bluuebunny/crossref_metadata_2025 huggingface dataset:
Second‐hand Smoke Increases Nitric Oxide and Alters the IgE Response in a Murine Model of Allergic Aspergillosis
Authors: Seymour Brian W. P. , Peake Janice L. , Pinkerton Kent E. , Kurup Viswanath P. , Gershwin Laurel J.
Published: September 2005

This study was performed to determine the effects of environmental
tobacco smoke (ETS) on nitric oxide (NO) and immunoglobulin (Ig) production in
a murine model of allergic bronchopulmonary aspergillosis (ABPA). Adult
BALB/c mice were exposed to aged and diluted sidestream cigarette smoke
from day 0 through day 43 to simulate “second‐hand
smoke”. During exposure,
mice were sensitized to soluble Aspergillus fumigatus (Af)
antigen intranasally
between day 14 and 24. All Af sensitized mice in ambient air (Af + AIR) made
elevated levels of IgE, IgG1, IgM, IgG2a and IgA. Af sensitized mice housed in
ETS (Af + ETS) made similar levels of immunoglobulins except for IgE that was
significantly reduced in the serum and bronchoalveolar lavage (BAL). However,
immunohistochemical evaluation of the lung revealed a marked accumulation of
IgE positive cells in the lung parenchyma of these Af + ETS mice. LPS stimulation
of BAL cells revealed elevated levels of NO in the Af + AIR group, which was further
enhanced in the Af+ETS group. In vitro restimulation of the BAL cells on day 45
showed a TH0 response with elevated levels of IL3, 4, 5, 10 and IFN‐γ. However,
by day 28 the response shifted such that TH2 cytokines increased while
IFN‐γ decreased. The Af + ETS group showed markedly reduced levels in all
cytokines tested, including the inflammatory cytokine IL6, when compared to
the Af+AIR group. These results demonstrate that ETS affects ABPA by further
enhancing the NO production and reduces
the TH2 and the inflammatory cytokines while altering the pattern of IgE responses.
Mold Hysteria: Origin of the Hoax
Authors: Chang Christopher , Gershwin M. Eric
Published: September 2005

The topic of building related illness came into the public′s eye as a major
health issue in the mid 1970s, when several cases of pneumonia were found to
be associated with an infectious agent in Philadelphia. This agent was
subsequently found to be a gram‐positive bacterium known as Legionella
pneumoniae. During the ensuing 30 years, a myriad of symptom constellations,
disorders, clinical syndromes and illnesses have been attributed to indoor living
or working environments. Over time, there appeared to be no limit to claims of
building related illness, and it was “reported” that almost any kind of clinical
symptom, real or imaginary, could be blamed on indoor environments. As society
became more and more litigious, many of these disorders were erroneously played
out in courtrooms rather than medical offices, creating a circus atmosphere
surrounding this class of disorders. With the advent of the internet, as well as
other advances in telecommunications, these issues eventually became part of
a media frenzy, and all truths could be thrown out the window as issues became
more and more decided upon by
emotions and unfounded beliefs, rather than scientific data and logical thinking.
Crohn′s Disease Associated with Sweet′s Syndrome and Sjögren′s Syndrome Treated with Infliximab
Authors: Foster Erina N. , Nguyen Khanh K. , Sheikh Rafiq A. , Prindiville Thomas P.
Published: September 2005

The association of Crohn′s disease (CD) and Sweet′s syndrome is rare and
the presence of Sjögren′s syndrome in Crohn′s disease is even rarer, with only
three reports found in the literature. We describe two cases of Crohn′s disease
associated with Sweet′s syndrome, one of which is the first case of CD and
Sweet′s concomitantly associated with Sjögren′s syndrome. Both cases
responded rapidly to Infliximab therapy with complete resolution of the skin lesions.
The Introduction of Voter Registration and Its Effect on Turnout
Authors: Ansolabehere Stephen , Konisky David M.
Published: September 2005

Studies of voter turnout across states find that those with more facilitative registration laws have higher turnout rates. Eliminating registration barriers altogether is estimated to raise voter participation rates by up to 10%. This article presents panel estimates of the effects of introducing registration that exploits changes in registration laws and turnout within states. New York and Ohio imposed registration requirements on all of their counties in 1965 and 1977, respectively. We find that the introduction of registration to counties that did not previously require registration decreased participation over the long term by three to five percentage points. Though significant, this is lower than estimates of the effects of registration from cross-sectional studies and suggests that expectations about the effects of registration reforms on turnout may be overstated.
Deletion of Dinucleotide Repeat (Δ14 Allele) in the Methylthioadenosine Phosphorylase (MTAP) Promoter and the Allelotype of MTAP Promoter in the Japanese Population
Authors: Kadariya Yuwaraj , Nishioka Junji , Nakatani Kaname , Nakashima Kunio , Nobori Tsutomu
Published: September 2005

