bluuebunny/crossref_metadata_2025 huggingface dataset:
Limits to gene flow in a cosmopolitan marine planktonic diatom
Authors: Casteleyn Griet , Leliaert Frederik , Backeljau Thierry , Debeer Ann-Eline , Kotaki Yuichi , Rhodes Lesley , Lundholm Nina , Sabbe Koen , Vyverman Wim
Published: July 2010

The role of geographic isolation in marine microbial speciation is hotly debated because of the high dispersal potential and large population sizes of planktonic microorganisms and the apparent lack of strong dispersal barriers in the open sea. Here, we show that gene flow between distant populations of the globally distributed, bloom-forming diatom speciesPseudo-nitzschia pungens(clade I) is limited and follows a strong isolation by distance pattern. Furthermore, phylogenetic analysis implies that under appropriate geographic and environmental circumstances, like the pronounced climatic changes in the Pleistocene, population structuring may lead to speciation and hence may play an important role in diversification of marine planktonic microorganisms. A better understanding of the factors that control population structuring is thus essential to reveal the role of allopatric speciation in marine microorganisms.
Presynaptic dopaminergic compartment determines the susceptibility to L-DOPA–induced dyskinesia in rats
Authors: Ulusoy Ayse , Sahin Gurdal , Kirik Deniz
Published: July 2010

Drug-induced dyskinesias in dopamine-denervated animals are known to depend on both pre- and postsynaptic changes of the nigrostriatal circuitry. In lesion models used thus far, changes occur in both of these compartments and, therefore, it has not been possible to dissect the individual contribution of each compartment in the pathophysiology of dyskinesias. Here we silenced the nigrostriatal dopamine neurotransmission without affecting the anatomical integrity of the presynaptic terminals using a short-hairpin RNA-mediated knockdown of tyrosine hydroxylase enzyme (shTH). This treatment resulted in significant reduction (by about 70%) in extracellular dopamine concentration in the striatum as measured by on-line microdialysis. Under these conditions, the animals remained nondyskinetic after chronic L-DOPA treatment, whereas partial intrastriatal 6-hydoxydopamine lesioned rats with comparable reduction in extracellular dopamine levels developed dyskinesias. On the other hand, apomorphine caused moderate to severe dyskinesias in both groups. Importantly, single-dose L-DOPA challenge in apomorphine-primed shTH animals failed to activate the already established abnormal postsynaptic responses. Taken together, these data provide direct evidence that the status of the presynaptic, DA releasing compartment is a critical determinant of both the induction and maintenance of L-DOPA–induced dyskinesias.
Correlated memory defects and hippocampal dendritic spine loss after acute stress involve corticotropin-releasing hormone signaling
Authors: Chen Yuncai , Rex Christopher S. , Rice Courtney J. , Dubé Céline M. , Gall Christine M. , Lynch Gary , Baram Tallie Z.
Published: July 2010

Stress affects the hippocampus, a brain region crucial for memory. In rodents, acute stress may reduce density of dendritic spines, the location of postsynaptic elements of excitatory synapses, and impair long-term potentiation and memory. Steroid stress hormones and neurotransmitters have been implicated in the underlying mechanisms, but the role of corticotropin-releasing hormone (CRH), a hypothalamic hormone also released during stress within hippocampus, has not been elucidated. In addition, the causal relationship of spine loss and memory defects after acute stress is unclear. We used transgenic mice that expressed YFP in hippocampal neurons and found that a 5-h stress resulted in profound loss of learning and memory. This deficit was associated with selective disruption of long-term potentiation and of dendritic spine integrity in commissural/associational pathways of hippocampal area CA3. The degree of memory deficit in individual mice correlated significantly with the reduced density of area CA3 apical dendritic spines in the same mice. Moreover, administration of the CRH receptor type 1 (CRFR
1
) blocker NBI 30775 directly into the brain prevented the stress-induced spine loss and restored the stress-impaired cognitive functions. We conclude that acute, hours-long stress impairs learning and memory via mechanisms that disrupt the integrity of hippocampal dendritic spines. In addition, establishing the contribution of hippocampal CRH–CRFR
1
signaling to these processes highlights the complexity of the orchestrated mechanisms by which stress impacts hippocampal structure and function.
Long-range function of an intergenic retrotransposon
Authors: Pi Wenhu , Zhu Xingguo , Wu Min , Wang Yongchao , Fulzele Sadanand , Eroglu Ali , Ling Jianhua , Tuan Dorothy
Published: July 2010

Retrotransposons including endogenous retroviruses and their solitary long terminal repeats (LTRs) compose >40% of the human genome. Many of them are located in intergenic regions far from genes. Whether these intergenic retrotransposons serve beneficial host functions is not known. Here we show that an LTR retrotransposon of ERV-9 human endogenous retrovirus located 40–70 kb upstream of the human fetal γ- and adult β-globin genes serves a long-range, host function. The ERV-9 LTR contains multiple CCAAT and GATA motifs and competitively recruits a high concentration of NF-Y and GATA-2 present in low abundance in adult erythroid cells to assemble an LTR/RNA polymerase II complex. The LTR complex transcribes intergenic RNAs unidirectionally through the intervening DNA to loop with and modulate transcription factor occupancies at the far downstream globin promoters, thereby modulating globin gene switching by a competitive mechanism.
Array comparative genomic hybridization-based characterization of genetic alterations in pulmonary neuroendocrine tumors
Authors: Voortman Johannes , Lee Jih-Hsiang , Killian Jonathan Keith , Suuriniemi Miia , Wang Yonghong , Lucchi Marco , Smith William I. , Meltzer Paul , Wang Yisong , Giaccone Giuseppe
Published: July 2010

