bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis of Hydroxyapatite by Interfacial Reaction in a Multiple Emulsion
Authors: Kimura Isao
Published: November 2007

Interfacial reaction in a multiple emulsion, which is one of methods for producing inorganic microspheres, was applied to synthesize hydroxyapatite. The multiple emulsion was a W/O/W emulsion, made of dipotassium hydrogen phosphate solution as an inner aqueous phase, benzene as an oil phase, and calcium nitrate solution as an outer aqueous phase. The reaction was carried out in the multiple emulsion for 24 hours at 323 K. The crystalline phase was varied with an initial pH of the inner aqueous phase, and single phase hydroxyapatite was synthesized at an initial pH of 12. The products were composed of porous microspheres of less than 3 μm in size. The microspheres were composed of nanospheres of less than 120 nm in size. By considering the mass balance, it was suggested that each nanosphere was formed in an inner aqueous phase droplet.
Peroxisome Proliferator Activated Receptors and Lipoprotein Metabolism
Authors: Kersten Sander
Published: November 2007

Plasma lipoproteins are responsible for carrying triglycerides and cholesterol in the blood and ensuring their delivery to target organs. Regulation of lipoprotein metabolism takes place at numerous levels including via changes in gene transcription. An important group of transcription factors that mediates the effect of dietary fatty acids and certain drugs on plasma lipoproteins are the peroxisome proliferator activated receptors (PPARs). Three PPAR isotypes can be distinguished, all of which have a major role in regulating lipoprotein metabolism. PPARα is the molecular target for the fibrate class of drugs. Activation of PPARα in mice and humans markedly reduces hepatic triglyceride production and promotes plasma triglyceride clearance, leading to a clinically significant reduction in plasma triglyceride levels. In addition, plasma high‐density lipoprotein (HDL)‐cholesterol levels are increased upon PPARα activation in humans. PPARγ is the molecular target for the thiazolidinedione class of drugs. Activation of PPARγ in mice and human is generally associated with a modest increase in plasma HDL‐cholesterol and a decrease in plasma triglycerides. The latter effect is caused by an increase in lipoprotein lipase‐dependent plasma triglyceride clearance. Analogous to PPARα, activation of PPARβ/δ leads to increased plasma HDL‐cholesterol and decreased plasma triglyceride levels. In this paper, a fresh perspective on the relation between PPARs and lipoprotein metabolism is presented. The emphasis is on the physiological role of PPARs and the mechanisms underlying the effect of synthetic PPAR agonists on plasma lipoprotein levels.
The Use of Telemedicine Access to Schools to Facilitate Expert Assessment of Children with Asthma
Authors: Bergman David A. , Sharek Paul J. , Ekegren Kathryn , Thyne Shannon , Mayer Michelle , Saunders Mara
Published: November 2007

Research has shown that access to an asthma specialist improves asthma outcomes. We hypothesized that we could improve access to expert asthma care through a telemedicine link between an asthma specialist and a school-based asthma program. We conducted a prospective cohort study in 3 urban schools to ascertain the feasibility of using an asthma-focused telemedicine solution. Each subject was seen by an asthma expert at 0, 8, and 32 weeks. The assessment and recommendations for care were sent to the primary care physician (PCP) and parents were told to contact their physician for follow-up care. Eighty three subjects participated in the study. Subjects experienced improvement (P<.05) in family social activities and the number of asthma attacks. Ninety four percent of subjects rated the program as good or excellent. This study demonstrates the feasibility and acceptance of a school-based asthma program using a telemedicine link to an asthma specialist.
The Hole in the Stomach
Authors: Bödeker Hans , Leinung Steffen , Wittenburg Henning , Fischer Julia , Schiefke Ingolf , Teich Niels
Published: November 2007

A 57 year old woman was presented to the emergency department with upper abdominal pain and left sided chest discomfort. No cardiac or pulmonary cause could be determined and the patient underwent upper gastrointestinal endoscopy. Inversion of the scope to the fundus and subsequent fluoroscopy revealed a diaphragmatic hernia with a large herniation of the gastric fundus. Immediate laparotomy showed a 3 cm orifice of the diaphragm. The orifice was widened and a partial necrosis of the incarcerated fundus was resected. The patient recovered fully and was discharged 12 days after laparotomy.
FOXO3a Turns the Tumor Necrosis Factor Receptor Signaling Towards Apoptosis Through Reciprocal Regulation of c-Jun N-Terminal Kinase and NF-κB
Authors: Lee Hae-Young , Youn Seock-Won , Kim Ju-Young , Park Kyung-Woo , Hwang Chang-Il , Park Woong-Yang , Oh Byung-Hee , Park Young-Bae , Walsh Kenneth , Seo Jeong-Sun , Kim Hyo-Soo
Published: November 2007

Objective—

We evaluated the full range effects of FOXO3a in endothelial cells (ECs) by microarray analysis and investigated the role of FOXO3a regulating TNF receptor signaling pathway.

