bluuebunny/crossref_metadata_2025 huggingface dataset:
Diversity of expressed cytochrome P450 genes in the adult Mediterranean Fruit Fly, Ceratitis capitata
Authors: Field L. M. , James A. A. , Danielson P. B. , Foster J. L. M. , Cooper S. K. , Fogleman J. C.
Published: September 2003

The cytochrome P450s comprise a superfamily of mostly microsomal haemoproteins which play a dominant role in the metabolism of a variety of endogenous and foreign compounds. The use of a degenerate PCR primer targeted to the haem‐binding decapeptide unique to the cytochrome P450 superfamily resulted in the identification of 14 novel cytochrome P450s in the Mediterranean Fruit Fly, Ceratitis capitata. Analysis of the relative frequency of individual isoforms within the pool of isolated sequences suggests that the CYP4 and CYP6 P450 families contain the most highly expressed isoforms in adult C. capitata. Phylogenetic analyses of the conceptual amino acid translations of PCR‐amplified cDNAs provides evidence that one of isolated sequences may represent a new P450 family.
Voltage‐gated sodium channel genes hscp and hDSC1 of Heliothis virescens F. genomic organization
Authors: Field L. M. , James A. A. , Park Y. , Taylor M. F. J. , Feyereisen R.
Published: September 2003

We report the genomic sequence of hscp, a sodium channel α subunit gene for Heliothis virescens. A 32‐kb genomic clone and six independent RT‐PCR products covering almost the entire coding region of the gene, contained thirty‐one deduced exons with a translation of 1695 residues. Overall amino acid similarity to the para locus of Drosophila melanogaster was 86%. The transcription of the gene was complex. Alternate splicing was evident for five optional exons and a pair of mutually exclusive exons. A number of alternatively spliced mRNA revealed a deduced translation product that included only the first homology domain. We also report the first partial sequence for hDSC1, a presumed orthologous of the DSC1 sodium channel α subunit gene of D. melanogaster.
A new aminopeptidase from diamondback moth provides evidence for a gene duplication event in Lepidoptera
Authors: Field L. M. , James A. A. , Chang W. X. Z. , Gahan L. J. , Tabashnik B. E. , Heckel D. G.
Published: September 2003

We screened a midgut cDNA library from diamondback moth, Plutella xylostella, with a probe generated using sequence information from an aminopeptidase N gene from Manduca sexta (MsAPN‐1). The sequence recovered (PxAPN‐A) encodes a protein of 988 resides with a 60% sequence identity to MsAPN‐1. The two proteins share a signal peptide which directs processing by the endoplasmic reticulum, a C‐terminal hydrophobic region satisfying the criterion for a GPI anchor and cleavage, and the possibility of an O‐glycosylated rigid stalk attached to the GPI anchor. PxAPN‐A is more closely related to MsAPN‐1 than it is to another aminopeptidase recently reported from P. xylostella. Sequence comparisons with other species suggests that at least one aminopeptidase gene duplication occurred in an ancestral lepidopteran.
Typing of sandflies from Greece and Cyprus by DNA polymorphism of 18S rRNA gene
Authors: Field L. M. , James A. A. , Aransay A. M. , Scoulica E. , Chaniotis B. , Tselentis Y.
Published: September 2003

A simple and reliable technique was developed to distinguish Phlebotomine sandflies by restriction fragment length polymorphism of PCR‐amplified (PCR‐RFLP) 18S rDNAs. Seven morphologically identified sandflies species from several localities of Greece and Cyprus were studied, and specific patterns were developed by double digesting amplified 18S rDNAs with Hpall and Rsal. Three additional species of the sub‐genus Larroussius were distinguished by a second double digestion with Accl and Banl. We have successfully applied the method on samples in which morphological characters were badly distinguished due to poor storage conditions and in larval stages.
Distribution and phylogeny of Wolbachia inducing thelytoky in Rhoditini and ‘Aylacini’ (Hymenoptera: Cynipidae)
Authors: Field L. M. , James A. A. , Plantard O. , Rasplus J.‐Y. , Mondor G. , Le Clainche I. , Solignac M.
Published: September 2003

Wolbachia are endosymbiotic bacteria responsible for thelytoky in several parasitoid hymenopteran genera. After finding these micro‐organisms in some populations of Diplolepis spinosissimae (Hymenoptera: Cynipidae) where they are responsible for thelytoky through gamete duplication, we searched for Wolbachia spp. using specific PCR primers in nineteen other species of the Rhoditini tribe (rose gallwasps) and eight species of the ‘Aylacini’ tribe (gallwasps associated with herbaceous plants). Wolbachia were found in twelve Rhoditini species and four ‘Aylacini’ species. The most infected species have very few males (spanandry) and the thelytoky of infected species/arrhenotoky of uninfected species is confirmed by previous research based on the sex of the offspring of virgin females. Phylogenetic analyses based on the partial Wolbachia ftsZ gene sequences indicate that some strains associated with closely related gallwasps are phylogenetically distant, suggesting that cynipids have been affected by several infection events. In contrast, the five infected European species of Diplolepis harbour the same strain of Wolbachia.
Structural polymorphism of the luciferase gene in the firefly, Luciola lateralis
Authors: Field L. M. , James A. A. , Cho Kook Ho , Lee Jong Seob , Choi Yang Do , Boo Kyung Saeng
Published: September 2003

