bluuebunny/crossref_metadata_2025 huggingface dataset:
Eruptive pruritic papular porokeratosis: A pruritic variant of porokeratosis
Authors: KANEKURA Takuro , YOSHII Noriko
Published: October 2006

Porokeratosis is a chronic skin disorder characterized clinically by the presence of crater‐like patches with an elevated thick keratotic border and central atrophy. Histology reveals cornoid lamellae. While porokeratosis is practically asymptomatic, a pruritic variant has been reported. We recently encountered an 82‐year‐old man with pruritic porokeratosis. He presented with erythematous papules and intensively itchy patches on his lower limbs that had been present for 6 months. Histopathological examination revealed the characteristic cornoid lamellae. We describe this case in detail and provide a review of the published work.
Hyperkeratotic lichenified skin lesion of gluteal region
Authors: NIIYAMA Shiro , SAKURAI Syugo , KATSUOKA Kensei
Published: October 2006

Elderly people may develop typical skin lesions on the superior part and both sides of the anal cleft. Although such skin changes have recently been reported as anosacral cutaneous amyloidosis in primary cutaneous amyloidosis, in our histopathological examinations no amyloid deposition has been detected at the sites of these lesions. Because hyperkeratosis, acanthosis and follicular plugging were seen in all of the cases histopathologically, we think that these skin manifestations are essentially a type of keratinization abnormality instead. We consider this disease to be a type of senile skin change that develops in response to prolonged mechanical irritation.
Functional analysis of the Alternaria brassicicola non‐ribosomal peptide synthetase gene AbNPS2 reveals a role in conidial cell wall construction
Authors: KIM KWANG‐HYUNG , CHO YANGRAE , LA ROTA MAURICIO , CRAMER ROBERT A. , LAWRENCE CHRISTOPHER B.
Published: October 2006

Alternaria brassicicola is a necrotrophic pathogen causing black spot disease on virtually all cultivated Brassica crops worldwide. In many plant pathosystems fungal secondary metabolites derived from non‐ribosomal peptide synthetases (NPSs) are phytotoxic virulence factors or are antibiotics thought to be important for niche competition with other micro‐organisms. However, many of the functions of NPS genes and their products are largely unknown. In this study, we investigated the function of one of the A. brassicicola NPS genes, AbNPS2. The predicted amino acid sequence of AbNPS2 showed high sequence similarity with A. brassicae, AbrePsy1, Cochliobolus heterostrophus, NPS4 and a Stagonospora nodorum NPS. The AbNPS2 open reading frame was predicted to be 22 kb in length and encodes a large protein (7195 amino acids) showing typical NPS modular organization. Gene expression analysis of AbNPS2 in wild‐type fungus indicated that it is expressed almost exclusively in conidia and conidiophores, broadly in the reproductive developmental phase. AbNPS2 gene disruption mutants showed abnormal spore cell wall morphology and a decreased hydrophobicity phenotype. Conidia of abnps2 mutants displayed an aberrantly inflated cell wall and an increase in lipid bodies compared with wild‐type. Further phenotypic analyses of abnps2 mutants showed decreased spore germination rates both in vitro and in vivo, and a marked reduction in sporulation in vivo compared with wild‐type fungus. Moreover, virulence tests on Brassicas with abnps2 mutants revealed a significant reduction in lesion size compared with wild‐type but only when aged spores were used in experiments. Collectively, these results indicate that AbNPS2 plays an important role in development and virulence.
Identification of transcripts modulated by ETV6 expression
Authors: Boily Gino , Larose Josiane , Langlois Sylvie , Sinnett Daniel
Published: October 2006

Deletions at chromosome 12p12‐13 are observed in 26–47% of childhood pre‐B acute lymphoblastic leukaemia (ALL) cases, suggesting the presence of a tumour suppressor gene (TSG). Accumulating genetic and functional evidence points to ETV6 as being the most probable TSG targeted by the deletions. ETV6 is a ubiquitously expressed transcription factor of the ETS family with very few known targets. To understand its function and to elucidate the impact of its absence in leukaemia, we conducted a study to identify targeted genes. Following the induction of ETV6 expression, global expression was evaluated at different time points. We identified 87 modulated genes, of which 10 (AKR1C1, AKR1C3, IL18, LUM, PHLDA1, PTGER4, PTGS2, SPHK1, TP53 and VEGF) were validated by real‐time quantitative reverse transcription‐polymerase chain reaction. To assess the significance of the validated candidate genes in leukaemia, their expression patterns were determined, as well as that of ETV6, in pre‐B ALL patients. The expression of IL18, LUM, PTGER4, SPHK1 and TP53 was significantly correlated with that of ETV6, further suggesting that ETV6 could regulate the expression of these genes in leukaemia. This work constitutes another step towards the understanding of the functions of ETV6 and the impact of its inactivation in childhood leukaemia.
Factors determining length distribution and abundance of the European eel, Anguilla anguilla, in the River Mondego (Portugal)
Authors: DOMINGOS ISABEL , COSTA JOSÉ LINO , COSTA MARIA JOSÉ
Published: October 2006

1. The distribution and abundance of the European eel, Anguilla anguilla, were investigated in the Mondego River, a strongly modified river in central Portugal. Nine freshwater sites, located below the first impassable obstacle to eel migration, were surveyed for 2 years. The study was conducted monthly during the first year and seasonally during the second year. Fish were caught by electric fishing and, physical and biotic variables potentially influencing eel length distribution and abundance were determined at each sampling session.