Methods and Results—

Human umbilical vein endothelial cells (HUVECs) were transfected with adenoviral vectors expressing constitutively active FOXO3a (Ad-TM-FOXO3a). Ad-TM-FOXO3a transfection caused remarkable apoptosis, which were accompanied with upregulation of genes related with TNF receptor signaling, such as TNF-α, TANK (TRAF-associated NF-κB activator), and TTRAP (TRAF and TNF receptor-associated protein). Furthermore, κB-Ras1 (IκB-interacting Ras-like protein-1) which is known to block IκB degradation was found increased, and intranuclear translocation of NF-κB was inhibited. GADD45β and XIAP, negative regulators of c-Jun N-terminal kinase (JNK), were suppressed and JNK activity was increased. Attenuation of TNF signaling pathway either by blocking antibody for TNF receptor or by blocking JNK with DMAP (6-dimethylaminopurine) or Ad-TAM67 (dominant negative c-Jun) cotransfection, significantly reduced FOXO3a-induced apoptosis. Finally, treatment of vasculature with heat shock, an activator of endogenous FOXO3a, resulted in EC apoptosis, which was completely rescued by Ad-TAM67.

Conclusion—

FOXO3a promotes apoptosis of ECs, through activation of JNK and suppression of NF-κB. These data identify a novel role of FOXO3a to turn TNF receptor signaling to a proapoptotic JNK-dependent pathway.
Florida's Sexually Violent Predator Program
Authors: Lucken Karol , Bales William
Published: November 2007

Sex offender civil commitment (SOCC) has been enacted in 16 states amid widespread controversy. A critical component of civil commitment is the risk assessment process that determines recommendations for civil confinement once a prison term has expired. This study analyzes the first stage of a two-stage risk assessment process that determines whether eligible sex offenders are referred for clinical evaluation in Florida's Sexually Violent Predator Program. A sample of 773 offenders referred to the program between July 2000 and August 2003 is examined to identify group differences between released and referred sex offenders and the unique effects of legal, clinical, and other factors on the decision to refer for clinical evaluation. Despite considerable discretion given evaluators in assessing risk, the findings indicate substantial and salient group differences between those released and referred and that the primary factors informing referral decisions are consistent with legislative intent, actuarial instruments, and sex offender recidivism research.
Effect of Roller Speed and Moisture Content of Cotton on Ginning Rate, Lint Quality and Electric Energy Consumption in Double Roller Gins
Authors: Prashantkumar Gulabrao Patil  , Pramod Madhukar Padole  , Jugalkishar Fulchand Agrawal  , Arun Bhimrao Dahake
Published: November 2007

To optimize the ginning parameters, a study was performed to investigate the influence of leather roller speed, fiber moisture on ginning rate, lint quality and electric energy consumption during ginning on double roller ginning machine. Seed cottons of different staple lengths were ginned at three levels of roller speed and moisture contents and experiments were performed on a specially designed experimental set up, which recorded roller revolutions per minute (RPM) and energy consumption. Duncan's Multiple Range Test revealed that the highest roller speed of 120 RPM coupled with 7 % moisture content of seed cotton showed the highest ginning rate with maximum saving in electric energy. Further, the highest ginning rate and maximum energy saving were observed for higher staple length of fibers as compared to lower staple length of fibers. Fiber properties of lint obtained from different treatments were measured using High Volume Instrument and Advanced Fiber Instrumentation System and results showed that the important fiber parameters remain unaffected.
Packing Density of Compact Yarns
Authors: Yilmaz Demet , Göktepe Fatma , Göktepe Özer , Kremenakova Dana
Published: November 2007

In this work, fiber distribution through the cross-sections of compact yarns and their packing density values was investigated to provide a better understanding of the internal structures of compact yarns produced by different compact spinning systems, since there is no information available so far regarding their internal structure. The results of packing density analysis indicated that compact yarns had nearly 15—30% higher packing density values compared to that of the conventional ring spun yarns. Also, the packing density values of compact yarns produced by three different compact yarn spinning systems, namely Rieter K44, Suessen Elite and Zinser Air-Com-Tex700, revealed that there were no significant differences among these systems in terms of yarn packing density values.
Synthesis and Application of Crosslinking Blue Anthraquinone Polyamine Dye with High Fixation
Authors: Yingling Li  , Yanfeng Tang  , Shufen Zhang  , Jinzong Yang
Published: November 2007

A polyamine crosslinking dye has been synthesized and applied to silk and cotton using crosslinking agents 2-chloro-4,6-di(amino-benzene-4'-β-sulfatoethylsulfone)-1,3,5- s-triazine and 2,4-dichloro-6-(aminobenzene-4'-β-sulfatoethylsulone)-1,3,5- s-triazine, respectively. The effect of pH, temperature and concentration of crosslinker on fixation of the dye has been discussed in detail. The fixation of this polyamine dye reached near 100 % and the dyeings were fast to dimethylformamide at the boil. This indicated that the dye fixation was achieved by covalent bonding with the crosslinking agent, which acted as a bridge between the fiber and the dye molecules. The dyed samples showed excellent fastness to washing and rubbing.
Single-end Sizing of Yarn using a Slot Applicator
Authors: Ok Hyunyoung , Carr Wallace W. , Park Heungsup
Published: November 2007

A single-end sizing process was developed to eliminate the problems associated with the traditional sizing method. By keeping the yarns separated in slots in the single-end sizing apparatus and drying them individually, less damage to the yarns occurred. In this paper, the process and device developed for single-end sizing are described and the properties of single-end sized yarn are compared with unsized yarn and commercially sized yarn.
Use of Electrospun Nanofiber Web for Protective Textile Materials as Barriers to Liquid Penetration
Authors: Lee Seungsin , Obendorf S. Kay
Published: November 2007