To study the structural features of genes for luciferase in Luciola lateralis, two different, functional luciferase genes, Luc1 and Luc2, were cloned and their nucleotide sequences were determined. The two genes were nearly identical, but a deletion of 128 bp was found in the 5′ upstream region of Luc2, and minor deletions and additions were also found in the 3′ downstream region. Seven base substitutions in the coding region produced two amino acid residue differences between the two genes. Southern blot analysis of genomic DNA isolated from 33 individual fireflies revealed that 26 were homozygous for the Luc1 gene and one was homozygous for Luc2. The others were heterozygotes with two, or even three, different alleles for the luciferase gene.
Moose, a new family of LTR‐retrotransposons in the mosquito Anopheles gambiae
Authors: Field L. M. , James A. A. , Biessmann H. , Walter M. F. , Le D. , Chuan S. , Yao J. G.
Published: September 2003

A novel LTR retrotransposable element called moose has been cloned and characterized from the malaria transmitting mosquito, Anopheles gambiae. This element has all the characteristic features of LTR retroelements and is related to retroelements from other insects and nematodes, belonging to a subgroup of retroelements distinct from the copia/Ty1 and gypsy/Ty3 groups. The moose element appears to be active in A. gambiae, and strong RNA expression is detected in the male and female gonads. The use of this retroelement as a potential vector for germ line transformation is discussed.
Genetic similarity among pheromone and voltinism races of Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae)
Authors: Field L. M. , James A. A. , Marçon P. C. R. G. , Taylor D. B. , Mason C. E. , Hellmich R. L. , Siegfried B. D.
Published: September 2003

The genetic variability of seven European corn borer populations, Ostrinia nubilalis, from North America and Europe was assessed by polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) analysis and DNA sequencing. The nuclear ribosomal internal transcribed spacer 1 (ITS‐1) region (≈ 500 base pair [bp]) and four mitochondrial (mtDNA) regions (1550 bp total) were examined. The smartweed borer, Ostrinia obumbratalis, and south‐Western corn borer, Diatraea grandiosella, were used for comparisons. Of 106 restriction sites identified (80 in mtDNA and 26 in ITS‐1), none differentiated geographical populations, pheromone races, or voltine ecotypes of the European corn borer. The lack of variation in the ITS‐1 of European corn borer was confirmed by DNA sequence analysis. The genetic similarity of European corn borer populations, despite their wide geographical range and physiological differences, may be explained by a relatively recent origin for the voltinism and pheromone races, gene flow among races, and/or expansion from genetic bottlenecks.
Proteins from nuclear extracts of two lepidopteran cell lines recognize the ends of TTAA‐specific transposons piggyBac and tagalong
Authors: Field L. M. , James A. A. , Bauser C. A. , Elick T. A. , Fraser M. J.
Published: September 2003

The transposons piggyBac and tagalong are Lepidopteran transposons that exhibit extreme site‐specificity for the tetranucleotide TTAA upon insertion and excise in a characteristic precise fashion, regenerating a single TTAA target site. The precise excision of both piggyBac and tagalong can occur in the absence of factors encoded by either transposon, possibly through the recruitment of host proteins and/or cross‐mobilizing transposons. In this report, we utilize mobility shift assays and exonuclease III protection analyses to identify DNA binding activities from IPLB‐SF21AE and TN‐368 cells that are specific for piggyBac and tagalong‐terminal repeats.
Identification of a non‐LTR retrotransposon from the gypsy moth
Authors: Field L. M. , James A. A. , Garner K. J. , Slavicek J. M.
Published: September 2003

A family of highly repetitive elements, named LDT1, has been identified in the gypsy moth, Lymantria dispar. The complete element is 5.4 kb in length and lacks long‐terminal repeats. The element contains two open reading frames with a significant amino acid sequence similarity to several non‐LTR retrotransposons. The first open reading frame contains a region that potentially encodes a polypeptide similar to DNA‐binding GAG‐like proteins. The second encodes a polypeptide resembling both endonuclease and reverse transcriptase sequences. All members of the LDT1 element family sequenced thus far have poly‐A tails or A‐rich tails of 12–18 nucleotides in length, but lack a poly‐A addition signal in the expected location. The amplification of retrotransposon insertion junction regions in different gypsy moth individuals indicates that polymorphisms exist at some of the insertion sites, suggesting that this element is or was, until recently, capable of transposition.
Microbe‐induced cytoplasmic incompatibility as a mechanism for introducing transgenes into arthropod populations
Authors: Field L. M. , James A. A. , Turelli M. , Hoffmann A. A.
Published: September 2003

Many arthropods are infected with maternally transmitted, intracellular bacteria of the genus Wolbachia. These infections often produce ‘cytoplasmic incompatibility’ (CI)—reduced egg‐hatch frequencies when uninfected females mate with infected males or when males and females carrying different Wolbachia strains mate. Because infected females often enjoy a fitness advantage—they are effectively immune to any effects from males carrying the same Wolbachia strain—Wolbachia and associated cytoplasmic elements can spread rapidly through natural populations. Wolbachia might therefore drive transgenes associated with disease control or pest abatement into populations. In this paper, simple mathematical analyses are presented of three alternative strategies for ‘CI drive’. The analyses reveal which parameters must be estimated in order to predict population dynamics, and they demonstrate stringent requirements for initially driving and/or maintaining transgenes in target populations.
Cloning and expression of ecto 5′‐nucleotidase from the cattle tick Boophilus microplus
Authors: Field L. M. , James A. A. , Liyou N. , Hamilton S. , Elvin C. , Willadsen P.
Published: September 2003

