bluuebunny/crossref_metadata_2025 huggingface dataset:
Pathogenesis of Autographa californica multiple nucleopolyhedrovirus in fifth-instar Anticarsia gemmatalis larvae
Authors: Chikhalya Aniska , Luu Dee Dee , Carrera Maggie , De La Cruz Alisa , Torres Marianne , Martinez Elisa N. , Chen Tiffany , Stephens Kimberly D. , Haas-Stapleton Eric J.
Published: July 2009

We have investigated infection and pathogenesis of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in Anticarsia gemmatalis (velvetbean caterpillar) larvae using a lacZ recombinant virus (AcMNPV-hsp70/lacZ) to track the temporal progression of infection in the midgut intestine and haemocoel. A. gemmatalis was highly resistant to fatal infection by occlusion bodies (OBs; LD50>5.5×105 OB) and budded virus (BV; LD50>3×105 BV) administered via oral and systemic routes, respectively. Orally administered occlusion-derived virus (ODV) efficiently attached and fused to midgut cells; however, high levels of infection-induced apoptosis limited infection in the midgut. Transcriptional analysis of AcMNPV genes expressed in the midgut of OB-inoculated A. gemmatalis larvae showed high levels of mRNA encoding the major capsid protein VP39 in the absence of immediate-early transactivator 1 (ie-1) expression. In the midgut, virus was efficiently transferred from infected midgut epithelial cells to nearby tracheolar cells and circulating haemocytes to initiate systemic infection in the haemocoel. However, haemocoelic BV did not efficiently disseminate infection and only cuticular epidermal cells displayed high levels of viral infection. Flow cytometry analysis of haemocytes isolated from BV-inoculated A. gemmatalis larvae showed low-level expression of the BV envelope protein GP64 on the cell surface, suggesting that A. gemmatalis haemocytes have a limited capacity for amplifying virus. These results show that AcMNPV is not an effective biological control agent for limiting crop damage caused by A. gemmatalis larvae.
Human immunodeficiency virus type 1 Vpr: functions and molecular interactions
Authors: Romani Bizhan , Engelbrecht Susan
Published: July 2009

Human immunodeficiency virus type 1 (HIV-1) viral protein R (Vpr) is an accessory protein that interacts with a number of cellular and viral proteins. The functions of many of these interactions in the pathogenesis of HIV-1 have been identified. Deletion of thevprgene reduces the virulence of HIV-1 dramatically, indicating the importance of this protein for the virus. This review describes the current findings on several established functions of HIV-1 Vpr and some possible roles proposed for this protein. Because Vpr exploits cellular proteins and pathways to influence the biology of HIV-1, understanding the functions of Vpr usually involves the study of cellular pathways. Several functions of Vpr are attributed to the virion-incorporated protein, but some of them are attributed to the expression of Vpr in HIV-1-infected cells. The structure of Vpr may be key to understanding the variety of its interactions. Due to the critical role of Vpr in HIV-1 pathogenicity, study of the interactions between Vpr and cellular proteins may help us to understand the mechanism(s) of HIV-1 pathogenicity.
Weighted parsimony phylogeny of the family Characidae (Teleostei: Characiformes)
Authors: Marcos Mirande J.
Published: July 2009

The family Characidae, including more than 1000 species, lacks a phylogenetic diagnosis, with many of its genera currently considered as incertae sedis. The aims of the present study are to propose a phylogenetic diagnosis and to assess higher‐level relationships of and within Characidae. In this regard, 360 morphological characters are studied for 160 species of Characidae and related families. Phylogenetic analyses under implied weighting and self‐weighted optimization are presented, exploring a broad range of parameters. The analysis under self‐weighted optimization is innovative for this size of matrices. Familial status of Serrasalmidae is supported, and Acestrorhynchidae and Cynodontidae are included in a monophyletic Characidae. Engraulisoma taeniatum is transferred from Characidae to Gasteropelecidae. Thus constituted, the monophyly of Characidae is supported by seven synapomorphies. A new subfamily, Heterocharacinae, is proposed, and the subfamilies Aphyocharacinae, Aphyoditeinae, Characinae, Gymnocharacinae, and Stevardiinae are redefined. The Glandulocaudinae are included in Stevardiinae together with remaining members of “clade A” (sensuMalabarba and Weitzman, 2003. Comun. Mus. Ciênc. Tecnol. PUCRS, Sér. Zool. 16, 67–151.) and the genera Aulixidens and Nantis. Most incertae sedis genera are assigned, at least tentatively, to a phylogenetically diagnosed clade.
Expression of Ret finger protein correlates with outcomes in endometrial cancer
Authors: Tsukamoto Hirohisa , Kato Takuya , Enomoto Atsushi , Nakamura Nobuhisa , Shimono Yohei , Jijiwa Mayumi , Asai Naoya , Murakumo Yoshiki , Shibata Kiyosumi , Kikkawa Fumitaka , Takahashi Masahide
Published: July 2009

