bluuebunny/crossref_metadata_2025 huggingface dataset:
Stn1–Ten1 is an Rpa2–Rpa3-like complex at telomeres
Authors: Sun Jia , Yu Eun Young , Yang Yuting , Confer Laura A. , Sun Steven H. , Wan Ke , Lue Neal F. , Lei Ming
Published: December 2009

In budding yeast, Cdc13, Stn1, and Ten1 form a heterotrimeric complex (CST) that is essential for telomere protection and maintenance. Previous bioinformatics analysis revealed a putative oligonucleotide/oligosaccharide-binding (OB) fold at the N terminus of Stn1 (Stn1N) that shows limited sequence similarity to the OB fold of Rpa2, a subunit of the eukaryotic ssDNA-binding protein complex replication protein A (RPA). Here we present functional and structural analyses of Stn1 and Ten1 from multiple budding and fission yeast. The crystal structure of the Candida tropicalis Stn1N complexed with Ten1 demonstrates an Rpa2N–Rpa3-like complex. In both structures, the OB folds of the two components pack against each other through interactions between two C-terminal helices. The structure of the C-terminal domain of Saccharomyces cerevisiae Stn1 (Stn1C) was found to comprise two related winged helix–turn–helix (WH) motifs, one of which is most similar to the WH motif at the C terminus of Rpa2, again supporting the notion that Stn1 resembles Rpa2. The crystal structure of the fission yeast Schizosaccharomyces pombe Stn1N–Ten1 complex exhibits a virtually identical architecture as the C. tropicalis Stn1N–Ten1. Functional analyses of the Candida albicans Stn1 and Ten1 proteins revealed critical roles for these proteins in suppressing aberrant telomerase and recombination activities at telomeres. Mutations that disrupt the Stn1–Ten1 interaction induce telomere uncapping and abolish the telomere localization of Ten1. Collectively, our structural and functional studies illustrate that, instead of being confined to budding yeast telomeres, the CST complex may represent an evolutionarily conserved RPA-like telomeric complex at the 3′ overhangs that works in parallel with or instead of the well-characterized POT1–TPP1/TEBPα–β complex.
Fusion of nearby inverted repeats by a replication-based mechanism leads to formation of dicentric and acentric chromosomes that cause genome instability in budding yeast
Authors: Paek Andrew L. , Kaochar Salma , Jones Hope , Elezaby Aly , Shanks Lisa , Weinert Ted
Published: December 2009

Large-scale changes (gross chromosomal rearrangements [GCRs]) are common in genomes, and are often associated with pathological disorders. We report here that a specific pair of nearby inverted repeats in budding yeast fuse to form a dicentric chromosome intermediate, which then rearranges to form a translocation and other GCRs. We next show that fusion of nearby inverted repeats is general; we found that many nearby inverted repeats that are present in the yeast genome also fuse, as does a pair of synthetically constructed inverted repeats. Fusion occurs between inverted repeats that are separated by several kilobases of DNA and share >20 base pairs of homology. Finally, we show that fusion of inverted repeats, surprisingly, does not require genes involved in double-strand break (DSB) repair or genes involved in other repeat recombination events. We therefore propose that fusion may occur by a DSB-independent, DNA replication-based mechanism (which we term “faulty template switching”). Fusion of nearby inverted repeats to form dicentrics may be a major cause of instability in yeast and in other organisms.
Divalent Metal Ions in Plant Mitochondria and Their Role in Interactions with Proteins and Oxidative Stress-Induced Damage to Respiratory Function
Authors: Tan Yew-Foon , O'Toole Nicholas , Taylor Nicolas L. , Millar A. Harvey
Published: December 2009

Understanding the metal ion content of plant mitochondria and metal ion interactions with the proteome are vital for insights into both normal respiratory function and the process of protein damage during oxidative stress. We have analyzed the metal content of isolated Arabidopsis (Arabidopsis thaliana) mitochondria, revealing a 26:8:6:1 molar ratio for iron:zinc:copper:manganese and trace amounts of cobalt and molybdenum. We show that selective changes occur in mitochondrial copper and iron content following in vivo and in vitro oxidative stresses. Immobilized metal affinity chromatography charged with Cu2+, Zn2+, and Co2+ was used to identify over 100 mitochondrial proteins with metal-binding properties. There were strong correlations between the sets of immobilized metal affinity chromatography-interacting proteins, proteins predicted to contain metal-binding motifs, and protein sets known to be oxidized or degraded during abiotic stress. Mitochondrial respiratory chain pathways and matrix enzymes varied widely in their susceptibility to metal-induced loss of function, showing the selectivity of the process. A detailed study of oxidized residues and predicted metal interaction sites in the tricarboxylic acid cycle enzyme aconitase identified selective oxidation of residues in the active site and showed an approach for broader screening of functionally significant oxidation events in the mitochondrial proteome.
Why does the biota of the Madagascar region have such a strong Asiatic flavour?
Authors: Warren Ben H. , Strasberg Dominique , Bruggemann J. Henrich , Prys‐Jones Robert P. , Thébaud Christophe
Published: December 2009

A corollary of island biogeographical theory is that islands are largely colonized from their nearest mainland source. Despite Madagascar’s extreme isolation from India and proximity to Africa, a high proportion of the biota of the Madagascar region has Asian affinities. This pattern has rarely been viewed as surprising, as it is consistent with Gondwanan vicariance. Molecular phylogenetic data provide strong support for such Asian affinities, but often not for their vicariant origin; most divergences between lineages in Asia and the Madagascar region post‐date the separation of India and Madagascar considerably (up to 87 Myr), implying a high frequency of dispersal that mirrors colonization of the Hawaiian archipelago in distance. Indian Ocean bathymetry and the magnitude of recent sea‐level lowstands support the repeated existence of sizeable islands across the western Indian Ocean, greatly reducing the isolation of Madagascar from Asia. We put forward predictions to test the role of this historical factor in the assembly of the regional biota.

