bluuebunny/crossref_metadata_2025 huggingface dataset:
Fish assemblages as influenced by environmental factors in streams in protected areas of the Czech Republic
Authors: Humpl M. , Pivnička K.
Published: February 2006

Abstract –  Three streams of comparable size located in different landscape‐protected areas were selected for studying the effect of environmental factors on fish assemblages using indirect (detrended correspondence analysis, DCA) and direct (canonical correspondence analysis, CCA) gradient analysis. DCA of species showed well a gradient of assemblage changes in the longitudinal profile. DCA of sites stressed the variability between the fish assemblages of the three streams. This pattern was then confirmed by the highly significant between‐stream CCA. In the within‐site CCA, environmental factors explained 50.7% variability for presence–absence data and 58.3% for the relative abundance data. The analysis revealed that number of ponds and land use are the most influential factors of the strongest environmental gradient. However, in the partial CCAs, factor substratum type explained the largest proportion of the variability affecting fish in their habitat choice. Generally, presence–absence and relative abundance data of fish gave similar results in both DCA and CCA analyses; the same environmental factors proved to be important in both data type analyses. The environmental factors explain more variability than the regional (between‐stream) one. The total proportion of variability explained by the presence–absence data analysis was 71.9% and in the relative abundance analysis even 80.8%. The environmental factors measured during the field survey explain 2.1‐ and 3.4‐times more assemblages’ variability than factors measured from a hydrological map.
The Monroe Doctrine: Meanings and Implications
Authors: GILDERHUS MARK T.
Published: February 2006

This article presents a brief history of the Monroe Doctrine since its articulation in 1823. First conceived as a statement in opposition to European intrusions in the Americas, it became under President Theodore Roosevelt a justification for U.S. intervention. To cultivate Latin American trade and goodwill during the Great Depression and the Second World War, Franklin Roosevelt's administration accepted the principle of nonintervention. Later with the onset of the Cold War, perceived international imperatives led to a series of new interventions in countries such as Guatemala, Cuba, the Dominican Republic, and Chile. Though typically couched in idealistic rhetoric emphasizing Pan‐American commitments to solidarity and democracy, the various versions of the Monroe Doctrine consistently served U.S. policy makers as a means for advancing what they understood as national strategic and economic interests.
The Roosevelt Corollary
Authors: RICARD SERGE
Published: February 2006

This article argues that in his annual message of December 6, 1904, Theodore Roosevelt enunciated not a corollary to the Monroe Doctrine but an entirely new principle which epitomized his “big stick” view of foreign relations: the United States was to act as sole policeman of the Western Hemisphere and deny the European powers any right of interference in what it regarded as its rightful “zone of influence,” given Washington's obvious supremacy in that part of the world at the dawn of the twentieth century. An initially defensive dictum had therefore been turned into an aggressive policy by a man who had long pondered over America's standing and role in the world. Strictly speaking, it was a “perversion” of Monroe's original intent, but fundamentally it reflected a new, innovative conception of security and defense, as well as changing geopolitical conditions.

The catalysts of that drastic mutation were the necessary protection of the projected isthmian canal and Germany's aggressiveness in the Caribbean. In time the United States would tighten its grip on some of its “sister republics” and incur the charge of imperialism by developing a “protectorate policy” that belied its oft‐proclaimed commitment to self‐determination. Repeated interventions in the name of law and order would leave an enduring legacy of anti‐Americanism in South America. The Roosevelt Corollary to the Monroe Doctrine would guide hemispheric diplomacy throughout World War II and during the Cold War, assurances to the contrary notwithstanding.
Securing the Middle East: The Eisenhower Doctrine of 1957
Authors: HAHN PETER L.
Published: February 2006

The Eisenhower Doctrine of 1957 declared that the United States would distribute economic and military aid and, if necessary, use military force to stop the spread of communism in the Middle East. Eisenhower found it difficult to convince leading Arab states or Israel to endorse the doctrine's purpose or usefulness. Nonetheless, he applied the doctrine in 1957‐58 by dispensing economic aid to shore up the Kingdom of Jordan, by encouraging Syria's neighbors to consider military operations against it, and by sending U.S. troops into Lebanon to prevent a radical revolution from sweeping over that country. The doctrine consisted of a major commitment by the United States to the security and stability of the Middle East and signaled a new level of U.S. resolve to exert influence in international affairs. By issuing the doctrine, Eisenhower raised the prospect that the United States would fight in the Middle East and accepted responsibilities in the region that the United States would retain for decades to come.
The Nixon Doctrine: A Saga of Misunderstanding
Authors: KIMBALL JEFFREY
Published: February 2006