Although 5′‐nucleotidases are ubiquitous in higher vertebrates, the arthropod enzymes have been little studied. The cDNA sequence of the mature 5′‐nucleotidase from the tick Boophilus microplus was therefore determined (GenBank accession number: U80634). The enzyme has 39–41% sequence identity with the vertebrate 5′‐nucleotidases and contains binuclear metal ion binding sites. There are no significant introns within the coding region of the genomic sequence. Southern blot analysis indicates the presence of multiple related genes encoding 5′‐nucleotidases. Recombinant tick 5′‐nucleotidase was expressed in both Escherichia coli and in baculovirus‐infected insect cells. The E. coli recombinant protein was truncated, inactive and produced in abundance. The enzyme was expressed in baculovirus‐infected insect cells as a secreted, soluble, glycosylated and enzymatically active protein. This represents the first successful expression and characterization of enzymatically active recombinant 5′‐nucleotidase from any organism. Supplementation of the culture medium with 25 μm zinc resulted in a twofold increase in the activity of the expressed protein. The enzyme was purified to homogeneity. It exists under non‐denaturing conditions as a homodimer, with an apparent molecular mass of 135 kDa. The Km for the hydrolysis of AMP was 0.37 μm and the kcat= 11.5/s, in agreement with data for the native enzyme.
Variation in the salivary peptide, maxadilan, from species in the Lutzomyia longipalpis complex
Authors: Field L. M. , James A. A. , Lanzaro G. C. , Lopes A. H. C. S. , Ribeiro J. M. C. , Shoemaker C. B. , Warburg A. , Soares M. , Titus R. G.
Published: September 2003

Maxadilan is an approximately 7kDa peptide that occurs in the saliva of the sand fly Lutzomyia longipalpis. This peptide is a potent vasodilator and may also have immunomodulatory effects related to the pathogenesis of leishmanial infections. Variation in the primary DNA and inferred amino acid sequence of maxadilan is reported. Differences were found within and among natural field populations as well as among sibling species. Extensive amino acid sequence differentiation, up to 23%, was observed among maxadilan from different populations. This is a remarkable degree of polymorphism considering the small size of this peptide. The vasodilatory activity of maxadilan was equivalent among recombinant maxadilan variants. All maxadilan variants induce interleukin‐6. Predicted secondary structure and hydrophobicity plots suggest that these characteristics are conserved among variant peptides. However, profiles based on the antigenic index do differ among peptides.
Identification and characterization of a putative sevenless homologue in the malaria vector Anopheles gambiae
Authors: Field L. M. , James A. A. , Kobayashi A. , Brey P. T. , Katsube K. , della Torre A. , Roth C. W. , Natori S. , Ollo R.
Published: September 2003

Tyrosine kinase sequences were identified and characterized in Anopheles gambiae, the major vector of malaria in subsaharan Africa. One of these sequences has the characteristics expected for a homologue of the Drosophila sevenless gene, which is necessary for R7 photoreceptor cell fate determination in the developing compound eye. The putative Anopheles seven‐less gene homologue is located in a telomeric region of the X chromosome and is expressed in the head of late larval and pupal stage mosquitoes. Identification of the Anopheles homologue of the sevenless gene is a first step towards the development of a dominant phenotypic marker that could be used for detecting transformed Anopheles mosquitoes in a wide variety of genetic backgrounds and, as such, could be used in the development of transgenic mosquitoes for the control of parasite transmission. Preliminary evidence for sevenless sequences were also found in DNA from blackfly, Mediterranean fruit fly and the honeybee.
Analysis of genetic variability in Anopheles arabiensis and Anopheles gambiae using microsatellite loci
Authors: Field L. M. , James A. A. , Kamau L. , Mukabana W. R. , Hawley W. A. , Lehmann T. , Irungu L. W. , Orago A. A. S. , Collins F. H.
Published: September 2003

We analysed genetic variability in Anopheles arabiensis and Anopheles gambiae populations using micro‐satellite loci to determine whether the Rift Valley restricts the flow of genes. Deviations from Hardy–Weinberg expectations were significant, and were most likely to be due to the high frequency of null alleles observed. An. arabiensis populations occurring between 40 and 700 km apart across the Eastern arm of the Rift Valley were not differentiated (pair‐wise Fst range: 0.0033–0.0265, P > 0.05). Neither were An. gambiae populations from Asembo Bay and Ghana (Fst: 0.0063, P > 0.05) despite a geographical separation of about 5000 km. In contrast, significant differentiation was observed between An. gambiae populations from Asembo Bay and Kilifi (about 700 km apart; FST= 0.1249, P < 0.01), suggesting the presence of a barrier to gene flow.
Cloning and sequence of a cDNA for a highly basic protease from the digestive juice of the silkworm, Bombyx mori
Authors: Field L. M. , James A. A. , Kotani E. , Niwa T. , Tokizane M. , Suga K. , Sugimura Y. , Oda K. , Mori H. , Furusawa T.
Published: September 2003

