bluuebunny/crossref_metadata_2025 huggingface dataset:
Cloning of novel LERGU mRNAs in GPR30 3′ untranslated region and detection of 2 bp‐deletion polymorphism in gastric cancer
Authors: Kakinuma Naoto , Sato Masaaki , Yamada Tatsuya , Kohu Kazuyoshi , Nakajima Motowo , Akiyama Tetsu , Ohwada Susumu , Shibanaka Yasuhiko
Published: March 2005

The improved IGCR (In–Gel Competitive Reassociation) method was applied to the analysis of human gastric cancer genomic DNA to identify its alterations, and it appeared that the IGCR library contained a fragment of 3′‐untranslated region (3′ UTR) of G‐protein coupled receptor 30 (GPR30) mRNA. When we searched genomic DNA pairs of gastric cancer patients with this IGCR clone, we found the deletion polymorphism with or without 2 bp (Cytosine and Thymine; CT). We confirmed the existence of a novel mRNA in GPR30 3′UTR by northern blotting, cloned this novel mRNA and named it Leucine Rich Protein in GPR30 3′UTR (LERGU). The EST database search gave one alternative splicing form in this 3′ UTR, which was named as LERGU‐1. A novel alternative splicing form of this mRNA was also identified from the stomach total RNA, which was named LERGU‐2. The LERGU mRNA was also detected in eight gastric cancer cell lines, but GPR30 mRNA scarcely existed. Furthermore, we detected the 2 bp‐deletion form in genomic DNAs and mRNAs derived from gastric cancers, but not in other type cancers. Since the 2 bp‐deletion position on LERGU corresponds to its alternative splicing site, this deletion may produce a frame‐shifted protein. Overall, our findings suggest that a mutation or disappearance of the normal LERGU protein may have a function in the development of gastric cancer. (Cancer Sci 2005; 96: 191–196)
A survey of sleep problems in autism, Asperger's disorder and typically developing children
Authors: Polimeni M. A. , Richdale A. L. , Francis A. J. P.
Published: March 2005

Background  Sleep problems are common in typically developing (TD) children and in children with autism, however, less is known about the sleep of children with Asperger's disorder (AD). The aim of this study was to compare sleep patterns of children with autism and AD to a TD group of children.

Methods  Sixty‐six parents of TD children, 53 parents of children with autism, and 52 parents of children with AD completed a survey on their child's sleep patterns, the nature and severity of any sleep problems and success of any treatment attempted.

Results  The results showed high prevalence of sleep problems with significantly more problems reported in the autism and AD groups (TD = 50%, autism = 73%, AD = 73%), with no significant differences between groups on severity or type of sleep problem. Children with AD were significantly more likely to be sluggish and disoriented after waking and had a higher Behavioral Evaluation of Disorders of Sleep (BEDS) total score compared to the other two groups. The autism and AD groups reported significantly better treatment success for medication compared to the TD group. The autism group reported significantly better success for behavioural treatment compared to the AD group.

Conclusions  In conclusion, children with AD may have more symptoms of sleep disturbance, and different types of sleep problems than children with autism. As this is the first study to compare autism and AD and to survey treatment outcomes, further research is needed to validate these findings.
Assessment of patients with intellectual disability using the International Classification of Functioning, Disability and Health to evaluate dental treatment tolerability
Authors: Maeda S. , Kita F. , Miyawaki T. , Takeuchi K. , Ishida R. , Egusa M. , Shimada M.
Published: March 2005

Background  Patients with serious intellectual disability (ID) are occasionally unable to tolerate dental treatment when intravenous sedation or general anaesthesia (IVS/GA) is involved. In order to make a decision regarding the application of IVS/GA, the International Classification of Functioning, Disability and Health (ICF) is useful. Therefore, in this study, a set of codes involved in dental problems were chosen from the ICF, and patients with ID who could tolerate dental treatments were compared with those who could not.

Methods  From preliminary interviews of six patients with ID, 16 codes were chosen, and an objective five‐rank scale was then constructed for use with all chosen codes. Forty‐nine ID patients who visited the Okayama University Hospital for dental treatment between January and April 2003 were evaluated. Facility workers were interviewed according to the code set chosen. The participants were then divided into two subgroups depending on their tolerability of dental treatment. The results of these groups for all 16 codes were then compared.

Results  Of the 49 patients interviewed, 23 were able to tolerate the dental treatment. In the ‘Activities & Participation’ section of the ICF, the tolerable group showed lower disability levels with regard to d110 Watching, d540 Dressing and d550 Eating. In other sections, there were no significant differences between the groups. The code set chosen in this study and the five‐rank scales in each code were useful as they enabled easy interviewing.

Conclusions  The ICF was raised as a possibility for considering the application of IVS/GA for dental treatment on patients with ID. For clinical use of the ICF, it is recommended that significant codes should be selected and that the five‐rank scale is used so that more objective results are obtained from interviews.
Measurement of perceived competence in Dutch children with mild intellectual disabilities
Authors: Elias C. , Vermeer A. , T Hart H.’
Published: March 2005

Background  Little research has been conducted on the perceived competence of children with mild intellectual disabilities (MID). One of the reasons for the marked absence of research appears to be the lack of reliable and clearly valid measurement instruments for this particular group of children. In the present study, it was examined whether a pictorial scale originally designed to measure perceived competence in typically developing children could successfully be used with children with MID.

Methods  The pictorial scale was administered to a group of 106 children with MID. The construct validity, reliability and stability of the scale were investigated.