2. Canonical correspondence analysis (CCA) was used to explain the relationships between abiotic and biotic habitat variables, and eel length distribution. Spatial variation was strongly associated with distance from the sea, number of obstacles, river width and percentage of instream cover. Depth, river flow, water temperature and cannibalism had weaker effects on the longitudinal distribution, but contributed as explanatory variables for the model. The influence of spatial variables outweighed the importance of temporal variables as predictors in this model.

3. Length distribution changed in space from a dominance of small eels (<100 mm) closer to the sea to larger eels (≥250 mm) in the upper reaches of the river. The smallest length class (eels <100 mm) was strongly associated with wider stretches where instream cover, mainly composed of aquatic submerged macrophytes, was abundant (>75% area coverage). On the contrary, larger eels (≥250 mm) preferred deeper stretches with stronger river flow and less instream cover.

4. The effect of biotic interactions with conspecifics as well as with the non‐native red swamp crayfish Procambarus clarkii, in sites with poor instream cover, modified this general pattern of distribution leading to fewer eels <100 mm at some sites closer to the sea. Therefore, despite the ubiquity and plasticity shown by this species, there were clear ontogenetic variations in habitat use such that the first two canonical axes of a CCA accounted for 58.4% of the spatial variability in size structure.
Population ecology, vertical migration and feeding of the Ponto‐Caspian invader Hemimysis anomala in a gravel‐pit lake connected to the River Rhine
Authors: BORCHERDING JOST , MURAWSKI SIMONE , ARNDT HARTMUT
Published: October 2006

1. The Ponto‐Caspian invader, Hemimysis anomala, was recently found in large numbers in a gravel‐pit lake connected to the Lower Rhine. Mysids were sampled with Perspex traps between September 2002 and April 2003 to study the population dynamics, vertical migration and feeding.

2. The abundance (as catch per unit effort, CPUE) of H. anomala declined from 270 individuals (ind.) trap−1 (4 h)−1 in December to below 4 ind. trap−1 (4 h)−1 in April. Average lengths ranged from 4.9 mm in autumn to 9.9 mm in March. The length–weight relationship of virgin females changed throughout the sampling period, from a size‐corrected wet weight of 5.1 mg in September to 16.6 mg in April for a female of 7.8 mm.

3. Successive monthly samples taken over 24 h revealed that H. anomala preferred the surface at twilight and night. During dawn the mysids migrated to the middle and bottom layers and were hardly found during day. Diel vertical migration clearly depends on the proximate factor light.

4. Stomach analyses revealed that larger H. anomala preferred zooplankton, whereas small individuals fed more on phytoplankton. The seasonal comparison showed an increasing percentage of zooplankton with increasing length. The proportion of zooplankton in the stomachs of large H. anomala individuals was highest during night and lowest during day.

5. The results suggest that H. anomala may become an important link between primary/secondary production and higher trophic levels in the food web of its new environments at the Lower Rhine.
Genomic and transcriptional analysis of protein heterogeneity of the honeybee venom allergen Api m 6
Authors: Peiren N. , De Graaf D. C. , Evans J. D. , Jacobs F. J.
Published: October 2006

Several components of honeybee venom are known to cause allergenic responses in humans and other vertebrates. One such component, the minor allergen Api m 6, has been known to show amino acid variation but the genetic mechanism for this variation is unknown. Here we show that Api m 6 is derived from a single locus, and that substantial protein‐level variation has a simple genome‐level cause, without the need to invoke multiple loci or alternatively spliced exons. Api m 6 sits near a misassembled section of the honeybee genome sequence, and we propose that a substantial number of indels at and near Api m 6 might be the root cause of this misassembly. We suggest that genes such as Api m 6 with coding‐region or untranslated region indels might have had a strong effect on the assembly of this draft of the honeybee genome.
A deficit of detoxification enzymes: pesticide sensitivity and environmental response in the honeybee
Authors: Claudianos C. , Ranson H. , Johnson R. M. , Biswas S. , Schuler M. A. , Berenbaum M. R. , Feyereisen R. , Oakeshott J. G.
Published: October 2006

The honeybee genome has substantially fewer protein coding genes (≈ 11 000 genes) than Drosophila melanogaster (≈ 13 500) and Anopheles gambiae (≈ 14 000). Some of the most marked differences occur in three superfamilies encoding xenobiotic detoxifying enzymes. Specifically there are only about half as many glutathione‐S‐transferases (GSTs), cytochrome P450 monooxygenases (P450s) and carboxyl/cholinesterases (CCEs) in the honeybee. This includes 10‐fold or greater shortfalls in the numbers of Delta and Epsilon GSTs and CYP4 P450s, members of which clades have been recurrently associated with insecticide resistance in other species. These shortfalls may contribute to the sensitivity of the honeybee to insecticides. On the other hand there are some recent radiations in CYP6, CYP9 and certain CCE clades in A. mellifera that could be associated with the evolution of the hormonal and chemosensory processes underpinning its highly organized eusociality.
Nuclear receptors of the honey bee: annotation and expression in the adult brain
Authors: Velarde Rodrigo A. , Robinson Gene E. , Fahrbach Susan E.
Published: October 2006