To examine the potential of electrospun nanofibrous webs as barriers to liquid penetration in protective clothing systems for agricultural workers, layered fabric systems with electrospun polyurethane fiber web layered on spunbonded nonwoven were developed. Barrier performance was evaluated for the layered fabric systems with different levels of electrospun web area density, using three pesticide mixtures that represent a range of surface tension and viscosity. Effects of electrospun web density on air permeability and water vapor transmission were assessed as indications of thermal comfort performance. Pore size distribution was measured to examine the effect of electrospun fiber web layers on pore size. Penetration testing showed that a very thin layer of electrospun polyurethane web significantly improved barrier performance for challenge liquids with a range of physicochemical properties. Air permeability decreased with increasing electrospun web area density, but was still higher than most of protective clothing materials currently available. No significant change was observed in moisture vapor transport of the system from electrospun nanofibrous web layers. Pore size distribution measurements indicated the feasibility of engineering pore size via the level of electrospun web area density, hence, controlling the level of protection and thermal comfort of layered fabric systems depending on their need.
Functionalization of Cotton Fabric with Vinyltrimethoxysilane
Authors: Abidi Noureddine , Hequet Eric , Tarimala Sowmitri
Published: November 2007

The surface of cotton fabric was successfully functionalized with vinyltrimethoxysilane in order to impart water repellency and wrinkle recovery and to introduce surface vinyl groups (—CH=CH2) to the fabric, which could then be initiated for copolymerization reactions with various monomers. The introduction of active groups onto the fabric surface was evidenced from the universal attenuated total reflectance Fourier transform infrared (UATR-FTIR) spectrum of the treated fabric. The spectrum shows two peaks located at 1410 and 1600 cm—1 (C=C stretch). An additional peak located at 756 cm—1 attributed to Si—O—Si symmetric stretch was also observed. Excellent water contact angle and wrinkle recovery angle values were obtained.
Variegated capitalism
Authors: Peck Jamie , Theodore Nik
Published: November 2007

The article critically engages with the `varieties of capitalism' school, which since its origins in the early 1990s has been consolidated into one of the most influential strands in comparative and heterodox political economy. While the `varieties' approach can be credited with the development of several of the most evocative stylized facts in heterodox political economy, having served as a potent foil against the orthodox globalization thesis, its alternative vision of a bipolar global economy comprising two competing capitalisms is found to be wanting. The approach is limited by its methodological nationalism, a tendency towards static analysis and latent institutional functionalism, and by an inability to adequately balance national specificity and path-dependency on the one hand with common underlying tendencies in capitalist restructuring on the other. Nevertheless, the varieties approach has spawned an influential account of the spatiality of advanced capitalism from which economic geography can certainly learn, and to which it has much to contribute.
Flying lessons: exploring the social and cultural geographies of global air travel
Authors: Adey Peter , Budd Lucy , Hubbard Phil
Published: November 2007

Geographic perspectives on civil aviation have traditionally been situated within the conceptual landscapes and languages of a transport geography in which quantitative methodologies have been to the fore. While such perspectives have shed light on the increasingly complex morphology of global air routes, this article argues such approaches tend to downplay crucial questions concerning the social production and consumption of airspace. Drawing on ideas from the newly-emergent `mobilities' paradigm, we use this article to flag up some alternative geographies of air travel, arguing that socially- and culturally-inflected perspectives can usefully reveal the iniquitous imprints of global air travel at a variety of spatial scales. We hence conclude that there is much to be gained by adopting such perspectives, and argue that work on the social dimensions of air travel is vital in a discipline where air transport is routinely described as an enabler of globalization, yet is often treated as an abstract, and oddly disembodied, space of flows.
The Relationship Between Career Variables and Occupational Aspirations and Expectations for Australian High School Adolescents
Authors: Patton Wendy , Creed Peter
Published: November 2007

This study surveyed 925 Australian high school students enrolled in grades 8 through 12 on measures of occupational aspirations, occupational expectations, career status aspirations, and career status expectations; it tested the association between these variables and career maturity, career indecision, career decision-making self-efficacy, and career barriers. Adolescents generally aspired to or expected to work within a small range of RIASEC (Realistic, Investigative, Artistic, Social, Enterprising, or Conventional) occupational categories. One third of students reported occupational aspiration and expectation discrepancies. These differed across gender and across age for female students but not for male students. Students who demonstrated both occupational and status aspiration and expectation discrepancies reported more career indecision, were less confident about making a career-related decision, and were less career mature. Students generally held higher occupational status aspirations than expectations, and male students were more likely to choose professional occupations than were female students. Age differences were found for status expectations but not for status aspirations.
The Relation of Secondary Students' Career-Choice Readiness to a Six-Phase Model of Career Decision Making
Authors: Hirschi Andreas , Läge Damian
Published: November 2007

Based on common aspects of recent models of career decision making (CDM), a six-phase model of CDM for secondary students is presented and empirically evaluated. The study tests the hypothesis that students who are in later phases possess more career-choice readiness and consider different numbers of career alternatives. Two hundred sixty-six Swiss secondary students completed measures tapping phase of CDM, career-choice readiness, and number of considered career options. Career-choice readiness showed an increase with phase of CDM. Later phases were associated with a larger increase in career-choice readiness. Number of considered career options showed a curve-linear development with fewer options considered at the beginning and at the end of the process. Male students showed a larger variability in their distribution among the process with more male than female students in the first and last phases of the process. Implications for theory and practice are presented.
Comparing the Chinese Career Key and the Self-Directed Search With High School Girls in Hong Kong
Authors: Ting Siu-Man Raymond
Published: November 2007