5′‐Deoxy‐5′‐methylthioadenosine phosphorylase (MTAP) is an enzyme involved in purine and polyamine metabolism and is ubiquitously expressed in normal human tissues and cells. However, this enzyme has been found to be deficient in a variety of human cancers. Although the enzyme deficiency is known to be caused by MTAP gene deletion, human diffuse histiocytic lymphoma cell line DHL‐9 without any detectable MTAP activity has been found to possess the intact MTAP gene. These lines of evidence suggested that promoter abnormality might cause the MTAP deficiency in DHL‐9. Therefore, we analyzed the MTAP promoter region of DHL‐9 and found the deletion of 14 bases in its sequence. We designated the allele lacking (GT)6GC as dinucleotide repeat deletion (Δ14 allele) and determined the effect of the Δ14 allele on the MTAP promoter activity by a luciferase reporter assay. We have also analyzed the distribution of the Δ14 allele and wild‐type (WT) allele in the Japanese population by PCR assay. A reporter plasmid harboring the Δ14 allele exhibited luciferase activity comparable to that of a plasmid containing the WT allele. Forty‐six (22%) out of 210 people were homozygous for WT allele in the MTAP promoter, whereas 43 (20.5%) were homozygous for Δ14 allele. The remaining 121 people (57.5%) possessed Δ14/WT alleles in the MTAP promoter region. These results indicated that the Δ14 allele has nothing to do with MTAP deficiency in DHL‐9. The Δ14 allele is distributed among the general population irrespective of gender.
Reversibility of Heterotopic Proliferative Glands in Glandular Stomach of Helicobacter pylori‐infected Mongolian Gerbils on Eradication
Authors: Nozaki Koji , Shimizu Nobuyuki , Tsukamoto Tetsuya , Inada Ken‐ichi , Cao Xueyuan , Ikehara Yuzuru , Kaminishi Michio , Sugiyama Atsushi , Tatematsu Masae
Published: September 2005

Helicobacter pylori (Hp) infection is an important factor in human gastric disorders. Mongolian gerbils can be easily infected with Hp and represent excellent experimental models to clarify the role of Hp in chronic active gastritis, peptic ulcers, intestinal metaplasia, and gastric carcinoma. We have proved the enhancing effects of Hp infection on all histological types of gastric cancers in Mongolian gerbils exposed to chemical carcinogens. Heterotopic proliferative glands (HPGs) also frequently develop with Hp infection in the glandular stomach of infected gerbils, with a slightly dysplastic change of constituent cells. Distinguishing reversible inflammatory lesions from true neoplasms upon eradication is necessary for further biological or histochemical investigations using this model. We employed an experimental model of long‐term Hp infection and eradication in gerbils. HPGs finally developed with a phenotypic shift of intestinalization with Paneth cells. After eradication, HPGs were obviously reduced, and gastric lesions in mucosa also improved with few remnants of the former injury. This shows that reversible HPGs are frequently induced solely by Hp infection in this annual species, and are related to severe gastritis, rather than being malignant in character. Thus, distinguishing reversible lesions from true neoplasms is necessary to investigate the relationship of Hp infection and gastric carcinogenesis in this animal model.
Frequent and Multiple Mutations at Minisatellite Loci in Sporadic Human Colorectal and Gastric Cancers—Possible Mechanistic Differences from Microsatellite Instability in Cancer Cells
Authors: Inamori Hideaki , Takagi Sachiyo , Tajima Rie , Ochiai Masako , Ubagai Tsuneyuki , Sugimura Takashi , Nagao Minako , Nakagama Hitoshi
Published: September 2005

Minisatellites (MNs), composed of 5 to 100 nucleotide repeat units, range from 0.5 to 30 kb in length, and have been reported to be mutated in various human malignancies. In this study, frequencies of MN mutations in sporadic human colorectal (34 cases) and gastric cancers (24 cases) at various clinicopathological stages were assessed by multilocus DNA fingerprint analysis with three MN probes, Pc‐1, 33.6 and 33.15. MN mutations were observed in both colorectal and gastric cancers, but at a significantly higher frequency in the former (56%) than in the latter (25%). Multiplicities of MN mutations were 1.50±1.81 and 0.46±1.10 in colorectal and gastric cancers, respectively, and the difference was also significant. Neither the presence nor multiplicity of MN mutations in either colorectal or gastric cancer cases had any correlation with the pathological stage, histological grading or the presence of microsatellite instability (MSI). Although the biological relevance of MN mutations still remains to be clarified, a subset of colorectal and gastric cancers could feature a new type of genomic instability, distinct from MSI.
Association of HLA‐DQB1★0301 and HLA‐DQB1★0602 with Different Subtypes of Gastric Cancer in Taiwan
Authors: Wu Ming‐Shiang , Hsieh Rhong‐Phong , Huang Shih‐Pei , Chang Yu‐Ting , Lin Ming‐Tsan , Chang Ming‐Chu , Shun Chia‐Tung , Sheu Jin‐Chuan , Lin Jaw‐Town
Published: September 2005

Gastric cancer (GC) is a heterogeneous disorder with multifactorial etiologies. Genetic predisposition, environmental factors, and Helicobacter pylori infection are thought to interact in the manifestation of GC. Particular human leukocyte antigen (HLA) alleles play a pivotal role in cellular immunity and may be an important genetically determined host trait. To elucidate the association between the genotype of HLA class II genes and the clinical phenotype of GC, polymorphisms of HLA‐DRB1 and HLA‐DQB1 were determined by polymerase chain reaction with sequence‐specific primers in 106 Taiwanese patients with GC and in 208 healthy controls. Comparison of allele frequencies between GC patients and healthy controls showed no significant difference at the HLA‐DRB1 locus. Patients with GC had a higher frequency of DQB1★0602 (9.4% vs. 3.6%, P<0.05, odds ratio 2.79, 95% confidence interval 1.41–5.47) and a lower frequency of DQB1★0301 (14.6% vs. 23.8%, P<0.05, odds ratio 0.55, 95% confidence interval 0.35–0.85) compared to healthy controls. Correlation of HLA‐DQB1 status with clinicopathologic features revealed predominance of male gender (16/3 vs. 50/37, P<0.05) and proximal location (12/7 vs. 28/59, P<0.05) in patients with positive HLA‐DBQ1★0602 compared to those with negative HLA‐DBQ1★0602. In contrast, a higher ratio of diffuse/intestinal subtype (20/10 vs. 30/46, P<0.05) and a lower seropositivity of Helicobacter pylori (14/30 vs. 58/76, P<0.005) were noted in patients with positive HLA‐DQB1★0301 compared to those with negative HLA‐DQB1★0301. In conclusion, HLA‐DQB1★0602 confers susceptibility to gastric cancer, especially for male Taiwanese and proximal tumor location, while HLA‐DQB1★0301 may have a protective effect on GC, probably through resistance to Helicobacter pyloriinfection. HLA‐DQB1 alleles are associated with susceptibility or resistance to GC and also influence its clinical features.
Enhanced Expression of Neuron‐specific Enolase (NSE) in Pyothorax‐associated Lymphoma (PAL)
Authors: Nakatsuka Shin‐ichi , Nishiu Mieko , Tomita Yasuhiko , Miwa Hideaki , Takakuwa Tetsuya , Luchi Keiji , Yamamoto Satoru , Aozasa Katsuyuki
Published: September 2005