The Drosophila genome encodes 18 canonical nuclear receptors. All of the Drosophila nuclear receptors are here shown to be present in the genome of the honey bee (Apis mellifera). Given that the time since divergence of the Drosophila and Apis lineages is measured in hundreds of millions of years, the identification of matched orthologous nuclear receptors in the two genomes reveals the fundamental set of nuclear receptors required to ‘make’ an endopterygote insect. The single novelty is the presence in the A. mellifera genome of a third insect gene similar to vertebrate photoreceptor‐specific nuclear receptor (PNR). Phylogenetic analysis indicates that this novel gene, which we have named AmPNR‐like, is a new member of the NR2 subfamily not found in the Drosophila or human genomes. This gene is expressed in the developing compound eye of the honey bee. Like their vertebrate counterparts, arthropod nuclear receptors play key roles in embryonic and postembryonic development. Studies in Drosophila have focused primarily on the role of these transcription factors in embryogenesis and metamorphosis. Examination of an expressed sequence tag library developed from the adult bee brain and analysis of transcript expression in brain using in situ hybridization and quantitative RT‐PCR revealed that several members of the nuclear receptor family (AmSVP, AmUSP, AmERR, AmHr46, AmFtz‐F1, and AmHnf‐4) are expressed in the brain of the adult bee. Further analysis of the expression of AmUSP and AmSVP in the mushroom bodies, the major insect brain centre for learning and memory, revealed changes in transcript abundance and, in the case of AmUSP, changes in transcript localization, during the development of foraging behaviour in the adult. Study of the honey bee therefore provides a model for understanding nuclear receptor function in the adult brain.
Expression of insulin pathway genes during the period of caste determination in the honey bee,Apis mellifera
Authors: Wheeler D. E. , Buck N. , Evans J. D.
Published: October 2006

Female honeybees have two castes, queens and workers. Developmental fate is determined by larval diet. Coding sequences made available through the Honey Bee Genome Sequencing Consortium allow for a pathway‐based approach to understanding caste determination. We examined the expression of several genes of the insulin signalling pathway, which is central to regulation of growth based on nutrition. We found one insulin‐like peptide expressed at very high levels in queen but not worker larvae. Also, the gene for an insulin receptor was expressed at higher levels in queen larvae during the 2nd larval instar. These results demonstrate that the insulin pathway is a compelling candidate for pursing the relationship between diet and downstream signals involved in caste determination and differentiation.
Comparative analysis of serine protease‐related genes in the honey bee genome: possible involvement in embryonic development and innate immunity
Authors: Zou Z. , Lopez Dawn L. , Kanost Michael R. , Evans Jay D. , Jiang Haobo
Published: October 2006

We have identified 44 serine protease (SP) and 13 serine protease homolog (SPH) genes in the genome ofApis mellifera. Most of these genes encode putative secreted proteins, but four SPs and three SPHs may associate with the plasma membrane via a transmembrane region. Clip domains represent the most abundant non‐catalytic structural units in these SP‐like proteins −12 SPs and six SPHs contain at least one clip domain. Some of the family members contain other modules for protein–protein interactions, including disulphide‐stabilized structures (LDLrA, SRCR, frizzled, kringle, Sushi, Wonton and Pan/apple), carbohydrate‐recognition domains (C‐type lectin and chitin‐binding), and other modules (such as zinc finger, CUB, coiled coil and Sina). Comparison of the sequences with those fromDrosophilaled to a proposed SP pathway for establishing the dorsoventral axis of honey bee embryos. Multiple sequence alignments revealed evolutionary relationships of honey bee SPs and SPHs with those inDrosophila melanogaster,Anopheles gambiae, andManduca sexta. We identified homologs ofD. melanogasterpersephone,M. sextaHP14, PAP‐1 and SPH‐1.A. melliferagenome includes at least five genes for potential SP inhibitors (serpin‐1 through ‐5) and three genes of SP putative substrates (prophenoloxidase, spätzle‐1 and spätzle‐2). Quantitative RT‐PCR analyses showed an elevation in the mRNA levels of SP2, SP3, SP9, SP10, SPH41, SPH42, SP49, serpin‐2, serpin‐4, serpin‐5, and spätzle‐2 in adults after a microbial challenge. The SP41 and SP6 transcripts significantly increased after an injection ofPaenibacillus larva, but there was no such increase after injection of saline orEscherichia coli. mRNA levels of most SPs and serpins significantly increased by 48 h after the pathogen infection in 1st instar larvae. On the contrary, SP1, SP3, SP19 and serpin‐5 transcript levels reduced. These results, taken together, provide a framework for designing experimental studies of the roles of SPs and related proteins in embryonic development and immune responses ofA. mellifera.
Characteristics of the nuclear (18S, 5.8S, 28S and 5S) and mitochondrial (12S and 16S) rRNA genes ofApis mellifera(Insecta: Hymenoptera): structure, organization, and retrotransposable elements
Authors: Gillespie J. J. , Johnston J. S. , Cannone J. J. , Gutell R. R.
Published: October 2006