Ret finger protein (RFP) is a nuclear protein with transcriptional repressive activity that is highly expressed in a variety of human and rodent tumor cell lines. We examined the expression of RFP in human endometrial cancer and assessed its clinical significance. Formalin‐fixed, paraffin‐embedded sections from endometrial cancer tissues were immunostained with the RFP antibody, and the staining intensity was evaluated. The clinicopathological factors examined were age, International Federation of Gynecology and Obstetrics stage, tumor grade, myometrial invasion, and pelvic lymph node metastasis. Overall survival (OS) and progression‐free survival (PFS) were evaluated using the Kaplan–Meier method, and multivariate analysis was performed using the Cox proportional hazard analysis. Of the 119 cancer tissues, 57 (47.9%) cases were positive for RFP immunoreactivity. RFP expression was not associated with any of the clinicopathological parameters examined. However, positive RFP expression significantly predicted poorer OS and PFS compared with negative expression (OS, P = 0.0011; PFS, P < 0.0001). In the multivariate analyses, positive RFP expression was an independent prognostic factor for survival in this study. RFP knockdown significantly impaired cancer cell migration and invasion in vitro with concomitant decreases of integrins β1 and α2. Positive RFP expression is a predictive marker for an unfavorable clinical outcome in patients with endometrial cancer. (Cancer Sci 2009; 100: 1895–1901)
Evaluation of radioiodinated vesamicol analogs for sigma receptor imaging in tumor and radionuclide receptor therapy
Authors: Ogawa Kazuma , Shiba Kazuhiro , Akhter Nasima , Yoshimoto Mitsuyoshi , Washiyama Kohshin , Kinuya Seigo , Kawai Keiichi , Mori Hirofumi
Published: July 2009

It has been reported that sigma receptors are highly expressed in a variety of human tumors. In this study, we selected (+)‐2‐[4‐(4‐iodophenyl)piperidino] cyclohexanol [(+)‐pIV] as a sigma receptor ligand and evaluated the potential of radioiodinated (+)‐pIV for tumor imaging and therapy. (+)‐[125/131I]pIV was prepared by an iododestannylation reaction under no‐carrier‐added conditions with radiochemical purity over 99% after HPLC purification. Biodistribution experiments were performed by the intravenous injection of (+)‐[125I]pIV into mice bearing human prostate tumors (DU‐145). Blocking studies were performed by intravenous injection of (+)‐[125I]pIV mixed with an excess amount of unlabeled sigma ligand into DU‐145 tumor‐bearing mice. For therapeutic study, (+)‐[131I]pIV was injected at a dose of 7.4 MBq followed by measurement of the tumor size. In biodistribution experiments, (+)‐[125I]pIV showed high uptake and long residence in the tumor. High tumor to blood and muscle ratios were achieved because the radioactivity levels of blood and muscle were low. However, the accumulations of radioactivity in non‐target tissues, such as liver and kidney, were high. The radioactivity in the non‐target tissues slowly decreased over time. Co‐injection of (+)‐[125I]pIV with an excess amount of unlabeled sigma ligand resulted in a significant decrease in the tumor/blood ratio, indicating sigma receptor‐mediated tumor uptake. In therapeutic study, tumor growth in mice treated with (+)‐[131I]pIV was significantly inhibited compared to that of an untreated group. These results indicate that radioiodinated (+)‐pIV has a high potential for sigma receptor imaging in tumor and radionuclide receptor therapy. (Cancer Sci 2009)
Prolonged low‐dose administration of the cyclooxygenase‐2 inhibitor celecoxib enhances the antitumor activity of irinotecan against neuroblastoma xenografts
Authors: Kaneko Michio , Kaneko Setsuko , Suzuki Kenshi
Published: July 2009

Cyclooxygenase (COX)‐2 is overexpressed in many human tumors including neuroblastoma (NB) and promotes tumor progression. We evaluated the antitumor effect of irinotecan (CPT‐11) treatment combined with prolonged very low‐dose administration of celecoxib, a selective COX‐2 inhibitor, against three human NB xenografts, TNB9, TS‐N‐2nu, and TS‐N‐5nu. In addition, the effects of the celecoxib‐combined treatment were examined on tumor cell proliferation, apoptosis, angiogenesis, and expression of vascular endothelial growth factor and apoptosis‐related proteins in xenografts. Celecoxib administered daily at 5 mg/kg body weight/day could not prevent the growth of any of the NB xenografts. However, the combination of daily low‐dose CPT‐11 (5.9 mg/kg body weight/day) and simultaneous very low‐dose celecoxib resulted in highly significant suppression of tumor growth in all three xenografts (P < 0.001) compared not only with low‐dose CPT‐11 therapy alone but also with the combination therapy of intermittent conventional‐dose CPT‐11 (59 mg/kg body weight) and celecoxib accompanied by decreased proliferation and increased induction of apoptosis in tumor cells. Induction of apoptosis by CPT‐11 with and without celecoxib was associated with the up‐regulation of Bax expression and the down‐regulation of Bcl‐2 expression. The enhanced antitumor effect of the combination of the two drugs against the NB xenografts might be partially COX‐2‐independent and was probably mediated through multiple factors including diminished expression of VEGF and activation of the caspase‐dependent mitochondrial apoptosis pathway. These findings demonstrate that prolonged low‐dose CPT‐11 treatment combined with very low‐dose celecoxib shows promising antitumor activity through the blockage of multiple critical targets related to NB tumor cell survival and proliferation. (Cancer Sci 2009; 00: 000–000)
Hepatocellular carcinoma development induced by conditional β‐catenin activation in Lkb1+/− mice
Authors: Miyoshi Hiroyuki , Deguchi Atsuko , Nakau Masayuki , Kojima Yasushi , Mori Akira , Oshima Masanobu , Aoki Masahiro , Taketo Makoto Mark
Published: July 2009