Pyothorax‐associated lymphoma (PAL) is a B‐cell lymphoma of mostly large cell type developing in the pleural cavity of patients with long‐standing pyothorax. Neuron‐specific enolase (NSE) is an enolase comprising γ subunit and is located at high levels in neuronal and neuroendocrine cells, together with their neoplasias. Expression of NSE at protein and mRNA levels was examined in PAL and other types (non‐PAL) of non‐Hodgkin's lymphomas. In PAL, serum levels of NSE were elevated (5.32 to 168.0, mean 42.6 ng/ml) and tended to decrease after incisional biopsy followed by chemotherapy (2.38 to 195.5, mean 34.1 ng/ml). Two cell lines established from two cases of PAL produced and secreted NSE in the culture medium. Immunohistochemistry revealed that the positive rate for NSE staining in PAL (10 of 14 cases, 71.4%) was significantly higher than that in non‐PAL cases (6 of 38 cases, 15.8%) (P<0.01). RT‐PCR analysis showed that the expression levels of NSE mRNA in two cell lines and a biopsy sample from PAL were rather similar to those of the control samples from non‐neoplastic lymph nodes. These findings suggest the posttranscriptional regulation of NSE in PAL. Thus, an elevation of serum NSE level in patients with chronic pyothorax may be an indicator of PAL development.
Macrophage Colony‐stimulating Factor (M‐CSF) Prevents Infectious Death Induced by Chemotherapy in Mice, While Granulocyte‐CSF Does Not
Authors: Hidaka Takao , Fujimura Masaki , Nakashima Akitoshi , Higuma Subaru , Yamagishi Naoko , Tsuda Hiroshi , Sakai Masatoshi , Saito Shigeru
Published: September 2005

To clarify the effect of granulocyte colony‐stimulating factor (G‐CSF) and macrophage colony‐stimulating factor (M‐CSF/CSF‐1) on chemotherapy‐induced infection, we estimated the effect of those CSFs on a mouse model under severe myelosuppression. First, we established an animal model in which 48.9% (22/45) of C3H/Hej mice died of sepsis related to severe myelosuppression after intraperitoneal administration of a single dose (9 mg/kg) of mitomycin C (MMC). G‐CSF or M‐CSF was administered to this model on various administration schedules after chemotherapy, and the effect of those CSFs on survival rates, peripheral blood granulocyte counts, expression of adhesion molecules (CD11a, CD11b, CD18) on granulocytes and granulocyte function (phagocytosis and superoxide anion production) were examined. In all G‐CSF administration groups, peripheral blood granulocyte counts were increased, but improvements in expression of adhesion molecules such as CD11a and CD18, and granulocyte function were less marked and survival rates were not unproved. Meanwhile, when M‐CSF was administered from 1 to 7 days after chemotherapy, granulocyte and platelet counts were increased, and moreover, expression of adhesion molecules and granulocyte function were markedly improved. Furthermore, the survival rate was significantly improved to 77.8% (28/36) compared with the MMC group (P<0.05). Positive rate of blood culture examination at 7 days after chemotherapy in the M group was 0%, and was significantly lower than that in the G group (40%) and the MMC group (40%) (P<0.05). These results demonstrated that it is important not only to increase the granulocyte counts, but also to improve granulocyte functions for preventing infection under myelosuppression after chemotherapy.
Tumor Necrosis Factor α‐Gene Therapy for an Established Murine Melanoma Using RGB (Arg‐Gly‐Asp) Fiber‐mutant Adenovirus Vectors
Authors: Okada Yuka , Okada Naoki , Nakagawa Shinsaku , Mizuguchi Hiroyuki , Takahashi Koichi , Mizuno Nobuyasu , Fujita Takuya , Yamamoto Akira , Hayakawa Takao , Mayumi Tadanori
Published: September 2005

Although adenovirus vectors (Ad) provide high‐level transduction efficacy to many cell types, extremely high doses of Ad are required for sufficient gene transduction into several tumors, including melanoma. Here, we demonstrated that the expression of coxsackie‐adenovirus receptor, a primitive Ad‐receptor, was very low in murine and human melanoma cells. We also found that fiber‐mutant Ad containing the Arg‐Gly‐Asp (RGD) sequence in the fiber knob remarkably augmented gene transduction efficacy in melanoma cells by targeting αv‐integrins. In addition, intratumoral injection of RGD fiber‐mutant Ad containing the tumor necrosis factor α gene (AdRGD‐TNFα) revealed dramatic anti‐tumor efficacy through hemolytic necrosis in an established murine B16 BL6 melanoma model. Ad‐RGD‐TNFα required one‐tenth the dosage of Ad‐TNFα to induce an equal therapeutic effect. These results suggest that αv‐integrin‐targeted Ad will be a very powerful tool for the advancement of melanoma gene therapy.
Elevated Level of Plasma Basic Fibroblast Growth Factor in Multiple Myeloma Correlates with Increased Disease Activity
Authors: Sato Norihide , Hattori Yutaka , Wenlin Du , Yamada Taketo , Kamata Tamihiro , Kakimoto Tsunayuki , Okamoto Shinichiro , Kawamura Chiharu , Kizaki Masahiro , Shimada Naoki , Ote Yukiko , Hata Jun‐ichi , Ikeda Yasuo
Published: September 2005