The goal of this study was to characterize and classify pulmonary neuroendocrine tumors based on array comparative genomic hybridization (aCGH). Using aCGH, we performed karyotype analysis of 33 small cell lung cancer (SCLC) tumors, 13 SCLC cell lines, 19 bronchial carcinoids, and 9 gastrointestinal carcinoids. In contrast to the relatively conserved karyotypes of carcinoid tumors, the karyotypes of SCLC tumors and cell lines were highly aberrant. High copy number (CN) gains were detected in SCLC tumors and cell lines in cytogenetic bands encoding JAK2, FGFR1, and MYC family members. In some of those samples, the CN of these genes exceeded 100, suggesting that they could represent driver alterations and potential drug targets in subgroups of SCLC patients. In SCLC tumors, as well as bronchial carcinoids and carcinoids of gastrointestinal origin, recurrent CN alterations were observed in 203 genes, including the RB1 gene and 59 microRNAs of which 51 locate in the DLK1-DIO3 domain. These findings suggest the existence of partially shared CN alterations in these tumor types. In contrast, CN alterations of the TP53 gene and the MYC family members were predominantly observed in SCLC. Furthermore, we demonstrated that the aCGH profile of SCLC cell lines highly resembles that of clinical SCLC specimens. Finally, by analyzing potential drug targets, we provide a genomics-based rationale for targeting the AKT-mTOR and apoptosis pathways in SCLC.
The emergence, structure and development of ethnic identity during childhood: The case of Roma identity
Authors: Pnevmatikos Dimitris , Geka Maria , Divane Maria
Published: July 2010

This study investigates the emergence, development and structure of ethnic identity during childhood. Forty Roma children living in Greece aged between 2.8 and 11.9 years answered questions about their awareness/recognition of four aspects of their ethnic identity—namely place of habitation, traditional costumes, the Roma language, and early betrothal of children—their identity and their sense of stability and constancy. The study also investigates how the children feel about the abandonment of those four aspects. The evidence from the current data supports the hypothesis that awareness of ethnic identity emerges before the age of 4. Moreover, this study offers direct empirical evidence of the multidimensionality of ethnic identity. A model of three concentric rings is proposed, extending from a core containing the most highly valued aspects of ethnic identity to the outer annulus that comprises the nonpermanent and nonstable aspects of ethnic identity. The aspects in each annulus differ in terms of the development of the sense of stability and constancy and the feelings associated with loss of the aspects in question. Even the youngest participants considered the aspects in the core to be stable and constant as well as emotionally charged; and even the 11‐year‐olds did not consider the aspects contained in the outer, more fluid annulus as stable and constant aspects of their ethnic identity. The development of an aspect is determined by what the majority of adults in a society, at a particular time in history, consider to be most important.
Separation and identification of mouse liver membrane proteins using a gel-based approach in combination with 2DnanoLC-Q-TOF-MS/MS
Authors: Tran The Thanh , Phan Van Chi
Published: July 2010

In this work, we present results of membrane proteome profiling from mouse liver tissues using a gel-based approach in combination with 2DnanoLC-Q-TOF-MS/MS. Following purification of the membrane fraction, SDS-PAGE was carried out as a useful separation step. After staining, gels with protein bands were cut, reduced, alkylated and trypsin-digested. The peptide mixtures extracted from each gel slice were fractionated by two-dimensional nano liquid chromatography (2DnanoLC) coupled online with tandem mass spectrometry analysis (NanoESI-Q-TOF-MS/MS). The proteins were identified by MASCOT search against a mouse protein database using a peptide and fragment mass tolerance of ±0.5 Da. Protein identification was carried out using a Mowse scoring algorithm with a confidence level of 95% and processed by MSQuant v1.5 software for further validation. In total, 318 verified membrane proteins from mouse liver tissues were identified; 66.67% of them (212 proteins) contained at least one or more transmembrane domains predicted by the SOSUI program and 43 were found to be unique microsome membranes. Furthermore, GRAVY values of membrane proteins varied in the range -1.1276 to 0.9016 and only 31 (9.76%) membrane proteins had positive values. The functions and subcellular locations of the identified proteins were categorized as well, according to universal GO annotations.
G-SQZ: compact encoding of genomic sequence and quality data
Authors: Tembe Waibhav , Lowey James , Suh Edward
Published: July 2010

SUMMARY: Large volumes of data generated by high-throughput sequencing instruments present non-trivial challenges in data storage, content access and transfer. We present G-SQZ, a Huffman coding-based sequencing-reads-specific representation scheme that compresses data without altering the relative order. G-SQZ has achieved from 65% to 81% compression on benchmark datasets, and it allows selective access without scanning and decoding from start. This article focuses on describing the underlying encoding scheme and its software implementation, and a more theoretical problem of optimal compression is out of scope. The immediate practical benefits include reduced infrastructure and informatics costs in managing and analyzing large sequencing data.

Availability:  http://public.tgen.org/sqz. Academic/non-profit: Source: available at no cost under a non-open-source license by requesting from the web-site; Binary: available for direct download at no cost. For-Profit: Submit request for for-profit license from the web-site.