The development of hepatocellular carcinomas (HCC) appears to be a multistep process that takes several decades in humans. However, the identities of specific gene alterations and their contribution to HCC pathogenesis remain poorly understood. We previously reported that Lkb1+/− mice spontaneously develop multiple hepatic nodular foci (NdFc) followed by HCC, and that the conditional activation of β‐catenin in Catnblox(ex3) mouse livers alone does not cause tumor formation. We show here that the conditional activation of β‐catenin accelerates HCC development in Catnb+/lox(ex3)Lkb1+/− compound mutant mice, affecting displastic hepatocytes in NdFc that suffered LOH at the Lkb1 locus. We further show that β‐catnin activation provides HCC with a growth advantage as well as transplantability. These results suggest that the loss of Lkb1 contributes to the formation of dysplastic NdFc, and that Wnt signaling activation is involved in ensuing progression toward HCC. A combination of these sequential changes can be a practical model for a subset of human HCC. (Cancer Sci 2009)
Systemic acquired resistance is induced by R gene‐mediated responses independent of cell death
Authors: LIU PO‐PU , BHATTACHARJEE SAIKAT , KLESSIG DANIEL F. , MOFFETT PETER
Published: July 2009

On infection by pathogens, plants initiate defence responses that are able to curtail infection locally. These responses are mediated either by receptor‐like proteins that recognize pathogen‐associated molecular patterns or by the protein products of disease resistance (R) genes. At the same time, primary defence responses often result in the generation of signals that induce what is known as systemic acquired resistance (SAR), such that defence responses are enhanced on secondary pathogen challenge in distal tissues. R protein‐mediated SAR induction is normally accompanied by a type of programmed cell death known as the hypersensitive response (HR) and, in some instances, cell death alone has been implicated in the induction of SAR. This has raised the question of whether R protein‐mediated signalling per se induces SAR or whether SAR is an indirect result of the induction of HR. Using the Rx gene of potato, which confers resistance to Potato Virus X in the absence of cell death, we have shown that the HR is dispensable for R protein‐mediated induction of SAR and that Rx‐induced SAR is mediated by the same salicylate‐dependent pathway induced by other R proteins.
Tobacco blue mould disease caused by Peronospora hyoscyami f. sp. tabacina
Authors: BORRÁS‐HIDALGO ORLANDO , THOMMA BART P. H. J. , SILVA YUSSUAN , CHACÓN OSMANY , PUJOL MERARDO
Published: July 2009

Blue mould [Peronospora hyoscyami f. sp. tabacina (Adam) Skalicky 1964] is one of the most important foliar diseases of tobacco that causes significant losses in the Americas, south‐eastern Europe and the Middle East. This review summarizes the current knowledge of the mechanisms employed by this oomycete pathogen to colonize its host, with emphasis on molecular aspects of pathogenicity. In addition, key biochemical and molecular mechanisms involved in tobacco resistance to blue mould are discussed.

Taxonomy: Kingdom: Chromista (Straminipila); Phylum: Heterokontophyta; Class: Oomycete; Order: Peronosporales; Family: Peronosporaceae; Genus: Peronospora; Species: Peronospora hyoscyami f. sp. tabacina.

Disease symptoms: The pathogen typically causes localized lesions on tobacco leaves that appear as single, or groups of, yellow spots that often coalesce to form light‐brown necrotic areas. Some of the leaves exhibit grey to bluish downy mould on their lower surfaces. Diseased leaves can become twisted, such that the lower surfaces turn upwards. In such cases, the bluish colour of the diseased plants becomes quite conspicuous, especially under moist conditions when sporulation is abundant. Hence the name of the disease: tobacco blue mould.

Infection process: The pathogen develops haustoria within plant cells that are thought to establish the transfer of nutrients from the host cell, and may also act in the delivery of effector proteins during infection.

Resistance: Several defence responses have been reported to occur in the Nicotiana tabacum–P. hyoscyami f. sp. tabacina interaction. These include the induction of pathogenesis‐related genes, and a correlated increase in the activities of typical pathogenesis‐related proteins, such as peroxidases, chitinases, β‐1,3‐glucanases and lipoxygenases. Systemic acquired resistance is one of the best characterized tobacco defence responses activated on pathogen infection.
Experimental analysis and modelling of in vitro proliferation of mesenchymal stem cells
Authors: Mancuso L. , Liuzzo M. I. , Fadda S. , Pisu M. , Cincotti A. , Arras M. , Desogus E. , Piras F. , Piga G. , La Nasa G. , Concas A. , Cao G.
Published: July 2009

Objectives:  Stem cell therapies based on differentiation of adult or embryonic stem cells into specialized ones appear to be effective for treating several human diseases. This work addresses the mathematical simulation of proliferation kinetics of stem cells.

Materials and methods:  Sheep bone marrow mesenchymal stem cells (phenotype characterized by flow cytometry analysis) seeded at different initial concentrations in Petri dishes were expanded to confluence. Sigmoid temporal profiles of total counts obtained through classic haemocytometry were quantitatively interpreted by both a phenomenological logistic equation and a novel model based on a one‐dimensional, single‐staged population balance approach capable of taking into account contact inhibition at confluence. The models’ parameters were determined by comparison with experimental data on population expansion starting from single seeding concentration. Reliability of the models was tested by predicting cell proliferation carried out starting from different seeding concentrations.