As an accompanying manuscript to the release of the honey bee genome, we report the entire sequence of the nuclear (18S, 5.8S, 28S and 5S) and mitochondrial (12S and 16S) ribosomal RNA (rRNA)‐encoding gene sequences (rDNA) and related internally and externally transcribed spacer regions ofApis mellifera(Insecta: Hymenoptera: Apocrita). Additionally, we predict secondary structures for the mature rRNA molecules based on comparative sequence analyses with other arthropod taxa and reference to recently published crystal structures of the ribosome. In general, the structures of honey bee rRNAs are in agreement with previously predicted rRNA models from other arthropods in core regions of the rRNA, with little additional expansion in non‐conserved regions. Our multiple sequence alignments are made available on several public databases and provide a preliminary establishment of a global structural model of all rRNAs from the insects. Additionally, we provide conserved stretches of sequences flanking the rDNA cistrons that comprise the externally transcribed spacer regions (ETS) and part of the intergenic spacer region (IGS), including several repetitive motifs. Finally, we report the occurrence of retrotransposition in the nuclear large subunit rDNA, as R2 elements are present in the usual insertion points found in other arthropods. Interestingly, functional R1 elements usually present in the genomes of insects were not detected in the honey bee rRNA genes. The reverse transcriptase products of the R2 elements are deduced from their putative open reading frames and structurally aligned with those from another hymenopteran insect, the jewel waspNasonia(Pteromalidae). Stretches of conserved amino acids shared betweenApisandNasoniaare illustrated and serve as potential sites for primer design, as target amplicons within these R2 elements may serve as novel phylogenetic markers for Hymenoptera. Given the impending completion of the sequencing of theNasoniagenome, we expect our report eventually to shed light on the evolution of the hymenopteran genome within higher insects, particularly regarding the relative maintenance of conserved rDNA genes, related variable spacer regions and retrotransposable elements.
Genome sequences of the honey bee pathogens Paenibacillus larvae and Ascosphaera apis
Authors: Qin X. , Evans J. D. , Aronstein K. A. , Murray K. D. , Weinstock G. M.
Published: October 2006

Genome sequences offer a broad view of host–pathogen interactions at the systems biology level. With the completion of the sequence of the honey bee, interest in the relevant pathogens is heightened. Here we report the genome sequences of two of the major pathogens of honey bees, the bacterium Paenibacillus larvae (causative agent for American foulbrood disease) and the fungus Ascosphaera apis. (causative agent for chalkbrood disease). Ongoing efforts to characterize the genomes of these species can be used to understand and mitigate the effects of two important pathogens, and will provide a contrast with pathogenic, benign and freeliving relatives.
Genes of the antioxidant system of the honey bee: annotation and phylogeny
Authors: Corona M. , Robinson G. E.
Published: October 2006

Antioxidant enzymes perform a variety of vital functions including the reduction of life‐shortening oxidative damage. We used the honey bee genome sequence to identify the major components of the honey bee antioxidant system. A comparative analysis of honey bee with Drosophila melanogaster and Anopheles gambiae shows that although the basic components of the antioxidant system are conserved, there are important species differences in the number of paralogs. These include the duplication of thioredoxin reductase and the expansion of the thioredoxin family in fly; lack of expansion of the Theta, Delta and Omega GST classes in bee and no expansion of the Sigma class in dipteran species. The differential expansion of antioxidant gene families among honey bees and dipteran species might reflect the marked differences in life history and ecological niches between social and solitary species.
Oxidatively Derived Volatile Compounds in Microencapsulated Fish Oil Monitored by Solid‐phase Microextraction (SPME)
Authors: Jónsdóttir Rósa , Bragadóttir Margrét , Arnarson GudmundurÖRN
Published: October 2006

The stability of microencapsulated fish oil was studied during storage at 4 °C for up to 20 wk. Different coating mixtures consisting of gelatin or caseinate in blends with carbohydrates (sucrose, lactose, maltodextrin) were investigated. Oxidative stability of the microencapsulated fish oil was monitored by analysis of volatile compounds using gas chromatography olfactometry (GC‐O) or GC flame ionization (GC‐FID) (SPME‐HS‐GC/O or GC/ FID and HS‐GC/MS), Oxipres test, thiobarbituric acid‐reactive substances (TBARS), and sensory analysis. Coating mixture of caseinate and lactose showed slightly better stability than the sucrose and maltodextrin caseinate mixtures. Combination of fish gelatin and maltodextrin did not show as good oxidative stability as the coating blend of caseinate, lactose, and lecithin. Hexanal, 2‐nonenal and 2,4‐decadienals were selected as quality indicators to monitor the lipid oxidation during storage of the samples. SPME‐GC‐O analysis of these indicators showed that they were representative for the oxidation occurring in the microencapsulated fish oil. SPME‐GC‐FID analysis was sensitive enough to detect oxidative changes during storage. Oxidative stability test, TBARS results, and sensory analysis were in agreement with the SPME, indicating that SPME (polydimethylsiloxane/divinylbenzene [PDMS/ DVB] fiber) can be a useful tool for rapid analysis of lipid oxidation in microencapsulated fish oil.
Fracture Analysis of Alginate Gels
Authors: Zhang Junhua , Daubert Christopher R. , Foegeding E. Allen
Published: October 2006

The fracture properties of alginate gels were investigated using torsion and compression. The gel fracture stress correlated with Ca2+ and alginate concentration, whereas the fracture strain was insensitive to composition. Considering the relationship of fracture stress with gel network crosslink density and the energy to break covalent and noncovalent bonds, the fracture of alginate gels is hypothesized to result from the disruption of junction zones. Consequently, the fracture stress was the stress required to overcome electrostatic forces that formed junction zones. The fracture stress‐strain relationship for alginate gels can be described by the Blatz, Sharda, adn Tschoegl (BST) equation, suggesting that for a given gel, the fracture strain can be predicted based on fracture stress, small‐strain shear modulus, and a fitted parameter describing nonlinearity of the gel. In addition, the fracture properties were affected by deformation rate. The influence of deformation rate on fracture was ascribed to structural changes among the alginate junction zones.
Influence of Surface Topography on the Effectiveness of Pulsed Light Treatment for the Inactivation of Listeria innocua on Stainless‐steel Surfaces
Authors: Woodling Sarah E. , Moraru Carmen I.
Published: October 2006