Contact:  wtembe@tgen.org
PERMORY: an LD-exploiting permutation test algorithm for powerful genome-wide association testing
Authors: Pahl Roman , Schäfer Helmut
Published: July 2010

Motivation: In genome-wide association studies (GWAS) examining hundreds of thousands of genetic markers, the potentially high number of false positive findings requires statistical correction for multiple testing. Permutation tests are considered the gold standard for multiple testing correction in GWAS, because they simultaneously provide unbiased type I error control and high power. At the same time, they demand heavy computational effort, especially with large-scale datasets of modern GWAS. In recent years, the computational problem has been circumvented by using approximations to permutation tests, which, however, may be biased.

Results: We have tackled the original computational problem of permutation testing in GWAS and herein present a permutation test algorithm one or more orders of magnitude faster than existing implementations, which enables efficient permutation testing on a genome-wide scale. Our algorithm does not rely on any kind of approximation and hence produces unbiased results identical to a standard permutation test. A noteworthy feature of our algorithm is a particularly effective performance when analyzing high-density marker sets.

Availability: Freely available on the web at http://www.permory.org

Contact:  rpahl@staff.uni-marburg.de
Determination and Comparison of Graded Dose–Response Curves for Epidural Bupivacaine and Ropivacaine for Analgesia in Laboring Nulliparous Women
Authors: Kee Warwick D. Ngan , Ng Floria F. , Khaw Kim S. , Lee Anna , Gin Tony
Published: July 2010

Background

The potencies of bupivacaine and ropivacaine have been compared using up-and-down methodology, but their complete dose-response curves have not been compared. The authors performed a random allocation-graded dose-response study of epidural bupivacaine and ropivacaine given epidurally for labor analgesia.

Methods

Three hundred laboring nulliparous patients were randomly given epidural bupivacaine (5, 10, 15, 20, 30, or 40 mg) or ropivacaine (7, 15, 20, 30, 45, or 60 mg) in 20 ml of saline. Visual Analog Scale pain scores were recorded for 30 min. Response was defined by the percentage decrease in pain score from baseline at 30 min, and dose-response data were analyzed by using nonlinear regression.

Results

Sigmoidal Emax model dose-response curves were fitted to the datasets for bupivacaine (R = 0.53) and ropivacaine (R = 0.59). The curves had similar steepness (Hill coefficient 2.02 [95% CI, 1.55-2.50] vs. 2.25 [1.70-2.79], P = 0.55). The ED50 (dose of the drug that reduces pain score to 50% of baseline at 30 min, also known as D50) of ropivacaine was greater than that of bupivacaine (15.3 [95% CI 13.7-17.1] mg vs. 11.3 [10.0-12.7] mg, P = 0.0003), but ED90 (D90) was similar (40.6 [32.4-51.1] mg vs. 33.4 [26.2-42.7] mg, P = 0.29). The potency ratio at ED50 for ropivacaine:bupivacaine was 0.75 (95% CI, 0.65-0.88).

Conclusions

Ropivacaine is less potent than bupivacaine, but otherwise they have similar dose-response characteristics. The difference in potency is not statistically significant at ED90 doses.
Volume Kinetics for Infusion Fluids
Authors: Hahn Robert G. , Warner David S.
Published: July 2010

Volume kinetics is a method used for analyzing and simulating the distribution and elimination of infusion fluids. Approximately 50 studies describe the disposition of 0.9% saline, acetated and lactated Ringer's solution, based on repeated measurements of the hemoglobin concentration and (sometimes) the urinary excretion. The slow distribution to the peripheral compartment results in a 50-75% larger plasma dilution during an infusion of crystalloid fluid than would be expected if distribution had been immediate. A drop in the arterial pressure during induction of anesthesia reduces the rate of distribution even further. The renal clearance of the infused fluid during surgery is only 10-20% when compared with that in conscious volunteers. Some of this temporary decrease can be attributed to the anesthesia and probably also to preoperative psychologic stress or dehydration. Crystalloid fluid might be allocated to "nonfunctional" fluid spaces in which it is unavailable for excretion. This amounts to approximately 20-25% during minor (thyroid) surgery.
Spinal α2-Adrenoceptor-mediated Analgesia in Neuropathic Pain Reflects Brain-derived Nerve Growth Factor and Changes in Spinal Cholinergic Neuronal Function
Authors: Hayashida Ken-ichiro , Eisenach James C.
Published: July 2010

Introduction

Spinal alpha2-adrenoceptor stimulation produces analgesia in neuropathic pain states, and this effect in animals is blocked by the inhibitors of brain-derived neurotrophic factor (BDNF) function. In rats, alpha2-adrenoceptor stimulation normally inhibits acetylcholine release, but it excites release after nerve injury. The authors examined the roles of BDNF and excitatory Gs-protein in this change.

Methods

Male rats underwent L5-L6 spinal nerve ligation (SNL), and their lumbar spinal dorsal horns with or without spinal BDNF infusion were used for either synaptosome preparation for acetylcholine release or immunostaining for choline acetyltransferase.

Results

SNL did not alter spontaneous release from synaptosomes or choline acetyltransferase immunoreactivity in the spinal dorsal horn, but it reduced KCl-evoked acetylcholine release. Dexmedetomidine inhibited KCl-evoked acetylcholine release in synaptosomes from normal rats, but it excited KCl-evoked release in synaptosomes from SNL rats, and both effects were blocked by the alpha2-adrenoceptor antagonist idazoxan. Spinal infusion of an antibody to BDNF reduced choline acetyltransferase immunoreactivity in the spinal dorsal horn in both normal and SNL rats and abolished facilitation of KCl-evoked acetylcholine release by dexmedetomidine in SNL rats. Dexmedetomidine facilitation of acetylcholine release was also blocked by the inhibitors of Gs function.