Recent reports that bone marrow angiogenesis is increased in multiple myeloma prompted us to examine plasma concentrations of angiogenic growth factors and to elucidate their clinical and biological significance. In 45 cases including 36 cases of multiple myeloma and 9 cases of monoclonal gammopathies of undetermined significance (MGUS), plasma concentrations of basic fibroblast growth factor (FGF‐2) and vascular endothelial growth factor (VEGF) were evaluated. FGF‐2 was significantly elevated in 25 out of 45 (56%) of the patients with multiple myeloma compared with control subjects (median 9.01 pg/ml vs. 1.58 pg/ml, P<0.0001). The 25 cases were all active multiple myeloma, and none of the non‐active myeloma and MGUS patients showed a high FGF‐2 level. VEGF level was also elevated in 26 out of 45 patients (58%) compared with control subjects (median 42.0 pg/ml vs. 15.8 pg/ml, P<0.0001 for VEGF). VEGF concentration was high in 20 active myelomas, but also in one non‐active myeloma and five MGUS. Elevation of FGF‐2 level was associated with β2‐microglobulin level, anemia and bone marrow plasma cell percentage, which represent disease activity. Interestingly, none of five Bence‐Jones type myelomas, including four clinically active cases, revealed a high plasma FGF‐2 level, while all of them showed a high VEGF level. In all five responders, the plasma FGF‐2 levels were significantly decreased after chemotherapy. FGF‐2 was immunohistochemically detected in the bone marrow myeloma cells of the patients with high plasma FGF‐2 level. We conclude that plasma concentration of FGF‐2 can be a useful indicator of disease activity.
Predictors of emotional outcomes of intensive care
Authors: Rattray J. E. , Johnston M. , Wildsmith J. A. W.
Published: September 2005

Negative emotional outcomes (anxiety, depression and post‐traumatic stress) have been identified in patients discharged from intensive care. The aims of this prospective, longitudinal study were to assess levels of and changes in emotional outcome after intensive care, and to explore how these relate to objective and subjective indicators of the intensive care experience. Emotional outcome was assessed using the Hospital Anxiety and Depression and Impact of Event Scales. Anxiety (p = 0.046) and depression (p = 0.001) were reduced subsequently, but not avoidance (p = 0.340) or intrusion (p = 0.419). Most objective (age, gender, length of ICU and hospital stay) and subjective indicators (as measured by the Intensive Care Experience Questionnaire) of the intensive care experience were related to negative emotional outcome. Subjective interpretation of the intensive care experience emerged as a consistent predictor of adverse emotional outcome, in both the short‐ and the long‐term.
Concomitant neutrophil JAK2V617F mutation screening and PRV‐1 expression analysis in myeloproliferative disorders and secondary polycythaemia
Authors: Tefferi Ayalew , Sirhan Shireen , Lasho Terra L. , Schwager Susan M. , Li Chin‐Yang , Dingli David , Wolanskyj Alexandra P. , Steensma David P. , Mesa Ruben , Gilliland D. G.
Published: September 2005

Polycythaemia vera (PV) is closely associated with both an acquired activating mutation of the JAK2 tyrosine kinase (JAK2V617F) in granulocyte‐derived DNA and increased granulocyte polycythaemia rubra vera‐1 (PRV‐1) expression. In order to explore the correlation between these two biological markers and compare their diagnostic utility, mutation analysis for JAK2V617F and quantitative measurement of granulocyte PRV‐1 expression were performed on the same study sample from 100 participants: 38 with PV, 22 with essential thrombocythaemia (ET), 10 with agnogenic myeloid metaplasia (AMM), 19 with secondary polycythaemia (SP) and 11 healthy volunteers. The respective overall (homozygous) JAK2V617F mutational frequencies were 95% (26%), 55% (0%), 30% (0%), 0% and 0%. The corresponding figures for increased PRV‐1 expression were 89%, 18%, 20%, 21% and 9%. In patients with either ET or AMM, the likelihood of detecting JAK2V617F was significantly higher in the presence of an increased PRV‐1 expression (83% vs. 38%; P = 0·05). Similarly, in patients with PV, homozygous as compared with heterozygous JAK2V617F correlated with higher levels of PRV‐1 expression (P = 0·11). The present study suggests an allele dose‐dependent effect of JAK2V617F on granulocyte PRV‐1 expression. However, compared with the PRV‐1 assay, mutation screening for JAK2V617F displayed greater accuracy in distinguishing PV from SP.
Factors influencing macroinvertebrate colonization of seasonal wetlands: responses to emergent plant cover
Authors: De Szalay Ferenc A. , Resh Vincent H.
Published: September 2005

1. We conducted field experiments to examine factors influencing macroinvertebrate colonization of seasonally flooded marshes. Few macroinvertebrate species were found aestivating in soils within non‐flooded wetlands indicating that most taxa colonize these marshes from other flooded habitats.  2. We manipulated amounts of salt grass (Distichlisspicata) to examine how emergent plant cover affects aerial colonization by macroinvertebrates. Areas mowed 3 weeks before flooding had low plant cover, areas mowed 5 and 9 weeks before flooding had medium and high plant cover, respectively, and non‐mowed control areas had the most plant cover. Macroinvertebrate numbers and biomass were generally higher in mowed treatment areas than in control areas, but overall diversity was generally higher in high plant cover and control areas than in low plant cover areas.  3. Mosquitoes (Culicidae), brine flies (Ephydridae) and hover flies (Syrphidae) were positively correlated with amount of plant cover, and waterboatmen (Corixidae), midges (Chironomidae) and water scavenger beetles (Hydrophilidae) were negatively correlated with plant cover. Species assemblages changed seasonally among treatment areas because these taxa colonize wetlands at different times in the year.  4. These results demonstrate that invertebrate communities may be different within plant stands with heterogeneous amounts of emergent cover, and management practices that alter the structure of wetland vegetation can influence macroinvertebrate communities colonizing seasonal marshes.
Silica and diatom growth in Lough Neagh: the importance of internal recycling
Authors: Gibson C. E. , Wang G. , Foy R. H.
Published: September 2005