Pulsed light (PL) treatment has been proven effective for killing a wide variety of microorganisms on foods and food contact materials. However, there is concern regarding how shading may impact the effectiveness of PL when applied to imperfect surfaces. The main objective of this work was to examine how surface properties, particularly topography, influence the microbicidal effect of PL. Four types of stainless‐steel surfaces were inoculated with Listeria innocua and treated with up to 12 pulses of light. The highest level of inactivation achieved was about a 4‐log reduction. Initially, an increase in inactivation with increasing treatment intensity was observed, but the inactivation curves tailed off above 3 light pulses. The differences in inactivation levels among the 4 finishes at specific treatment levels were rather insignificant, but some interesting trends were observed. At low treatment levels, inactivation on the smoothest finish was slightly lower than for the other surfaces, due to clustering of the cells on the highly hydrophobic smooth surface and to its reflective nature. For the roughest surface, scanning electron microscopy (SEM) imaging confirmed the preferential location of the cells inside surface features, which also promoted a relatively uniform distribution of the cells across the surface. This counter balanced to some extent of the shading effects, and as a result inactivation on the roughest surface was comparable to inactivation on the smoother surfaces. These results demonstrate that PL can be effective on both smooth and rough surfaces, but also indicate a complex effect of various surface properties on inactivation.
Headspace Gas Chromatography‐Mass Spectrometry and Electronic Nose Analysis of Volatile Compounds in Canned Alaska Pink Salmon Having Various Grades of Watermarking
Authors: Oliveira Alexandra C.M. , Crapo Charles A. , Himelbloom Brian , Vorholt Carey , Hoffert Jennifer
Published: October 2006

Volatiles in canned pink salmon, produced from different degrees of skin watermarked raw material and stored for 2 and 9 mo, were characterized and compared using static headspace gas chromatography analysis coupled to a mass spectrometer (SHGCMS). Sulfur‐containing compounds comprised 30% to 50% of the total volatiles and tended to decrease with increasing degrees of skin watermarking, and dimethyl sulfide was the most abundant compound of this class of molecules. A few alcohols, aldehydes, ketones, and furans were also identified. Forward stepwise general discriminant analysis (FSGDA) was used to investigate prediction models based on degree of skin watermarking. The 2‐ and 9‐mo models using SHGCMS showed 92.5% and 93.75% correct classifications, respectively. The ability of the Cyranose 320, a hand‐held electronic nose (EN), to differentiate these grades of watermarking in the canned samples was also tested. EN analysis using FSGDA resulted in models with 90% and 92.5% correct classifications for the 2‐ and 9‐mo samples, respectively. Overall, results indicate that the watermarking grades studied are not readily distinguishable from each other by either method of analysis.
Influence of Flavor Variability in Skim Milk Powder on Consumer Acceptability of Ingredient Applications
Authors: Caudle Alissa D. , Yoon Youngmo , Drake MaryAnne
Published: October 2006

Flavor variability in skim milk powder (SMP) has been documented by descriptive sensory analysis and instrumental analysis. However, research has not addressed how or if SMP flavor variability impacts consumer acceptance of ingredient applications. Twenty reconstituted low‐heat SMPs were screened using a previously established defined sensory language. Two SMPs free of off‐flavors (flavor similar to fresh fluid skim milk) and 2 SMPs exhibiting off‐flavors (exhibiting flavors not found in fluid skim milk) were selected for further study. Volatile compounds were determined using solid phase microextraction (SPME) with gas chromatography‐mass spectrometry (GC‐MS). Each SMP was subsequently directly reconstituted or incorporated into standard formulas for vanilla ice cream, strawberry yogurt, hot cocoa mix, and white chocolate bars. Consumer acceptance testing was conducted. Descriptive panelists documented sweet aromatic and cooked flavors in the fresh‐flavored SMP and these flavors as well as fatty/fryer oil and animal‐like flavors in the off‐flavored SMP. Concurrently, instrumental volatile analysis revealed higher relative abundances of lipid oxidation compounds in the off‐flavored SMP. Consumer acceptance scores were lower for the off‐flavored SMP than for the fresh‐flavored SMP when evaluated directly reconstituted (P< 0.05) and for ingredient applications made using off‐flavored SMP (P< 0.05). Off‐flavors in SMP documented by descriptive analysis and trained panelists can carry through into ingredient applications with SMP and negatively affect consumer acceptability.
Formulation of Corn Zein Chewing Gum and Evaluation of Sensory Properties by the Time‐Intensity Method
Authors: McGowan Bridget A. , Padua Graciela W. , Lee Soo‐Yeun
Published: October 2006