Results and discussion:  It was found that the proposed population balance modelling approach was successful in predicting the experimental data over the whole range of initial cell numbers investigated, while prediction capability of phenomenological logistic equation was more limited.
Gender differences in pulmonary regenerative response to naphthalene‐induced bronchiolar epithelial cell injury
Authors: Oliver J. R.  , Kushwah R.  , Wu J.  , Cutz E.  , Yeger H.  , Waddell T. K.  , Hu J. 
Published: July 2009

Objectives: Accumulating evidence suggests that gender affects the incidence and severity of several pulmonary diseases. Previous studies on mice have shown gender differences in susceptibility to naphthalene‐induced lung injury, where Clara cell damage was found to occur earlier and to be more extensive in females than in males. However, very little is known about whether there are any gender differences in subsequent lung repair responses. The aim of this study was to investigate whether gender plays an important role in pulmonary regenerative response to naphthalene‐induced Clara cell ablation.

Materials and methods: Adult male and female mice were injected with a low, medium, or high dose of naphthalene, and lung tissue regeneration was examined by immunohistochemical staining for cell proliferation marker (Ki‐67) and mitosis marker (phosphohistone‐3).

Results: Histopathological analysis showed that naphthalene‐induced Clara cell necrosis was more prominent in the lungs of female mice as compared to male mice. Cell proliferation and mitosis in both the distal bronchiolar airway epithelium and peribronchiolar interstitium of female mice was significantly greater than that of male mice after treatment with the low and medium doses. However, after treatment with high dose of naphthalene, lung regeneration was delayed in female mice, while male mice mounted a timely regenerative response.

Conclusions: These findings show that there are clear gender differences in naphthalene‐induced lung injury and repair.
‘I can see parents being reluctant’: perceptions of parental involvement using child and family teams in schools
Authors: Taliaferro Jocelyn DeVance , DeCuir‐Gunby Jessica , Allen‐Eckard Kara
Published: July 2009

The purpose of this study is to examine the attitudes beliefs, and perceptions of school and community personnel regarding parental involvement via the implementation of child and family team meetings. Interviews were conducted with 10 school and community personnel in a high school in a small county in the south‐eastern region of the USA. Several themes emerged from the data, including the definition of parental involvement, parental work and life circumstances, and parental esteem and position within schools. Findings suggest that school and community personnel hold conflicting beliefs regarding parents' desire and ability to be involved in their children's schooling. Recommendations for social work practitioners' implementation of child and family team meetings in the school context are provided.
Participatory social work and report writing
Authors: Roose Rudi , Mottart Andre , Dejonckheere Nele , Van Nijnatten Carol , De Bie Maria
Published: July 2009

Participation of children and parents is a central notion in child and family social work. This paper reports on a research dealing with how the participative paradigm comes to the fore in the practice of report writing in the work with looked‐after children. As social work is essentially a language‐centred activity, report writing is a core skill in social work. A participative practice of report writing would imply that the perspective of the children and parents is present in the reports. The results of our research show that a participatory approach to writing reports is not self‐evident. On the one hand, the practice of report writing shows a big diversity in the way the client's perspectives are given attention. On the other hand, the incorporation of the client's voice does not necessarily stand for a participatory approach because it may be used to strengthen professional views rather than clarify clients' perspectives. The case is made in such a way that a participatory approach of child and family social work demands more attention to the practice of writing reports.
Response of juvenile Atlantic salmon, Salmo salar, to the introduction of salmon carcasses in upland streams
Authors: WILLIAMS K. L. , GRIFFITHS S. W. , NISLOW K. H. , McKELVEY S. , ARMSTRONG J. D.
Published: July 2009

Abstract  The role of nutrients in the production of Atlantic salmon, Salmo salar L., in oligotrophic upland catchments is poorly understood. Atlantic salmon carcasses were introduced to 12 upland streams in the River Conon catchment, Scotland, UK, to assess potential nutrient limitation and evaluate a potential role for migratory Atlantic salmon in nutrient import. The objective was to test the hypothesis that nutrient supplementation would increase juvenile Atlantic salmon biomass. Streams received one of six treatment levels up to a maximum of one carcass per 15 m2 of wetted area with duplicates at each level, and study areas were sampled to measure salmon biomass in treated and reference reaches. There was a significant positive relationship between treatment addition level and response of juvenile salmon biomass. At higher carcass addition levels, salmon biomass in treatment sections was approximately twice that in paired reference sections. The results are discussed in the light of management methods in upland regions and the likely role of cultural oligotrophication in suppressing current levels of salmon production.
A comparison of gastrically and surgically implanted telemetry transmitters in shovelnose sturgeon
Authors: NEELY B. C. , STEFFENSEN K. D. , PEGG M. A.
Published: July 2009

Abstract  Transmitter retention and the effects gastrically and surgically implanted telemetry transmitters had on condition and mortality of shovelnose sturgeon, Scaphirhynchus platorhynchus (Rafinesque), >450‐mm fork length were examined. Four, 35‐day trials were conducted with approximately equal numbers of control fish, fish receiving dummy transmitters gastrically and fish receiving dummy transmitters surgically. Four sturgeon expelled gastrically implanted transmitters during the observation period. Two mortalities occurred among fish receiving gastrically implanted transmitters during the study period and one mortality was recorded for control fish. No mortalities were recorded for fish receiving surgically implanted transmitters. Condition change (ΔKn) was significantly different between fish receiving surgical implantation (mean ΔKn ± SE; −5.17 ± 0.94), fish receiving gastric implantation (−2.98 ± 0.72) and control fish (−0.65 ± 1.09). Surgical transmitter implantation is recommended for telemetry studies on shovelnose sturgeon because of increased transmitter expulsion and occurrence of mortality in fish receiving gastric implantation.
The effect of land use on dissolved organic carbon and nitrogen uptake in streams
Authors: JOHNSON LAURA T. , TANK JENNIFER L. , ARANGO CLAY P.
Published: July 2009