© The Willi Hennig Society 2009.
Membrane rafts: a potential gateway for bacterial entry into host cells
Authors: Hartlova Anetta , Cerveny Lukas , Hubalek Martin , Krocova Zuzana , Stulik Jiri
Published: December 2009

Pathogenic bacteria have developed various mechanisms to evade host immune defense systems. Invasion of pathogenic bacteria requires interaction of the pathogen with host receptors, followed by activation of signal transduction pathways and rearrangement of the cytoskeleton to facilitate bacterial entry. Numerous bacteria exploit specialized plasma membrane microdomains, commonly called membrane rafts, which are rich in cholesterol, sphingolipids and a special set of signaling molecules which allow entry to host cells and establishment of a protected niche within the host. This review focuses on the current understanding of the raft hypothesis and the means by which pathogenic bacteria subvert membrane microdomains to promote infection.
Initiation of malignancy by duodenal contents reflux and the role of ezrin in developing esophageal squamous cell carcinoma
Authors: Ling Zhi‐Qiang , Mukaisho Ken‐ichi , Yamamoto Hiroto , Chen Kuan‐Hao , Asano Shinji , Araki Yoshio , Sugihara Hiroyuki , Mao Wei‐Min , Hattori Takanori
Published: December 2009

(Cancer Sci 2010; 101: 624–630)

Gastroesophageal reflux has recently been implicated as a causative factor in upper aerodigestive tract carcinogenesis. Esophageal squamous cell carcinomas (ESCCs) have developed in duodenal‐content reflux animals without any known carcinogen present. We established a cell line, designated ESCC‐DR, from a thoracic metastatic tumor in a reflux animal. To gain insight into the genomic alterations associated with duodenal content reflux‐induced carcinogenesis, we first performed comparative genomic hybridization using an Agilent rat 244K array in ESCC‐DR and identified many chromosomal gains and losses. Of the many genes identified, we detected an interesting ezrin amplicon that has been recently reported in human ESCC. Ezrin, which cross‐links the cytoskeleton and plasma membrane, is involved in the growth and metastatic potential of cancer cells. Overexpression of ezrin protein in ESCC‐DR was confirmed by Western blotting. We also compared ezrin protein expression levels and patterns in hyperplastic, dysplastic, ESCC, and metastatic sites developed in two distinct reflux models using immunohistochemistry. Immunohistochemical staining of ezrin revealed overexpression in the nucleus, and the cytoplasm as well as plasma membrane of ESCC cells. Phosphorylated ERM (ezrin, radixin, moesin) was expressed at the leading edge, or invasive front, of larger metastatic sites. Taken together, duodenal reflux has a great potential for initiating malignancy, and thus likely plays a role in development of ESCC. Ezrin probably influences the growth and invasiveness of ESCC cells, and phosphorylation is only required in metastatic behavior of tumor cells at the leading edge and invasive front.
Identification of increased NBS1 expression as a prognostic marker of squamous cell carcinoma of the oral cavity
Authors: Hsu Dennis Shin‐Shian , Chang Shyue‐Yih , Liu Chung‐Ji , Tzeng Cheng‐Hwai , Wu Kou‐Juey , Kao Jung‐Yie , Yang Muh‐Hwa
Published: December 2009

Oral squamous cell carcinoma (OSCC) is one of the most prevalent cancers worldwide; however, accurate molecular markers to predict its prognosis are still limited. We previously demonstrated that overexpression of the DNA double‐strand break repair protein NBS1 is a prognostic marker of advanced head and neck squamous cell carcinoma (HNSCC). Therefore, we aimed to investigate the feasibility of using NBS1 as a biomarker in OSCC. In this study, we enrolled 148 OSCC for immunohistochemical (IHC) and clinical analysis. Data from 58 advanced non‐oral‐cavity HNSCC (NO‐HNSCC) cases were also included for comparison due to the biological and clinical discrepancy between OSCC and HNSCC originated from the other sites (e.g. pharynx or larynx). First, we validated the NBS1 IHC results by real‐time RT‐PCR analysis, and an excellent correlation between the results of these two assays confirmed the reliability and robustness of IHC procedures and interpretation. NBS1 overexpression was an independent prognostic marker in both OSCC and NO‐HNSCC cases. In OSCC, the prognostic significance of NBS1 was shown regardless of T stage and lymph node status. Increased NBS1 expression correlated with advanced T stage and recurrence/metastasis. NBS1 overexpression correlated with the phosphorylation levels of Akt and its downstream target mammalian target of rapamycin (mTOR). These results clearly illustrate the expression profile of NBS1 in OSCC and NO‐HNSCC, and highlight the role of NBS1 in HNSCC irrespective of the primary sites. It also indicates the practicability of application of NBS1 as a marker in OSCC.