Results  The results of the exploratory factor analyses and the confirmatory factor analyses supported the conceptual framework proposed. The construct validity was also supported by the pattern of intercorrelations between the subscales. The scale had adequate internal consistency and the stability analyses showed sufficient stability across a 4‐month period.

Conclusions  The findings show the psychometric properties of the pictorial scale to justify its use with children with MID.
Determining self‐incompatibility genotypes in Belgian wild cherries
Authors: DE CUYPER B. , SONNEVELD T. , TOBUTT K. R.
Published: March 2005

The self‐incompatibility (S) genotypes of a collection of 65 Belgian accessions of wild cherry, selected within two populations and planted in a seed orchard, were determined using polymerase chain reaction (PCR) methods. Initially, DNA extracts were amplified with consensus primers that amplify across the second intron of the S‐ribonuclease gene which shows considerable length polymorphism. The provisional genotypes deduced were checked with the appropriate allele‐specific primers for the known alleles S1 to S16. Putative new alleles were subjected to PCR with consensus primers amplifying across the first intron. Six new alleles, S17 to S22, were thus indicated on the basis of the estimated lengths of the first and second intron PCR products. Examples of these alleles were partially sequenced and were indeed mutually distinct and different from the known alleles. The incompatibility genotypes of all 65 accessions were determined and one triploid individual was found. Seventeen alleles were detected in all. Allele frequencies differed between samples and the expected total number of alleles in the underlying populations was estimated. The wild cherry populations differed significantly with respect to allelic frequencies from sweet cherry cultivars; alleles S4 and S5, which are moderately frequent in sweet cherry, were absent from the wild cherry accessions. The knowledge of the S genotypes will be useful for studying the gene flow within the seed orchard and these approaches should also be informative in wild populations.
Origin and diversification ofHibiscus glaber, species endemic to the oceanic Bonin Islands, revealed by chloroplast DNA polymorphism
Authors: TAKAYAMA KOJI , OHI‐TOMA TETSUO , KUDOH HIROSHI , KATO HIDETOSHI
Published: March 2005

Two woodyHibiscusspecies co‐occur in the Bonin Islands of the northwestern Pacific Ocean:Hibiscus glaberMatsum. is endemic to the islands, and its putative ancestral species,Hibiscus tiliaceusL., is widely distributed in coastal areas of the tropics and subtropics. To infer isolating mechanisms that led to speciation ofH. glaberand the processes that resulted in co‐occurrence of the two closely related species on the Bonin Islands, we conducted molecular phylogenetic analyses on chloroplast DNA (cpDNA) sequences. Materials collected from a wide area of the Pacific and Indian Oceans were used, and two closely related species,Hibiscus hamaboSiebold Zucc. andHibiscus macrophyllusRoxb., were also included in the analyses. The constructed tree suggested thatH. glaberhas been derived fromH. tiliaceus,and that most of the modern Bonin populations ofH. tiliaceusdid not share most recent ancestry withH. glaber.Geographic isolation appears to be the most important mechanism in the speciation ofH. glaber.The co‐occurrence of the two species can be attributed to multiple migrations of different lineages into the islands. While a wide and overlapping geographical distribution of haplotypes was found inH. tiliaceus, localized geographical distribution of haplotypes was detected inH. glaber. It is hypothesized that a shift to inland habitats may have affected the mode of seed dispersal from ocean currents to gravity and hence resulted in geographical structuring ofH. glaberhaplotypes.
Range expansions in the flightless longhorn cactus beetles,Moneilema gigasandMoneilema armatum, in response to Pleistocene climate changes
Authors: SMITH CHRISTOPHER IRWIN , FARRELL BRIAN D.
Published: March 2005

Pollen cores and plant and animal fossils suggest that global climate changes at the end of the last glacial period caused range expansions in organisms indigenous to the North American desert regions, but this suggestion has rarely been investigated from a population genetic perspective. In order to investigate the impact of Pleistocene climate changes and glacial/interglacial cycling on the distribution and population structure of animals in North American desert communities, biogeographical patterns in the flightless, warm‐desert cactus beetles,Moneilema gigasandMoneilema armatum, were examined using mitochondrial DNA (mtDNA) sequence data from the cytochrome oxidase I (COI) gene. Gene tree relationships between haplotypes were inferred using parsimony, maximum‐likelihood, and Bayesian analysis. Nested clade analysis and coalescent modelling using the programsmdivandfluctuatewere used to identify demographically independent populations, and to test the hypothesis that Pleistocene climate changes caused recent range expansions in these species. A sign test was used to evaluate the probability of observing concerted population growth across multiple, independent populations. The phylogeographical and nested clade analyses reveal a history of northward expansion in both of these species, as well as a history of past range fragmentation, followed by expansion from refugia. The coalescent analyses provide highly significant evidence for independent range expansions from multiple refugia, but also identify biogeographical patterns that predate the most recent glacial period. The results indicate that widespread desert environments are more ancient than has been suggested in the past.
Local genetic structure in a clonal dioecious angiosperm
Authors: RUGGIERO M. V. , REUSCH T. B. H. , PROCACCINI G.
Published: March 2005