A serine protease of the silkworm, Bombyx mori, with an isoelectric point of pH 10–11 and a pH optimum for succinyl‐Leu‐Leu‐Val‐Tyr‐MCA degrading activity of about 10, was found in a 0.33 m NaCI‐eluted fraction obtained from cation‐exchange chromatography of digestive juice. The activity of the enzyme was strongly inhibited by chymostatin and PMSF, indicating that the protease is a chymotrypsin‐like serine protease. The N‐terminal amino acid sequence of the protease was determined, and a full‐length cDNA clone (0.92 kbp) which was isolated from a midgut cDNA library was sequenced. The cDNA encodes a pre‐proenzyme of 284 amino acids with a pro‐segment of 50 amino acids and mature protein of 234 amino acids. From its primary structure, the predicted molecular mass of the mature protein is 24.5 kDa. A sequence comparison of the Bombyx highly basic protease with other serine proteases revealed that this enzyme is a mammalian‐type serine protease with a catalytic triad consisting of His45, Asp92 and Ser186. A large number of Arg residues are encoded by the cDNA which may be responsible for its stability and/or function in the alkaline condition, by remaining charged at high pH.
cDNA cloning and transcript distribution of two novel members of the neuroparsin family in the desert locust, Schistocerca gregaria
Authors: Claeys I. , Simonet G. , Van Loy T. , De Loof A. , Vanden Broeck J.
Published: September 2003

This study describes the identification and distribution of two novel neuroparsin precursor transcripts (Scg‐NPP3/Scg‐NPP4) in the desert locust, Schistocerca gregaria. Unlike Scg‐NPP1 and Scg‐NPP2, both transcripts were not only detected in the brain, but also in various other tissues, such as fat body, ventral nerve cord, testis and male accessory glands. Northern analysis showed that the levels of these transcripts are regulated during larval development, as well as during moulting and reproductive cycles. A significant increase in both mRNAs was observed during the period that just precedes the initial sexual activity of adult females and males.

In silico analysis of sequence databases revealed the existence of several other neuroparsin‐like peptides in a variety of arthropod species, including crustaceans and chelicerates. Neuroparsins also display similarities with vertebrate IGFBP.
A mammalian‐like lipase gene is expressed in the female reproductive accessory glands of the sand fly Phlebotomus papatasi (Diptera, Psychodidae)
Authors: Rosetto M. , Belardinelli M. , Fausto A. M. , Marchini D. , Bongiorno G. , Maroli M. , Mazzini M.
Published: September 2003

A lipase‐like protein (PhpaLIP) was identified as the major protein component in the secretion of the female reproductive accessory glands of the sand fly Phlebotomus papatasi. The full‐length cDNA encoding this protein was isolated and its nucleotide sequence determined. The deduced translational product of the gene contains a GFSFG motif, consistent with a GXSXG consensus, which is shared by most bacterial and eukaryotic hydrolases. Transcriptional analysis of the PhpaLIP gene showed that its expression is female‐specific, and is also detectable in districts other than accessory glands, suggesting that it might play different functions. Taken together with the observation of sequence similarity shared by PhpaLIP and mammalian lipases, the demonstration of the presence of lipase activity in the accessory gland secretion suggests a possible biological role of PhpaLIP gene product.
Molecular characterization of prothoracicotropic hormone and diapause hormone in Heliothis virescens during diapause, and a new role for diapause hormone
Authors: Xu W.‐H. , Denlinger D. L.
Published: September 2003

cDNAs encoding prothoracicotropic hormone (PTTH) and diapause hormone (DH) were isolated from the tobacco budworm, Heliothis virescens (Hvi). Hvi‐PTTH cDNA reveals key structural features known from other PTTHs, yet there is a > 30% amino acid difference between the PTTH sequences found in this noctuid when compared with the PTTHs known from Bombycoidea. Hvi‐DH‐PBAN cDNA encodes a precursor protein including a DH‐like peptide, pheromone biosynthesis‐activating neuropeptide (PBAN) and three additional neuropeptides with an FXPRL sequence at the C terminus. PTTH and DH‐PBAN transcripts are most highly expressed in the brain and subesophageal ganglion (SG), respectively. Expression of both the PTTH and the DH‐PBAN transcripts is high in larvae and remains high in nondiapausing pupae, but in larvae programmed for pupal diapause, expression declines sharply at the onset of larval wandering behaviour and remains low during pupal diapause. This pattern implies that both of these genes are shut down during diapause. These results are not consistent with a role for DH in promoting the entry into diapause as noted in Bombyx mori. Instead, the higher expression of DH in nondiapausing pupae suggests a possible role for DH in promoting continuous development. The injection of DH into diapausing pupae did indeed successfully terminate diapause, thus suggesting a possible new role for this neuropeptide.
A genetic linkage map for Tribolium confusum based on random amplified polymorphic DNAs and recombinant inbred lines
Authors: Yezerski A. , Stevens L. , Ametrano J.
Published: September 2003

Tribolium beetles provide an excellent and easily manipulated model system for the study of genetics. However, despite significant increases in the availability of molecular markers for the study of genetics in recent years, a significant genetic linkage map for these beetles remains undeveloped. We present the first molecular genetic linkage map for Tribolium confusum using random amplified polymorphic DNA markers. The linkage map contains 137 loci mapped on to eight linkage groups totaling 968.5 cM.
The effects on the quality of Cavendish bananas, which have been treated with ethylene, of exposure to 1‐methylcyclopropene
Authors: Bagnato N. , Barrett R. , Sedgley M. , Klieber Andreas
Published: September 2003