Discussion

The increased reliance of spinal alpha2 adrenoceptors on cholinergic stimulation to cause analgesia after nerve injury reflects in part a shift from direct inhibition to direct excitation of spinal cholinergic neurons. The authors' results suggest that this shift relies on an interaction with Gs-proteins and BDNF.
Multicenter, Randomized, Comparative Cost-effectiveness Study Comparing 0, 1, and 2 Diagnostic Medial Branch (Facet Joint Nerve) Block Treatment Paradigms before Lumbar Facet Radiofrequency Denervation
Authors: Cohen Steven P. , Williams Kayode A. , Kurihara Connie , Nguyen Conner , Shields Cynthia , Kim Peter , Griffith Scott R. , Larkin Thomas M. , Crooks Matthew , Williams Necia , Morlando Benny , Strassels Scott A.
Published: July 2010

Background

Among patients presenting with axial low back pain, facet arthropathy accounts for approximately 10-15% of cases. Facet interventions are the second most frequently performed procedures in pain clinics across the United States. Currently, there are no uniformly accepted criteria regarding how best to select patients for radiofrequency denervation.

Methods

A randomized, multicenter study was performed in 151 subjects with suspected lumbar facetogenic pain comparing three treatment paradigms. Group 0 received radiofrequency denervation based solely on clinical findings; group 1 underwent denervation contingent on a positive response to a single diagnostic block; and group 2 proceeded to denervation only if they obtained a positive response to comparative blocks done with lidocaine and bupivacaine. A positive outcome was predesignated as > or =50% pain relief coupled with a positive global perceived effect persisting for 3 months.

Results

In group 0, 17 patients (33%) obtained a successful outcome at 3 months versus eight patients (16%) in group 1 and 11 (22%) patients in group 2. Denervation success rates in groups 0, 1, and 2 were 33, 39, and 64%, respectively. Pain scores and functional capacity were significantly lower at 3 months but not at 1 month in group 2 subjects who proceeded to denervation compared with patients in groups 0 and 1. The costs per successful treatment in groups 0, 1, and 2 were $6,286, $17,142, and $15,241, respectively.

Conclusions

Using current reimbursement scales, these findings suggest that proceeding to radiofrequency denervation without a diagnostic block is the most cost-effective treatment paradigm.
Pharmacokinetic-Pharmacodynamic Modeling of Propofol in Children
Authors: Rigouzzo Agnes , Servin Frederique , Constant Isabelle
Published: July 2010

Background

The aim of this study was to identify the best model to describe pharmacokinetics and pharmacodynamics in prepubertal children and therefore to calculate the corresponding pharmacodynamic parameters. In addition, and to confirm our method, a group of postpubertal subjects was also studied.

Methods

Sixteen children (9.5 yr, range 6-12) and 13 adults (22 yr, range 13-35) were included. Induction was performed by plasma target-controlled infusion of propofol (6 microg/ml) based on the Kataria model in children and on the Schnider model in adults. The relationship of bispectral index to predicted concentrations was studied during induction using the Kataria, pediatric Marsh, Schüttler, and Schnider models in children. Because the best performance was obtained, strangely enough, with the Schnider model, the two groups were pooled to investigate influence of puberty on pharmacodynamic parameters (kE0 [plasma effect-site equilibration rate constant] and Ce50 [effect-site concentration corresponding with 50% of the maximal effect]). The time-to-peak effect was calculated, and the kE0 was determined for the Kataria model (nonlinear mixed-effects modeling; pkpdtools).

Results

In children, the predicted concentration/effect relationship was best described using the Schnider model. When the whole population was considered, a significant improvement in this model was obtained using puberty as a covariate for kE0 and Ce50. The time to peak effect, Tpeak (median, 0.71 [range, 0.37-1.64] and 1.73 [1.4-2.68] min), and the Ce50 (3.71 [1.88-4.4] and 3.07 [2.95-5.21] microg/ml) were shorter and higher, respectively, in children than in adults. The kE0 linked to the Kataria model was 4.6 [1.4-11] min.

Conclusions

In children, the predicted concentration/effect relationships were best described using the Schnider model described for adults compared with classic pediatric models. The study suggests that the Schnider model might be useful for propofol target-control infusion in children.
Virus-Induced Gene Silencing of Plastidial Soluble Inorganic Pyrophosphatase Impairs Essential Leaf Anabolic Pathways and Reduces Drought Stress Tolerance inNicotiana benthamiana
Authors: George Gavin M. , van der Merwe Margaretha J. , Nunes-Nesi Adriano , Bauer Rolene , Fernie Alisdair R. , Kossmann Jens , Lloyd James R.
Published: July 2010