(Cancer Sci 2010; 101: 1029–1037)
Subsidence history from a backstripping analysis of the Permo‐Mesozoic succession of the Central Southern Alps (Northern Italy)
Authors: Berra Fabrizio , Carminati Eugenio
Published: December 2009

Seven tectonic subsidence curves, based on outcrop data, have been calculated in order to constrain the geodynamic evolution of the Permian–Mesozoic sedimentary succession (up to 10 km thick) of the Central Southern Alps basin (Italy). The analysis of the tectonic subsidence curves, covering a time span of about 200 Ma, allowed us to quantify the subsidence rates, to document the activity of syndepositional fault systems and calculate their slip rates. Different stages, in terms of duration and magnitude of subsidence‐uplift trends, have been identified in the evolution of the basin. The fault activity, reconstructed by comparing subsidence curves from adjacent sectors, resulted as highly variable both temporally and spatially. Strike‐slip tectonics was coeval to Permian sedimentation, as suggested by the strong differences in the subsidence rates in the sections. The evolution and subsidence rates suggest a continental shelf deposition from Early Triassic to Carnian, when subsidence came to a stop. A rapid resumption of subsidence is observed from the Norian, with a subsidence pulse in the Late Norian, followed by the regional uplift, in the Late Rhaetian. The following Early Jurassic subsidence is characterized by tectonic subsidence similar to that of the Norian. The Norian and Early Jurassic pulses were characterized by the highest slip rates along growth faults and are identified as two distinct tectonic events. The Norian–Rhaetian event is tentatively related to transtensional tectonics whereas the Early Jurassic event is related to crustal extension. The Early Jurassic subsidence records a shift in space an time of the beginning of the extensional stage, from Late Hettangian to the east to Late Pliensbachian–Toarcian to the west. From the Toarcian to the Aptian, the curves are compatible with regional thermal subsidence, later followed (Albian–Cenomanian) by uplift pulses in a retrobelt foreland basin (from Cenomanian onward).
Indispensable function for embryogenesis, expression and regulation of the nonspecific form of the 5‐aminolevulinate synthase gene in mouse
Authors: Okano Satoshi , Zhou Lingyun , Kusaka Toshimasa , Shibata Kazuhide , Shimizu Kazuhiro , Gao Xu , Kikuchi Yuko , Togashi Yoshiyuki , Hosoya Tomonori , Takahashi Satoru , Nakajima Osamu , Yamamoto Masayuki
Published: December 2009

The first step of heme biosynthesis in animals is catalyzed by 5‐aminolevulinate synthase (ALAS), which controls heme supply in various tissues. To clarify the roles that the nonspecific isoform of ALAS (ALAS‐N) plays in vivo, we prepared a green fluorescent protein (GFP) knock‐in mouse line in which the Alas1 gene (encoding ALAS‐N) is replaced with a gfp gene. We found that mice bearing a homozygous knock‐in allele (Alas1GFP/GFP) were lethal by embryonic day 8.5, demonstrating that ALAS‐N is essential for early embryogenesis. Fluorescence microscopic and flow cytometric analyses of heterozygous mouse (Alas1+/GFP) tissues showed that the Alas1 expression level differs substantially in tissues; Alas1 is highly expressed in testis Leydig cells, exocrine glands (including submandibular and parotid glands), endocrine glands (such as adrenal and thyroid glands) and hematopoietic lineage cells (including neutrophils and eosinophils). Quantitative analyses of GFP mRNA and ALAS‐N mRNA in various tissues of Alas1+/GFP mice suggested that the destabilization of ALAS‐N mRNA was not uniform in the various tissues. These results thus lay bare that elaborate control of the endogenous heme supply operates in various mouse tissues through regulation of the ALAS‐N expression level and that this control is essential for heme homeostasis in animals.
Survival of plants of common sainfoin (Onobrychis viciifolia Scop.) in competition with two companion grass species
Authors: Liu Z. , Baines R. N. , Lane G. P. F. , Davies W. P.
Published: December 2009

A 2‐year experiment on competition between sainfoin (Onobrychis viciifolia Scop.), meadow fescue (Festuca pratensis) and tetraploid perennial ryegrass (Lolium perenne) was conducted using plants grown in plastic containers outdoors. Root and shoot systems of sainfoin and the grasses were separated in order to investigate full and no competition of root and shoots, at two planting ratios (0·33 grass:0·66 sainfoin and 0·66 grass:0·33 sainfoin). Survival of sainfoin plants was lower at the higher grass:sainfoin ratio. More sainfoin plants died in the winter than during the growing season. Root competition had no effect on survival of sainfoin plants, but shoot competition reduced survival of sainfoin plants during one growing season. Companion grass species had no effect on survival of sainfoin plants, except in the first winter, when fewer sainfoin plants survived when grown with meadow fescue than perennial ryegrass.
Reseeding Mediterranean dryland cereal fallows using Stipa lagascae R. & Sch.: influence of cutting regime during the establishment phase
Authors: Visser M. , Ouled Belgacem A. , Neffati M.
Published: December 2009

When grass species are used to reseed depleted drylands, grazing is often prescribed during the establishment phase. Total protection from grazing often leads to the presence of persistent weed species and is hard to accept by land users keen to graze reseeded land as soon as possible. The particular case of reseeding arid Mediterranean cereal fallows with one native grass species, Stipa lagascae (Stipa), in Tunisia was tested. Seedlings, derived from two different seed sources (selected and bulk), and volunteer annual weed species underwent four different cutting treatments to ground level (early cut, late cut, early and late cuts and no cuts). No effect of any of the treatments on seedling survival or dry matter mass of Stipa seedlings was found. It was concluded that Stipa seedlings are indifferent to the presence of annual weed species and can tolerate more than one cut during the year of establishment. In the context of the study, early grazing of reseeded fallows is worth investigating further.
Influence of different forage grasses on nitrate capture and leaching loss from a pumice soil
Authors: Popay A. J. , Crush J. R.
Published: December 2009