1. Agricultural and urban land use may increase dissolved inorganic nitrogen (DIN) concentrations in streams and saturate biotic nutrient demand, but less is known about their impacts on the cycling of organic nutrients. To assess these impacts we compared the uptake of DIN (as ammonium, NH4+), dissolved organic carbon (DOC, as acetate), and dissolved organic nitrogen (DON, as glycine) in 18 low‐gradient headwater streams in southwest Michigan draining forested, agricultural, or urban land‐use types. Over 3 years, we quantified uptake in two streams in each of the three land‐use types during three seasons (spring, summer and autumn).

2. We found significantly higher NH4+ demand (expressed as uptake velocity, Vf) in urban compared to forested streams and NH4+Vf was greater in spring compared to summer and autumn. Acetate Vf was significantly higher than NH4+ and glycine Vf, but neither acetate nor glycine Vf were influenced by land‐use type or season.

3. We examined the interaction between NH4+ and acetate demand by comparing simultaneous short‐term releases of both solutes to releases of each solute individually. Acetate Vf did not change during the simultaneous release with NH4+, but NH4+Vf was significantly higher with increased acetate. Thus, labile DOC Vf was not limited by the availability of NH4+, but NH4+Vf was limited by the availability of labile DOC. In contrast, neither glycine nor NH4+Vf changed when released simultaneously indicating either that overall N‐uptake was saturated or that glycine and NH4+ uptake were controlled by different factors.

4. Our results suggest that labile DOC and DON uptake can be equivalent to, or even higher than NH4+ uptake, a solute known to be highly bioreactive, but unlike NH4+ uptake, may not differ among land‐use types and seasons. Moreover, downstream export of nitrogen may be exacerbated by limitation of NH4+ uptake by the availability of labile DOC in headwater streams from the agricultural Midwestern United States. Further research is needed to identify the factors that influence cycling of DOC and DON in streams.
Shredder–tadpole facilitation of leaf litter decomposition in a tropical stream
Authors: IWAI NORIKO , PEARSON RICHARD G. , ALFORD ROSS A.
Published: July 2009

1. Leaf litter decomposition is one of the most important ecosystem processes in streams. Recent studies suggest that facilitation, in which litter is processed by a succession of species with differing abilities and requirements, may be important in making the nutrients bound in litter available to the stream assemblage.

2. We predicted that stream invertebrates that feed on terrestrial leaf litter (shredders) and tadpoles would facilitate leaf litter decomposition by changing the quality of leaf material directly via physical contact or indirectly via nutrient release. We experimentally examined the ability of shredders and tadpoles to break down leaves, independently and together, in artificial streams beside a natural forest stream.

3. The decomposition rate was greater when shredders and tadpoles were together than was expected from rates in single‐species treatments, indicating that facilitation occurred. This facilitation operated in one direction only: the rate of leaf breakdown by tadpoles was higher when leaves had been partly processed by shredders, but there was no similar effect when leaves previously occupied by tadpoles were processed by shredders. We did not detect facilitation caused by indirect nutrient release.

4. Shredders may have benefited tadpoles by roughening leaf surfaces, making them easier for the tadpoles to consume and enhancing leaf breakdown in the presence of both taxa. This indicates that the loss of a single species can have impacts on ecosystem functioning that go beyond the loss of its direct contribution.
Palmitoylation of the sphingosine 1‐phosphate receptor S1P1 is involved in its signaling functions and internalization
Authors: Ohno Yusuke , Ito Ayako , Ogata Ren , Hiraga Yuki , Igarashi Yasuyuki , Kihara Akio
Published: July 2009

The lipid mediator sphingosine 1‐phosphate (S1P) regulates several cellular processes through binding to its receptors (S1P1–S1P5), which are heterotrimeric G protein‐coupled receptors. Here, we report that all S1P receptors are palmitoylated. In S1P1, three Cys residues in the cytoplasmic tail are palmitoylated. We examined the roles of palmitoylation of S1P1 using model cells in which wild‐type S1P1 or a non‐palmitoylated mutant S1P1 was overproduced. Compared with wild‐type S1P1, the non‐palmitoylated S1P1 exhibited binding affinity similar to the natural ligand S1P but lower to the synthetic ligand FTY720 phosphate (FTY720‐P), the active form of the immunomodulator FTY720. However, downstream signaling of non‐palmitoylated S1P1 was similarly affected by S1P and FTY720‐P stimulation. Moreover, upon stimulation with S1P, internalization of the mutant non‐palmitoylated S1P1 was retarded, compared with that of the wild‐type protein. This effect was much more pronounced with FTY720‐P stimulation. Similar differences were observed for the phosphorylation of S1P1 and its mutant. These findings may provide insights into the molecular mechanisms of the pharmacological effects of FTY720. Finally, palmitoylation of wild‐type S1P1 increased upon treatment with S1P, suggesting that S1P1 undergoes a palmitoylation/depalmitoylation cycle after stimulation by its ligands.
Progression of cutaneous squamous cell carcinoma in immunosuppressed patients is associated with reduced CD123+ and FOXP3+ cells in the perineoplastic inflammatory infiltrate
Authors: Mühleisen Beda , Petrov Ivaylo , Gächter Tamara , Kurrer Michael , Schärer Leo , Dummer Reinhard , French Lars E , Hofbauer Günther F L
Published: July 2009

Aims:  Squamous cell carcinoma of the skin (SCC) increases dramatically in organ transplant recipients (OTRs). The aim was to determine whether qualitative and quantitative differences in perineoplastic inflammation in OTRs contribute to the increased carcinogenesis.