A career interest inventory, the Chinese Career Key (CCK) adapted from the Career Key based on Holland's theory of vocational choice, was studied. The purpose of the study was to further examine the psychometric qualities and user satisfaction of the CCK by comparing it to the Self-Directed Search. Students at a girls' public high school ( N = 130) in Hong Kong participated in the study. The research findings provided further evidence of validity, reliability, and user satisfaction among students in Hong Kong. Cultural differences, limitations, and implications for future studies and counseling practice are discussed.
Pandimensional Field Pattern Changes in Healers and Healees
Authors: Smith Dorothy Woods , Broida John P.
Published: November 2007

Rogers's Science of Unitary Human Beings framed this study of pandimensional pattern changes in healers and healees paired for an 8-week series of Therapeutic Touch (TT) sessions. Comparison of healee patterns before and after 141 TT sessions supported the hypotheses that healees would manifest decreased pulse and blood pressure, and reduced pain and stress ( p < .05). Duration of TT sessions was not preset but determined by healers according to energy cues. TT time was not related to pattern changes, consistent with the clinical practice of TT and the atemporal nature of Rogers's conceptual framework. Healers and healees showed parallel changes after the TT series. Participants manifested greater spirituality ( p <.05). Although increases in three of the four dimensions of power were significant ( p <.05), changes in power measured as a whole were not. Contrary to expectations, manifestations of diversity were decreased in healers and healees.
Spirituality in Nursing and Health-Related Literature
Authors: Sessanna Loralee , Finnell Deborah , Jezewski Mary Ann
Published: November 2007

S pirituality has become an increasingly popular concept among the nursing and health-related literature. The purpose for conducting this concept analysis, guided by Walker and Avant's methodology, was to (a) examine how spirituality has been used within the current body of nursing and health-related literature, (b) clarify the meaning of spirituality by discovering what this concept's current critical attributes/characteristics are, and (c) propose a definition of spirituality based on the concept analysis findings. A total of 90 references were reviewed, including 73 nursing and health-related references. Concept analysis findings revealed that spirituality was defined within four main themes in the nursing and health-related literature: (a) spirituality as religious systems of beliefs and values (spirituality = religion); (b) spirituality as life meaning, purpose, and connection with others; (c) spirituality as nonreligious systems of beliefs and values; and (d) spirituality as metaphysical or transcendental phenomena.
The Effect of Freezing Modes and Frozen Storage on Aroma, Enzyme and Micro-organism in Hami Melon
Authors: Ma Yongkun , Hu Xiaosong , Chen Jiluan , Chen Fang , Wu Jihong , Zhao Guanghua , Liao Xiaojun , Wang Zhengfu
Published: November 2007

The effects of freezing modes and time in frozen storage on aroma, enzymes and micro-organisms in Hami melon, also called cantaloupe melon (Cucumis melo. var. reticulatus Hami melon), were determined. Freezing process and frozen storage could not significantly reduce either micro-organisms counts or the enzymatic activities of polyphenoloxidase (PPO), peroxidase (POD) and lipoxidase (LOX) in melon. Liquid nitrogen ultra-rapid freezing maintained most aromatic components of the melon as compared with slow freezing. The mechanism of aromatic change in frozen melon was mainly ascribed to esters changing continuously in variety and concentration because of enzymolysis or synthetic reactions of esters catalysed by LOX. Meanwhile, increasing concentration of linoleic acid and linolenic acid in frozen melon cells promoted the biochemical reaction of unsaturated fatty acids in LOX system, so that enzymolysis of the double bonds in the ninth and tenth carbon of linoleic acid and linolenic acid was accelerated. Consequently, with the prolongation of time in frozen storage more and more unsaturated alcohols and aldehydes with 6 and 9 carbons were produced, the green notes of frozen storage melon became more and more intense, and the ester fragrance became more and more feeble. The above findings can benefit the development of cantaloupe melon process industry.
Mediation of apoptosis by and antitumor activity of lumiliximab in chronic lymphocytic leukemia cells and CD23+ lymphoma cell lines
Authors: Pathan Nuzhat I. , Chu Peter , Hariharan Kandasamy , Cheney Carolyn , Molina Arturo , Byrd John
Published: November 2007

Lumiliximab is a chimeric macaque-human monoclonal antibody to CD23, a protein expressed on virtually all chronic lymphocytic leukemia (CLL) cells. We examined the ability of lumiliximab to mediate apoptosis, antibody-dependent cellular cytotoxicity, and complement-dependent cytotoxicity against primary CLL cells and CD23-expressing B-cell lines. Our data suggest that lumiliximab kills CLL cells and CD23-expressing B cells predominantly by apoptosis, which occurs through the intrinsic pathway. Lumiliximab-induced apoptosis was accompanied by the down-regulation of antiapoptotic proteins Bcl-2, Bcl-XL, and XIAP, activation of Bax, and release of cytochrome c from the mitochondria. We also found that the addition of lumiliximab to rituximab or fludarabine results in synergistic cytotoxicity of primary CLL cells and CD23-expressing B-cell lines. We investigated the in vivo activity of lumiliximab in a human disseminated CD23+ B-cell lymphoma SCID mouse model and found greater antitumor activity with it than with control antibody. We also found that paralysis-free survival was greater with lumiliximab plus rituximab or fludarabine than with any of those agents alone. These results suggest that lumiliximab may be an effective treatment alone or in combination with rituximab or chemotherapy agents in CLL or other CD23-overexpressing B-cell malignancies.
Snoring in preschool children: prevalence, severity and risk factors
Authors: Kuehni C. E. , Strippoli M-P. F. , Chauliac E. S. , Silverman M.
Published: November 2007

Epidemiological data on snoring from preschool children are scarce, although habitual snoring (snoring on almost all nights) has been associated with poor long-term outcomes.