When President Richard Nixon met with reporters in Guam on July 25, 1969 to discuss the U.S. role in Asia, he did not intend his comments to be understood as constituting a new policy doctrine. After some reporters began referring to key elements of his remarks as the “Nixon Doctrine,” Nixon came to appreciate the political value of the phrase and publicly began using it himself, even though his policies toward some of the nations of Asia were inconsistent with the “new” policy, and as he was also about to embark on a course of action in Vietnam that contradicted the “doctrine.”
The Polls: Public Opinion and Secretary of Defense Donald Rumsfeld
Authors: PANAGOPOULOS COSTAS
Published: February 2006

Secretary of Defense Donald Rumsfeld has been characterized simultaneously as a central political figure in the executive branch as well as one of the most controversial and contentious cabinet members of the Bush administration. This article assesses Americans’ impressions of Secretary Rumsfeld's job performance and reveals that evaluations of Secretary Rumsfeld's performance have steadily declined since 2001. Over the same period, President Bush's approval ratings have dwindled, although the evidence I present indicates no relationship exists between the two. The broader implications of this research suggest that public evaluations of a president's key subordinates may not necessarily influence levels of presidential approval.
Effects of sequestration on signal transduction cascades
Authors: Blüthgen Nils , Bruggeman Frank J. , Legewie Stefan , Herzel Hanspeter , Westerhoff Hans V. , Kholodenko Boris N.
Published: February 2006

The building blocks of most signal transduction pathways are pairs of enzymes, such as kinases and phosphatases, that control the activity of protein targets by covalent modification. It has previously been shown [Goldbeter A & Koshland DE (1981) Proc Natl Acad Sci USA78, 6840–6844] that these systems can be highly sensitive to changes in stimuli if their catalysing enzymes are saturated with their target protein substrates. This mechanism, termed zero‐order ultrasensitivity, may set thresholds that filter out subthreshold stimuli. Experimental data on protein abundance suggest that the enzymes and their target proteins are present in comparable concentrations. Under these conditions a large fraction of the target protein may be sequestrated by the enzymes. This causes a reduction in ultrasensitivity so that the proposed mechanism is unlikely to account for ultrasensitivity under the conditions present in most in vivo signalling cascades. Furthermore, we show that sequestration changes the dynamics of a covalent modification cycle and may account for signal termination and a sign‐sensitive delay. Finally, we analyse the effect of sequestration on the dynamics of a complex signal transduction cascade: the mitogen‐activated protein kinase (MAPK) cascade with negative feedback. We show that sequestration limits ultrasensitivity in this cascade and may thereby abolish the potential for oscillations induced by negative feedback.
Cloning and characterization of CBL‐CIPK signalling components from a legume (Pisum sativum)
Authors: Mahajan Shilpi , Sopory Sudhir K , Tuteja Narendra
Published: February 2006

The studies on calcium sensor calcineurin B‐like protein (CBL) and CBL interacting protein kinases (CIPK) are limited to Arabidopsis and rice and their functional role is only beginning to emerge. Here, we present cloning and characterization of a protein kinase (PsCIPK) from a legume, pea, with novel properties. The PsCIPK gene is intronless and encodes a protein that showed partial homology to the members of CIPK family. The recombinant PsCIPK protein was autophosphorylated at Thr residue(s). Immunoprecipitation and yeast two‐hybrid analysis showed direct interaction of PsCIPK with PsCBL, whose cDNA and genomic DNA were also cloned in this study. PsCBL showed homology to AtCBL3 and contained calcium‐binding activity. We demonstrate for the first time that PsCBL is phosphorylated at its Thr residue(s) by PsCIPK. Immunofluorescence/confocal microscopy showed that PsCBL is exclusively localized in the cytosol, whereas PsCIPK is localized in the cytosol and the outer membrane. The exposure of plants to NaCl, cold and wounding co‐ordinately upregulated the expression of PsCBL and PsCIPK genes. The transcript levels of both genes were also coordinately stimulated in response to calcium and salicylic acid. However, drought and abscisic acid had no effect on the expression of these genes. These studies show the ubiquitous presence of CBL/CIPK in higher plants and enhance our understanding of their role in abiotic and biotic stress signalling.
Activation of theTorpedonicotinic acetylcholine receptor
Authors: Kapur Ankur , Davies Martin , Dryden William F. , Dunn Susan M.J.
Published: February 2006