We used seven microsatellite loci to characterize genetic structure and clonal architecture at three different spatial scales (from meters to centimetres) of aCymodocea nodosapopulation. C. nodosaexhibits both sexual reproduction and vegetative propagation by rhizome elongation. Seeds remain buried in the sediment nearby the mother plant in a dormant stage until germination. Seed dispersal potential is therefore expected to be extremely restricted. High clonal diversity (up to 67% of distinct genotypes) and a highly intermingled configuration of genets at different spatial scales were found. No significant differences in genetic structure were found among the three spatial scales, indicating that genetic diversity is evenly distributed along the meadow. Autocorrelation analyses of kinship estimates confirmed the absence of spatial clumping of genets at small spatial scale and the expectations of a very restricted seed dispersal (observed dispersal range 1–21 m) in this species.
On the verge of extinction: genetics of the critically endangered Iberian plant species,Borderea chouardii(Dioscoreaceae) and implications for conservation management
Authors: SEGARRA‐MORAGUES J. G. , PALOP‐ESTEBAN M. , GONZÁLEZ‐CANDELAS F. , CATALÁN P.
Published: March 2005

Borderea chouardiiis a relictual and dioecious, strictly sexually reproducing, long‐living geophyte of the Dioscoreaceae family. Previous biological and demographic studies have indicated the existence of a uniformly distributed panmictic population of this taxon at the southernmost Spanish pre‐Pyrenean mountain ranges where it occurs in rather inaccessible crevices of a single limestone cliff. However, individuals ofB. chouardiiare spatially subdivided into two subpopulations located, respectively, on the upper and lower parts of the cliff, and vertically separated 150 m. Because of its extreme rarity,B. chouardiiwas the first Iberian taxon to have a specific conservation plan and has been included in several red lists under the category of critically endangered (CR). However, no previous attempts have been conducted to analyse the fine scale evolutionary mechanisms involved in its present microspatial distribution. Genetic diversity and population structure have been investigated through the analysis of neutral hypervariable markers such as simple sequence repeats (SSRs) and randomly amplified polymorphic DNAs (RAPDs) to unravel the impact of life history traits in the differentiation of the two subpopulations. Both types of molecular markers were unequivocal in distinguishing two genetically distinct groups of individuals corresponding to their spatial separation. However, SSRs detected a higher level of subpopulation differentiation (FST = 0.35,RST = 0.32) than RAPDs (FST = 0.21). SSR data indicated significant deviation from random dispersal of genes and genotypes between the two groups, suggesting that mating occurs mainly among individuals within subpopulations, thus, favouring the divergence between the two groups. This microevolutionary differentiation scenario might have been caused by a coupled effect of past genetic drift and reproductive isolation, as a result of strong glacial age bottlenecks and inefficient dispersal system of pollen and seeds, respectively. The identification of such genetic structure in this narrow endemic prompts a modification of the management strategies of its single extant population.
Complex mating system and dispersal patterns in a social lizard,Egernia whitii
Authors: CHAPPLE DAVID G. , KEOGH J. SCOTT
Published: March 2005

In contrast to the polygynous mating systems typically displayed by most reptilian taxa, long‐term genetic monogamy appears to be widespread within a lineage of group‐living Australian scincid lizards, theEgerniagroup. We have recently shown that White's skink,Egernia whitii, lives in small but temporally stable social aggregations. Here, we examine the mating system, spatial organization, and dispersal patterns ofE. whitiiusing behavioural field studies and data from four microsatellite loci. Parentage analysis ofE. whitiilitters revealed that its mating system is characterized by both polygyny and monogamy. Polygyny was the predominant mating system but within‐season social and genetic monogamy was common (36–45% of breeding pairs). The incidence of between‐season monogamy inE. whitiiwas rare compared to that reported for its congeners. Low levels of multiple paternity (12% of litters) and extra‐group paternity (16%) were detected. Social groups are generally comprised of closely related individuals, but breeding pairs were not more closely related compared to other potential mates. Spatial autocorrelation analyses revealed significant positive local genetic structure over 50 m, which was consistent for all age–sex classes. There was no clear and consistent evidence for sex‐biased dispersal, with assignment tests (mean assignment index) and relatedness analyses suggesting female‐biased dispersal, but spatial autocorrelation analyses indicating a trend for male‐biased dispersal. We discuss the implication of our results in regard to the factors promoting the evolution of monogamy within theEgerniagroup.
Molecular phylogenetics of the Macaronesian‐endemic genus Bystropogon (Lamiaceae): palaeo‐islands, ecological shifts and interisland colonizations
Authors: TRUSTY JENNIFER L. , OLMSTEAD RICHARD G. , SANTOS‐GUERRA ARNOLDO , SÁ‐FONTINHA SUSANA , FRANCISCO‐ORTEGA JAVIER
Published: March 2005

A molecular phylogenetic study of Bystropogon L’Hèr. (Lamiaceae) is presented. We performed a cladistic analysis of nucleotide sequences of the internal transcribed spacers (ITS), of the nuclear ribosomal DNA, and of the trnL gene and trnL‐trnF intergenic spacer of the chloroplast DNA. Bystropogon odoratissimus is the only species endemic to the Canary Islands that occurs in the three palaeo‐islands of Tenerife. This species is not part of an early diverging lineage of Bystropogon and we suggest that it has a recent origin. This phylogenetic pattern is followed by most of the species endemic to the palaeo‐islands of Tenerife. The two sections currently recognized in Bystropogon form two monophyletic groups. Taxa belonging to the section Bystropogon clade show interisland colonization limited to the Canary Islands with ecological shifts among three ecological zones. Taxa from the section Canariense clade show interisland colonization both within the Canary Islands and between the Canary Islands and Madeira. Speciation events within this clade are mostly limited to the laurel forest. The genus has followed a colonization route from the Canaries towards Madeira. This route has also been followed by at least five other plant genera with species endemic to Macaronesia. Major incongruences were found between the current infrasectional classification and the molecular phylogeny, because the varieties of Bystropogon origanifolius and Bystropogon canariensis do not form two monophyletic groups. The widespread B. origanifolius appears as progenitor of the other species in section Bystropogon with a more restricted distribution.
Genetic and epigenetic consequences of recent hybridization and polyploidy in Spartina (Poaceae)
Authors: SALMON ARMEL , AINOUCHE MALIKA L. , WENDEL JONATHAN F.
Published: March 2005