In a population survey of 6,811 children aged 1–4 yrs (from Leicestershire, UK) the present authors determined prevalence, severity and risk factors for snoring, especially habitual snoring.

In 59.7% of the children, parents reported snoring in the previous 12 months, including 7.9% with habitual snoring and 0.9% with habitual snoring and sleep disturbance. Prevalence of habitual snoring increased with age from 6.6% in 1-yr-olds to 13.0% in 4-yr-olds. Habitual snoring was associated with: one and both parents smoking (adjusted odds ratio (OR) 1.46 and 2.09, respectively); road traffic (OR 1.23); single parent (OR 1.60); and in White but not South Asian children, socioeconomic deprivation (OR 1.25 and 2.03 for middle and upper thirds of Townsend score, respectively). Respiratory tract symptoms related to atopic disorders and to respiratory infections were strongly associated with snoring; however, body mass index was not.

In conclusion, habitual snoring is common in preschool children with one-third of cases attributable to avoidable risk factors. The strong association with atopic disorders, viral infections and environmental exposures suggests a complex aetiology, based on a general vulnerability of the respiratory tract.
Genome-wide transcriptional analysis of grapevine berry ripening reveals a set of genes similarly modulated during three seasons and the occurrence of an oxidative burst at vèraison
Authors: Pilati Stefania , Perazzolli Michele , Malossini Andrea , Cestaro Alessandro , Demattè Lorenzo , Fontana Paolo , Dal Ri Antonio , Viola Roberto , Velasco Riccardo , Moser Claudio
Published: November 2007

Background

Grapevine (Vitis species) is among the most important fruit crops in terms of cultivated area and economic impact. Despite this relevance, little is known about the transcriptional changes and the regulatory circuits underlying the biochemical and physical changes occurring during berry development.

Results

Fruit ripening in the non-climacteric crop species Vitis vinifera L. has been investigated at the transcriptional level by the use of the Affymetrix Vitis GeneChip® which contains approximately 14,500 unigenes. Gene expression data obtained from berries sampled before and after véraison in three growing years, were analyzed to identify genes specifically involved in fruit ripening and to investigate seasonal influences on the process. From these analyses a core set of 1477 genes was found which was similarly modulated in all seasons. We were able to separate ripening specific isoforms within gene families and to identify ripening related genes which appeared strongly regulated also by the seasonal weather conditions. Transcripts annotation by Gene Ontology vocabulary revealed five overrepresented functional categories of which cell wall organization and biogenesis, carbohydrate and secondary metabolisms and stress response were specifically induced during the ripening phase, while photosynthesis was strongly repressed. About 19% of the core gene set was characterized by genes involved in regulatory processes, such as transcription factors and transcripts related to hormonal metabolism and signal transduction. Auxin, ethylene and light emerged as the main stimuli influencing berry development. In addition, an oxidative burst, previously not detected in grapevine, characterized by rapid accumulation of H2O2 starting from véraison and by the modulation of many ROS scavenging enzymes, was observed.

Conclusion

The time-course gene expression analysis of grapevine berry development has identified the occurrence of two well distinct phases along the process. The pre-véraison phase represents a reprogramming stage of the cellular metabolism, characterized by the expression of numerous genes involved in hormonal signalling and transcriptional regulation. The post-véraison phase is characterized by the onset of a ripening-specialized metabolism responsible for the phenotypic traits of the ripe berry. Between the two phases, at véraison, an oxidative burst and the concurrent modulation of the anti-oxidative enzymatic network was observed. The large number of regulatory genes we have identified represents a powerful new resource for dissecting the mechanisms of fruit ripening control in non-climacteric plants.
Squalestatin alters the intracellular trafficking of a neurotoxic prion peptide
Authors: Wilson Rona , Bate Clive , Boshuizen Ronald , Williams Alun , Brewer James
Published: November 2007

Background

Neurotoxic peptides derived from the protease-resistant core of the prion protein are used to model the pathogenesis of prion diseases. The current study characterised the ingestion, internalization and intracellular trafficking of a neurotoxic peptide containing amino acids 105–132 of the murine prion protein (MoPrP105-132) in neuroblastoma cells and primary cortical neurons.

Results

Fluorescence microscopy and cell fractionation techniques showed that MoPrP105-132 co-localised with lipid raft markers (cholera toxin and caveolin-1) and trafficked intracellularly within lipid rafts. This trafficking followed a non-classical endosomal pathway delivering peptide to the Golgi and ER, avoiding classical endosomal trafficking via early endosomes to lysosomes. Fluorescence resonance energy transfer analysis demonstrated close interactions of MoPrP105-132 with cytoplasmic phospholipase A2 (cPLA2) and cyclo-oxygenase-1 (COX-1), enzymes implicated in the neurotoxicity of prions. Treatment with squalestatin reduced neuronal cholesterol levels and caused the redistribution of MoPrP105-132 out of lipid rafts. In squalestatin-treated cells, MoPrP105-132 was rerouted away from the Golgi/ER into degradative lysosomes. Squalestatin treatment also reduced the association between MoPrP105-132 and cPLA2/COX-1.