This study was conducted to determine the feasibility of using corn zein as a natural gum base to be an alternative to the currently used synthetic gum base. The objectives were to (1) develop a corn zein chewing gum and (2) evaluate the taste, texture, and aroma qualities compared with synthetic gum base using a time‐intensity (T‐I) method. Four corn zein gum samples, each made with a different plasticizer (oleic acid, glycerin, propylene glycol, and 1 without any plasticizer), were included in the study along with 2 synthetic gum‐base samples. Nine panelists participated in the T‐I study. Thirteen attributes were evaluated. Maximum intensity, time to maximum intensity, and duration were parameters extracted from the T‐I curve. The synthetic gum‐base samples were rated higher for the maximum intensity of sweet taste and cinnamon aroma‐by‐mouth and lower for the maximum intensity of bitter taste. The propylene glycol corn zein sample was rated the highest for cinnamon aroma and the lowest for stale, rancid, and cheesy aromas. Out of the corn zein gums, the oleic acid sample was rated the lowest for the maximum intensity of hardness. It was also rated highest for the maximum intensity of bitter taste and the lowest for sweet taste. This study showed that it is feasible to use corn zein as a gum base, but future work is needed to develop an acceptable product for consumers.
Lipid Oxidations in Ordinary and Dark Muscles of Fish: Influences on Rancid Off‐odor Development and Color Darkening of Yellowtail Flesh During Ice Storage
Authors: Sohn Jeong‐Ho , Taki Yusuke , Ushio Hideki , Kohata Tomoko , Shioya Itaru , Ohshima Toshiaki
Published: October 2006

We investigated the role of lipid oxidation in the development of a rancid off‐odor in the early stage of ice storage in both ordinary and dark muscles of cultured yellowtail (Seriola quinqueradiata). The total lipid hydroperoxide content and thiobarbituric acid‐reactive substances (TBARS) of the dark muscle were significantly higher (P < 0.01) than those of the ordinary muscle during 2 d of ice storage. The increase in the metmyoglobin content of the dark muscle was accompanied by gradual darkening of the muscle color during ice storage. To distinguish the changes in odor characteristics of the muscles, sensory evaluation was carried out. Slight changes in the intensity of 7 different odors were noted in the ordinary muscles, and there was a significant increase in the overall, fishy, spoiled, and rancid smells of the dark muscle during 2 d of ice storage. No correlation was found between the total content of lipid hydroperoxide and the odor intensities in ordinary muscle; however, there was a significant correlation between the total lipid hydroperoxide content and rancid off‐odor and overall smell intensities in the dark muscle. The rate of lipid oxidation of the yellowtail dark muscle was significantly faster than that of the ordinary muscle. Lipid oxidation of the dark muscle was closely related to meat darkening and development of the rancid off‐odor during the early stage of ice storage.
Antioxidant Capacity of Fruit Extracts of Blackberry (Rubus sp.) Produced in Different Climatic Regions
Authors: Reyes‐Carmona Josefina , Yousef Gad G. , Martínez‐Peniche Ramon A. , Lila Mary Ann
Published: October 2006

A range of blackberry genotypes harvested in different seasons and regions in Mexico (Michoacan) and in the United States (Pacific Northwest) were collected to determine their antioxidant capacity using oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays. Total acidity, ascorbic acid, soluble solids, total phenols, and total anthocyanins, as well as the correlation between all these parameters, were determined for all treatments. Total acidity ranged from 4.22% in wild blackberry from Patzcuaro, Mexico, to 1.02% in ‘Evergreen’ from Woodburn, Oreg. These treatments were also the outliers in terms of ascorbic acid content. Cultivar ‘Brazos’ did not exhibit any significant differences in acidity or ascorbic acid as a consequence of the geographic origin or harvest season. The highest concentration of soluble solids was recorded for ‘Evergreen’ from Woodburn and relatively low soluble solids levels were recorded for all the Mexican treatments. Wild blackberry from Patzcuaro exhibited the highest values for ORAC, FRAP, total phenolic and anthocyanin content. Other relatively high antioxidant capacity values were detected for ‘Marion’ and ‘Evergreen’, both produced in Oregon. Different cultivars grown in the same region/season consistently showed differences in antioxidant capacity. There was little effect of harvest season on phenolic levels. We conclude that levels of total acidity, ascorbic acid, soluble solids, antioxidant capacity, and polyphenols mainly depended on the genotype and not on the climate or the season. ORAC and FRAP values were both highly correlated with each other, and with total phenols and anthocyanin content.
Characterisation of the Mouse Vasoactive Intestinal Peptide Receptor Type 2 Gene,Vipr2, and Identification of a Polymorphic LINE‐1‐like Sequence That Confers Altered Promoter Activity
Authors: Steel G. , Lutz E. M.
Published: October 2006

The VPAC2receptor is a seven transmembrane spanning G protein‐coupled receptor for two neuropeptides, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase‐activating polypeptide (PACAP). It has a distinct tissue‐specific, developmental and inducible expression that underlies an important neuroendocrine role. Here, we report the characterisation of the gene that encodes the mouse VPAC2receptor (Vipr2), localisation of the transcriptional start site and functional analysis of the promoter region. TheVipr2gene contains 12 introns within its protein‐coding region and spans 68.6 kb. Comparison of the 5′ untranslated region sequences for cloned 5′‐RACE products amplified from different tissues showed they all were contained within the same exon, with the longest extending 111 bp upstream of the ATG start site. Functional analysis of the 3.2‐kb 5′‐flanking region using sequentially deleted sequences cloned into a luciferase gene reporter vector revealed that this region is active as a promoter in mouse AtT20 D16:16 and rat GH4C1 cell lines. The core promoter is located within a 180‐bp GC‐rich region proximal to the ATG start codon and contains potential binding sites for Sp1 and AP2, but no TATA‐box. Further upstream, in two out of three mice strains examined, we have discovered a 496‐bp polymorphic DNA sequence that bears a significant identity to mouse LINE‐1 DNA. Comparison of the promoter activity between luciferase reporter gene constructs derived from the BALB/c (which contains this sequence) and C57BL/6J (which lacks this sequence)Vipr2promoter regions has shown three‐fold difference in luciferase gene activity when expressed in mouse AtT20 D16:16 and αT3‐1 cells, but not when expressed in the rat GH4C1 cells or in COS 7 cells. Our results suggest that the mouseVipr2gene may be differentially active in different mouse strains, depending on the presence of this LINE‐1‐like sequence in the promoter region.
Presence in the 'silent’terminus region of the Escherichia coli K12 chromosome of cryptic gene(s) encoding a new nitrate reductase
Authors: Bonnefoy V. , Burinl J.‐F. , Giordano G. , Pascal M.‐C. , Chippaux M.
Published: October 2006