After 48 h of ethylene treatment (300 μL L−1), to induce ripening of mature, green bananas (Musa sp., AAA type, Cavendish subgroup, cv. Williams), fruit were exposed to 0 (control), 3, 300 nL L−1, or 30 μL L−1 1‐methylcyclopropene (1‐MCP) for 24, 48 or 72 h at 20 °C. Bananas treated with 300 nL L−1 1‐MCP had a 6‐day shelf‐life compared with 3 days for non‐treated fruit, and 4 days for fruit treated with 3 nL L−1 1‐MCP. Increased shelf‐life (half‐ripe to over‐ripe) occurred without affecting the green life (unripe to half‐ripe) of bananas, peel appearance, pulp texture, soluble solid concentrations or aroma profiles. Fruit treated with 30 μL L−1 were externally and internally commercially unacceptable, as fruit developed crown rot prior to ripening. Application of 1‐MCP at suitable concentrations could extend banana shelf‐life, by enhancing marketing and consumer expectations without compromising banana quality.
Effect of location and soaking treatments on the cooking quality of some chickpea breeding lines
Authors: Coşkuner Yalçın , Karababa Erşan
Published: September 2003

The effects on some physical criteria and on cooking quality of soaking chickpeas (of different genotypic origins) in sodium chloride or sodium bicarbonate were investigated. The material was grown at five locations in Çukurova, Turkey. The seed size, hundred seed weight, seed density, hydration capacity, swelling capacity and cooking time values differed significantly among locations. Cooking time differed significantly among locations and treatments, but not with cultivars. Soaking with NaCl and NaHCO3was effective for decreasing the cooking time.
Aspartame: studies on UV spectral characteristics
Authors: Dattatreya B. S. , Usha D. V. , Susheelamma N. S. , Bhat K. K.
Published: September 2003

Model experiments were done by using aspartame (APM) solutions containing organic acids, such as malic, tartaric and citric acids, before and after heating at 80 °C for 1 h. Ultra violet spectral data was gathered and indicated an increase in λmax as well as the general absorbance with increasing concentration of APM. Spectra were recorded for both APM and its degradation products in water and 0.01% HCl and also before and after heating. The quantitation of APM by this spectral method was found to be comparable with that obtained by an HPLC method. Spectra were recorded for concentrations of 0.25–5 mm APM solutions between 200 and 400 nm. Linear, semi‐log and polynomial equations were found to give good fits to the observed data with major and minor peaks in both normal and derivative spectral modes. Beverages containing lemon juice showed higher λmax and absorbance values than normal with APM present. The results indicated that UV spectrophotometry could be used to monitor changes in APM concentration during processing and/or storage, and could indicate the presence or absence of APM polymorphism in solution.
Use of Australian cardoon extract in the manufacture of ovine milk cheese – a comparison with commercial rennet preparations
Authors: Chen Shaojiang , Agboola Samson , Zhao Jian
Published: September 2003

Aqueous extract of Australian cardoon (Cynara cardunculus L.) flowers was used to produce Pecorino cheese from ovine milk and the chemical, biochemical and sensory properties of the cheese were compared with cheeses made using two commercial (calf and microbial) rennet preparations. The cheeses contained 49.8–54.6% moisture, 28.2–31.2% fat, 2.1–2.3% NaCl and 2.6–3.1% total nitrogen. Very few differences in chemical composition were observed between cheeses manufactured with the three different coagulants. Biochemically, however, cheese manufactured with cardoon extract contained significantly (P < 0.05) higher levels of water‐soluble nitrogen but lower levels of phosphotungstic acid soluble nitrogen, while the levels of trichloroacetic acid soluble nitrogen were similar among the three types of cheese. Compared with cheeses made with commercial rennet preparations, cheese made with cardoon extract was perceived to be softer, creamier and less bitter, and was rated significantly (P < 0.05) higher in overall liking by both trained and consumer panels.
Strength of health‐related claims and their perceived advantage
Authors: Urala Nina , Arvola Anne , Lähteenmäki Liisa
Published: September 2003

Consumers (n = 958) were asked to evaluate whether product‐related health claims in foods were either advantageous or disadvantageous, in their opinion. Claims were made for six functional components and two control products, namely low salt content and a high‐pressure technique and the strength of claims varied between four intensity levels. In general, all claims were perceived as neutral or as advantageous. Increasing the strength of the claim did not automatically increase the perceived benefit. Gender, trust in different information sources and the frequency of use of so‐called functional foods affected the perceived benefit. Women perceived the claims to be more beneficial than men did. Trustful respondents perceived the claims as more advantageous than did sceptical respondents, and the users of functional foods perceived health claims to be more advantageous than did non‐users. In addition, personal motivation affected the perception of the claims. With less familiar functional components, the strength of the claim increased the perceived benefit, whereas with familiar components, claims mentioning the reduced risk or prevention of a disease did not increase the perceived advantage.
State diagrams to help describe wheat bread processing
Authors: Cuq Bernard , Abecassis Joel , Guilbert Stéphane
Published: September 2003

Physical changes in wheat bread processing can be described by the use of state diagrams and phase transition. State diagrams were constructed from real experimental results that described the glass transition and melting curves for starch and proteins as a function of water content. Wheat gluten protein is the main component responsible for bread dough structural formation, while starch is mainly involved in final textural properties and stability. During processing, hydration of wheat flour induces the glass transition of wheat gluten proteins. The input of mechanical energy during the mixing allows the development of continuous viscoelastic dough due to interactions between rubbery proteins. During baking, irreversible modifications of starch and proteins at high temperatures (>200 °C) and superficial dehydration are responsible for bread crust formation. In the central part of the mixture, starch gelatinization and protein coagulation induce breadcrumb formation. After cooling, the low water content of the crust (3–7%) is sufficient for the transition of bread components from the rubbery to the glassy state. Consequently, structural hardening occurs and is responsible for crust crispness. The higher final water content of the crumb (35–40%) is responsible for rubbery behaviour, which gives structural mobility, smooth breadcrumb behaviour and explains the sensitiveness to starch retrogradation during storage.
Genetic services for people with intellectual disability and their families
Authors: Raymond F. L.
Published: September 2003