Methods and results:  We studied the perineoplastic inflammatory infiltrate in SCC, assessing depth, density and phenotype (CD3, 4, 8, FOXP3, CD123 and STAT1) by immunohistochemistry in paired biopsy specimens of intraepithelial and invasive SCC in immunocompetent patients and OTRs. Considerable inflammation was observed in all intraepithelial SCC (inflammatory infiltrate depth 2.80 ± 2.21 mm immunocompetent patients, 2.15 ± 2.95 mm OTRs). Inflammation was more pronounced in invasive SCC of immunocompetent patients (4.60 ± 4.67 mm) and OTRs (3.30 ± 5.90 mm) (P < 0.005). The density of perineoplastic inflammatory infiltrates increased from intraepithelial to invasive SCC (P = 0.005). OTRs showed a lower density of perineoplastic inflammatory infiltrate (P = 0.041). OTRs also showed reduced CD3+ T‐lymphocyte and CD8+ cytotoxic T‐lymphocyte proportions in intraepithelial SCC (P = 0.025 and 0.027, respectively). FOXP3+ regulatory T‐lymphocyte proportions in OTRs’ invasive SCC were markedly diminished (P = 0.048). CD123+ plasmacytoid dendritic cells increased in the progression from intraepithelial to invasive SCC in immunocompetent patients (P = 0.040). CD123+ cells were reduced in all SCC of OTRs (P = 0.036).

Conclusions:  Perineoplastic inflammation in intraepithelial SCC is pronounced both in immunocompetent patients and OTRs. Inflammation increases further in invasive SCC. OTRs show reduced proportions of FOXP3+ regulatory T cells and CD123+ plasmacytoid dendritic cells. This distinct inflammatory infiltrate may result in increased cutaneous carcinogenesis and more aggressive behaviour of SCC in OTRs.
Tracking with virtual slides: a tool to study diagnostic error in histopathology
Authors: Treanor Darren , Lim Chee Hooi , Magee Derek , Bulpitt Andy , Quirke Phil
Published: July 2009

Aims:  To determine the reasons for diagnostic error by virtual slides which allow unsupervised study of diagnosis and error.

Methods and results:  Software was developed to produce visualizations of the diagnostic track followed by pathologists as they viewed virtual slides. These showed the diagnostic path in four dimensions (x, y, time and zoom), areas studied for >1000 ms, and included pathologists’ comments about the areas viewed. The system was used to study two trainee and two expert pathologists diagnosing 60 Barrett’s oesophageal biopsy specimens. Comparisons of the diagnostic tracks showed the reason for errors. Forty‐six cases had an expert consensus diagnosis. The trainees made errors in 21 and 15 cases, respectively, of which 11 and nine were clinically significant. Errors were made across the whole spectrum of diagnoses from negative to intramucosal carcinoma. Detailed examination of the tracks showed that in all errors there was incorrect interpretation of information; in three errors there was an additional failure to identify diagnostic features.

Conclusions:  Tracking with virtual slides is a useful tool in studying diagnosis and error, which has the potential for use in training and assessment.
Observer agreement in the diagnosis of serrated polyps of the large bowel
Authors: Wong Newton A C S , Hunt Linda P , Novelli Marco R , Shepherd Neil A , Warren Bryan F
Published: July 2009

Aims:  To assess observer agreement in the diagnosis of colorectal serrated polyps, in particular, serrated adenomas and admixed polyps (i.e. ‘polyps with admixed hyperplastic and adenomatous glands’).

Methods and results:  Sixty cases of large bowel polyps were assessed by four specialist gastrointestinal histopathologists and allocated into one of five categories: serrated adenoma, hyperplastic polyp, conventional adenoma, admixed polyp, and other polyps with serration. Complete agreement amongst all four assessors was seen with only two‐fifths of the cases. The overall κ value for all the assessors distinguishing between all five categories was 0.49. The κ values for diagnosing serrated adenoma versus all other polyps, and admixed polyp versus all other polyps were 0.38 and 0.3, respectively.

Conclusions:  Specialist gastrointestinal histopathologists show only moderate concordance when distinguishing between serrated colorectal polyps. There is only fair interobserver agreement in the diagnosis of serrated adenomas and admixed polyps of the large bowel.
The helicase‐like transcription factor is a strong predictor of recurrence in hypopharyngeal but not in laryngeal squamous cell carcinomas
Authors: Capouillez Aurélie , Debauve Gael , Decaestecker Christine , Filleul Olivier , Chevalier Dominique , Mortuaire Geoffrey , Coppée Frederic , Leroy Xavier , Belayew Alexandra , Saussez Sven
Published: July 2009

Aims:  To examine the immunohistochemical expression of helicase‐like transcription factor (HLTF) in relation to the prognosis of hypopharyngeal (HSCCs) and laryngeal (LSCCs) squamous cell carcinomas, and to characterize the HLTF protein variants expressed in biopsy specimens of head and neck squamous cell carcinoma (HNSCC) as well as the HeLa cell line.