TheTorpedonicotinic acetylcholine receptor is a heteropentamer (α2βγδ) in which structurally homologous subunits assemble to form a central ion pore. Viewed from the synaptic cleft, the likely arrangement of these subunits is α–γ–α–δ–β lying in an anticlockwise orientation. High affinity binding sites for agonists and competitive antagonists have been localized to the α–γ and α–δ subunit interfaces. We investigated the involvement of amino acids lying at an adjacent interface (γ–α) in receptor properties. RecombinantTorpedoreceptors, expressed inXenopusoocytes, were used to investigate the consequences of mutating αArg55 and γGlu93, residues that are conserved in most species of the peripheral nicotinic receptors. Based on homology modeling, these residues are predicted to lie in close proximity to one another and it has been suggested that they may form a salt bridge in the receptor's three‐dimensional structure (Sineet al. 2002J Biol Chem277,29 210–29 223). Although substitution of αR55 by phenylalanine or tryptophan resulted in approximately a six‐fold increase in the EC50value for acetylcholine activation, the charge reversal mutation (αR55E) had no significant effect. In contrast, the replacement of γE93 by an arginine conferred an eight‐fold increase in the potency for acetylcholine‐induced receptor activation. In the receptor carrying the double mutations, αR55E‐γE93R or αR55F‐γE93R, the potency for acetylcholine activation was partially restored to that of the wild‐type. The results suggest that, although individually these residues influence receptor activation, direct interactions between them are unlikely to play a major role in the stabilization of different conformational states of the receptor.
Peptides corresponding to helices 5 and 6 of Bax can independently form large lipid pores
Authors: García‐Sáez Ana J. , Coraiola Manuela , Serra Mauro Dalla , Mingarro Ismael , Müller Peter , Salgado Jesús
Published: February 2006

Proteins of the B‐cell lymphoma protein 2 (Bcl2) family are key regulators of the apoptotic cascade, controlling the release of apoptotic factors from the mitochondrial intermembrane space. A helical hairpin found in the core of water‐soluble folds of these proteins has been reported to be the pore‐forming domain. Here we show that peptides including any of the two α‐helix fragments of the hairpin of Bcl2 associated protein X (Bax) can independently induce release of large labelled dextrans from synthetic lipid vesicles. The permeability promoted by these peptides is influenced by intrinsic monolayer curvature and accompanied by fast transbilayer redistribution of lipids, supporting a toroidal pore mechanism as in the case of the full‐length protein. However, compared with the pores made by complete Bax, the pores made by the Bax peptides are smaller and do not need the concerted action of tBid. These data indicate that the sequences of both fragments of the hairpin contain the principal physicochemical requirements for pore formation, showing a parallel between the permeabilization mechanism of a complex regulated protein system, such as Bax, and the much simpler pore‐forming antibiotic peptides.
Insulin‐dependent phosphorylation of DPP IV in liver
Authors: Bilodeau Nicolas , Fiset Annie , Poirier Guy G. , Fortier Suzanne , Gingras Marie‐Claude , Lavoie Josée N. , Faure Robert L
Published: February 2006

Dipeptidyl peptidase IV (DPP IV, CD26, EC 3.4.14.5) serves as a model aimed at elucidating protein sorting signals. We identify here, by MS, several tyrosine‐phosphorylated proteins in a rat liver Golgi/endosome (G/E) fraction including DPP IV. We show that a pool of DPP IV is tyrosine‐phosphorylated. Maximal phosphorylation was observed after 2 min following intravenous insulin injection. DPP IV coimmunoprecipitated with the cellular tyrosine kinase Src (c‐Src) with maximal association also observed after 2 min following insulin injection. DPP IV was found phosphorylated after incubation of nonsolubilized G/E membranes with [γ‐32P]ATP. The c‐Src inhibitor PP2 inhibited DPP IV phosphorylation. Oriented proteolysis experiments indicate that a large pool of c‐Src is protected in G/E fractions. Following injection of the protein‐tyrosine phosphatase inhibitor bpV(phen), DPP IV levels markedly decreased by 40% both in plasma membrane and G/E fractions. In the fraction designated Lh, DPP IV levels decreased by 50% 15 min following insulin injection. Therefore, a pool of DPP IV is tyrosine‐phosphorylated in an insulin‐dependent manner. The results suggest the presence of a yet to be characterized signalling mechanism whereby DPP IV has access to c‐Src‐containing signalling platforms.
The enzyme‐binding region of human GM2‐activator protein
Authors: Wendeler Michaela , Werth Norbert , Maier Timm , Schwarzmann Guenter , Kolter Thomas , Schoeniger Maike , Hoffmann Daniel , Lemm Thorsten , Saenger Wolfram , Sandhoff Konrad
Published: February 2006

