MNGIE‐associated gastrointestinal dysmotility are still widely unknown. As yet, though their critical role in proper gastrointestinal transit in terms of spontaneous pacemaker activity and enteric neurotransmission is well established, the population of the interstitial cells of Cajal (ICC) has not been investigated in MNGIE. Therefore, we examined small bowel samples of a well‐characterized MNGIE patient by using conventional histology and immunohistochemistry techniques. The ICC network was studied by immunohistochemistry for the tyrosine kinase Kit (CD117), known to reliably detect ICCs, while mucosal mast cells served as an internal and normal small bowel specimen as external controls. At a light microscopic level, no gross structural alteration of the bowel wall composition and its neuromuscular elements was noted. However, a complete absence of Kit immunoreactive cells could be demonstrated in regions where ICCs are normally abundant, while internal and external controls retained strong Kit positivity. In conclusion, our preliminary results provide a first evidence for an alteration of the ICC network in MNGIE, and support the notion that ICC loss might be an early pathogenetic event in MNGIE‐associated gut motor dysfunction before significant myopathic and/or neuropathic structural changes occur.
Characterization of gastric volume responses and liquid emptying in functional dyspepsia and health by MRI or barostat and simultaneous 13C‐acetate breath test
Authors: Fruehauf H. , Steingoetter A. , Fox M. R. , Kwiatek M. A. , Boesiger P. , Schwizer W. , Fried M. , Thumshirn M. , Goetze O.
Published: March 2009

Abstract  The assessment of gastric accommodation and emptying by different methodologies provides inconsistent results. We aimed to compare magnetic resonance imaging (MRI), barostat and 13C‐acetate breath test (BT) for the assessment of gastric volume responses and emptying in healthy controls (HC) and patients with functional dyspepsia (FD). Eight HC and eight FD patients underwent: (i) continuous BT with simultaneous MRI in the upright position after ingestion of isocaloric, 300 kcal, 200 and 800 mL meals, both labelled with 100 mg of 13C‐acetate; and (ii) BT with gastric barostat after ingestion of the 200 mL meal. MRI measured total gastric volume and gastric content volume (GCV) at baseline, after filling and during emptying. Meal emptying half‐times (T½) for MRI and BT were calculated (mean ± SD). We found: (i) Initial GCV was lower in FD than in HC (762 ± 22 vs 810 ± 52 mL, P < 0.04) after the 800 mL meal but not the 200 mL meal. T½MRI was shorter for the 800 mL than the 200 mL meal (P < 0.001), but similar in HC and FD (200 mL: HC 117 ± 30 min vs FD 138 ± 42 min, ns; 800 mL: HC 71 ± 16 min vs FD 78 ± 27 min, ns). In contrast, T½BT was similar between meals and groups (200 mL: HC 111 ± 11 min vs FD 116 ± 19 min; 800 mL: HC 114 ± 14 min vs FD: 113 ± 17 min). (ii) Barostat measurements showed similar postprandial volume increases between groups. We conclude that direct measurements by MRI provide a sensitive, non‐invasive assessment of gastric accommodation and emptying after a meal. In contrast to MRI, BT did not detect faster emptying of high‐volume compared to low‐volume liquid nutrient meals in HC or FD.
Multi‐proxy geoarchaeological study redefines understanding of the paleocoastlines and ancient harbours of Liman Tepe (Iskele, Turkey)
Authors: Goodman Beverly N. , Reinhardt Eduard G. , Dey Hendrik W. , Boyce Joseph I. , Schwarcz Henry P. , Sahoğlu Vasif , Erkanal Hayat , Artzy Michal
Published: March 2009

Determining the position of Liman Tepe's (ancient `Clazomenae') archaeological features relative to the coastline is important for understanding their intended function and reconstructing the character of Aegean maritime activities and sea‐based trade. Previous attempts at reconstructing harbour locations at Liman Tepe relied on extrapolating paleoenvironments based on modern surface topography. In light of this, samples from a sediment coring survey and terrestrial and underwater archaeological excavations were analysed using multi‐proxy geoarchaeological methods to determine paleoenvironmental facies. Micropaleontological (foraminifera), sedimentological (grain‐size analysis) and geochemical (δ13C/δ18O) analyses resulted in the reconstruction of the coastal paleogeomorphology, including the presence and absence of ancient harbouring areas. Neither of the previous coastal reconstructions was supported by the new results. Instead, two separate harbouring areas were recognized, one coincident with the Early Bronze Age (4800–3900 years bp) and a second during the archaic and classical periods (c. 2800–2400 years bp). These results emphasize the necessity for multi‐proxy geoarchaeological studies when approaching coastal archaeological sites as a means to reconstruct paleocoastal geomorphology and understand ancient maritime development better.
A new HLA‐C allele: Cw*0819
Authors: Johnson J. , Street J. , Hammond L. , Pepperall J. , Darke C.
Published: March 2009