1. Lough Neagh is a large eutrophic lake with a high dissolved silica (SiO2) concentration arising from the basaltic rock in parts of the catchment and the normal winter maximum concentration is over 8 mg L−1. Based on frequent observations between 1974 and 1997, the annual cycles of SiO2 input, uptake and release are explored.  2. Large spring blooms of the planktonic diatoms Aulacoseira subarctica Haworth and Stephanodiscus astraea occur every year and are terminated by SiO2 depletion. Although there are periods when one diatom species has dominated the other, over the period as a whole, the two diatoms appear to be in a stable coexistence.

 3. A model of the effect of river inputs on lake concentration shows that without SiO2 release from the sediment, the maximum spring dissolved SiO2 concentration (and hence diatom crop) would be only about one‐third of that actually observed.

 4. It is concluded that within‐lake processes play a large role and are potentially more variable than catchment processes in determining the available SiO2 in Lough Neagh.

 5. The role of benthic animals, especially Chironomus anthracinus, in the sediment SiO2 release process is discussed. Field data suggest the SiO2 release rate is highly sensitive to temperature, but this could be partly caused by an interaction between temperature and animal activity.
Prediction and assessment of local stream habitat features using large‐scale catchment characteristics
Authors: Davies Nerida M. , Norris Richard H. , Thoms Martin C.
Published: September 2005

1. Knowledge of what a habitat should be like, in the absence of the effects of human activities, is fundamental to local stream habitat assessment. It has been suggested that stream habitats are influenced by large‐scale catchment features. This study aimed to identify these relationships so that local‐scale habitat features could be predicted from larger‐scale characteristics.  2. Fifty‐one reference sites from the Upper Murrumbidgee River catchment, south‐eastern Australia, were classified on the basis of the local features of their stream habitat. Large‐scale variables, namely catchment area, stream length, relief ratio, alkalinity, percentage of volcanic rocks, percentage of metasediments, dominant geology and dominant soil type, provided sufficient information for classifying 69% of reference sites into appropriate reference site groups.  3. A model created using these large‐scale catchment variables was able to predict the local habitat features that were expected (E) to occur at a site in the absence of the effects of human activities. These were compared with observed (O) local habitat features to provide an observed‐to‐expected (O/E) ratio, an assessment score of the habitat at a site. The departure of this ratio from 1 enables identification of those sites that may be impacted. A list of habitat features that are expected at a site can provide targets for habitat restoration or enhancement.  4. For impacted sites, when habitat assessment from the habitat predictive model was compared with biological assessment from the Australian River Assessment System (AUSRIVAS) predictive model, it was possible to identify whether habitat degradation or water quality degradation was the cause of biological impairment. Such assessment may make it possible to identify rehabilitation goals relevant to the biota.
Release and bioavailability of dissolved organic matter from floodplain litter: influence of origin and oxygen levels
Authors: O'Connell M. , Baldwin D. S. , Robertson A. I. , Rees G.
Published: September 2005

1. We determined the rate of release and microbial uptake of dissolved organic carbon (DOC) leached from three components (leaves, bark and twigs) of river red gum (Eucalyptus camaldulensis) forest litter originating from different parts of a floodplain and under different oxygen levels.  2. Preliminary experiments showed that substantially more DOC was released from leaves than from bark or twigs; there was relatively little DOC release from coarse particulate matter or soil.  3. Both the amount of DOC released from each litter component and the amount metabolized by the microbial community were independent of position on the flood‐plain or amount of oxygen available to microbes.  4. Although the bioavailability of DOC was independent of oxygen concentration, the microbial utilization of DOC under aerobic and anaerobic conditions differed. Under aerobic conditions, leaves were colonized by fungi, while bacteria were dominant under anoxic conditions.  5. Phospholipid fatty acid profiles of the microbial communities growing on leaf extracts showed that different microbial communities developed in each oxygen concentration treatment suggesting that, irrespective of flood conditions, a microbial community will develop to utilize a significant proportion of the DOC leached from litter.
Gene expression in insecticide resistant and susceptible Anopheles gambiae strains constitutively or after insecticide exposure
Authors: Vontas J. , Blass C. , Koutsos A. C. , David J.‐P. , Kafatos F. C. , Louis C. , Hemingway J. , Christophides G. K. , Ranson H.
Published: September 2005

A microarray containing approximately 20 000 expressed sequence tags (ESTs; 11 760 unique EST clusters) from the malaria vector, Anopheles gambiae, was used to monitor differences in global gene expression in two insecticide resistant and one susceptible strains. Statistical analysis identified 77 ESTs that were differentially transcribed among the three strains. These include the cytochrome P450 CYP314A1, over‐transcribed in the DDT resistant ZAN/U strain, and many genes that belong to families not usually associated with insecticide resistance, such as peptidases, sodium/calcium exchangers and genes implicated in lipid and carbohydrate metabolism. Short‐term (6 and 10 h) effects of exposure of the pyrethroid resistant RSP strain to permethrin were also detected. Several genes belonging to enzyme families already implicated in insecticide or xenobiotic detoxification were induced, including the carboxylesterase COEAE2F gene and members of the UDP‐glucuronosyl transferase and nitrilase families.
Inactivated mariner‐like elements (MLE) in pink bollworm, Pectinophora gossypiella
Authors: Wang J. , Staten R. , Miller T. A. , Park Y.
Published: September 2005