Methods and results:  HLTF expression was determined by immunohistochemistry on a series of 100 hypopharyngeal (stage IV) and 56 laryngeal SCCs (stages I, II and IV). The HLTF variants were defined using reverse transcriptase‐polymerase chain reaction and Western blots in 13 fresh HNSCC biopsies and in HeLa cells. High levels of HLTF expression were associated with rapid recurrence rates in a subgroup of 81 stage IV hypopharyngeal SCCs (with complete follow‐up). A 95‐kDa HLTF variant truncated at the carboxyl‐terminal domain was detected in addition to the 115‐kDa full‐size protein in HNSCC biopsies, while six variants were observed in HeLa cells.

Conclusions:  Our results demonstrate, for the first time, that hypopharyngeal SCCs presenting high levels of HLTF have a worse prognosis. The quantitative determination of HLTF in hypopharyngeal SCCs was an independent prognostic marker alongside tumour node metastasis staging. HNSCCs expressed the truncated HLTF variant lacking the domains involved in DNA repair.
The assessment of prognosis of surgically resected oesophageal cancer is dependent on the number of lymph nodes examined pathologically
Authors: Twine Christopher P , Lewis Wyn G , Morgan Matthew A , Chan David , Clark Geoffrey W B , Havard Tim , Crosby Tom D , Roberts S Ashley , Williams Geriant T
Published: July 2009

Aims: The prognosis in surgically resected oesophageal carcinoma (OC) is dependent on the number of regional lymph nodes (LN) involved, but no guidance exists on how many LNs should be examined histopathologically to give a reliable pN status. The aim of this study was to determine whether the number of LNs examined after OC resection has a significant effect on the assessment of prognosis.

Methods and results: Routinely generated pathology reports from 237 consecutive patients undergoing oesophagectomy for OC were examined and analysed in relation to survival. The main outcome measure was survival from date of diagnosis. Lymph node count (LNC) correlated strongly with survival; a plateau was reached after a count of 10. Median and 2‐year survival was 30 months and 42%, respectively, if <10 nodes were examined (n = 88), compared with 51 months and 61% if >10 nodes were examined (P = 0.005). This effect was greatest in pN0 cases. The prognostic value of the absolute number of LN metastases (<4) and LN ratio (<0.4) was strongly dependent on a LNC of >10.

Conclusions: These results demonstrate the importance of careful pathological examination and lymph node retrieval after OC resection. At least 10 nodes should be examined to designate an OC as pN0.
Advanced glycation end products accumulate in the reproductive tract of men with diabetes
Authors: Mallidis C. , Agbaje I. M. , Rogers D. A. , Glenn J. V. , Pringle R. , Atkinson A. B. , Steger K. , Stitt A. W. , McClure N.
Published: July 2009

Light microscopic studies comparing sperm parameters show little association between diabetes and male fertility. However, with the introduction of new analytical techniques, evidence is now emerging of previously undetectable effects of diabetes on sperm function. Specifically, a recent study has found a significantly higher sperm nuclear DNA fragmentation in diabetic men. As advanced glycation end products (AGEs) are important instigators of oxidative stress and cell dysfunction in numerous diabetic complications, we hypothesized that these compounds could also be present in the male reproductive tract. The presence and localization of the most prominent AGE, carboxymethyl‐lysine (CML), in the human testis, epididymis and sperm was determined by immunohistochemistry. Parallel ELISA and Western blot analyses were performed to ascertain the amount of CML in seminal plasma and sperm from 13 diabetic and nine non‐diabetic subjects. CML immunoreactivity was found throughout the seminiferous epithelium, the nuclei of spermatogonia and spermatocytes, in the basal and principle cells cytoplasm and nuclei of the caput epididymis and on most sperm tails, mid pieces and all cytoplasmic droplets. The acrosomal cap, especially the equatorial band, was prominently stained in diabetic samples only. The amount of CML was significantly higher (p = 0.004) in sperm from non‐diabetic men. Considering the known detrimental actions of AGEs in other organs, the presence, location and quantity of CML, particularly the increased expression found in diabetic men, suggest that these compounds may play a hitherto unrecognized role in male infertility.
Effect of nanoparticles on the male reproductive system of mice
Authors: Yoshida S. , Hiyoshi K. , Ichinose T. , Takano H. , Oshio S. , Sugawara I. , Takeda K. , Shibamoto T.
Published: July 2009

The effects of nanoparticles toward on the male reproductive system of mice were investigated. Three sizes (14, 56 and 95 nm) of carbon black nanoparticles were intratracheally administered (0.1 mg/mouse for 10 times every week) to ICR male mice to investigate their adverse effects on the reproductive function. The serum testosterone levels were elevated significantly in the 14‐ and 56‐nm carbon nanoparticles‐exposed groups. Histological examination showed partial vacuolation of the seminiferous tubules. In addition, the effects of particle number towards the male reproductive system were investigated. The particle number controlled 14‐nm nanoparticles‐exposed group (14 N group, which has approximately the same particle number per unit volume as the 56‐nm nanoparticles) showed fewer effects than did the 56‐nm nanoparticles‐exposed groups. These results suggest that carbon nanoparticle‐exposure has adverse effects on the mouse male reproductive function. Furthermore, the effects of nanoparticles on the male reproductive system depend on particle mass rather than particle number.
Modelling the distribution of a threatened habitat: the California sage scrub
Authors: Riordan Erin C. , Rundel Philip W.
Published: July 2009

Aim Using predictive species distribution and ecological niche modelling our objectives are: (1) to identify important climatic drivers of distribution at regional scales of a locally complex and dynamic system – California sage scrub; (2) to map suitable sage scrub habitat in California; and (3) to distinguish between bioclimatic niches of floristic groups within sage scrub to assess the conservation significance of analysing such species groups.