The role of pyrophosphate in primary metabolism is poorly understood. Here, we report on the transient down-regulation of plastid-targeted soluble inorganic pyrophosphatase in Nicotiana benthamiana source leaves. Physiological and metabolic perturbations were particularly evident in chloroplastic central metabolism, which is reliant on fast and efficient pyrophosphate dissipation. Plants lacking plastidial soluble inorganic pyrophosphatase (psPPase) were characterized by increased pyrophosphate levels, decreased starch content, and alterations in chlorophyll and carotenoid biosynthesis, while constituents like amino acids (except for histidine, serine, and tryptophan) and soluble sugars and organic acids (except for malate and citrate) remained invariable from the control. Furthermore, translation of Rubisco was significantly affected, as observed for the amounts of the respective subunits as well as total soluble protein content. These changes were concurrent with the fact that plants with reduced psPPase were unable to assimilate carbon to the same extent as the controls. Furthermore, plants with lowered psPPase exposed to mild drought stress showed a moderate wilting phenotype and reduced vitality, which could be correlated to reduced abscisic acid levels limiting stomatal closure. Taken together, the results suggest that plastidial pyrophosphate dissipation through psPPase is indispensable for vital plant processes.
Cooperation of LPA3 and LPA2 Is Essential for Photosystem II Assembly in Arabidopsis
Authors: Cai Wenhe , Ma Jinfang , Chi Wei , Zou Meijuan , Guo Jinkui , Lu Congming , Zhang Lixin
Published: July 2010

Photosystem II (PSII) is a multisubunit membrane protein complex that is assembled in a sequence of steps. However, the molecular mechanisms responsible for the assembly of the individual subunits into functional PSII complexes are still largely unknown. Here, we report the identification of a chloroplast protein, Low PSII Accumulation3 (LPA3), which is required for the assembly of the CP43 subunit in PSII complexes in Arabidopsis (Arabidopsis thaliana). LPA3 interacts with LPA2, a previously identified PSII CP43 assembly factor, and a double mutation of LPA2 and LPA3 is more deleterious for assembly than either single mutation, resulting in a seedling-lethal phenotype. Our results indicate that LPA3 and LPA2 have overlapping functions in assisting CP43 assembly and that cooperation between LPA2 and LPA3 is essential for PSII assembly. In addition, we provide evidence that LPA2 and LPA3 interact with Albino3 (Alb3), which is essential for thylakoid protein biogenesis. Thus, the function of Alb3 in some PSII assembly processes is probably mediated through interactions with LPA2 and LPA3.
Lake‐specific variation in growth, migration timing and survival of juvenile sockeye salmon Oncorhynchus nerka: separating environmental from genetic influences
Authors: Reed T. E. , Martinek G. , Quinn T. P.
Published: July 2010

Time series on juvenile life‐history traits obtained from sockeye salmon Oncorhynchus nerka were analysed to assess lake‐specific environmental influences on juvenile migration timing, size and survival of fish from a common gene pool. Every year for the past two decades, O. nerka have been spawned at a hatchery facility, and the progeny released into two lakes that differ in average summer temperatures, limnological attributes and growth opportunities. Juveniles reared in the warmer, more productive Crosswind Lake were larger and heavier as smolts compared to those from the cooler, less productive Summit Lake and had higher in‐lake and subsequent marine survival. Crosswind Lake smolts migrated from the lake to sea slightly earlier in the season but the migration timing distributions overlapped considerably across years. Fry stocking density had a negative effect on smolt length for both lakes, and a negative effect on in‐lake survival in Summit Lake. Taken together, the results revealed a strong effect of lake‐rearing environment on the expression of life‐history variation in O. nerka. The stocking of these lakes each year with juveniles from a single mixed‐source population provided a large‐scale reverse common‐garden experiment, where the same gene pool was exposed to different environments, rather than the different gene pools in the same environment approach typical of evolutionary ecology studies. Other researchers are encouraged to seek and exploit similar serendipitous situations, which might allow environmental and genetic influences on ecologically important traits to be distinguished in natural or semi‐natural settings.
New system for multilocus variable‐number tandem‐repeat analysis of the enterohemorrhagic Escherichia coli strains belonging to three major serogroups: O157, O26, and O111
Authors: Izumiya Hidemasa , Pei Yingxin , Terajima Jun , Ohnishi Makoto , Hayashi Tetsuya , Iyoda Sunao , Watanabe Haruo
Published: July 2010

Enterohemorrhagic Escherichia coli (EHEC), a food‐ and waterborne pathogen, causes diarrhea, hemorrhagic colitis, and life‐threatening HUS. MLVA is a newly developed and widely accepted genotyping tool. An MLVA system for EHEC O157 involving nine genomic loci has already been established. However, the present study revealed that the above‐mentioned MLVA system cannot analyze EHEC O26 and O111 isolates—the second and third most dominant EHEC serogroups in Japan, respectively. Therefore, with several modifications to the O157 system and the use of nine additional loci, we developed an expanded MLVA system applicable to EHEC O26, O111, and O157. Our MLVA system had a relatively high resolution power for each of the three serogroups: Simpson's index of diversity was 0.991 (95% CI = 0.989–0.993), 0.988 (95% CI, 0.986–0.990), and 0.986 (95% CI, 0.979–0.993) for O26, O111, and O157, respectively. This system also detected outbreak‐related isolates; the isolates collected during each of the 12 O26 and O111 outbreaks formed unique clusters, and most of the repeat copy numbers among the isolates collected during the same outbreak exhibited no or single‐locus variations. These results were comparable to those of cluster analyses based on PFGE profiles. Therefore, our system can complement PFGE analysis—the current golden method. Because EHEC strains of three major serogroups can be rapidly analyzed on a single platform with our expanded MLVA system, this system could be widely used in molecular epidemiological studies of EHEC infections.
Nobiletin, a citrus polymethoxyflavonoid, suppresses multiple angiogenesis‐related endothelial cell functions and angiogenesis in vivo
Authors: Kunimasa Kazuhiro , Ikekita Masahiko , Sato Mayumi , Ohta Toshiro , Yamori Yukio , Ikeda Megumi , Kuranuki Sachi , Oikawa Tsutomu
Published: July 2010