The GM2‐activator protein (GM2AP) is an essential cofactor for the lysosomal degradation of ganglioside GM2 by β‐hexosaminidase A (HexA). It mediates the interaction between the water‐soluble exohydrolase and its membrane‐embedded glycolipid substrate at the lipid–water interface. Functional deficiencies in this protein result in a fatal neurological storage disorder, the AB variant of GM2 gangliosidosis. In order to elucidate this cofactor's mode of action and identify the surface region of GM2AP responsible for binding to HexA, we designed several variant forms of this protein and evaluated the consequences of these mutations for lipid‐ and enzyme‐binding properties using a variety of biophysical and functional studies. The point mutants D113K, M117V and E123K showed a drastically decreased capacity to stimulate HexA‐catalysed GM2 degradation. However, surface plasmon resonance (SPR) spectroscopy showed that the binding of these variants to immobilized lipid bilayers and their ability to solubilize lipids from anionic vesicles were the same as for the wild‐type protein. In addition, a fluorescence resonance energy transfer (FRET)‐based assay system showed that these variants had the same capacity as wild‐type GM2AP for intervesicular lipid transfer from donor to acceptor liposomes. The concentration‐dependent effect of these variants on hydrolysis of the synthetic substrate 4‐methylumbelliferyl‐2‐acetamido‐2‐deoxy‐6‐sulfo‐β‐d‐glucopyranoside (MUGS) indicated a weakened association with the enzyme's α subunit. This identifies the protein region affected by these mutations, the single short α helix of GM2AP, as the major determinant for the interaction with the enzyme. These results further confirm that the function of GM2AP is not restricted to a biological detergent that simply disrupts the membrane structure or lifts the substrate out of the lipid plane. In contrast, our data argue in favour of the critical importance of distinct activator–hexosaminidase interactions for GM2 degradation, and corroborate the view that the activator/lipid complex represents the true substrate for the degrading enzyme.
Identification and characterization of two dipeptidyl‐peptidase III isoforms in Drosophila melanogaster
Authors: Mazzocco Claire , Gillibert‐Duplantier Jennifer , Neaud Veronique , Fukasawa Kayoko M. , Claverol Stéphane , Bonneu Marc , Puiroux Jacques
Published: February 2006

Dipeptidyl‐peptidase III (DPP III) hydrolyses small peptides with a broad substrate specificity. It is thought to be involved in a major degradation pathway of the insect neuropeptide proctolin. We report the purification and characterization of a soluble DPP III from 40 g Drosophila melanogaster. Western blot analysis with anti‐(DPP III) serum revealed the purification of two proteins of molecular mass 89 and 82 kDa. MS/MS analysis of these proteins resulted in the sequencing of 45 and 41 peptide fragments, respectively, confirming ≈ 60% of both annotated D. melanogaster DPP III isoforms (CG7415‐PC and CG7415‐PB) predicted at 89 and 82 kDa. Sequencing also revealed the specific catalytic domain HELLGH in both isoforms, indicating that they are both effective in degrading small peptides. In addition, with a probe specific for D. melanogaster DPP III, northern blot analysis of fruit fly total RNA showed two transcripts at ≈ 2.6 and 2.3 kb, consistent with the translation of 89‐kDa and 82‐kDa DPP III proteins. Moreover, the purified enzyme hydrolyzed the insect neuropeptide proctolin (Km≈ 4 µm) at the second N‐terminal peptide bound, and was inhibited by the specific DPP III inhibitor tynorphin. Finally, anti‐(DPP III) immunoreactivity was observed in the central nervous system of D. melanogaster larva, supporting a functional role for DPP III in proctolin degradation. This study shows that DPP III is in actuality synthesized in D. melanogaster as 89‐kDa and 82‐kDa isoforms, representing two native proteins translated from two alternative mRNA transcripts.
TNF‐857 polymorphism in Israeli Jewish patients with inflammatory bowel disease
Authors: Fidder Herma H. , Heijmans Roel , Chowers Yehuda , Bar‐Meir Simon , Avidan Benjamin , Pena A. Salvador , Crusius J. Bart A.
Published: February 2006