To study the consequences of hybridization and genome duplication on polyploid genome evolution and adaptation, we used independently formed hybrids (Spartina × townsendii and Spartina × neyrautii) that originated from natural crosses between Spartina alterniflora, an American introduced species, and the European native Spartina maritima. The hybrid from England, S. × townsendii, gave rise to the invasive allopolyploid, salt‐marsh species, Spartina anglica. Recent studies indicated that allopolyploid speciation may be associated with rapid genetic and epigenetic changes. To assess this in Spartina, we performed AFLP (amplified fragment length polymorphism) and MSAP (methylation sensitive amplification polymorphism) on young hybrids and the allopolyploid. By comparing the subgenomes in the hybrids and the allopolyploid to the parental species, we inferred structural changes that arose repeatedly in the two independently formed hybrids. Surprisingly, 30% of the parental methylation patterns are altered in the hybrids and the allopolyploid. This high level of epigenetic regulation might explain the morphological plasticity of Spartina anglica and its larger ecological amplitude. Hybridization rather than genome doubling seems to have triggered most of the methylation changes observed in Spartina anglica.
Positional cues for the starch/lipid balance in maize kernels and resource partitioning to the embryo
Authors: Rolletschek Hardy , Koch Karen , Wobus Ulrich , Borisjuk Ljudmilla
Published: March 2005

This study tests the hypotheses that in vivo oxygen levels inside developing maize grains locally affect assimilate partitioning and ATP distribution within the kernel. These questions were addressed through combined topographical analysis (O2‐ and ATP‐mapping), metabolite profiling, and isotope flux analysis. Internal and external oxygen levels were also experimentally altered. Under ambient conditions, mean O2 concentration immediately inside starchy endosperm dropped to only 1.4% of atmospheric saturation (approximately 3.8 μm), but was 10‐fold higher in the oil‐storing embryo. Increasing the O2 supply to intact kernels stimulated their O2 demand, shifted ATP localization within the kernel, and elevated their ATP/ADP ratio. Enhanced O2 availability also increased steady‐state levels of glycolytic intermediates and those of the citric acid cycle, as well as some related pools of free amino acids. Subsequent analyses indicated that starch formation within endosperm, but not lipid biosynthesis within embryo, was adapted to the endogenous low oxygen. Increasing the O2 supply did not change ADP‐glucose levels, activity of ADP‐glucose pyrophosphorylase, 13C‐labeling of ADP‐glucose, or flux of 14C‐sucrose into starch. In contrast, enhanced O2 availability increased 14C‐label uptake into the embryo, 13C‐labeling of acetyl‐coenzyme A, and finally 14C‐incorporation into lipids. Lipid accumulation in embryo appeared highest in regions with higher ATP. Consistent with labeling data, a decrease in O2 supply most strongly affected the embryo, whereas rising O2 levels expanded ATP‐rich zones toward the starch‐storing endosperm and the scutellar part of embryo. The latter might be responsible for higher 14C‐label uptake into the embryo and flux toward lipid. Collectively, data indicate that the in vivo oxygen distribution in maize kernels markedly affects ATP gradients, metabolite levels, and favors assimilate partitioning toward starch within the O2‐depleted endosperm. Clear advantages are thus evident for peripheral localization of the protein and lipid storing structures in maize kernels.
A role of Toc33 in the protochlorophyllide‐dependent plastid import pathway of NADPH:protochlorophyllide oxidoreductase (POR) A†
Authors: Reinbothe Steffen , Pollmann Stephan , Springer Armin , James Richard Johari , Tichtinsky Gabrielle , Reinbothe Christiane
Published: March 2005

NADPH:protochlorophyllide oxidoreductase (POR) A is a key enzyme of chlorophyll biosynthesis in angiosperms. It is nucleus‐encoded, synthesized as a larger precursor in the cytosol and imported into the plastids in a substrate‐dependent manner. Plastid envelope membrane proteins, called protochlorophyllide‐dependent translocon proteins, Ptcs, have been identified that interact with pPORA during import. Among them are a 16‐kDa ortholog of the previously characterized outer envelope protein Oep16 (named Ptc16) and a 33‐kDa protein (Ptc33) related to the GTP‐binding proteins Toc33 and Toc34 of Arabidopsis. In the present work, we studied the interactions and roles of Ptc16 and Ptc33 during pPORA import. Radiolabeled Ptc16/Oep16 was synthesized from a corresponding cDNA and imported into isolated Arabidopsis plastids. Crosslinking experiments revealed that import of 35S‐Oep16/Ptc16 is stimulated by GTP. 35S‐Oep16/Ptc16 forms larger complexes with Toc33 but not Toc34. Plastids of the ppi1 mutant of Arabidopsis lacking Toc33, were unable to import pPORA in darkness but imported the small subunit precursor of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (pSSU), precursor ferredoxin (pFd) as well as pPORB which is a close relative of pPORA. In white light, partial suppressions of pSSU, pFd and pPORB import were observed. Our results unveil a hitherto unrecognized role of Toc33 in pPORA import and suggest photooxidative membrane damage, induced by excess Pchlide accumulating in ppi1 chloroplasts because of the lack of pPORA import, to be the cause of the general drop of protein import.
microRNA‐directed cleavage of ATHB15 mRNA regulates vascular development in Arabidopsis inflorescence stems
Authors: Kim Joonki , Jung Jae‐Hoon , Reyes Jose L. , Kim Youn‐Sung , Kim Sun‐Young , Chung Kyung‐Sook , Kim Jin A. , Lee Minsun , Lee Yoontae , Narry Kim V. , Chua Nam‐Hai , Park Chung‐Mo
Published: March 2005