This paper reviews the advances in molecular genetics over the recent years and discusses the impact it may have on those with intellectual disability and their families. The aim is not to present a comprehensive scientific treatise but rather to use illustrations from genetics to highlight our current thinking and draw attention to areas of uncertainty and misinformation. As our knowledge and understanding of the genetic basis of disease increases over the years, there may be significant benefits to some families, but the potential for discrimination against individuals on genetic grounds will also increase.
The Human Genome Project: considerations for people with intellectual disabilities
Authors: Holland A. , Clare I. C. H.
Published: September 2003

The paper discusses the impact which recent advances in gene technology may have for people with intellectual disabilities. It highlights a conflict between the apparent benefits of advances in genetics for the population in general and the negative effects on persons with intellectual disabilities in particular. This conflict is illustrated in the paper through the use of examples, including the implications of a person's lack of capacity to consent to investigations and treatment; the issue of individual responsibility for antisocial behaviour when this is considered to have a genetic component; and the issue of life and death decisions about treatment in the face of a progressive, genetically determined, disorder. The authors adopt a human rights perspective to discuss these examples because it provides a richness to articulate and justify moral concerns in this area which is lacking in much of the current debate as this is dominated by the primacy of autonomy.
Reported symptoms and clinical findings in relation to serum cobalamin, folate, methylmalonic acid and total homocysteine among elderly Swedes: a population‐based study
Authors: Björkegren K. , Svärdsudd K.
Published: September 2003

Abstract Björkegren K, Svärdsudd K, (Department of Public Health and Caring Sciences, Uppsala University, Uppsala, Sweden). Reported symptoms and clinical findings in relation to serum cobalamin, folate, methylmalonic acid and total homocysteine among elderly Swedes: a population‐based study. J Intern Med 2003; 254: 343–352.

Objectives. The early stages of tissue B12 or folate deficiency often cause diagnostic problems. In this report, the levels of serum cobalamin, folate, methylmalonic acid (MMA) and total homocysteine (tHcy), and their relationships with clinical findings and reported symptoms in a representative random population sample are presented.

Design. Cohort study.

Setting. A general central Swedish population 70 years or older.

Subjects and methods.  A 20% random sample of persons 70 years or older in a defined geographical area were invited to a survey. A total of 235 (85%) persons responded, of whom 161 had no interfering diseases or medication. Blood specimens for serum cobalamin, folate, MMA and tHcy analyses were drawn.

Main outcome measures. Presence of anaemic, gastrointestinal, neurological and psychiatric symptoms, obtained by questionnaire, and vibration sense measurement and findings at a physical and Mini Mental State Examination.

Results. Among a large number of symptoms and clinical findings that traditionally have been linked to vitamin B12 or folate tissue deficiency, only changes in the tongue mucosa and mouth angle stomatitis turned out to be significantly associated with abnormal serum tHcy and serum folate levels. There were no relationships to serum cobalamin and serum MMA.

Conclusions. Changes in the oral mucosa were the only signs and symptoms found in this study, indicating that these may be the very early markers of metabolic defects. The traditional symptoms of vitamin deficiency may appear later in the course.
Antibodies to myelin basic protein, myelin oligodendrocytes peptides, α‐β‐crystallin, lymphocyte activation and cytokine production in patients with multiple sclerosis
Authors: Vojdani A. , Vojdani E. , Cooper E.
Published: September 2003

Abstract Vojdani A, Vojdani E, Cooper E (Laboratory of Comparative Neuroimmunology, David Geffen School of Medicine at UCLA, University of California Los Angeles, Los Angeles; Immunosciences Lab., Inc., Beverly Hills; and University of California Berkeley, Berkeley; CA, USA). Antibodies to myelin basic protein, myelin oligodendrocytes peptides, α‐β‐crystallin, lymphocyte activation and cytokine production in patients with multiple sclerosis. J Intern Med 2003; 254: 363–374.

Objective. To measure neurone‐specific humoral and cellular immune parameters in MRI‐positive patients with multiple sclerosis (MS).

Background. It has been postulated from animal models for MS and in situ evidence in MS patients that antibodies, activated T cells and proinflam‐ matory cytokines are involved in the destruction of myelin sheaths and loss of oligodendrocytes in active areas.

Subjects and methods. Blood samples were obtained from 20 healthy control subjects and 20 patients with abnormal MRI and clinical diagnosis of MS. Sera were tested for levels of IgG, IgM and IgA against myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG) peptides, and a small heat‐shock protein, α‐β‐crystallin. Lympho‐ cytes were isolated and cultured in the presence or absence of MBP, MOG peptides and α‐β‐crystallin, measured for stimulated T cells, cytokine production and compared with controls.

Results. Patients with MS showed the highest levels of IgG, IgM or IgA antibodies against one or all three tested antigens. Moreover, in the presence of MBP, MOG peptides or α‐β‐crystallin, a significant percent‐ age of lymphocytes from MS patients underwent blast transformation, which resulted in high levels of interferon γ (IFN‐γ), tumour necrosis factor alpha (TNF‐α) and tumour necrosis factor beta (TNF‐β) production. Sensitivity of these assays was 60–80% and specificity, 65–70%.