We isolated multiple copies of mariner‐like element (MLE) from the pink bollworm (PBW), Pectinophora gossypiella, a key lepidopteran cotton pest. Although all the MLE sequences contain multiple mutations accumulated in their transposase coding region, the consensus sequence revealed a putative ancestral transposase encoding 339 amino acid residues with a D,D(34)D motif, and 36 bp inverted terminal repeats, belonging to the cecropia subfamily, and most similar to the MLE found in Antheraea species. Examining six different pink bollworm populations, we conclude that the MLE in PBW described in this study are ancient and are undergoing the process of accumulating inactivating mutations. This conclusion is supported by the patterns of polymorphisms revealed by genomic Southern hybridization, transposable element displays, and sequences from multiple MLE.
Immunohistochemical estimation of cell cycle entry and phase distribution in astrocytomas: applications in diagnostic neuropathology
Authors: Scott Ian S. , Morris Lesley S. , Rushbrook S. M. , Bird Kate , Vowler Sarah L. , Burnet Neil G. , Coleman Nicholas
Published: September 2005

An immunohistochemical method for assessing cell cycle phase distribution in neurosurgical biopsies would enable such data to be incorporated into diagnostic algorithms for the estimation of prognosis and response to adjuvant chemotherapy in glial neoplasms, without the requirement for flow cytometric analysis. Paraffin‐embedded sections of intracerebral gliomas (n = 48), consisting of diffuse astrocytoma (n = 9), anaplastic astrocytoma (n = 8) and glioblastoma (n = 31), were analysed by immunohistochemistry using markers of cell cycle entry, Mcm‐2 and Ki67, and putative markers of cell cycle phase, cyclins D1 (G1‐phase), cyclin A (S‐phase), cyclin B1 (G2‐phase) and phosphohistone H3 (Mitosis). Double labelling confocal microscopy confirmed that the phase markers were infrequently coexpressed. Cell cycle estimations by immunohistochemistry were corroborated by flow cytometric analysis. There was a significant increase in Mcm‐2 (P < 0.0001), Ki67 (P < 0.0001), cyclin A (P < 0.0001) and cyclin B1 (P = 0.002) expression with increasing grade from diffuse astrocytoma through anaplastic astrocytoma to glioblastoma, suggesting that any of these four markers has potential as a marker of tumour grade. In a subset of glioblastomas (n = 16) for which accurate clinical follow‐up data were available, there was a suggestion that the cyclin A:Mcm‐2 labelling fraction might predict a relatively favourable response to radical radiotherapy. These provisional findings, however, require confirmation by a larger study. We conclude that it is feasible to obtain detailed cell cycle data by immunohistochemical analysis of tissue biopsies. Such information may facilitate tumour grading and may enable information of prognostic value to be obtained in the routine diagnostic laboratory.
Characterization of the caspase cascade in a cell culture model of SOD1‐related familial amyotrophic lateral sclerosis: expression, activation and therapeutic effects of inhibition
Authors: Sathasivam S. , Grierson A. J. , Shaw P. J.
Published: September 2005

There is increasing evidence that apoptosis or a similar programmed cell death pathway is the mechanism of cell death responsible for motor neurone degeneration in amyotrophic lateral sclerosis. Knowledge of the relative importance of different caspases in the cell death process is at present incomplete. In addition, there is little information on the critical point of the death pathway when the process of dying becomes irreversible. In this study, using the well‐established NSC34 motor neurone‐like cell line stably transfected with empty vector, normal or mutant human Cu‐Zn superoxide dismutase (SOD1), we have characterized the activation of the caspase cascade in detail, revealing that the activation of caspases‐9, ‐3 and ‐8 are important in motor neurone death and that the presence of mutant SOD1 causes increased activation of components of the apoptotic cascade under both basal culture conditions and following oxidative stress induced by serum withdrawal. Activation of the caspases identified in the cellular model has been confirmed in the G93A SOD1 transgenic mice. Furthermore, investigation of the effects of anti‐apoptotic neuroprotective agents including specific caspase inhibitors, minocycline and nifedipine, have supported the importance of the mitochondrion‐dependent apoptotic pathway in the death process and revealed that the upstream caspase cascade needs to be inhibited if useful neuro‐protection is to be achieved.
Genetic alteration and expression of the phosphoinositol‐3‐kinase/Akt pathway genes PIK3CA and PIKE in human glioblastomas
Authors: Knobbe C. B. , Trampe‐Kieslich A. , Reifenberger G.
Published: September 2005

Glioblastomas frequently carry genetic alterations resulting in an aberrant activation of the phosphoinositol‐3‐kinase (Pi3k)/protein kinase B (Akt) signalling pathway, including most notably phosphatase and tensin homolog (PTEN) mutation, epidermal growth factor receptor (EGFR) amplification and rearrangement, as well as carboxyl‐terminal modulator protein (CTMP) hypermethylation [Knobbe et al., (2004) Hypermethylation and transcriptional downregulation of the carboxyl‐terminal modulator protein gene in glioblastomas. J Natl Cancer Institute, 96, 483–486]. Here, we investigated two further Pi3k/Akt pathway genes, namely PIK3CA (3q26.3) and phosphatidylinositol‐3‐kinase enhancer (PIKE) (CENTG1, 12q14), for genetic alteration and aberrant expression in a series of 97 primary glioblastomas. Single strand conformation polymorphism (SSCP) analysis of PIK3CA revealed somatic mutations in five tumours (5%). Twelve glioblastomas (12%) showed amplification of PIKE with invariable co‐amplification of the adjacent CDK4 gene. All tumours with PIKE amplification as well as the vast majority of glioblastomas without amplification demonstrated increased expression of PIKE‐A but not PIKE‐S/L transcripts as compared with non‐neoplastic brain tissue. Taken together, our data support an important role of PIK3CA and PIKE gene aberrations in the molecular pathogenesis of primary glioblastomas.
Novel ‘inflammatory plaque’ pathology in presenilin‐1 Alzheimer's disease
Authors: Shepherd C. E. , Gregory G. C. , Vickers J. C. , Halliday G. M.
Published: September 2005