Conclusion

As the observed shift in peptide trafficking was accompanied by increased cell survival these studies suggest that the neurotoxicity of this PrP peptide is dependent on trafficking to specific organelles where it activates specific signal transduction pathways.
A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm
Authors: Philibert Pascal , Stoessel Audrey , Wang Wei , Sibler Annie-Paule , Bec Nicole , Larroque Christian , Saven Jeffery G , Courtête Jérôme , Weiss Etienne , Martineau Pierre
Published: November 2007

Background

Intrabodies are defined as antibody molecules which are ectopically expressed inside the cell. Such intrabodies can be used to visualize or inhibit the targeted antigen in living cells. However, most antibody fragments cannot be used as intrabodies because they do not fold under the reducing conditions of the cell cytosol and nucleus.

Results

We describe the construction and validation of a large synthetic human single chain antibody fragment library based on a unique framework and optimized for cytoplasmic expression. Focusing the library by mimicking the natural diversity of CDR3 loops ensured that the scFvs were fully human and functional. We show that the library is highly diverse and functional since it has been possible to isolate by phage-display several strong binders against the five proteins tested in this study, the Syk and Aurora-A protein kinases, the αβ tubulin dimer, the papillomavirus E6 protein and the core histones. Some of the selected scFvs are expressed at an exceptional high level in the bacterial cytoplasm, allowing the purification of 1 mg of active scFv from only 20 ml of culture. Finally, we show that after three rounds of selection against core histones, more than half of the selected scFvs were active when expressedin vivoin human cells since they were essentially localized in the nucleus.

Conclusion

This new library is a promising tool not only for an easy and large-scale selection of functional intrabodies but also for the isolation of highly expressed scFvs that could be used in numerous biotechnological and therapeutic applications.
Altering coenzyme specificity of Pichia stipitis xylose reductase by the semi-rational approach CASTing
Authors: Liang Ling , Zhang Jingqing , Lin Zhanglin
Published: November 2007

Background

The NAD(P)H-dependentPichia stipitisxylose reductase (PsXR) is one of the key enzymes for xylose fermentation, and has been cloned into the commonly used ethanol-producing yeastSaccharomyces cerevisiae. In order to eliminate the redox imbalance resulting from the preference of this enzyme toward NADPH, efforts have been made to alter the coenzyme specificity of PsXR by site-directed mutagenesis, with limited success. Given the industrial importance of PsXR, it is of interest to investigate further ways to create mutants of PsXR that prefers NADH rather than NADPH, by the alternative directed evolution approach.

Results

Based on a homology model of PsXR, six residues were predicted to interact with the adenine ribose of NAD(P)H in PsXR and altered using a semi-rational mutagenesis approach (CASTing). Three rounds of saturation mutagenesis were carried to randomize these residues, and a microplate-based assay was applied in the screening. A best mutant 2-2C12, which carried four mutations K270S, N272P, S271G and R276F, was obtained. The mutant showed a preference toward NADH over NADPH by a factor of about 13-fold, or an improvement of about 42-fold, as measured by the ratio of the specificity constantkcat/Kmcoenzyme. Compared with the wild-type, thekcatNADHfor the best mutant was only slightly lower, while thekcatNADPHdecreased by a factor of about 10. Furthermore, the specific activity of 2-2C12 in the presence of NADH was 20.6 U·mg-1, which is highest among PsXR mutants reported.

Conclusion

A seemingly simplistic and yet very effective mutagenesis approach, CASTing, was applied successfully to alter the NAD(P)H preference forPichia stipitisxylose reductase, an important enzyme for xylose-fermenting yeast. The observed change in the NAD(P)H preference for this enzyme seems to have resulted from the altered active site that is more unfavorable for NADPH than NADH in terms of bothKmandkcat. There are potentials for application of our PsXR in constructing a more balanced XR-XDH pathway in recombinant xylose-fermentingS. cerevisiaestrains.
Recruitment of dendritic cells and macrophages during T cell-mediated synovial inflammation
Authors: Moghaddami Mahin , Cleland Leslie G , Radisic Gorjana , Mayrhofer Graham
Published: November 2007