Nobiletin is a citrus polymethoxyflavonoid that suppresses tumor growth and metastasis, both of which depend on angiogenesis. We recently identified nobiletin as a cell differentiation modulator. Because cell differentiation is a critical event in angiogenesis, it might be possible that nobiletin could exhibit antiangiogenic activity, resulting in suppression of these tumor malignant properties. To verify this possibility, we examined the antiangiogenic effects of nobiletin in vitro and in vivo. Nobiletin had concentration‐dependent inhibitory effects on multiple functions of angiogenesis‐related endothelial cells (EC); it suppressed the proliferation, migration and tube formation on matrigel of human umbilical vein EC (HUVEC) stimulated with endothelial cell growth supplement (ECGS), a mixture of acidic and basic fibroblast growth factors (FGFs). Gelatin zymography and northern blotting revealed that nobiletin suppressed pro‐matrix metalloproteinase‐2 (proMMP‐2) production and MMP‐2 mRNA expression in ECGS‐stimulated HUVEC. Nobiletin also downregulated cell‐associated plasminogen activator (PA) activity and urokinase‐type PA mRNA expression. Furthermore, nobiletin inhibited angiogenic differentiation induced by vascular endothelial growth factor and FGF, an in vitro angiogenesis model. This inhibition was accompanied by downregulation of angiogenesis‐related signaling molecules, such as extracellular signal‐regulated kinase 1/2 and c‐Jun N‐terminal kinase, and transcriptional factors (c‐Jun and signal transducer and activator of transcription 3), and activation of the caspase pathway. In a chick embryo chorioallantoic membrane assay, nobiletin showed an antiangiogenic activity, the ID50 value being 10 μg (24.9 nmol) per egg. These results indicate that nobiletin is a novel antiangiogenic compound that exhibits its activity through combined inhibition of multiple angiogenic EC functions. (Cancer Sci 2010; 101: 2462–2469)
Momordica charantia leaf extract suppresses rat prostate cancer progression in vitro and in vivo
Authors: Pitchakarn Pornsiri , Ogawa Kumiko , Suzuki Shugo , Takahashi Satoru , Asamoto Makoto , Chewonarin Teera , Limtrakul Pornngarm , Shirai Tomoyuki
Published: July 2010

Cancer metastasis is a major cause of death in cancer patients, with invasion as a first step greatly contributing to the failure of clinical treatments. Any compounds with an inhibitory influence on this process are therefore of prime interest. Momordica charantia (bitter melon) is widely consumed as a vegetable and especially as a folk medicine in Asia. Here, we investigated the anti‐invasive effects of bitter melon leaf extract (BMLE) on a rat prostate cancer cell line (PLS10) in vitro and in vivo. The results indicated that non‐toxic concentrations of BMLE significantly inhibited the migration and invasion of cells in vitro. The results of zymography showed that BMLE inhibited the secretion of MMP‐2, MMP‐9 and urokinase plasminogen activator (uPA) from PLS10. Real‐time RT‐PCR revealed that BMLE not only significantly decreased gene expression of MMP‐2 and MMP‐9, but also markedly increased the mRNA level of TIMP‐2, known to have inhibitory effects on the activity of MMP‐2. An EnzChek gelatinase/collagenase assay showed that collagenase type IV activity was partially inhibited by BMLE. In the in vivo study, intravenous inoculation of PLS10 to nude mice resulted in a 100% survival rate in the mice given a BMLE‐diet as compared with 80% in the controls. The incidence of lung metastasis did not show any difference, but the percentage lung area occupied by metastatic lesions was slightly decreased in the 0.1% BMLE treatment group and significantly decreased with 1% BMLE treatment as compared with the control. Thus, the results indicate for the first time an anti‐metastatic effect of BMLE both in vitro and in vivo. (Cancer Sci 2010)
Amphiregulin regulates the activation of ERK and Akt through epidermal growth factor receptor and HER3 signals involved in the progression of pancreatic cancer
Authors: Yotsumoto Fusanori , Fukami Tatsuya , Yagi Hiroshi , Funakoshi Akihiro , Yoshizato Toshiyuki , Kuroki Masahide , Miyamoto Shingo
Published: July 2010

Pancreatic cancer is one of the most lethal malignancies. Epidermal growth factor receptor (EGFR), HER3, Akt, and amphiregulin have been recognized as targets for pancreatic cancer therapy. Although gemcitabine + erlotinib has been the recommended chemotherapy for pancreatic cancer, the prognosis is extremely poor. The development of molecularly targeted therapies has been required for patients with pancreatic cancer. To assess the validation of amphiregulin as a target for pancreatic cancer therapy, we examined its expression in pancreatic cancer using real‐time PCR analyses and ELISA. We also measured the apoptotic cell rate using TUNEL assays. In addition, alterations in signaling pathways were detected by immunoblotting analyses. Treatment with gemcitabine, which reduced the cell viability and augmented the cell apoptotic rate, activated and subsequently attenuated ERK and EGFR signals. However, gemcitabine, paclitaxel, or cisplatin treatment enhanced the Akt activation, heterodimer formation of EGFR with HER3, and secretion of amphiregulin, indicating that the presence of gemcitabine promoted the activity of targeted molecules including amphiregulin, Akt, and HER3 for pancreatic cancer therapy. Combined treatment with an inhibitor for amphiregulin and gemcitabine, paclitaxel, or cisplatin induced synergistic antitumor effects, accompanied by the suppression of Akt and ERK activation. Blockade of amphiregulin suppressed the activities of EGFR, HER3, and Akt and the expression of amphiregulin itself. According to this evidence, combination chemotherapy of conventional anticancer drugs plus an inhibitor for amphiregulin would allow us to provide more favorable clinical outcomes for patients with pancreatic cancer. (Cancer Sci 2010; 101: 2351–2360)
Expression of C4.4A at the invasive front is a novel prognostic marker for disease recurrence of colorectal cancer
Authors: Konishi Ken , Yamamoto Hirofumi , Mimori Koshi , Takemasa Ichiro , Mizushima Tsunekazu , Ikeda Masataka , Sekimoto Mitsugu , Matsuura Nariaki , Takao Toshifumi , Doki Yuichiro , Mori Masaki
Published: July 2010