Nitrogen (N) uptake and loss in leachate from a pulse of 15N‐labelled synthetic cow urine applied to a pumice soil were compared in a glasshouse lysimeter experiment among the grass species, Agrostis capillaris, Dactylis glomerata, Phalaris aquatica, Lolium multiflorum, L. perenne and a L. multiflorum/A. capillaris mixture. In addition, four L. perenne treatments investigated the effect of infection by Neotyphodium lolii strains. Leachate volumes and leachate nitrate concentrations were measured. At final harvest 24 weeks after sowing, dry matter (DM) mass of shoots and roots, plant N and 15N contents, soil residual mineral N, and root diameters and lengths were measured. Endophyte had no effect on plant or nitrate variables. Lysimeters planted with L. perenne (pooled endophyte treatments) leached 48 mg of NO3‐N compared with <3 mg N for the other grasses. Recovery of 15N was highest in A. capillaris (0·99), followed by D. glomerata and P. aquatica (0·89), the L. multiflorum/A. capillaris mixture (0·87), L. multiflorum (0·60) and L. perenne (0·44). Low 15N recoveries and high leaching losses from L. perenne were associated with low plant and root masses of DM and low rooting depth. High aerial mass of DM, root systems extending below 20 cm and high root masses of DM in lysimeters with A. capillaris, P. aquatica and D. glomerata contributed to a high rate of nitrate interception by these species and low leachate losses. The L. multiflorum/A. capillaris mixture was intermediate between the two species for most of the variables measured.
harrow: new Drosophila hAT transposons involved in horizontal transfer
Authors: Mota N. R. , Ludwig A. , Da Silva Valente V. L. , Loreto E. L. S.
Published: December 2009

In this study we characterize the transposable elements harrow, which belong to the hAT superfamily of DNA transposons. Searches for harrow sequences were performed in 65 Drosophilidae species, mainly representing Neotropical and cosmopolitan groups from the genus Drosophila. The nucleotide divergence among elements found in these species suggests that harrow sequences could be clustered in a subfamily. The patchy distribution throughout the genus Drosophila and the high similarity presented between all harrow sequences indicate that horizontal transfer could play a major role in the evolution of harrow elements. The results obtained suggest an evolutionary scenario in which harrow would have undergone multiple horizontal transfer events in the Neotropics, involving D. tripuncatata, D. mojavensis (Subgenus Drosophila) and several species of the willistoni and saltans groups (subgenus Sophophora).
Analysis of Rickettsia typhi‐infected and uninfected cat flea (Ctenocephalides felis) midgut cDNA libraries: deciphering molecular pathways involved in host response to R. typhi infection
Authors: Dreher‐Lesnick S. M. , Ceraul S. M. , Lesnick S. C. , Gillespie J. J. , Anderson J. M. , Jochim R. C. , Valenzuela J. G. , Azad A. F.
Published: December 2009

Murine typhus is a flea‐borne febrile illness that is caused by the obligate intracellular bacterium, Rickettsia typhi. The cat flea, Ctenocephalides felis, acquires R. typhi by imbibing a bloodmeal from a rickettsemic vertebrate host. To explore which transcripts are expressed in the midgut in response to challenge with R. typhi, cDNA libraries of R. typhi‐infected and uninfected midguts of C. felis were constructed. In this study, we examined midgut transcript levels for select C. felis serine proteases, GTPases and defence response genes, all thought to be involved in the fleas response to feeding or infection. An increase in gene expression was observed for the serine protease inhibitors and vesicular trafficking proteins in response to feeding. In addition, R. typhi infection resulted in an increase in gene expression for the chymotrypsin and rab5 that we studied. Interestingly, R. typhi infection had little effect on expression of any of the defence response genes that we studied. We are unsure as to the physiological significance of these gene expression profiles and are currently investigating their potential roles as it pertains to R. typhi infection. To our knowledge, this is the first report of differential expression of flea transcripts in response to infection with R. typhi.
Inferring spatial structure from time‐series data: using multivariate state‐space models to detect metapopulation structure of California sea lions in the Gulf of California, Mexico
Authors: Ward Eric J. , Chirakkal Haridas , González‐Suárez Manuela , Aurioles‐Gamboa David , Holmes Elizabeth E. , Gerber Leah
Published: December 2009

1. Understanding spatial structure and identifying subpopulations are critical for estimating population growth rates and extinction risk, and as such essential for effective conservation planning. However, movement and spatiotemporal environmental data are often unavailable, limiting our ability to directly define subpopulations and their level of asynchrony.

2. This study applies a recently developed statistical technique using time‐series analysis of abundance data to identify subpopulations. The approach uses multivariate state‐space models and Akaike’s Information Criterion‐based model selection to quantify the data support for different subpopulation numbers and configurations. This technique is applied to the population of California sea lionsZalophuscalifornianusin the Gulf of California, Mexico, distributed across 13 breeding sites.

3. The abundance of California sea lions in the Gulf of California has declined over the last decade, though not all areas have been equally affected. In light of this variation, it is important to understand the population structure to ensure accurate viability assessments and effective management.

4. Our data support the hypothesis that the Gulf of California sea lion population has four subpopulations, each with 2–5 breeding sites. The dynamics between several adjacent subpopulations were correlated, suggesting that they experience similar environmental variation. For each subpopulation, we estimated long‐term growth rates, as well as the environmental and observation variation.

5. For most of the subpopulations, our estimates of growth rates were considerably lower than those previously reported. In addition, we found considerable variability across subpopulations in their projected risk of severe decline over the next 50 years.