Adoptive transfer of adjuvant-induced arthritis was used in this study to examine local macrophages and dendritic cells (DCs) during T cell-mediated synovial inflammation. We studied the influx of CD11b+CD11c+ putative myeloid DCs and other non-lymphoid CD45+ cells into synovium-rich tissues (SRTs) of the affected hind paws in response to a pulse of autoreactive thoracic duct cells. Cells were prepared from the SRTs using a collagenase perfusion-digestion technique, thus allowing enumeration and phenotypic analysis by flow cytometry. Numbers of CD45+ cells increased during the first 6 days, with increases in CD45+MHC (major histocompatibility complex) II+ monocyte-like cells from as early as day 3 after transfer. In contrast, typical MHC II- monocytes, mainly of the CD4- subset, did not increase until 12 to 14 days after cell transfer, coinciding with the main influx of polymorphonuclear cells. By day 14, CD45+MHC IIhi cells constituted approximately half of all CD45+ cells in SRT. Most of the MHC IIhi cells expressed CD11c and CD11b and represented putative myeloid DCs, whereas only approximately 20% were CD163+ macrophages. Less than 5% of the MHC IIhi cells in inflamed SRT were CD11b-, setting a maximum for any influx of plasmacytoid DCs. Of the putative myeloid DCs, a third expressed CD4 and both the CD4+ and the CD4- subsets expressed the co-stimulatory molecule CD172a. Early accumulation of MHC IIhiCD11c+ monocyte-like cells during the early phase of T cell-mediated inflammation, relative to typical MHC II- blood monocytes, suggests that recruited monocytes differentiate rapidly toward the DC lineage at this stage in the disease process. However, it is possible also that the MHC IIhiCD11c+ cells originate from a specific subset of DC-like circulating mononuclear cells.
Inflammation, endothelium, and coagulation in sepsis
Authors: Schouten Marcel , Wiersinga Willem Joost , Levi Marcel , van der Poll Tom
Published: November 2007

Sepsis is a systemic response to infection, and symptoms are produced by host defense systems rather than by the invading pathogens. Amongst the most prominent features of sepsis, contributing significantly to its outcome, is activation of coagulation with concurrent down-regulation of anticoagulant systems and fibrinolysis. Inflammation-induced coagulation on its turn contributes to inflammation. Another important feature of sepsis, associated with key symptoms such as hypovolemia and hypotension, is endothelial dysfunction. Under normal conditions, the endothelium provides for an anticoagulant surface, a property that is lost in sepsis. In this review, data about the interplay between inflammation and coagulation in sepsis are summarized with a special focus on the influence of the endothelium on inflammation-induced coagulation and vice versa. Possible procoagulant properties of the endothelium are described, such as expression of tissue factor (TF) and von Willebrand factor and interaction with platelets. Possible procoagulant roles of microparticles, circulating endothelial cells and endothelial apoptosis, are also discussed. Moreover, the important roles of the endothelium in down-regulating the anticoagulants TF pathway inhibitor, antithrombin, and the protein C (PC) system and inhibition of fibrinolysis are discussed. The influence of coagulation on its turn on inflammation and the endothelium is described with a special focus on protease-activated receptors (PARs). We conclude that the relationship between endothelium and coagulation in sepsis is tight and that further research is needed, for example, to better understand the role of activated PC signaling via PAR-1, the role of the endothelial PC receptor herein, and the role of the glycocalyx.
Two distinct activation states of plasmacytoid dendritic cells induced by influenza virus and CpG 1826 oligonucleotide
Authors: Iparraguirre Amaya , Tobias John W , Hensley Scott E , Masek Katherine S , Cavanagh Lois L , Rendl Michael , Hunter Christopher A , Ertl Hildegund C , von Andrian Ulrich H , Weninger Wolfgang
Published: November 2007

There is growing evidence that plasmacytoid dendritic cells (pDC) are involved in the innate recognition of various microbes. However, the precise consequences of pathogen recognition on pDC activation and function are incompletely understood. Using a novel transgenic mouse model that facilitates the isolation of highly pure pDC populations, we found that influenza virus PR/8, a TLR7 ligand, and CpG 1826 oligonucleotide, a TLR9 ligand, induced surprisingly divergent activation programs in these cells. pDC stimulated with PR/8 produced large amounts of type I IFNs, and CpG 1826-stimulated pDC expressed higher levels of costimulatory molecules and proinflammatory cytokines and induced stronger proliferation of T cells. Transcriptome analysis uncovered the differential regulation in pDC of 178 and 1577 genes by PR/8 and CpG 1826, respectively. These differences may relate to the activation of discrete signaling pathways, as evidenced by distinct ERK1/2 and p38 MAPK phosphorylation kinetics. Finally, pDC isolated ex vivo during PR/8 infection or after i.v. CpG 1826 injection resembled their in vitro counterparts, corroborating that these cells can adopt specialized phenotypes in vivo. Thus, pDC display remarkable functional flexibility, which emphasizes their versatile functions in antimicrobial immunity and inflammatory processes.
A systemic granulomatous response toSchistosoma mansonieggs alters responsiveness of bone marrow-derived macrophages to Toll-like receptor agonists
Authors: Joshi Amrita D , Raymond Tracy , Coelho Ana L , Kunkel Steven L , Hogaboam Cory M
Published: November 2007

Macrophages play a pivotal role in innate and acquired immune responses to Schistosoma mansoni. Classical (M1) or alternative (M2) activation states of these cells further delineate their roles in tissue damage through innate immunity or fibrotic remodeling, respectively. In the present study, we addressed the following question: Does systemic Th2-type cytokine polarization evoked by S. mansoni affect macrophage differentiation and activation? To this end, we analyzed bone marrow-derived macrophages from mice with S. mansoni egg-induced pulmonary granulomas and unchallenged (or naïve) mice to determine their activation state and their response to specific TLR agonists, including S. mansoni egg antigens. Unlike naïve macrophages, macrophages from Th2-polarized mice constitutively expressed significantly higher “found in inflammatory zone-1” (FIZZ1) and ST2 (M2 markers) and significantly lower NO synthase 2, CCL3, MIP-2, TNF-α, and IL-12 (M1 markers). Also, compared with naïve macrophages, Th2-polarized macrophages exhibited enhanced responses to the presence of specific TLR agonists, which consistently induced significantly higher levels of gene and protein levels for M2 and M1 markers in these cells. Together, these data show that signals received by bone marrow precursors during S. mansoni egg-induced granuloma responses dynamically alter the development of macrophages and enhance the TLR responsiveness of these cells, which may ultimately have a significant effect on the pulmonary granulomatous response.
Dialogue on Reverse‐Engineering Assessment and Methods
Authors: STOLOVITZKY GUSTAVO , MONROE DON , CALIFANO ANDREA
Published: November 2007