Class III homeodomain‐leucine zipper proteins regulate critical aspects of plant development, including lateral organ polarity, apical and lateral meristem formation, and vascular development. ATHB15, a member of this transcription factor family, is exclusively expressed in vascular tissues. Recently, a microRNA (miRNA) binding sequence has been identified in ATHB15 mRNA, suggesting that a molecular mechanism governed by miRNA binding may direct vascular development through ATHB15. Here, we show that miR166‐mediated ATHB15 mRNA cleavage is a principal mechanism for the regulation of vascular development. In a gain‐of‐function MIR166a mutant, the decreased transcript level of ATHB15 was accompanied by an altered vascular system with expanded xylem tissue and interfascicular region, indicative of accelerated vascular cell differentiation from cambial/procambial cells. A similar phenotype was observed in Arabidopsis plants with reduced ATHB15 expression but reversed in transgenic plants overexpressing an miR166‐resistant ATHB15. ATHB15 mRNA cleavage occurred in standard wheat germ extracts and in Arabidopsis and was mediated by miR166 in Nicotiana benthamiana cells. miR166‐assisted ATHB15 repression is likely to be a conserved mechanism that regulates vascular development in all vascular plants.
The atypical resistance gene, RPW8, recruits components of basal defence for powdery mildew resistance in Arabidopsis
Authors: Xiao Shunyuan , Calis Ozer , Patrick Elaine , Zhang Guangmin , Charoenwattana Piyavadee , Muskett Paul , Parker Jane E. , Turner John G.
Published: March 2005

Genetic studies have identified a number of components of signal transduction pathways leading to plant disease resistance and the accompanying hypersensitive response (HR) following detection of pathogens by plant resistance (R) genes. In Arabidopsis, the majority of R proteins so far characterized belong to a plant superfamily that have a central nucleotide‐binding site and C‐terminal leucine‐rich‐repeats (NB‐LRRs). Another much less prevalent class comprises RPW8.1 and RPW8.2, two related proteins that possess a putative N‐terminal transmembrane domain and a coiled‐coil motif, and confer broad‐spectrum resistance to powdery mildew. Here we investigated whether RPW8.1 and RPW8.2 engage known pathway(s) for defence signalling. We show that RPW8.1 and RPW8.2 recruit, in addition to salicylic acid and EDS1, the other NB‐LRR gene‐signalling components PAD4, EDS5, NPR1 and SGT1b for activation of powdery mildew resistance and HR. In contrast, NDR1, RAR1 and PBS3 that are required for function of certain NB‐LRR R genes, and COI1 and EIN2 that operate, respectively, in the jasmonic acid and ethylene signalling pathways, do not contribute to RPW8.1 and RPW8.2‐mediated resistance. We further demonstrate that EDR1, a gene encoding a conserved MAPKK kinase, exerts negative regulation on HR cell death and powdery mildew resistance by limiting the transcriptional amplification of RPW8.1 and RPW8.2. Our results suggest that RPW8.1 and RPW8.2 stimulate a conserved basal defence pathway that is negatively regulated by EDR1.
Morphoquantitative Aspects of Nitrergic Myoenteric Neurons from the Stomach of Diabetic Rats Supplemented with Acetyl‐L‐Carnitine
Authors: Fregonesi C. E. P. T. , Molinari S. L. , Alves Â. M. P. , Defani M. A. , Zanoni J. N. , Bazotte R. B. , De Miranda Neto M. H.
Published: March 2005

The NADPH‐diaphorase (NADPH‐d) positive myoenteric neurons from the body of the stomach of rats with streptozotocin‐induced diabetes with or without supplementation with acetyl‐L‐carnitine (ALC) were evaluated. At the age of 105 days the animals were divided into four groups: normoglycaemic (C), normoglycaemic supplemented with ALC (CC), diabetic (D) and diabetic supplemented with ALC (DC). The supplementation with ALC (200 mg/kg body weight/day) to groups CC and DC was made during 105 days. After this period the animals were killed and the stomach removed and subjected to the histochemical technique of NADPH‐d for the staining of the neurons of the myoenteric plexus. The area of 500 neurons of each group was investigated, as well as the neuronal density in an area of 23.84 mm2 in each stomach. ALC promoted reduction (P < 0.05) of fasting glycaemia, water ingestion and areas of the profiles of the cell bodies of the NADPH‐d neurons in the diabetic animals. The density of these neurons was not statistically different in the groups studied. It is suggested, therefore, a moderate neuroprotective effect of ALC, because the diminishment of the areas of the neuronal profiles in the supplemented diabetic animals, although being statistically significant relative to the non‐supplemented diabetics, was not sufficient to equal the values from the non‐diabetic controls.
Histological Basis of the Porcine Femoral Artery for Vascular Research
Authors: Solanes N. , Rigol M. , Ramírez J. , Segalés J. , Roqué M. , Marimon J.M. , Pérez‐Villa F. , Roig E. , Sanz G. , Heras M.
Published: March 2005