6. Synthesis and applications. We illustrate a new multivariate state‐space modelling technique that uses time series of abundance to quantify the data support for different subpopulation configurations. Our analysis of the California sea lion population in the Gulf of California indicates that the population is spatially structured into four subpopulations, each exhibiting distinct risks of extinction. Based on our results, we recommend that conservation and management efforts in the Gulf of California focus on the two subpopulations with high probabilities of extinction within the next 50 years (Northern Midriff, Southern Midriff). Multivariate state‐space models provide a practical approach to determine the spatial structure of virtually any species; they may be particularly useful for species of conservation concern for which data on dispersal and environmental drivers are likely to be scarce.
Biogeography and diversification of the Pacific ant genus Lordomyrma Emery
Authors: Lucky Andrea , Sarnat Eli M.
Published: December 2009

Aim  This study addresses the origins of terrestrial biodiversity of the Fijian islands using the ant genus Lordomyrma (Hymenoptera: Formicidae: Myrmicinae) as a model system. We derive the evolution of the genus and determine its closest extra‐Fijian relatives from geological data, molecular phylogenetic reconstruction and divergence estimates.

Location  Ant taxa were sampled in the Southwest Pacific, Melanesia, Southeast Asia, Australia and mainland China.

Methods  Phylogeny and divergence estimates of the ant genus Lordomyrma based on four nuclear genes (28S, ArgK, LW Rh, CAD) plus data on Indo‐Pacific geological history are used to address current hypotheses regarding the origins of the Fijian biota.

Results  The genus Lordomyrma probably originated in mainland Asia, with subsequent colonization of Australia and the Pacific. The Fijian Lordomyrma clade is monophyletic, and originated c. 8.8 Ma, when it diverged from a sister group in Papua New Guinea.

Main conclusions  The colonization of Fiji by Lordomyrma is probably a result of long‐distance dispersal from New Guinea, possibly aided by island hopping across the Vitiaz Arc. The timeline of diversification in Lordomyrma is broadly congruent with the Miocene fragmentation of the Vitiaz Arc and the Pliocene emergence of Vanua Levu. The biotic shuttle hypothesis, which posits ‘Eua Island as the source of Fijian endemics, is rejected based on the sister relationship of Fiji and New Guinea lineages, as well as on the Miocene submergence of the terrane below sea level. The diversity of Fijian Lordomyrma results from the radiation of a single lineage, which diverged from a New Guinea sister group. The genus appears to have originated in Asia rather than in Australia.
Developmental Iodine Deficiency and Hypothyroidism Impair Neural Development, Up‐Regulate Caveolin‐1 and Down‐Regulate Synaptophysin in Rat Hippocampus
Authors: Gong J. , Dong J. , Wang Y. , Xu H. , Wei W. , Zhong J. , Liu W. , Xi Q. , Chen J.
Published: December 2009

Developmental iodine deficiency leads to inadequate thyroid hormone, which damages the hippocampus. In the present study, we implicate hippocampal caveolin‐1 and synaptophysin in developmental iodine deficiency and hypothyroidism. Two developmental rat models were established: pregnant rats were administered either an iodine‐deficient diet or propylthiouracil (PTU)‐adulterated (5 p.p.m. or 15 p.p.m.) drinking water from gestational day 6 until postnatal day (PN) 28. Nissl staining and the levels of caveolin‐1 and synaptophysin in several hippocampal subregions were assessed on PN14, PN21, PN28 and PN42. The results obtained show that surviving cells in the iodine‐deficient and PTU‐treated rats were lower than in controls. Up‐regulation of caveolin‐1 and down‐regulation of synaptophysin were observed in the iodine‐deficient and PTU‐treated rats. Our findings implicate decreases in the number of surviving cells and alterations in the levels of caveolin‐1 and synaptophysin in the impairments in neural development induced by developmental iodine deficiency and hypothyroidism.
A real‐time (TaqMan) PCR assay to differentiate Monilinia fructicola from other brown rot fungi of fruit crops
Authors: Van Brouwershaven I. R. , Bruil M. L. , Van Leeuwen G. C. M. , Kox L. F. F.
Published: December 2009

To prevent the entry and spread of the brown rot fungus Monilinia fructicola in Europe, a fast and reliable method for detection of this organism is essential. In this study, an automated DNA extraction method combined with a multiplex real‐time PCR based on TaqMan chemistry was developed for fast, convenient and reliable detection of both the EU quarantine organism Monilinia fructicola and the three other brown rot fungi M. fructigena, M. laxa and Monilia polystroma. Using the internal transcribed spacer (ITS) region of the nuclear ribosomal RNA gene repeat, a Monilinia genus‐specific primer pair and two differently labelled fluorogenic probes specific for M. fructicola and the group M. fructigena/M. laxa/Monilia polystroma were developed. The analytical specificity of the assay was assessed by testing 33 isolates of the four brown rot fungi and 13 isolates of related fungal species or other fungal species that can be present on stone and pome fruit. No cross‐reactions were observed. The assay was found to have a detection limit of 0·6 pg of DNA, corresponding to 27 haploid genomes or four conidia. Comparison of a manual DNA isolation followed by a conventional PCR with an automated DNA isolation combined with the presently developed real‐time PCR showed that the latter method gave improved results when tested with 72 naturally infected stone fruit samples. The detection rate increased from 65 to 97%.
Striking Activation of NALT and Nasal Passages Lymphocytes Induced by Intranasal Immunization with Cry1Ac protoxin
Authors: Rodriguez‐Monroy M. A. , Moreno‐Fierros L.
Published: December 2009