Tumour necrosis factor (TNF)‐α is an important pro‐inflammatory cytokine that has been implicated in the pathogenesis of inflammatory bowel disease (IBD). The promoter TNF‐857 C→T single nucleotide polymorphism (SNP) is functional through the binding to the transcription factor octamer transcription factor‐1 (OCT‐1). In order to investigate the frequency of this SNP in Israeli Jewish IBD patients, we analysed a cohort of well‐characterized patients, 153 with Crohn's disease (CD) and 78 with ulcerative colitis (UC) and 188 healthy controls individually matched for age, sex and ethnicity. Forty‐one per cent of the patients were of Ashkenazi and 48% were of non‐Ashkenazi background. The remaining 11% were of mixed Ashkenazi–non‐Ashkenazi background. Patients and controls were genotyped for the TNF‐857 SNP by Taqman technology. Stratification for the CARD15 Arg702Trp, Gly908Arg and Leu1007fsinsC mutations took place in 136 CD patients.

Carrier frequency of TNF‐857T between CD and controls (36% vs. 40%; P = 0.556; OR: 1.18, 95% CI 0.74–1.88), or between UC and controls (41% vs. 37%; P = 0.743; OR: 0.85, 95% CI 0.45–1.62) did not differ significantly. Neither did stratifying for the presence of at least one of the common CARD15 mutations result in a significant difference between CD and controls. No associations were found between TNF‐857T and CD phenotype as defined by the Vienna classification, perianal disease or extra‐intestinal disease irrespective of CARD15 carrier status. In conclusion, it appears that TNF‐857 SNP does not contribute to susceptibility of IBD, neither does it define the phenotype of CD in Israeli Jewish IBD patients.
THE STUDY OF OPTIMUM OPERATING CONDITIONS OF EXTRUDED SNACK FOOD WITH TOMATO POWDER
Authors: HUANG RONG‐CHENG , PENG JINCHYAU , LU FUNG‐JOU , LUI WAI‐BUN , LIN JENSHINN
Published: February 2006

Tomatoes are an integral part of diets worldwide. Many population studies have established a link between dietary intake of tomato, a major source of a carotenoid antioxidant, lycopene, and reduced risk of chronic diseases. Response surface methodology was used to analyze the effect of screw speed (X1), moisture content (X2) and tomato powder content (X3) on the objective attributes (radial expansion ratio [Y1], bulk density [Y2], degree of gelatinization [DG], water absorption index [WAI] and hardness [Y5]) of an extruded snack food with tomato powder. A Box‐Behnken design was used to develop models for the objective responses. The experiments were performed at 100C with the following: feed rate, 19.2 kg/h; X1, 250 rpm; and die diameter, 3.98 mm. Individual contour plots of the different responses were overlaid. Regions meeting 3.22 optimum Y1, 0.13‐g/cm Y2, 81.25% DG, 5.12‐g gel/g dry weight WAI and 35.43‐N Y5 were identified at 12.5 and 25% X2 and X3, respectively.
FACTORS AFFECTING THE ULTRAVIOLET INACTIVATION OF ESCHERICHIA COLI K12 IN APPLE JUICE AND A MODEL SYSTEM*
Authors: MURAKAMI EDGAR G. , JACKSON LAUREN , MADSEN KEVIN , SCHICKEDANZ BRIAN
Published: February 2006

Ultraviolet (UV) light has been used successfully for years to sterilize water and was recently approved as an acceptable irradiation treatment for the processing of juice. Although there is considerable information on the efficacy of UV processing in the treatment of water, limited data are available on its efficacy in fluid food systems. The objectives of this work were to determine the effects of apple‐juice properties on the UV inactivation of Escherichia coli K12 and the interdependence of intensity and time on the efficacy of UV light. Results showed that absorbance (A) and suspended solids (SS) affected UV inactivation, while pH and dissolved solids did not. Concerning the interdependence of intensity and time, intensity levels can only be changed without sacrificing effectiveness at a limited range of intensity and dose levels. This means that the range of the intensity level of the actual UV reactor must be considered in process‐parameter determination.
OPTIMIZATION OF NATTOKINASE PRODUCTION CONDUCTION USING RESPONSE SURFACE METHODOLOGY
Authors: WANG DJA‐SHIN , TORNG CHAU‐CHEN , LIN I‐PING , CHENG BOR‐WEN , LIU HUI‐RONG , CHOU CHAO‐YU
Published: February 2006