The similarities between the porcine and human cardiovascular systems make the pig a useful animal model for the study of vascular biology. However, a standardized method is needed to describe the normal histological properties of porcine arteries in order to evaluate pathologic lesions in future studies. Descriptive and morphometric analyses were done on 16 porcine femoral arteries. For these purposes, three histological stains (haematoxylin eosin, Masson's trichrome, and orcein), four immunohistochemical methods (using antibodies anti‐α‐actin, anti‐CD3, anti‐L1 and anti‐lysozyme), and a glycohistochemical method (usingDolichos bifloruslectin) were performed. The porcine femoral arteries evaluated had a mean total area of 6.25 ± 1.99 mm2and a diameter of 2.79 ± 0.41 mm. The majority of the total area was occupied by the medial layer (42.97 ± 5.38%) and was mainly constituted by smooth muscle cells (94.58 ± 2.65%). All the cell markers used reacted with porcine paraffin‐embedded tissue. However, the anti‐lysozyme antibody was excluded from this histological analysis because of cytoplasmatic reactivity in smooth muscle cells. In summary, this study proposes histological methods to describe the normal characteristics of the porcine femoral artery and raises the possibility of applying this methodology in future studies on porcine vascular research.
Ultrastructure and Histochemical Study of Glycoconjugates in the Gills of the White Croaker (Micropogonias furnieri)
Authors: Díaz A. O. , García A. M. , Devincenti C. V. , Goldemberg A. L.
Published: March 2005

The ultrastructure of the primary and secondary lamellae of gills was investigated in a marine teleost, the white croaker. The following cells were identified and briefly described: pavement cells, mucous cells, mitochondria‐rich cells and rodlet cells. These cell types are present throughout the length of the lamellae. They are studied by means of a series of carbohydrate histochemical methods, including lectin procedures. Neutral sugars and substituted sialic acid were detected by means of periodic acid–borohydride reduction–saponification–periodic acid Schiff reaction (PA/Bh/KOH/PAS), saponification–selective periodic acid Schiff reaction (KOH/PA*S) and saponification–selective periodic acid–borohydride reduction–periodic acid Schiff reaction (KOH/PA*/Bh/PAS) histochemical techniques. A battery of seven lectins was used to study binding on tissue sections at the light microscopic level to characterize glycoconjugates in gills. The reaction to Canavalia ensiformis agglutinin (Con‐A), Triticum vulgaris agglutinin (WGA), and Ricinus cummunis agglutinin‐1 (RCA‐1) was weak in pavement cells; unlike Con‐A, the reaction to WGA and RCA‐1 was more intense in mucous cells. Arachis hypogaea agglutinin (PNA) lectin showed a strong reaction in mucous cells. Ulex europaens agglutinin‐1 (UEA‐1) lectin was negative in all cell types. The lectin pattern was similar for both primary and secondary lamellae, except for PNA reaction, which was weak in the pavement cells of the secondary lamella and negative in the pavement cells of the primary lamella.
Immune response to a laminin‐binding protein (Lmb) in group A streptococcal infection
Authors: Wahid Rezwanul M. , Yoshinaga Masao , Nishi Junichiro , Maeno Nobuaki , Sarantuya Jav , Ohkawa Toshiya , Jalil Abdul M. , Kobayashi Keiko , Miyata Koichiro
Published: March 2005

Abstract 
Background
: A laminin‐binding protein (Lmb) similar to that of group B streptococcus is conserved in group A streptococcus (GAS) and has a role in adhesion of GAS to epithelial cells. The role of this protein is yet to be clarified in disease process and thus, it is important to know its role in binding of GAS to laminin and the immunogenic response against it in patients related with GAS infection.

Methods
: A recombinant protein (rGAS‐Lmb) was purified using the lmb gene from M1 GAS and tested for its role in binding of GAS with laminin. The antibody response against rGAS‐Lmb in patient sera related with GAS infection was measured by ELISA.

Results
: The rGAS‐Lmb bound with laminin directly and inhibited the binding of GAS to laminin. The antibody response against rGAS‐Lmb in patients with uncomplicated streptococcal infection (U. Strep) and those with rheumatic fever (RF) were significantly higher than those in the control group (P < 0.0001 and P < 0.001, respectively). No difference of anti‐rGAS‐Lmb antibody titer could be found between these two disease groups.

Conclusion
: The higher antibody response in patients with GAS infection implies that the protein is well expressed during the period of infection and may be related with the colonization and infection of GAS in pharyngeal mucosa.
Neutrophil‐mediated inflammation in respiratory syncytial viral bronchiolitis
Authors: Yasui Kozo , Baba Atsushi , Iwasaki Yasushi , Kubo Tetsuo , Aoyama Kohki , Mori Tetsuo , Yamazaki Takashi , Kobayashi Norimoto , Ishiguro Akira
Published: March 2005

Abstract 
Background
: The involvement of neutrophil‐mediated inflammation may play an important role in the pathogenesis of acute respitory syncytial virus bronchiolitis. However, no measurable marker is sensitive enough to assess neutrophil‐mediated inflammation in the airways. Released neutrophil elastase (NE) in intraluminal airways has been reported to induce pulmonary inflammation. The aim of this study was to determine whether the amount of urinary trypsin inhibitor (UTI) in serum, a degenerate induced by NE, reflects the degree of airway inflammation in children with respiratory syncytial viral (RSV) bronchiolitis and whether the severity of inflammation is evaluated. The pre‐α‐/inter‐α‐trypsin inhibitor is assumed to be precursors of the UTI. When NE degrades these inhibitors, UTI is liberated.