Cry1Ac protoxin from Bacillus thuringiensis is a potent mucosal immunogen and adjuvant. When delivered intranasally (i.n.) Cry1Ac elicits significant antibody response and is able to improve vaccination against Naegleria fowleri infection, but the functional effects occurring in nasal lymphocytes when this protein is administered alone have not been determined. Here, we investigated the effects of i.n. immunization with Cry1Ac on antibody production, lymphocyte activation and cytokine production in lymphocytes from nasal‐associated lymphoid tissue (NALT) and nasal passages (NP). Our results show that i.n. immunization with Cry1Ac induced significant specific IgA and IgG cell responses, especially in NP. Besides, it increased the proportion of lymphocytes expressing the activation markers CD25 and CD69 in both nasal tissues, but differently. CD25 was increased in B cells along with CD4 and CD8 T cells from NALT and NP, while CD69 was increased in B cells from both tissues but only in CD4 T cells from NP. Finally, we found that Cry1Ac augmented especially a Th2 profile of cytokines, as the proportion of T cells that spontaneously produced IL‐4, IL‐5 and IL‐10 was increased and this effect was higher in NP than in NALT. These data contribute to explain the potent immunogenicity of Cry1Ac via i.n. route.
Antitumour Response of a Double Mutant of Staphylococcal Enterotoxin C2 with the Decreased Affinity for MHC Class II Molecule
Authors: Cheng X. , Cao P. , Ji X. , Lu W. , Cai X. , Hu C. , Wang Z. , Zhang S.
Published: December 2009

Staphylococcal enterotoxin C2 (SEC2) is one of the most potent known activators of human T lymphocytes, and recombinant SEC2 shows promising clinical values, but SEC2 can cause food poisoning and toxic shock syndrome in vivo. In this study, site‐directed mutagenesis has been used to introduce alanine substitutions at Phe144 and Leu45 in the molecule. The mutant genes were cloned and expressed, and the corresponding proteins were purified by nickel agarose affinity chromatography. We found that the SEC2 mutant proteins could stimulate the proliferation of human peripheral blood lymphocytes and inhibit the growth of tumour cells as native SEC2. Furthermore, flow cytometry assay showed that mSEC2(F44A, L45A) drastically reduced the ability of the toxin to bind to MHC class II. Physiological parameters revealed that mSEC2(F44A, L45A) reduced significantly rat temperature compared with native SEC2 in vivo. Our results clearly suggest that this genetically modified SEC2 protein is less toxic and justifies its further development as a new, safer antitumour superantigen to prevent SEC2 intoxication.
IL‐23 in the Pathogenesis of Rheumatoid Arthritis
Authors: Paradowska‐Gorycka A. , Grzybowska‐Kowalczyk A. , Wojtecka‐Lukasik E. , Maslinski S.
Published: December 2009

Interleukin‐23 (IL‐23) is a heterodimeric cytokine belonging to the IL‐6/IL‐12 family that plays a key role in several of autoimmune and inflammatory disorders. This family contains the 34 type I cytokine receptor chains and 27 ligands, which share structural and functional similarities, but on the other hand they display distinct roles in shaping Th cells responses. IL‐12 family cytokines have not only proinflammatory effects but they also promote inflammatory responses. IL‐23 is composed of the p40 subunit in common with IL‐12, and with a unique p19 subunit. IL‐23 binding to an IL‐23 receptor expressed on dendritic cells, macrophages and monocytes triggers the activation of Jak2 and Tyk2, which in turn phosphorylates STAT1, STAT3, STAT4 and STAT5 as well as induce formation of STAT3‐STAT4 heterodimers. IL‐23 is one of the essential factors required for the survival and/or expansion of Th17 cells, which produce IL‐17, IL‐17F, IL‐6 and TNF‐α. Th17 cells stimulated by the IL‐23 promote osteoclastogenesis through production of IL‐17, which induce receptor activator of NF‐kappa B ligand on mesenchymal cells. The IL‐23‐IL‐17 axis includes Th17 cells and plays a key role in the development of autoimmune arthritis.
Immune surveillance of human cancer: if the cytotoxic T‐lymphocytes play the music, does the tumoral system call the tune?
Authors: Hamaï A. , Benlalam H. , Meslin F. , Hasmim M. , Carré T. , Akalay I. , Janji B. , Berchem G. , Noman M. Z. , Chouaib S.
Published: December 2009

Abstract

Accumulating evidence indicates that the innate and adaptive immune systems participate in the recognition and destruction of cancer cells by a process known as cancer immunosurveillance. Tumor antigen‐specific cytotoxic T‐lymphocytes (CTL) are the major effectors in the immune response against tumor cells. The identification of tumor‐associated antigen (TAA) recognized primarily by CD 8+ T‐lymphocytes has led to the development of several vaccination strategies that induce or potentiate specific immune responses. However, large established tumors, which are associated with the acquisition of tumor resistance to specific lysis, are usually not fully controlled by the immune system. Recently, it has become clear that the immune system not only protects the host against tumor development but also sculpts the immunogenic phenotype of a developing tumor and can favor the emergence of resistant tumor cell variants. Moreover, it has become obvious that the evasion of immunosurveillance by tumor cells is under the control of the tumor microenvironment complexity and plasticity. In this review, we will focus on some new mechanisms associated with the acquisition of tumor resistance to specific lysis during tumor progression, involving genetic instability, structural changes in cytoskeleton, and hypoxic stress. We will also discuss the interaction between CTLs and tumor endothelial cells, a major component of tumor stroma.
Relationship between osteology and aquatic locomotion in birds: determining modes of locomotion in extinct Ornithurae
Authors: HINIĆ‐FRLOG S. , MOTANI R.
Published: December 2009