Metastasis‐associated gene C4.4A is a glycolipid‐anchored membrane protein expressed in several human malignancies. The aim of this study was to explore the expression and clinical relevance of C4.4A in colorectal cancer. By quantitative RT‐PCR, 154 colorectal cancer tissues were examined for C4.4A mRNA. We examined 132 colorectal cancer tissues by immunohistochemistry using a new polyclonal antibody that recognizes the C4.4A protein C‐terminus containing the glycosylphosphatidyl‐inositol anchor signaling sequence. A significant difference in 5‐year overall survival was found between samples with high and low expression of C4.4A mRNA (P = 0.0005). Immunohistochemistry showed strong membranous staining of C4.4A at the invasive front of colorectal cancer tumors and at the frontier of metastatic lesions to lymph node and lung. The membranous staining with enhanced intensity at the invasive front of the primary colorectal cancer (Type A: 34/132, 25.6%) was associated with depth of invasion (P = 0.033) and venous invasion (P = 0.003), and was a significant independent prognostic factor (5‐year overall survival in the entire series [n = 132; P = 0.004] and disease‐free survival in stage II and III colorectal cancers [n = 82; P = 0.003]). Moreover, Type A C4.4A expression was linked to shorter liver metastasis‐free survival rate, lung metastasis‐free survival rate, or hematogenous metastasis‐free survival (P = 0.0279, P = 0.0061, and P = 0.0006, respectively). Our data indicate that expression of the C4.4A protein at the invasive front acts as a novel prognostic marker in colorectal cancer, possibly through invasion‐related mechanisms. (Cancer Sci 2010)
Underutilization of gastroprotection for at‐risk patients undergoing percutaneous coronary intervention: Spain compared with the United States
Authors: Casado‐Arroyo R. , Scheiman J. M. , Polo‐Tomas M. , Saini S. D. , Del Rio A. , Guastello E. , Lanas A.
Published: July 2010

Aliment Pharmacol Ther 2010; 32: 689–695

Background  Proton pump inhibitors (PPIs) are the preferred agents for the prevention of aspirin‐associated upper gastrointestinal bleeding (UGIB). Data are limited to determine whether PPIs are being used to reduce UGIB risk.

Aim  To evaluate the implementation of PPI treatment to reduce the GI risk in two cardiology centres from Europe and the United States.

Methods  A retrospective cross‐sectional study was carried out at the University of Michigan and University Hospital‐Zaragoza in 429 consecutive patients hospitalized for percutaneous coronary intervention (PCI) on dual antiplatelet therapy.

Results  Admission for PPI co‐therapy was similar (34% vs. 30%) in both centres. At discharge, the proportion of high‐risk patients receiving PPI therapy in the Spanish centre (75.4%) was higher than their American peers (55.6%) (OR: 2.5; 95% CI; 1.3–4.7). No differences in PPI prescription rates were found among Spanish patients with/without GI risk factors. The opportunity to initiate PPI co‐therapy in high‐risk patients was missed in 81.8% (36/44) of those not on PPI at admission in US patients vs. 24.1% (19/79) (P < 0.0001) in Spanish patients.

Conclusions  There are important differences concerning PPI prescription and risk stratification in the two centres when managing PCI patients. Efforts to stratify risks and utilize appropriate strategies for UGIB prophylaxis in high‐risk patients are warranted.
Specific effects exerted by B‐lymphoproliferative diseases on peripheral T‐lymphocyte protein expression
Authors: Borro Marina , Gentile Giovanna , De Luca Ottavia , Torre Maria Simona , Aimati Laura , Tatarelli Caterina , Antonietta Aloe Spiriti Maria , Christina Cox Maria , Simmaco Maurizio
Published: July 2010

A proteomic approach was applied to study the protein expression profile of peripheral T‐cells derived from patients at the onset of different B‐lymphoproliferative diseases, because a rising interest in specific actions played by T‐cells in such pathologies has emerged. Decreased levels of profilin‐1 and cofilin‐1 and increased levels of coronin1A and prohibitin were found in patients, compared with healthy controls. The protein‐protein interaction network of these proteins was studied using a web‐based bioinformatics tool, highlighting the actin cytoskeleton regulation as the main biological process involved in peripheral T‐cells of such patients. Unsupervised cluster analysis of protein expression data shows that the recorded alteration of T‐cell proteome was specifically induced by B‐cell pathologies.
Dietary polyunsaturated fatty acids in the prevention of allergic inflammatory conditions: progress and perspectives
Authors: West C. E. , Videky D. J. , Prescott S. L.
Published: July 2010

Cite this as: C. E. West, D. J. Videky and S. L. Prescott, Clinical & Experimental Allergy, 2010 (40) 1113–1115.
Increased serum FGF21 levels in patients with nonalcoholic fatty liver disease
Authors: Yilmaz Yusuf , Eren Fatih , Yonal Oya , Kurt Ramazan , Aktas Bilge , Celikel Cigdem Ataizi , Ozdogan Osman , Imeryuz Nese , Kalayci Cem , Avsar Erol
Published: July 2010

Eur J Clin Invest 2010; 40 (10): 887–892

Background  The fibroblast growth factor 21 (FGF21) hormonal pathway is a metabolic signalling cascade and has been recently identified as the master hormonal regulator of glucose, lipids and overall energy balance. In this observational, case–control study, we assayed serum levels of FGF21 in patients with nonalcoholic fatty liver disease (NAFLD), a hepatic manifestation of the metabolic syndrome, and examined their association with clinical, biochemical and histological phenotypes.