Location Coastal mediterranean‐type shrublands of southern and central California.

Methods Using point localities from georeferenced herbarium records, we modelled the potential distribution and bioclimatic envelopes of 14 characteristic sage scrub species and three floristic groups (south‐coastal, coastal–interior disjunct and broadly distributed species) based upon current climate conditions. Maxentwas used to map climatically suitable habitat, while principal components analysis followed by canonical discriminant analysis were used to distinguish between floristic groups and visualize species and group distributions in multivariate ecological space.

Results Geographical distribution patterns of individual species were mirrored in the habitat suitability maps of floristic groups, notably the disjunct distribution of the coastal–interior species. Overlap in the distributions of floristic groups was evident in both geographical and multivariate niche space; however, discriminant analysis confirmed the separability of floristic groups based on bioclimatic variables. Higher performance of floristic group models compared with sage scrub as a whole suggests that groups have differing climate requirements for habitat suitability at regional scales and that breaking sage scrub into floristic groups improves the discrimination between climatically suitable and unsuitable habitat.

Main conclusions The finding that presence‐only data and climatic variables can produce useful information on habitat suitability of California sage scrub species and floristic groups at a regional scale has important implications for ongoing efforts of habitat restoration for sage scrub. In addition, modelling at a group level provides important information about the differences in climatic niches within California sage scrub. Finally, the high performance of our floristic group models highlights the potential a community‐level modelling approach holds for investigating plant distribution patterns.
Nursing staff turnover at a Swedish university hospital: an exploratory study
Authors: Sellgren Stina F , Kajermo Kerstin N , Ekvall Göran , Tomson Göran
Published: July 2009

Aim.  The aim was to explore opinions on individual needs and other factors that may influence nursing staff turnover.

Background.  High staff turnover is a great problem for many hospitals. It is shown to have a negative effect on the quality of nursing care and to increase hospital costs.

Design.  In 2004 in a large university hospital in Sweden five focus group discussions (FGDs) including department heads (1), nursing managers (2) and members of nursing staff (2) were carried out. The questions to be addressed were ‘Why do nurses leave?’ and ‘Why do nurses stay?’ In addition, register data of staff turnover for 2002–2003 were analysed in relation to different facts about the units, such as number of employees, type of care and medical specialty. Categories of opinions identified in the FGDs were compared with results of the statistical analyses on the relationship between staff turnover and unit parameters to identify overall factors that may influence on nurse staff turnover.

Findings.  Four major factors were identified as having a possible influence on staff turnover: ‘intrinsic values of motivation’, ‘work load’, ‘unit size ‘and ‘leadership’. Smaller units had lower staff turnover as well as outpatient units and day care. It was not possible to compare statements from participants from smaller units with those from participants from larger units. Two factors had diverging data, ‘salary’ and ‘spirit of the time’. A surprising finding was the little mention of patient care in relation to staff turnover.

Relevance to clinical practice.  It is important for managers to ensure that intrinsic values of nurses are met to minimise the risk for high turnover rates. Inpatient care must receive adequate staffing and nursing care could be organised into smaller units or work teams to avoid dissatisfaction and high turnover.
Nurse prescribing by children’s nurses: views of doctors and clinical leads in one specialist children’s hospital
Authors: Courtenay Molly , Carey Nicola
Published: July 2009

Aim.  To explore the views of doctors and clinical leads (CLs), who care for children and young people, on nurse prescribing in one specialist children’s hospital.

Background.  Nearly 14,000 nurses in the UK have virtually the same prescribing rights as doctors. Benefits of nurses adopting this role have been reported, but doctors do have some concerns. Increasing numbers of specialist nurses involved in the care of children in the hospital setting are undertaking prescribing training. No research has explored the views of health care professionals on nurse prescribing in this setting.

Design.  A subset of qualitative data taken from a larger study that adopted an intrinsic case study design.

Method.  Interviews were conducted between October 2006–July 2007 with 11 doctors and three clinical leads in one specialist children’s hospital. A thematic analysis was conducted on the interview data.

Results.  Nurse prescribing improved access to medicines and continuity of care. Concerns included the need for doctors to have confidence in the ability of the nurse who wanted to become a prescriber, the selection of nurses for prescribing training, the effects of nurse prescribing on the individual roles and responsibilities of doctors and nurses’ clinical skills.

Conclusion.  Doctors and clinical leads working in a specialist children’s hospital in the UK recognise that nurse prescribing makes a positive contribution to the delivery of services. However, it is important that nurses have the appropriate clinical skills and doctors understand nurse prescribing. This will only take place if there is good communication across professional boundaries.

Relevance to clinical practice.  Selection processes for the prescribing programmes must ensure that students have the necessary course prerequisites. Communication across professional boundaries is crucial to the successful implementation of nurse prescribing in the care of children and young people in the hospital setting.