The evolutionary history of aquatic invasion in birds would be incomplete without incorporation of extinct species. We show that aquatic affinities in fossil birds can be inferred by multivariate analysis of skeletal features and locomotion of 245 species of extant birds. Regularized discriminant analyses revealed that measurements of appendicular skeletons successfully separated diving birds from surface swimmers and flyers, while also discriminating among different underwater modes of swimming. The high accuracy of this method allows detection of skeletal characteristics that are indicative of aquatic locomotion and inference of such locomotion in bird species with insufficient behavioural information. Statistical predictions based on the analyses confirm qualitative assessments for both foot‐propelled (Hesperornithiformes) and wing‐propelled (Copepteryx) underwater locomotion in fossil birds. This is the first quantitative inference of underwater modes of swimming in fossil birds, enabling future studies of locomotion in extinct birds and evolutionary transitions among locomotor modes in avian lineage.
A cancer‐causing gene is positively correlated with male aggression in Xiphophorus cortezi
Authors: FERNANDEZ A. A.
Published: December 2009

The persistence of seemingly maladaptive genes in organisms challenges evolutionary biological thought. In Xiphophorus fishes, certain melanin patterns form malignant melanomas because of a cancer‐causing gene (Xiphophorus melanoma receptor kinase; Xmrk), which arose several millions years ago from unequal meiotic recombination. Xiphophorus melanomas are male biased and induced by androgens however male behaviour and Xmrk genotype has not been investigated. This study found that male X. cortezi with the spotted caudal (Sc) pattern, from which melanomas originate, displayed increased aggression in mirror image trials. Furthermore, Xmrk males (regardless of Sc phenotype) bit and performed more agonistic displays than Xmrk deficient males. Male aggressive response decreased when males viewed their Sc image as compared with their non‐Sc image. Collectively, these results indicate that Xmrk males experience a competitive advantage over wild‐type males and that intrasexual selection could be an important component in the evolutionary maintenance of this oncogene within Xiphophorus.
Paternal effects on seed mass in Arabidopsis thaliana
Authors: De Jong T. J. , Hermans C. M. , Van Der Veen‐van Wijk K. (C.) A. M.
Published: December 2009

Who is in control of seed size, and do some fathers sire bigger seeds than others? We used isogenic male‐sterile genotypes of the Arabidopsis thaliana accessions Col and Ler. By fertilising flowers side‐by‐side with either pollen from the same accession (‘self‐pollination’) or pollen from another accession (outcrossing), we compared, on the same mother plant, seed set of flowers that were very similar in resource status. Some paternal genotypes had a significant effect on seed mass, with the most extreme father siring seeds 15.3% heavier than seeds resulting from ‘self‐pollination’. There was no correlation between seed mass of paternal parents and the seeds they sired. We discuss the evolution of seed size as a tug‐of‐war between parent and offspring.
Fitness costs of Cry1F resistance in laboratory‐selected European corn borer (Lepidoptera: Crambidae)
Authors: Pereira E. J. G. , Storer N. P. , Siegfried B. D.
Published: December 2009

The study of fitness costs associated with resistance to toxins fromBacillus thuringiensisBerliner (Bt) is important for understanding resistance evolution and for evaluating resistance management practices that prevent or mitigate resistance. A strain of European corn borer,Ostrinia nubilalis(Hübner) (Lepidoptera: Crambidae) obtained from field collections throughout the U.S. Corn Belt in 1996, was selected in the laboratory for resistance to Cry1F by exposure to the toxin incorporated into artificial diet. The selected strain developed more than 3000‐fold resistance to Cry1F after 35 generations of selection and readily consumed Cry1F expressing maize tissue. Using this resistant strain, a susceptible strain with similar genetic background and reciprocal crosses between them, we estimated fitness costs and their dominance by measuring fitness components and population parameters determined by fertility life tables. Comparison of life‐history traits and population growth rates of genotypes homozygous and heterozygous for resistance relative to susceptible genotypes indicated existence of weak and recessive fitness costs associated with resistance. The significance of these results in relation to current resistance management practices is discussed.
Interrater, intra‐rater and internal consistency reliability of the Theory Application Assessment Instrument
Authors: Ikiugu Moses N. , Smallfield Stacy
Published: December 2009

Aim:  To investigate the interrater, intra‐rater and internal consistency reliability of the Theory Application Assessment Instrument, a tool for testing competency in combining multiple theoretical conceptual practice models in client evaluation and treatment planning.

Methods:  The assessment was administered to 27 graduate students in two phases. The principal investigator rated the participants on the assessment scale on two separate occasions.

Results:  Interrater and intra‐rater reliability were good for Phase 2 data. Internal consistency reliability for Phase 2 data was in the ‘questionable’ range.

Conclusion:  The instrument shows promise of being an efficient tool. However, this was an exploratory study with a small sample. Its replication with a larger more representative sample is needed to increase external validity.
Happy and healthy only if occupied? Perceptions of health sciences students on occupation in later life
Authors: Zecevic Aleksandra , Magalhaes Lilian , Madady Mona , Halligan Michelle , Reeves Alicia
Published: December 2009

Background/aim: In this study, we bring attention to the university education of health science students with respect to occupation in later life. Our goal was to provide descriptive data from narratives of a group of undergraduate students and initiate discussion about the place of occupation in the context of ageing to answer the following questions: (i) How young people perceive successful ageing in relation to occupation? and (ii) can spirituality‐related activities be considered occupations in later life?

Methods: Based on a thematic selection, the quality of photographs and reflective narratives, 60 Photovoice assignments created by health sciences students were analysed using content analysis.