Materials and methods  Serum levels of FGF21 were assayed by ELISA in 82 patients with biopsy‐proven NAFLD and 77 controls. We analysed associations between FGF21 and the characteristics of patients with NAFLD by multiple linear regression analysis.

Results  Levels of FGF21 were significantly higher in patients with NAFLD (median 200 pg mL−1; interquartile range: 87–410 pg mL−1) than in healthy controls (median 93 pg mL−1; interquartile range: 70–180 pg mL−1, Mann–Whitney U‐test, P < 0·001). There was a stepwise increase in serum FGF21 levels according to the liver steatosis score (median level in subjects with score 1: 170 pg mL−1; score 2: 220 pg mL−1; score 3: 280 pg mL−1, P for trend < 0·01). After stepwise linear regression analysis, serum FGF21 levels were the only independent predictor of hepatic steatosis scores in patients with NAFLD (β = 0·26; t = 2·659, P < 0·01).

Conclusions  Serum FGF21 levels are increased in patients with NAFLD regardless of potential confounders and represent an independent predictor of liver steatosis. These findings support further investigation of this molecule in metabolic liver diseases.
Extremely high prevalence of metabolic syndrome manifestations among Arab youth: a call for early intervention
Authors: Al‐Daghri Nasser M.
Published: July 2010

Eur J Clin Invest 2010; 40 (12): 1063–1066

Background  Epidemiological studies have suggested an increased prevalence of metabolic syndrome (MS) among adults in the Middle‐East. This study aims to determine the prevalence of MS in a large cohort of Saudi children and adolescents.

Materials and methods  In this cross‐sectional observational study, a total of 1231 randomly selected Saudi children and adolescents aged 10–18 years were recruited. Subjects’ information was generated from a database of more than 10 000 Saudi citizens from the existing Biomarkers Screening in Riyadh Program (RIYADH Cohort), Saudi Arabia. Anthropometrics included body mass index, blood pressure, as well as waist and hip circumferences. Fasting blood glucose and lipid profile were determined using routine laboratory procedures. The modified definition of ATP‐III (NHANES III) was used for the diagnosis of MS.

Results  Overall prevalence of complete MS was 9·4% [confidence interval (CI) 7·8–11·0]. Age‐adjusted prevalence according to the European standard population is 5·8%. Boys had a comparable MS prevalence with girls [10·3% (CI 8·2–12·4) vs. 8·1% (CI 5·7–10·5), respectively (P = 0·20)]. Low HDL‐cholesterol was the most prevalent of all MS risk factors, affecting 86% (CI 85·0–88·6) and hypertriglyceridemia, the second most prevalent, affecting 33% (CI 30·6–35·8) of the subjects.

Conclusions  The prevalence of MS manifestations among Arab children is extremely high, with dyslipidemia being the most common MS abnormality. Screening for dyslipidemia among Saudi children is warranted especially among those most at risk. Scientific inquiry into the molecular causes of these manifestations should be pursued as a first step in the discovery of aetiological therapies.
Influence of rearing and experimental temperatures on predator avoidance behaviour in a freshwater pulmonate snail
Authors: DALESMAN SARAH , RUNDLE SIMON D.
Published: July 2010

1. Predicted increases in the temperature of freshwaters is likely to affect how prey species respond to predators. We investigated how the predator avoidance behaviour of the freshwater pulmonate snail Lymnaea stagnalis is influenced by the temperature at which it was reared and that at which behavioural trials were carried out.

2. Crawl‐out behaviour of juvenile snails from two populations (high predation risk versus low predation risk) reared at either 15 or 20 °C was assessed in response to predation cues (predatory fish kairomones and conspecific alarm cues) in behavioural trials at both 15 and 20 °C.

3. Trial temperature had a significant effect on the time that snails spent in avoidance, regardless of their population of origin. Crawl‐out behaviour was greater during behavioural trials at 15 °C, but there was no effect of trial temperature on the speed with which animals showed avoidance behaviour.

4. There was no interactive effect of rearing temperature (RT) and trial temperature, but the effect of RT on avoidance behaviour did differ between populations. For an RT of 15 °C, snails from the South Drain (high risk) population showed a more rapid and longer avoidance response than those from the Chilton Moor (low risk) population. In contrast, for snails reared at 20 °C, there was no difference between populations for the duration of the avoidance response and snails from Chilton Moor crawled out faster than those from South Drain.

5. Hence, whilst (predictable) differences relative to natural predation threat in crawl‐out behaviour were apparent at 15 °C, raising the developmental temperature to 20 °C eliminated or, in the case of latency, reversed these differences. This suggests that L. stagnalis populations that cohabit with predatory fish and experience high developmental temperatures may have a reduced ability to respond to fish predation risk.