Human leucocyte antigen‐Cw*0819 differs from Cw*0802 by two nucleotides resulting in an amino acid change of 99tyrosine to 99phenylalanine and a synonymous substitution.
HLA‐G polymorphism and in vitro fertilization failure in a Polish population
Authors: Sipak‐Szmigiel O. , Cybulski C. , Wokołorczyk D. , Lubiński J. , Kurzawa R. , Bączkowski T. , Radwan M. , Radwan P. , Ronin‐Walknowska E.
Published: March 2009

To investigate whether human leukocyte antigen‐G (HLA‐G) gene polymorphism is associated with in vitro fertilization (IVF) failure, we sequenced exons 2–4 of the HLA‐G gene in 50 couples with three or more IVFs (including 10 couples with five or more IVFs) and 58 control fertile couples from a Polish population. Of the 10 different HLA‐G alleles identified in our study subjects, neither allele was found to be associated with IVF. We also genotyped 50 couples with IVF and 71 control couples for the −725C>G variant in the promoter region and the 14 bp insertion or deletion polymorphism in the 3′ untranslated region of the HLA‐G gene. The frequency of −725GG or GC genotype in women with IVF and in control fertile women was similar [26% vs 25.3%; odds ratio (OR) = 1.0; P = 1.0]. The 14 bp ins/ins or ins/del genotype was more common in women with IVF than in control women (76.9% vs 59.1%; OR 2.4; P = 0.03), but the difference was not significant after Bonferroni correction for multiple comparisons. The frequency of the ins/ins or ins/del genotype was particularly high (90%) in women who experienced five or more IVFs (OR = 6.2; P = 0.08), but again, the excess was not statistically significant, possibly because of small sample sizes. These results are in line with functional studies that show lower levels of HLA‐G mRNA and protein related to the HLA‐G allele including the 14 bp sequence and suggest that the insertion allele may be associated with an increased risk of IVF.
HLA‐B51 and haplotypic diversity of B‐Cw associations: implications for matching in unrelated hematopoietic stem cell transplantation
Authors: Bettens F. , Nicoloso de Faveri G. , Tiercy J.‐M.
Published: March 2009

In unrelated hematopoietic stem cell transplantation (HSCT), human leukocyte antigen (HLA)‐C locus incompatibilities occur frequently and are associated with increased risk of posttransplant complications. Because HLA‐B51 is associated with a high rate of Cw disparities, we performed a comprehensive four‐digit typing analysis of 140 ABCDRB1 B51 genotypes proven by pedigree analysis and 311 unrelated donors selected for 75 B51‐positive patients. In addition, 145 A1/Ax‐B8/B51‐DR3/DRx donors were HLA typed at a high‐resolution level and tested for three microsatellite (Msat) polymorphisms located in the HLA class I and III regions. Based on these data sets, 182 different ABCDR haplotypes with 14 different B‐Cw associations were detected. Rates of Cw mismatches were shown to be highly correlated with the ABDRB1 haplotypes. We have computed 21 B51 haplotypes that disclose a high probability of HLA‐C allele matching and 30 haplotypes with a low (<25%) probability. The HLA‐C allele frequency profiles were quite different in these two groups, with a more heterogeneous distribution in the low matching probability group. HLA‐Cw*1502 was inversely correlated with the likelihood to identify a Cw‐mismatched donor: it was present in 61% of the high vs 18% of the low probability group (P < 0.0001). The analysis of three Msats in the class I and III regions showed a higher allelic diversity in B51‐positive haplotypes compared with the conserved A1‐B8‐DR3 haplotype. HLA‐B51 haplotypes therefore exhibit a high diversity at the level of B‐Cw associations and of non‐HLA polymorphisms in the class I and III regions. Such heterogeneity negatively impacts on overall matching in HSCT.
Autoimmune disease association signals in CIITA and KIAA0350 are not involved in celiac disease susceptibility
Authors: Dema B. , Martínez A. , Fernández‐Arquero M. , Maluenda C. , Polanco I. , Figueredo M. Ángeles , De La Concha E. G. , Urcelay E. , Núñez C.
Published: March 2009