Methods
: Serum UTI concentrations in infants admitted with RSV bronchiolitis, other viral infections, bacterial pneumonia and control subjects were measured by means of one‐step sandwich‐type enzyme immunoassay.

Results
: Serum UTI concentrations in 25 patients on admission were significantly higher than the 15 infantile control values (mean ± SEM, 22.126 ± 2.317 and 6.701 ± 0.719 U/mL, respectively; P < 0.0001). The elevated levels returned to baseline values with improvement in the respiratory symptoms. Higher levels of serum UTI with RSV infection were consistently associated with clinical symptoms and artificial ventilation. Serum NE concentrations of patients were elevated in some patients but not significantly different from controls in the patients who showed only upper respiratory symptoms with RSV infections.

Conclusion
: The findings strongly suggested that neutrophil‐mediated events are involved in the pathogenesis of RSV bronchiolitis, and the monitoring of UTI concentrations might be useful for evaluating the neutrophil‐mediated airway inflammation.
Vitamin D deficiency rickets caused by improper lifestyle in Japanese children
Authors: Miyako Kenichi , Kinjo Saori , Kohno Hitoshi
Published: March 2005

Abstract 
Background
: Nutritional rickets is considered rare in developed countries. However, reports on vitamin D deficiency rickets caused by improper lifestyle have recently increased. The clinical and laboratory characteristics of patients with vitamin D deficiency rickets treated at Fukuoka Children's Hospital, Fukuoka, Japan, were evaluated to clarify current causes and ways to prevent this disease.

Methods
: Clinical records were reviewed, and obtained information and data were summarized.

Results
: Eight patients with vitamin D deficiency rickets (five boys and three girls) were treated during the past 10 years (January 1992 to December 2001). Two infants were referred to the hospital for hypocalcemia and convulsion, and six toddlers (1–2 years old) for bowlegs. One patient lacked exposure to sunlight, and six had an unbalanced diet. The cause of rickets could not be established in one patient. Anthropometric and laboratory data did not indicate malnutrition. Serum alkaline phosphatase was 2518.3 ± 1401.7 IU/l, calcium was 8.2 ± 2.6 mg/dL (including 4.7 mg/dL in one infant and 4.8 mg/dL in another), and phosphorus was 4.9 ± 1.0 mg/dL. High sensitive parathyroid hormone was 1393.1 ± 321.7 pg/mL (reference range, 180–560), 1,25‐dihydroxyvitamin D was 86.0 ± 61.5 pg/mL (reference range, 20–70), and 25‐hydroxyvitamin D was 11.6 ± 5.6 ng/mL (reference range, 10–30). The patients recovered with a change to a balanced diet, the promotion of weaning, and/or an increase in sunlight exposure.

Conclusion
: Vitamin D deficiency rickets remains a common condition that is best managed by education and disease prevention.
Continuity and change from early childhood to adolescence in autism
Authors: McGovern Corina W. , Sigman Marian
Published: March 2005

Background:  This longitudinal study of 48 children diagnosed with autism at 2–5 years of age was designed to test the hypothesis that diagnosis would remain stable for most of the sample but that there would be improvements in symptom severity, adaptive behavior, and emotional responsiveness in adolescence.

Methods:  A sample of children with autism assessed in both early and middle childhood were observed in late adolescence with the Autism Diagnostic Observation Scale (ADOS) and their parents were administered the Autism Diagnostic Interview‐Revised (ADI‐R) and the Vineland Adaptive Behavior Scale.

Results:  All but 2 adolescents (46 of 48) met lifetime criteria for autism according to the ADI‐R, and all but 4 adolescents (40 of 44) met criteria for autism spectrum disorder on the ADOS. In contrast to the continuity in diagnosis, parents described improvements in social interactions, repetitive/stereotyped behaviors, adaptive behaviors, and emotional responsiveness to others’ distress in adolescence compared to middle childhood. High‐functioning adolescents with autism showed more improvement in these domains than low‐functioning adolescents with autism. The extent to which the adolescents were observed to be socially engaged with their peers in school in middle childhood predicted adaptive behavior skills even when intelligence level was statistically constrained.

Conclusions:  The developmental trajectory of children with autism appears to show both continuity and change. In this sample, most individuals continued to be diagnosed in the autism spectrum but parents reported improvements in adolescence. The results suggest that social involvement with peers improves adaptive behavior skills, and this argues for focusing intervention programs in this area. In addition, it is clear that high‐functioning adolescents improve more than low‐functioning individuals not only in cognitive abilities but also in social interaction skills. Thus, any early intervention that impacts the cognitive abilities of young children with autism is likely to have a parallel influence on their social skills as they mature into late adolescence and early adulthood.
Reduced expression of frataxin extends the lifespan of Caenorhabditis elegans
Authors: Ventura Natascia , Rea Shane , Henderson Samuel T. , Condo Ivano , Johnson Thomas E. , Testi Roberto
Published: March 2005