Conclusions. Detection of antibodies against MBP, MOG peptides, α‐β‐crystallin, lymphocyte stimula‐ tion and production of proinflammatory cytokines in response to these antigens could be used as surrogate markers for the confirmation of MS diagnosis. A combination of antibodies, lymphocyte activation or cytokine production with abnormal MRI may significantly increase the sensitivity and specificity of MS diagnosis.
High mobility group 1 B‐box mediates activation of human endothelium
Authors: Treutiger C. J. , Mullins G. E. , Johansson A.‐S. M. , Rouhiainen A. , Rauvala H. M. E. , Erlandsson‐Harris H. , Andersson U. , Yang H. , Tracey K. J. , Andersson J. , Palmblad J. E. W.
Published: September 2003

Abstract Treutiger CJ, Mullins GE, Johansson A‐SM, Rouhiainen A, Rauvala HME, Erlandsson‐Harris H, Andersson U, Yang H, Tracey KJ, Andersson J, Palmblad JEW (Center for Infectious Medicine; Center for Inflammation and Haematology Research; Karolinska Institute at Huddinge University Hospital, Stockholm, Sweden, Finnish Red Cross Blood Transfusion Service; Institute of Biotechnology, University of Helsinki; Helsinki, Finland, Center for Molecular Medicine, Karolinska Institutet, Stockholm, Sweden and North Shore‐Long Island Jewish Research Institute, NY, USA). High mobility group 1 B‐box mediates activation of human endothelium. J Intern Med 2003; 254: 375–385.

Objectives. Severe sepsis and septic shock is a consequence of a generalized inflammatory systemic response because of an invasive infection that may result in acute organ dysfunction. Mortality is high despite access to modern intensive care units. The nuclear DNA binding protein high mobility group 1 (HMGB1) protein has recently been suggested to act as a late mediator of septic shock via its function as a macrophage‐derived pro‐inflammatory cytokine (J Exp Med 2000; 192: 565, Science1999; 285: 248). We investigated the pro‐inflammatory activities of the A‐box and the B‐box of HMGB1 on human umbilical venular endothelial cells (HUVEC).

Design. The HUVEC obtained from healthy donors were used for experiments. Recombinant human full‐length HMGB1, A‐box and B‐box were cloned by polymerase chain reaction (PCR) amplification from a human brain quick‐clone cDNA. The activation of HUVEC was studied regarding (i) upregulation of adhesion molecules, (ii) the release of cytokines and chemokines, (iii) the adhesion of neutrophils to HUVEC, (iv) the activation of signalling transduction pathways and (v) the involvement of the receptor for advanced glycation end‐products (RAGE).

Results. The full‐length protein and the B‐box of HMGB1 dose‐dependently activate HUVEC to upregulate adhesion molecules such as ICAM‐1, VCAM‐1 and E‐selectin and to release IL‐8 and G‐CSF. The activation of HUVEC could be inhibited to 50% by antibodies directed towards the RAGE. HMGB1‐mediated HUVEC stimulation resulted in phosphorylation of the ELK‐1 signal transduction protein and a nuclear translocation of p65 plus c‐Rel, suggesting that HMGB1 signalling is regulated in endothelial cells through NF‐κB.

Conclusions. The HMGB1 acts as a potent pro‐inflammatory cytokine on HUVEC and the activity is mainly mediated through the B‐box of the protein. HMGB1 may be a key factor mediating part of the pro‐inflammatory response occurring in septic shock and severe inflammation.
Lesions of the Diagonal Band of Broca Enhance Drinking in the Rat
Authors: Sullivan M. J. , Cunningham J. T. , Mazzella D. , Allen A. M. , Nissen R. , Renaud L. P.
Published: September 2003

This study examined the role of the diagonal band of Broca (DBB) in drinking behaviour and vasopressin release. Adult male rats were anaesthetized (pentobarbital 50 mg/kg) and received DBB injections of either ibotenic acid (0.5 µl of 5 µg/µl) or vehicle (0.5 µl of phosphate‐buffered saline). Although baseline drinking and urine output were not affected, drinking to 30% polyethylene glycol (MW 8000; 1 ml/100 g s.c.) and angiotensin II (0, 1.5 and 3.0 mg/kg s.c.) were significantly increased in ibotenic acid in phosphate‐buffered saline (DBBX) rats. Drinking to hypertonic saline (0.9, 4 and 6%; 1 ml/100 g), and water deprivation were not significantly affected. DBBX rats had significantly lower basal heart rates than controls but the cardiovascular responses to infusions of angiotensin II (100 ng/kg/min i.v. for 45 min) were not affected. DBBX rats had significantly higher basal vasopressin, but angiotensin‐stimulated vasopressin release was not significantly different. Although the DBB is not involved in basal water intake, it is involved in dipsogenic responses to hypovolemic stimuli and possibly basal autonomic function and basal vasopressin release.
Corticotropin‐Releasing Factor Type‐1 Receptor mRNA is Not Induced in Mouse Hypothalamus By Either Stress or Osmotic Stimulation
Authors: Imaki T. , Katsumata H. , Konishi S.‐I. , Kasagi Y. , Minami S.
Published: September 2003