Celiac disease (CD) is a multifactorial disease characterized by intestinal inflammation after gluten exposure in genetically susceptible individuals. A strong influence of certain human leukocyte antigen (HLA) alleles (those coding the HLA‐DQ2 and DQ8 heterodimers) is well established, but they cannot explain the overall genetic risk. CIITA could be a good candidate gene for CD because it is mainly transcriptionally regulated, and it encodes the master regulator of major histocompatibilty complex class II gene transcription. CIITA is located in 16p13, a region also containing KIAA0350 (CLEC16A), associated with two autoimmune diseases in genome‐wide association studies. We aimed at studying the involvement of polymorphisms in CIITA and KIAA0350 in CD susceptibility, with special attention to evaluate the possible presence of more than one risk factor in the region. We performed a case–control study with 607 CD patients and up to 794 healthy controls, all Spaniards. All samples were genotyped for five single nucleotide polymorphisms: rs3087456 (−168A/G) and rs4774 in CIITA and rs7203459, rs6498169 and rs2903692 in KIAA0350. No significant results were obtained when comparing genotypic, allelic or haplotypic frequencies between patients and controls. Our results seem to discard the influence in CD susceptibility of CIITA and KIAA0350 markers previously associated with other autoimmune diseases.
A real‐time polymerase chain reaction assay for the rapid identification of the autoimmune disease‐associated allele HLA‐DQB1*0602
Authors: Gersuk V. H. , Nepom G. T.
Published: March 2009

Many autoimmune diseases share a genetic association with the presence or absence of human leukocyte antigen (HLA)‐DQB1*0602, including type I diabetes, multiple sclerosis, and narcolepsy. High‐resolution HLA typing to determine the presence of this allele is cumbersome and expensive by currently available techniques. We present a real‐time polymerase chain reaction (PCR) assay for the identification of HLA‐DQB1*0602, using sequence‐specific primers and probes, that provides rapid and sensitive identification of this allele, involves minimal hands‐on time, and provides major cost savings compared with existing methods. The assay allows the simultaneous determination of both the presence and the number of copies of this allele. Because there is no post‐PCR handling, the risk of contamination is avoided. We have validated the assay using 44 blinded and 32 unblinded samples, previously typed by standard techniques, which were identified with 100% accuracy, sensitivity, and specificity. Furthermore, using a narcolepsy cohort of 722 subjects, we demonstrated the robustness of the assay to analyze DNA isolated from buccal swabs, demonstrating the applicability of this assay as a suitable approach for population‐based studies.
A new HLA‐B*51 variant, B*5158, identified by sequence‐based typing in a Korean individual
Authors: Roh E. Y. , Shin S. , Yoon J. H. , Ahn B. M. , Chang J. Y.
Published: March 2009

A novel human leukocyte antigen (HLA)‐B*51 allele, officially named HLA‐B*5158, was identified in the cord blood from Korean. HLA‐B*5158 allele shows single nucleotide difference from B*510101 in exon 2 at nucleotide position 214 (C/T), resulting in an amino acid substitution, Trp48Arg.
Identification of a novel HLA‐B allele, B*460102, in three Taiwanese individuals
Authors: Chu C.‐C. , Lee H.‐L. , Lin M.
Published: March 2009

We here report a new human leukocyte antigen (HLA)‐B allele, B*460102, which differs from B*460101 by synonymous substitution at the third nucleotide of codon 74 (GAC→GAT) and independently found in three Taiwanese individuals.
Assessment of the amino acid substitution in HLA‐B*5719 for allorecognition
Authors: Figueiredo C. , Mbulaiteye S. , Ndugwa C. M. , Schulz T. , Eiz‐Vesper B.
Published: March 2009

The novel allele human leukocyte antigen (HLA)‐B*5719 differs from HLA‐B*5701 by a synonymous nucleotide exchange at position 539 in exon 3 (C→T), replacing Leucine by Arginine at amino acid position 156 in the α2 domain.