Inflammation, in the form of reactive astrocytes and microglia, is thought to play an important role in Alzheimer's disease (AD) pathogenesis where it correlates with brain atrophy and disease severity. The Aβ protein, which comprises senile plaques, is thought to be responsible for initiating this inflammatory response. Despite having a more aggressive disease process and greater Aβ deposition, few studies have investigated inflammation in early onset AD cases with mutations in the presenilin‐1 (PS‐1) gene. In fact, many researchers place importance on a variant plaque pathology in PS‐1 cases, known as cotton wool plaques, which lack significant inflammatory infiltrate. We investigated the association between inflammation and plaque pathology in PS‐1 AD. Classic cored, cotton wool and diffuse Aβ plaques were observed in all cases. PS‐1 cases also exhibited a novel plaque pathology with a significantly greater inflammatory response in the form of reactive microglia and astrocytes. These ‘inflammatory plaques’ consisted of a dense cresyl violet‐, silver‐, and thioflavin S‐positive, but Aβ‐, tau‐, apolipoprotein E (ApoE)‐, non‐Aβ component of Alzheimer's disease amyloid (NAC)‐ and PS‐1‐negative core. These findings indicate potent stimulator(s) of inflammation that are not typical of the Aβ that accumulates in the pathological hallmarks of sporadic AD. Identification of this substance may be important for the development of future therapeutic strategies.
Attenuation of Aβ deposition in the entorhinal cortex of normal elderly individuals associated with tobacco smoking
Authors: Court J. A. , Johnson M. , Religa D. , Keverne J. , Kalaria R. , Jaros E. , McKeith I. G. , Perry R. , Naslund J. , Perry E. K.
Published: September 2005

Investigating correlates of tobacco smoking provides the only currently available opportunity of examining effects of long‐term exposure of nicotinic receptors on a specific nicotinic agonist in human. Alzheimer‐type pathology (Aβ and abnormally phosphorylated tau assessed on the basis of AT8 immunoreactivity) together with vascular markers has been compared in age‐matched groups of normal elderly smokers and non‐smokers in the entorhinal cortex, an area of noted age‐related pathology. The density of total Aβ and diffuse Aβ immunoreactivity, together with formic acid‐extractable Aβ42 but not Aβ40, was reduced in smokers (n = 10–18) compared with non‐smokers (n = 10–20) (P < 0.05). There was also a reduced percentage of cortical and leptomeningeal vessels with associated Aβ immunoreactivity in smokers (n = 13) compared with non‐smokers (n = 14) (P < 0.005 and 0.05, respectively). There was a significant inverse correlation between formic acid‐extractable Aβ42 and pack years (n = 34, r = −0.389, P = 0.025), with a similar trend for total Aβ immunoreactivity which did not reach statistical significance (n = 30, r = −0.323, P = 0.082). In contrast, there were no significant group differences for vascular markers (collagen IV, α‐actin or glucose transporter 1), AT8 immunoreactivity or phosphate‐buffered saline‐soluble Aβ peptides, and no significant associations with gender for any of the measured parameters. These findings are consistent with previously reported reductions in histologically assessed amyloid plaques in aged human brain associated with tobacco use and dramatic lessening of Aβ deposits in APPsw mice after nicotine treatment. Development of nicotinic drugs to protect against β‐amyloidosis as one of the principal pathological hallmarks of brain ageing and Alzheimer's disease is indicated.
Characterization of matrix metalloproteinases in denervated muscle
Authors: Demestre M. , Orth M. , Wells G. M. , Gearing A. J. , Hughes R. A. C. , Gregson N. A.
Published: September 2005

In a nerve crush model of denervation, we examined muscle matrix metalloproteinase (MMP) expression, localization and activity. In normal muscle, MMP mRNA levels were low, and immunohistochemically MMPs were distributed around the muscle fibre with MMPs‐3, ‐7 and ‐9 also staining at the neuromuscular junction. Seven days after nerve crush, muscle MMP immunoreactivity, especially MMP‐12 and MMP‐14, became irregularly distributed. At 20 days reinnervation of the muscle was observed, and some restitution of the normal pattern of immunoreactivity was noted concomitant with a higher level of MMP mRNA expression. In situ zymography showed that MMP activity was very weak in normal muscle whereas it was increased up to 40 days following denervation. Our results suggest that MMPs in muscle are involved in the tissue changes following denervation. Further experiments are required to test the hypothesis that MMP inhibition may be beneficial in protecting muscle from excessive remodelling following denervation and therefore improve reinnervation.
Continuous expression in tobacco leaves of a Brassica napus PEND homologue blocks differentiation of plastids and development of palisade cells
Authors: Wycliffe Paul , Sitbon Folke , Wernersson Jonny , Ezcurra Inés , Ellerström Mats , Rask Lars
Published: September 2005