In rats, acute stress substantially increases corticotropin‐releasing factor (CRF) type 1 receptor (CRFR‐1) mRNA expression in the paraventricular nucleus (PVN) and osmotic stimulation induces both CRF and CRFR‐1 mRNA in magnocellular PVN and supraoptic nucleus (SON). However, these phenomena have not been analysed in other species. We compared CRF and CRFR‐1 expression in rat and mouse hypothalamus. Male C57BL/6 mice and Wistar rats were exposed to acute restraint stress for 3 h, or to hypertonic saline ingestion for 7 days. Restraint stress increased CRF and c‐fos mRNA expression in both rat and mouse PVN. CRFR‐1 mRNA was barely detectable in controls, whereas restraint stress substantially increased CRFR‐1 mRNA in rat PVN, but not in mouse. Hypertonic saline ingestion induced CRF mRNA in magnocellular PVN and SON of the rat, but did not alter CRF mRNA levels in mouse hypothalamus. CRFR‐1 mRNA was also induced in magnocellular PVN and SON of the rat in response to osmotic stimulation, but not in mouse. Immunohistochemistry demonstrated that CRFR‐1‐like immunoreactivity (ir) was distributed within parvocellular and magnocellular PVN of mouse and rat. CRFR‐1‐ir in rat PVN was increased by acute stress and osmotic stimulation. By contrast, these treatments did not alter CRFR‐1‐ir in mouse PVN. Combined immunohistochemistry and in situ hybridization revealed that CRFR‐1‐ir was most frequently colocalized to CRF in mouse PVN, whereas only a small percentage of oxytocin and vasopressin‐producing cells coexpressed CRFR‐1‐ir. These results indicate that (i) by contrast to rats, neither acute stress nor osmotic stimulation induces CRFR‐1 mRNA expression in the mouse PVN; (ii) osmotic stimulation does not alter CRF mRNA expression in parvocellular and magnocellular neurones of mouse PVN; and (iii) acute stress increases c‐fos and CRF mRNA to a similar degree in mouse and rat PVN. Thus, differences may exist between mouse and rat in the regulation of CRF and CRFR‐1 gene expression in hypothalamus following stress and osmotic stimulation.
Influence of Monoamines on Differentiating Gonadotropin‐Releasing Hormone Neurones in Foetal Mice
Authors: Pronina T. , Ugrumov M. , Calas A. , Seif I. , Tramu G.
Published: September 2003

This study evaluated the influence of monoamines, serotonin (5‐hydroxytryptamine, 5‐HT) and noradrenaline, on differentiating gonadotropin‐releasing hormone (GnRH)‐producing neurones in foetal mice. The differentiation and migration of GnRH neurones were compared in Tg8 mice (the knocked‐out gene encoding monoamine oxidase A) with increased levels of 5‐HT and noradrenaline and in C3H mice with normal metabolism of monoamines in C3H mice. To achieve this, immunocytochemistry for GnRH combined with quantitative and semiquantitative image analysis were employed. GnRH neurones in foetuses at the 18th embryonic day were detected in the forebrain along the trajectory of their migration from the olfactory bulbs to the hypothalamic retrochiasmatic region. The total number of GnRH neurones in the forebrain in knockout mice was significantly lower compared to C3H mice, suggesting an inhibiting influence of monoamines on the proliferation of precursor cells. The fraction of GnRH neurones in the caudal part of the trajectory of their migration in Tg8 mice exceeded significantly those in C3H foetuses, whereas there was a reverse in the rostral part of the trajectory. These data suggest that an excess of 5‐HT and noradrenaline served to accelerate the GnRH neurone migration in Tg8 mice. Moreover, an excess of 5‐HT and noradrenaline provided a minor effect on the area and optical density of GnRH neurones (i.e. on GnRH neurone differentiation). Thus, an excess of 5‐HT and noradrenaline appears to inhibit the proliferation of the precursor cells of GnRH neurones and stimulates the GnRH neurone migration to the place of their final location in the septo‐preoptic region.
Histamine and Prostaglandin Interaction in Regulation of Oxytocin and Vasopressin Secretion
Authors: Knigge U. , Kjær A. , Kristoffersen U. , Madsen K. , Toftegaard C. , Jørgensen H. , Warberg J.
Published: September 2003

Prostaglandins and histamine in the hypothalamus are involved in the regulation of oxytocin and vasopressin secretion, and appear to be involved in the mediation of pituitary hormone responses to immunochallenges. Therefore, we investigated in conscious male rats: (i) whether blockade of H1 or H2 receptors affected the oxytocin and vasopressin responses to prostaglandins and (ii) whether blockade of prostaglandin synthesis affected the oxytocin and vasopressin responses to histamine or to Escherichia coli lipopolysaccharide (LPS), in order to determine any interaction between prostaglandins and histamine in the hypothalamus. Oxytocin secretion was dose‐dependently stimulated by intracerebroventricular infusion of 1 or 5 µg of PGE1, PGE2 or PGF2α, with PGE2 being the most potent of the compounds used. Prior central infusion of the H1 receptor antagonist mepyramine or the H2 receptor antagonist cimetidine significantly inhibited the oxytocin response to all three prostaglandins by approximately 50%. Vasopressin secretion was increased by PGE1 but not by PGE2 or PGF2α. The stimulatory effect of PGE1 was almost annihilated by prior administration of mepyramine or cimetidine. Central infusion of histamine or immunochallenge with LPS administered intraperitoneally increased oxytocin and vasopressin secretion four‐ and two‐fold, respectively. Pretreatment with systemic injection of the prostaglandin synthesis inhibitor indomethacin dose‐dependently reduced the oxytocin response and prevented the vasopressin response to histamine or LPS. We conclude that histamine and PGE1, PGE2 or PGF2α interact in the regulation of oxytocin secretion, whereas histamine and only PGE1 interact in the regulation of vasopressin secretion. Furthermore, histamine as well as LPS may affect oxytocin and vasopressin neurones via activation of prostaglandins, probably in the hypothalamic supraoptic nucleus.