Results: The findings of this study indicate that students seem to neglect the benefits of ‘being’ through spiritual engagement, and instead emphasise the importance of ‘doing’, and perpetuate pervasive successful ageing discourses in Western societies.

Conclusions: Occupational therapists have potential to take an active role in undergraduate health science education and to inform the development of holistic models that would include spirituality as an avenue to live late life to its fullest potential. Photovoice emerged as a powerful teaching method to increase awareness, empathy and compassion of young adults towards ageing.
Repeated stent thrombosis after DES implantation and localized hypersensitivity to a stent implanted in the distal portion of a coronary aneurysm thought to be a sequela of Kawasaki disease: Autopsy report
Authors: Yokouchi Yuki , Oharaseki Toshiaki , Ihara Fumie , Naoe Shiro , Sugawara Shigetada , Takahashi Kei
Published: December 2009

The patient was a 40‐year‐old Japanese woman. At 37 years of age she underwent stent implantation in LAD#7 for an acute myocardial infarction. Subsequently, coronary intervention was performed four times because of occlusion of the stent. Sudden death occurred at 40 years of age due to ventricular tachycardia. Clinically, the patient had had no history of collagen disease, anti‐phospholipid antibody syndrome or coagulation disorder. The autopsy revealed only very mild atherosclerotic changes in the aorta and various other organs, but concentric thickening of the intima was observed in all three branches of the coronary arteries. Also, aneurysms accompanied by calcification were observed at each of LAD #6, LCx #11 and RCA #4PD. The stent was occluded with a thrombus, and the vascular walls showed infiltration by lymphocytes, plasma cells and numerous eosinophils. The eosinophil infiltration was confined to the site of the stent. It was surmised that the patient had experienced late stent thrombosis due to a hypersensitivity reaction to the DES on the basis of a development of a state of high susceptibility to thrombus formation because of a coronary aneurysm. The aneurysm was suspected of being a post‐inflammatory change of Kawasaki disease.
USEFULNESS OF DOUBLE BALLOON ENTEROSCOPY AND VIDEO CAPSULE ENDOSCOPY FOR THE DIAGNOSIS AND MANAGEMENT OF PRIMARY FOLLICULAR LYMPHOMA OF THE GASTROINTESTINAL TRACT IN ITS EARLY STAGES
Authors: Akamatsu Taiji , Kaneko Yasunori , Ota Hiroyoshi , Miyabayashi Hideharu , Arakura Norikazu , Tanaka Eiji
Published: December 2009

Aim:  The aim of this study is to evaluate the usefulness of double balloon enteroscopy (DBE) and video capsule endoscopy (VCE) in patients with primary follicular lymphoma (FL) of the gastrointestinal (GI) tract. Furthermore, we estimate the effectiveness of chemotherapy (cyclophosphamide, doxorubicin, vincristine, and prednisone) including rituximab for them.

Methods:  Thirteen consecutive patients who were diagnosed of having FL in the duodenum between July 2005 and September 2008 were studied. All patients were given the conventional staging examinations, including total enteroscopy using DBE and/or VCE procedures. Chemotherapy was performed after written informed consent. Response assessment was performed every 6‐12 months. The median follow‐up period was 30.2 months.

Results:  FL was diagnosed in each patient as low grade (grade 1, n = 7; 2, n = 6) and, in all but 4 patients, localized lymphoma (stage I, n = 8; II1, n = 1; II2, n = 4). DBE revealed multifocal lesions in the jejunum in 10 of the patients, and in the ileum in 6. VCE showed similar findings in the jejunum in the recent 2 patients. Eleven of 13 patients finally received chemotherapy, and all of them achieved complete regression. They showed no evidence of recurrence after that.

Conclusion:  Total examination of the small intestine using DBE should be performed before treatment to choose a suitable treatment procedure for primary FL of the GI tract. On the other hand, VCE is useful for screening and following the small intestine in the patients with it. Chemotherapy is effective to achieve complete regression of primary FL of the GI tract.
VALIDITY OF ACTIVITY INDICES IN ULCERATIVE COLITIS: COMPARISON OF CLINICAL AND ENDOSCOPIC INDICES
Authors: Hirai Fumihito , Matsui Toshiyuki , Aoyagi Kunihiko , Inoue Nagamu , Hibi Toshifumi , Oshitani Nobuhide , Fujii Hisao , Kobayashi Kiyonori , Suzuki Yasuo , Tanaka Shinji
Published: December 2009

Aim:  Various different clinical indices (CI) and endoscopic indices (El) have been used to evaluate the clinical disease activity of ulcerative colitis (UC). However, no standardized CI or El of UC has been established. The aim of the present study was to determine whether or not any of the CI and/or EI for assessing the disease activity of UC could be established as a standard.

Methods:  The most frequently used CI and EI were identified from 100 published clinical trials dealing with UC, and representative CI and El were selected. Seventy‐four patients were enrolled in this study and their CI and El were assessed prior to treatment and at 2, 4 and 8 weeks after treatment. Furthermore, changes over time and relationships among the indices were analyzed. In this study, the clinical activity index (CAI), the disease activity index (DAI), the Lichtiger index (LI) and the Seo index were selected as the representative CI, and the Baron score and the Rachmilewitz endoscopic index (REI) were selected as the representative EI.

Results:  A significant decrease in all the CI and El was observed after treatment, as compared with the baseline values. Moreover, there were positive relationships among the CI and between the CI and El.

Conclusion:  Our results demonstrated that all the CI and El examined were almost equally useful for evaluating disease activity in UC patients. Further studies may help to determine which of the indices is the most suitable for use in UC clinical trials.