Defects in the expression of the mitochondrial protein frataxin cause Friedreich's ataxia, an hereditary neurodegenerative syndrome characterized by progressive ataxia and associated with reduced life expectancy in humans. Homozygous inactivation of the frataxin gene results in embryonic lethality in mice, suggesting that frataxin is required for organismic survival. Intriguingly, the inactivation of many mitochondrial genes in the nematode Caenorhabditis elegans by RNAi extends lifespan. We therefore investigated whether inactivation of frataxin by RNAi‐mediated suppression of the frataxin homolog gene (frh‐1) would also prolong lifespan in the nematode. Frataxin‐deficient animals have a small body size, reduced fertility and altered responses to oxidative stress. Importantly, frataxin suppression by RNAi significantly extends lifespan in C. elegans.
Aging induces cardiac diastolic dysfunction, oxidative stress, accumulation of advanced glycation endproducts and protein modification
Authors: Li Shi‐Yan , Du Min , Dolence E. Kurt , Fang Cindy X. , Mayer Gabriele E. , Ceylan‐Isik Asli F. , LaCour Karissa H. , Yang Xiaoping , Wilbert Christopher J. , Sreejayan Nair , Ren Jun
Published: March 2005

Evidence suggests that aging, per se, is a major risk factor for cardiac dysfunction. Oxidative modification of cardiac proteins by non‐enzymatic glycation, i.e. advanced glycation endproducts (AGEs), has been implicated as a causal factor in the aging process. This study was designed to examine the role of aging on cardiomyocyte contractile function, cardiac protein oxidation and oxidative modification. Mechanical properties were evaluated in ventricular myocytes from young (2‐month) and aged (24–26‐month) mice using a MyoCam® system. The mechanical indices evaluated were peak shortening (PS), time‐to‐PS (TPS), time‐to‐90% relengthening (TR90) and maximal velocity of shortening/relengthening (± dL/dt). Oxidative stress and protein damage were evaluated by glutathione and glutathione disulfide (GSH/GSSG) ratio and protein carbonyl content, respectively. Activation of NAD(P)H oxidase was determined by immunoblotting. Aged myocytes displayed a larger cell cross‐sectional area, prolonged TR90, and normal PS, ± dL/dt and TPS compared with young myocytes. Aged myocytes were less tolerant of high stimulus frequency (from 0.1 to 5 Hz) compared with young myocytes. Oxidative stress and protein oxidative damage were both elevated in the aging group associated with significantly enhanced p47phox but not gp91phox expression. In addition, level of cardiac AGEs was ∼2.5‐fold higher in aged hearts than young ones determined by AGEs‐ELISA. A group of proteins with a molecular range between 50 and 75 kDa with pI of 4–7 was distinctively modified in aged heart using one‐ or two‐dimension SDS gel electrophoresis analysis. These data demonstrate cardiac diastolic dysfunction and reduced stress tolerance in aged cardiac myocytes, which may be associated with enhanced cardiac oxidative damage, level of AGEs and protein modification by AGEs.
Reduced insulin/IGF‐1 signalling and human longevity
Authors: Van Heemst Diana , Beekman Marian , Mooijaart Simon P. , Heijmans Bastiaan T. , Brandt Bernd W. , Zwaan Bas J. , Slagboom P. Eline , Westendorp Rudi G. J.
Published: March 2005

Evidence is accumulating that aging is hormonally regulated by an evolutionarily conserved insulin/IGF‐1 signalling (IIS) pathway. Mutations in IIS components affect lifespan in Caenorhabditis elegans, Drosophila melanogaster and mice. Most long‐lived IIS mutants also show increased resistance to oxidative stress. In D. melanogaster and mice, the long‐lived phenotype of several IIS mutants is restricted to females. Here, we analysed the relationship between IIS signalling, body height and longevity in humans in a prospective follow‐up study. Based on the expected effects (increased or decreased signalling) of the selected variants in IIS pathway components (GHRHR, GH1, IGF1, INS, IRS1), we calculated composite IIS scores to estimate IIS pathway activity. In addition, we analysed the relative impact on lifespan and body size of the separate variants in multivariate models. In women, lower IIS scores are significantly associated with lower body height and improved old age survival. Multivariate analyses showed that these results were most pronounced for the GH1 SNP, IGF1 CA repeat and IRS1 SNP. In females, for variant allele carriers of the GH1 SNP, body height was 2 cm lower (P = 0.007) and mortality 0.80‐fold reduced (P = 0.019) when compared with wild‐type allele carriers. Thus, in females, genetic variation causing reduced IIS activation is beneficial for old age survival. This effect was stronger for the GH1 SNP than for variation in the conserved IIS genes that were found to affect longevity in model organisms.
Metric capabilities of low‐cost digital cameras for close range surface measurement
Authors: Chandler Jim H. , Fryer John G. , Jack Amanda
Published: March 2005

This paper examines the potential of low‐cost digital cameras for close range surface measurement using feature‐based image matching methods. This is achieved through extracting digital elevation models (DEMs) and comparing accuracies between three low‐cost consumer‐grade digital cameras (Sony DSC‐P10, Olympus C3030, Nikon Coolpix 3100) and the proven Kodak DCS460. Surprisingly, the tests revealed that the highest accuracies were achieved using the Sony DSC‐P10, not the Kodak DCS460, whilst the other two cameras certainly proved suitable for most close range surface measurement tasks. Lens modelling was found to provide a limiting constraint on final accuracies, with very small systematic error surfaces caused by residual imperfections in lens modelling. The IMAGINE OrthoBASE Pro software and an independent self‐calibrating bundle adjustment were used to process these data. These tests identified an inaccuracy in the self‐calibrating capability of IMAGINE OrthoBASE Pro version 8·6 and Leica Geosystems LPS 8·7, which will be rectified in subsequent software releases. The study has demonstrated that cheaper consumer‐grade digital cameras have potential for routine surface measurement provided lens modelling is considered. The lead author is maintaining a web‐based repository for camera calibration data, which may assist other users.