A cosmid complementing narG mutants defective in nitrate reductase activity was isolated from a genomic library of Escherichia coli. The restriction map of the insert differed from that of the narGHI operon. The new enzyme, termed NarZ, required molybdenum for activity. The expression of narZ was not affected by the factors controlling narGHI. Insertion mutations indicated that the narZ locus covered about 8 kb of DNA; narZ is located at 32.5 U on the chromosome, in the cotransduction gap near the replication terminus. Southern blot experiments under stringent conditions using narGHI or narZ DNA as probes revealed a large extent of homology, with a small area of very high homology. We propose that narZ and narGHI have descended from a common ancestor by gene duplication.
Localization of a conserved epitope and an azurin‐like domain in the H.8 protein of pathogenic Neisseria
Authors: Kawula T. H. , Spinota S. M. , Klapper D. G. , Cannon J. G.
Published: October 2006

The pathogenic neisseriae, Neisseria gonorrhoeae and Neisseria meningitidis, possess an outer membrane protein, H.8, which contains a conserved monoclonal antibody (MAb)‐binding epitope in all strains tested. We have cloned and sequenced a meningococcal H.8 gene, and determined the characteristics of the predicted protein. The predicted signal peptide has features characteristic of a prokaryotic lipoprotein. The region at the N‐terminal end of the mature protein (39 amino acids) is primarily composed of alanine, glutamate and prollne residues arranged in imperfect repeats with the consensus sequence AAEAP. The epitope for H.8 MAb‐binding was localized to a 20‐amino acid sequence within this region. The remainder of the predicted amino acid sequence shows extensive homology to azurins, which are small blue copper‐binding proteins found in a limited number of species of pathogenic bacteria.
Identification, cloning and sequence analysis of the Bacillus sphaericus 1593 41.9 kD larvicidal toxin gene
Authors: Hindley J. , Berry C.
Published: October 2006

A number of strains of the widespread aerobic soil bacterium, Bacillus sphaericus, possess crystalline inclusions of a toxin lethal to a variety of insect (larvae) which are vectors of major tropical diseases. Partlal amino acid sequence data from one strain, B. sphaerlcus 2362 have permitted us to design oLigo‐nucleotide probes for identifying the toxin gene in the closely related B. sphaericus 1593. The gene was found to be contained within an Eco RI‐Hin dIII fragment and was cloned in its entirety in the bacterial plasmid pUC12. The DNA sequence was determined together with the upstream and downstream controlling elements, and a sequence of 370 amino acids was deduced for the toxin protein. This is the first reported sequence of a B. sphaericus toxin gene and will facilitate further work in characterizing the genes from other strains of different virulence and host range. The data do not support the suggestion that the toxin is derived by proteolysis of a protoxin precursor.
Distinct control sites located upstream from the levansucrase gene of Bacillus subtilis
Authors: Klier A. , Fouet. A. , Debarbouille. M. , Kunst F. , Rapoport G.
Published: October 2006

The sacR regulatory region, which modulates the expression of sacB, the structural gene for ievansucrase, was separated into two parts: an upstream region which carries a constitutive promoter and a downstream region which carries a palindromic structure. Three types of fusions were constructed in which the aphA3 gene coding for kanamycin resistance of Streptococcus faecalis was placed downstream from different deleted sacR regions. Other fusions were constructed by inserting a promoter from phage SPO1 upstream from the sacB gene and part of the sacA region. A third kind of fusion was constructed in which the palindromic structure was flanked by a heterologous promoter and a heterologous structural gene. After introduction of these fusions into the chromosomal DNA of mutants affected in sacB regulation, it was possible to reveal different targets for the regulatory genes sacU, sacQ and sacS: the sacU and sacQ genes act on a region located near or just upstream from the promoter, and the sacS gene, which is involved in the Induction process, acts on the palindromic structure.
Common mechanism controlling phase and antigenic variation in pathogenic Neisseriae
Authors: Stern A. , Meyer T. F.
Published: October 2006

The expression of the Neisseria gonorrhoeae opacity protein (Op, protein II), a major antigenic determinant of the outer membrane, is subject to frequent phase transitions. At least nine expression loci (opaE) are involved in the production of a large number of serologically distinct Op types. Using opa‐specific oligonucleotides as probes in genomic blots, we detect Op‐related gene sequences (opr) in N. meninglitidis as well as in N. lactamica. DNA sequence analysis of such opr genes derived from N. meninglitidis reveals distinct regions of homology with gonococcal opa E genes. As shown in the immunoblot, the proteins encoded by ops and opr are serologically related. Like the opa E genes, the 5′‐coding sequences of the opr genes include a repetitive sequence composed of pentameric CTCTT units. The number of these coding repeat (CR) units is variable. This finding, together with the observation that all opr genes are constitutively transcribed, regardless of the status of protein production, suggests a translational control mechanism identical to that of the opa genes in gonococci. The related structures and control mechanisms of opa and opr genes imply a general significance of their gene products for the pathogenic character of the investigated Neisseria species.
An 18 amino acid amphiphilic helix forms the membrane‐anchoring domain of the Escherichia coli penicillin‐binding protein 5
Authors: Jackson M. E. , Pratt J. M.
Published: October 2006