Brassica napus complementary deoxyribonucleic acid (cDNA) clones encoding a DNA‐binding protein, BnPEND, were isolated by Southwestern screening. A distinctive feature of the protein was a bZIP‐like sequence in the amino‐terminal portion, which, after expression in Escherichia coli, bound DNA. BnPEND transcripts were present in B. napus roots and flower buds, and to a lesser extent in stems, flowers and young leaves. Treatment in the dark for 72 h markedly increased the amount of BnPEND transcript in leaves of all ages. Sequence comparison showed that BnPEND was similar to a presumed transcription factor from B. napus, GSBF1, a protein deduced from an Arabidopsis thaliana cDNA (BX825084) and the PEND protein from Pisum sativum, believed to anchor the plastid DNA to the envelope early during plastid development. Homology to expressed sequence tag (EST) sequences from additional species suggested that BnPEND homologues are widespread among the angiosperms. Transient expression of BnPEND fused with green fluorescent protein (GFP) in Nicotiana benthamiana epidermal cells showed that BnPEND is a plastid protein, and that the 15 amino acids at the amino‐terminal contain information about plastid targeting. Expression of BnPEND in Nicotiana tabacum from the Cauliflower Mosaic Virus 35S promoter gave stable transformants with different extents of white to light‐green areas in the leaves, and even albino plants. In the white areas, but not in adjacent green tissue, the development of palisade cells and chloroplasts was disrupted. Our data demonstrate that the BnPEND protein, when over‐expressed at an inappropriate stage, functionally blocks the development of plastids and leads to altered leaf anatomy, possibly by preventing the release of plastid DNA from the envelope.
Identification of the conserved serine/threonine residues important for gibberellin‐sensitivity of Arabidopsis RGL2 protein
Authors: Hussain Alamgir , Cao Dongni , Cheng Hui , Wen Zilong , Peng Jinrong
Published: September 2005

The DELLA proteins GAI, RGA, RGL1 and RGL2 in Arabidopsis are plant growth repressors, repressing diverse developmental processes. Studies have shown that gibberellin (GA) attenuates the repressive function of DELLA proteins by triggering their degradation via the proteasome pathway. However, it is not known if GA‐induced protein degradation is the only pathway for regulating the bioactivity of DELLA proteins. We show here that tobacco BY2 cells represent a suitable system for studying GA signaling. RGL2 exists in a phosphorylated form in BY2 cells. RGL2 undergoes GA‐induced degradation, and this process is blocked by proteasome inhibitors and serine/threonine phosphatase inhibitors; however, serine/threonine kinase inhibitors had no detectable effect, suggesting that dephosphorylation of serine/threonine is probably a prerequisite for degradation of RGL2 via the proteasome pathway. Site‐directed substitution of all 17 conserved serine and threonine residues showed that six mutants (RGL2S441D, RGL2S542D, RGL2T271E, RGL2T319E, RGL2T411E and RGL2T535E) mimicking the status of constitutive phosphorylation are resistant to GA‐induced degradation. This suggests that these sites are potential phosphorylation sites. A functional assay based on the expression of GA 20‐oxidase revealed that RGL2T271E is probably a null mutant, RGL2S441D, RGL2S542D, RGL2T319E and RGL2T411E only retained about 4–17% of the activity of the wild type RGL2, whereas RGL2T535E retained about 66% of the activity of the wild type RGL2. However, expression of GA 20‐oxidase in BY2 cells expressing these mutant proteins is still responsive to GA, suggesting that the stabilization of RGL2 protein is not the only pathway for regulating its bioactivity.
Pseudomonas syringae pv. tomato type III effectors AvrPto and AvrPtoB promote ethylene‐dependent cell death in tomato
Authors: Cohn Jonathan R. , Martin Gregory B.
Published: September 2005

The type III secretion system (TTSS) of Pseudomonas syringae pv. tomato (Pst) injects into the plant cell effector proteins that play an essential role in the formation of bacterial speck disease. To investigate the molecular roles of TTSS effectors in disease formation, we used a cDNA microarray to analyze the expression of approximately 8600 random tomato genes in response to wild‐type Pst strain DC3000 and a mutant lacking a functional TTSS. Many of the differentially expressed genes identified encode proteins associated with hormone response or hormone biosynthesis pathways. Using isogenic mutant strains of DC3000, we monitored host transcriptional changes in response to the TTSS effector proteins AvrPto and AvrPtoB, both of which are important virulence factors on susceptible tomato lines. We found that AvrPto and AvrPtoB induce a set of host genes involved in ethylene biosynthesis and signaling, and in particular they regulate the expression of two genes, LeACO1 and LeACO2, encoding the ethylene‐forming enzyme ACC oxidase. Analysis of transgenic tomato lines with diminished ACC oxidase activity revealed that ethylene production by the host is required for the full virulence activity of both AvrPto and AvrPtoB. AvrPto and AvrPtoB therefore appear to promote enhanced disease in tomato leaves, in part, by upregulating genes involved in ethylene production.
Predicting longer‐term changes in weed populations under GMHT crop management
Authors: HEARD M S , ROTHERY P , PERRY J N , FIRBANK L G
Published: September 2005

The UK Farm Scale Evaluations (FSE) compared the effects on biodiversity of management of genetically modified herbicide‐tolerant (GMHT) crops and conventional crops over the shorter term. We simulated population changes over seven 4‐year rotations (28 years) for weeds in crop rotations that included cereals and spring‐sown GMHT and conventional oilseed rape and beet, using FSE data and assuming the continuation of the weed management systems practised in the FSE. The weed density dependence that was modelled integrated change caused by population dynamics and farmers’ responses to changes in weed density. Predicted weed seed populations decreased under conventional management and at a greater rate under GMHT. Total seed densities were lower for GMHT cropping by a factor of 0.7–0.8. The predicted distributions of weeds had more fields with lower weed densities under GMHT cropping. Such changes could affect animal populations on farmland, depending on the scale of uptake of GMHT crop cultivars.