Abstract:  The biotechnological advances of the last decade have confronted us with an explosion of genetics, genomics, transcriptomics, proteomics, and metabolomics data. These data need to be organized and structured before they may provide a coherent biological picture. To accomplish this formidable task, the availability of an accurate map of the physical interactions in the cell that are responsible for cellular behavior and function would be exceedingly helpful, as these data are ultimately the result of such molecular interactions. However, all we have at this time is, at best, a fragmentary and only partially correct representation of the interactions between genes, their byproducts, and other cellular entities. If we want to succeed in our quest for understanding the biological whole as more than the sum of the individual parts, we need to build more comprehensive and cell‐context–specific maps of the biological interaction networks. DREAM, the Dialogue on Reverse Engineering Assessment and Methods, is fostering a concerted effort by computational and experimental biologists to understand the limitations and to enhance the strengths of the efforts to reverse engineer cellular networks from high‐throughput data. In this chapter
we will discuss the salient arguments of the first DREAM conference. We will highlight both the state of the art in the field of reverse engineering as well as some of its challenges and opportunities.
Dose-Dependent Regulation of High-Density Lipoprotein Metabolism with Rosuvastatin in the Metabolic Syndrome
Authors: Ooi Esther M. M. , Watts Gerald F. , Nestel Paul J. , Sviridov Dmitri , Hoang Anh , Barrett P. Hugh R.
Published: November 2007

Background: Low plasma concentration of high-density lipoprotein (HDL) cholesterol is a risk factor for cardiovascular disease and a feature of the metabolic syndrome. Rosuvastatin has been shown to increase HDL cholesterol concentration, but the mechanisms remain unclear.

Methods and Results: Twelve men with the metabolic syndrome were studied in a randomized, double-blind, crossover trial of 5-wk therapeutic periods with placebo, 10 mg/d rosuvastatin, or 40 mg/d rosuvastatin, with 2-wk placebo washout between each period. Compared with placebo, there was a significant dose-dependent increase in HDL cholesterol, HDL particle size, and concentration of HDL particles that contain apolipoprotein A-I (LpA-I). The increase in LpA-I concentration was associated with significant dose-dependent reductions in triglyceride concentration and LpA-I fractional catabolic rate, with no changes in LpA-I production rate. There was a significant dose-dependent reduction in the fractional catabolic rate of HDL particles containing both apolipoprotein A-I and A-II (LpA-I:A-II), with concomitant reduction in LpA-I:A-II production rate, and hence no change in LpA-I:A-II concentration.

Conclusions: Rosuvastatin dose-dependently increased plasma HDL cholesterol and LpA-I concentrations in the metabolic syndrome. This could relate to reduction in plasma triglycerides with remodeling of HDL particles and reduction in LpA-I fractional catabolism. The findings contribute to understanding mechanisms for the HDL-raising effect of rosuvastatin in the metabolic syndrome with implications for reduction in cardiovascular disease.
The Risk of Second Primary Malignancies up to Three Decades after the Treatment of Differentiated Thyroid Cancer
Authors: Brown Aaron. P. , Chen Jergin , Hitchcock Ying J. , Szabo Aniko , Shrieve Dennis C. , Tward Jonathan. D.
Published: November 2007

Background: The 10-yr survival rate of patients with differentiated thyroid cancer exceeds 90%. These patients may be at elevated risk for secondary cancers.

Methods: The risk of nonthyroid second primary malignancies after differentiated thyroid cancer was determined in 30,278 patients diagnosed between 1973 and 2002 from centers participating in the National Cancer Institute’s Surveillance, Epidemiology, and End Results program. Median follow-up was 103 months (range, 2–359 months). Risk was further assessed for the addition of radioisotope therapy, gender, latency to development of secondary cancer, and age at thyroid cancer diagnosis.

Results: There were 2158 patients who developed a total of 2338 nonthyroid second primary malignancies, significantly more than that expected in the general population [observed/expected (O/E) = 1.09; 95% confidence interval (CI), 1.05–1.14; P < 0.05; absolute excess risk per 10,000 person-years (AER) = 6.39]. A significantly greater risk of second primary malignancies over that expected in the general population was for patients treated with radioisotopes (O/E = 1.20; 95% CI, 1.07–1.33; AER = 11.8) as well as for unirradiated patients (O/E = 1.05; 95% CI, 1.00–1.10; AER = 3.53). However, the increased risk was greater for the irradiated vs. the unirradiated cohort (relative risk = 1.16; 95% CI, 1.05–1.27; P < 0.05). Gender did not affect risk. The greatest risk of second primary cancers occurred within 5 yr of diagnosis and was elevated for younger patients.

Conclusions: The overall risk of second primary malignancies is increased for thyroid cancer survivors and varies by radioisotope therapy, latency, and age at diagnosis.