Small (10 residue) C‐terminal deletions of PBP5 cause release of this Inner membrane protein into the periplasm, indicating disruption of the membrane binding domain. To define the extent of the membrane anchoring domain, oligonucleotide‐directed mutagenesis was used to introduce both single amino acid changes and novel restriction sites into the DN A, allowing subsequent construction of precise internal deletions. The 10 C‐terminal amino acid residues possess very weak membrane anchoring potential. By extending the sequence to 18 residues membrane binding equivalent to that of authentic PBP5 was achieved. A proline substitution in this region, breaking a potential α‐helix, also disrupts the membrane binding domain. These results are discussed with respect to the amphi‐philicity of the C‐terminal sequence when arranged in an α‐helix.
Cloning and nucleotide sequence of the thrB gene from the cyanobacterium Calothrix PCC 7601
Authors: Parsot C. , Mazel D.
Published: October 2006

The cyanobacterium Calothrix PCC 7601 thrB gene, encoding homoserine kinase (EC 2.7.1.39), was cloned via complementation of an Escherichia coli threonine auxotroph, and its nucleotide sequence was determined. The comparison of the homoserine kinase amino acid sequences from Calothrix PCC 7601, E. coli K12 and Bacillus subtilis 168 indicates a closer relationship between cyanobacteria and bacillaceae than between cyanobacteria and enterobacteriaceae. Sequence analysis of the 5′ and 3′ flanking regions of the Calothrix thrB gene revealed the existence of a 169‐codon‐long open reading frame downstream from thrB: this sequence may be the second gene of a Calothrix thr operon. Two types of tandemly repeated sequences, sharing similarities with other prokaryotic transcriptional regulatory elements, were detected in the region upstream from the thrB gene.
Exotoxin A of Pseudomonas aeruginosa: evidence that Domain I functions In receptor binding
Authors: Guidi‐Rontani C. , Collier R. J.
Published: October 2006

We have constructed defined deletions in the structural gene of Pseudomonas aeruginosa exotoxin A (ETA) In order to probe the function of Domain t of this protein. Three forms of the gene containing specific deletions were expressed in a strain of Escherichia coli K12 with lesions in the htpR and ion genes; extracts containing the gene products were tested for ADP‐ribosylation activity, cytotoxicity, and ability to protect sensitive cells from the cytotoxic action of authentic ETA. Two of the mutant ETAs gave concentration‐dependent protection against authentic ETA, and protection correlated with the presence of the bulk of Domain I. The results support the notion that Domain I functions in binding the toxin to specific cell‐surface receptors.
DNA sequence analysis of three missense mutations affecting colicin E3 bactericidal activity
Authors: Escuyer V. , Mock M.
Published: October 2006

Summary

We have determined the nucleotide sequence changes caused by three missense mutations leading to the production of inactive colicin E3 proteins. The ceaC1 mutation, affecting the transiocation of colicin E3 through bacterial membranes, is caused by a serine to phenylalanine change at position 37 within the glycine‐rich region at the N‐terminal part of colicin E3. This confirms previous results suggesting a role for this domain in colicin uptake by sensitive cells. The ceaC2 and ceaC3 mutations, abolishing colicin E3 RNase activity, affect the C‐terminal enzymatic domain of the molecule, in the ceaC2 mutant, serine at position 529 was converted to leucine. The ceaC3 mutation replaced a glycine residue at position 524 with an aspartic acid residue. The two mutations ceaC2 and ceaC3 yieid information on the amino acid residues involved in the RNase activity of colicin E3.
Export and secretion of the lipoprotein Pullulanase by Klebsiella pneumoniae
Authors: D'Enfert C. , Chapon C. , Pugsley A. P.
Published: October 2006

Pullulanase, a secreted lipoprotein of Klebsiella pneumoniae, is initially localized to the outer face of the outer membrane, as shown by protease and substrate accessibility and by immunofluorescence tests. Freeze‐thaw disruption of these cells released both membrane‐associated and apparently soluble forms of Pullulanase. Membrane‐associated Pullulanase co‐fractionated with authentic outer membrane vesicles upon isopycnic sucrose‐gradient centrifugation, whereas the quasi‐soluble form had the same equilibrium density as inner membrane vesicles and extracellular Pullulanase aggregates. The latter also contained outer membrane maltoporin, but were largely devoid of other membrane components including LPS and lipids. K. pneumoniae carrying multiple copies of the Pullulanase structural gene (pulA) produced increased amounts of cell‐associated and secreted Pullulanase, but a large proportion of the enzyme was neither exposed on the cell surface nor released into the medium, even after prolonged incubation. This suggests that factors necessary for Pullulanase secretion were saturated by the over‐produced Pullulanase. When pulA was expressed under lacZ promoter control, the Pullulanase which was produced was not exposed on the cell surface at any time, suggesting that Pullulanase secretion genes are not expressed constitutively, and raising the possibility that they, like puM, may be part of the maltose regulon.