Natto has attracted worldwide attention because of its health benefits and long history in Japanese food. It has been found that a potent fibrinolytic enzyme named nattokinase, which is extracted from natto, is able to prevent atherosclerosis. The production of nattokinase may be influenced by various factors such as temperature, shaking speed, volume of medium, fermentation time and so forth. Three‐step response surface methodology was applied to obtain the optimal operation conditions of the fermentation process in order to maximize the nattokinase yield. The three major steps are described as follows. First, the important factors for fermentation were identified by L8 orthogonal array experiment. The chosen factors were temperature (37 or 45C), shaking speed (110 or 150 rpm), volume of medium (80 or 120 mL), Brix of wheat bran extract (1.5 or 3°), Brix of soy meal extract (1 or 2°), glucose concentration (0.6 or 1.2%) and fermentation time (24 or 36 h). Second, a regression equation was established between the response (i.e., the enzyme activity) and the two statistically significant factors (i.e., the volume of medium and fermentation time). Third, the optimal solutions for the volume of medium and fermentation time were obtained based on the response surface of the regression equation. According to the response surface analysis, the optimal operation conditions for the fermentation process should be 80 mL and 37.0817 h for the volume of medium and the fermentation time, respectively, which resulted in 459.11 FU/mL as the predicted enzyme activity.
Improving the Dietary Patterns of Adolescents Using a Computer‐Based Approach
Authors: Casazza Krista , Ciccazzo Michele
Published: February 2006

Abstract: The Centers for Disease Control and Prevention (CDC) recently reported that 16‐33% of children and adolescents in the United States are overweight or obese, making this the number one nutritional disease of this group. The prevalence appears to be increasing annually, as are the associated consequences. Moreover, the CDC estimates that less than 50% of adolescents are physically active on a regular basis. In order to improve the health of these individuals, an intervention must be focused to modify these behaviors. Facilitating the understanding of proper nutrition and need for physical activity among adolescents will likely prevent health problems such as overweight and obesity and the development of chronic diseases later in life. Despite these concerns, adolescents remain one of the most underserved populations in terms of preventive services. In the past, health education interventions have shown increases in knowledge; however, the ultimate goal of these programs is to positively impact behavior. To accomplish this, we must focus on the learning styles of “today’s” adolescents. Computer‐based interventions allow for individualized strategies to promote healthy lifestyles. A computer‐based intervention program has the potential to elicit a greater behavior change in comparison to a traditional didactic intervention program. (J Sch Health. 2006;76(2):43‐46)
Medication Adherence Among Adolescents in a School‐Based Health Center
Authors: Mears Cynthia J. , Charlebois Nicole M. , Holl Jane L.
Published: February 2006

Abstract: School‐based health centers are an integral part of the health care delivery system for low‐income children. Medication adherence for these patients may be challenging because the student is often responsible for bringing home the prescription and receiving the instructions. This study assesses medication fill, initiation, and adherence rates among adolescents in a school‐based health center to identify major barriers to medication compliance. Students enrolled in a school‐based health center, ≥10 years old, able to read and write English, and whose parent had provided consent for participation, were eligible for the study. Eligible students who received a prescription from the health center were invited to return to the clinic a week later to complete a questionnaire (with verbal assent). Primary outcome measures included medication fill rates, medication initiation rates, medication adherence rates, and reasons for nonadherence. Eighty‐one students completed the questionnaire: 45 students (55.6%) filled their prescription. Of the students who filled their prescriptions, 75.6% reported always taking their medication at the appropriate time, 22.2% reported sometimes forgetting to take their medication, and 2.2% reported never taking the medication. However, many discrepancies were found between reported medication‐taking behavior and the instructions provided to the student. Medication fill, initiation, and adherence rates among students receiving prescriptions for medications in school‐based health centers are suboptimal. Interventions that address key identified barriers need to be developed and evaluated in order to achieve optimal fill and adherence rates. (J Sch Health. 2006;76(2):52‐56)
Internet Versus Paper Mode of Health and Health Behavior Questionnaires in Elementary Schools: Asthma and Fruit as Examples
Authors: Mangunkusumo Resiti T. , Duisterhout Joop S. , De Graaff Nijs , Maarsingh Erik J. , De Koning Harry J. , Raat Hein
Published: February 2006

Abstract: The Internet has become an inevitable tool for collecting health and health behavior questionnaires. This study compared the feasibility, presence of score differences, and subjective evaluations by children between Internet and identical paper (asthma/fruit) questionnaires in elementary schools. A randomized crossover design was applied, with children starting with one administration mode before completing (5 minutes later) the other mode. Ten Dutch elementary schools with 270 school children (fifth grade, 10‐12 years) were approached to participate. Response was 92%. The Internet mode had significantly less missing/nonunique answers than the paper mode (p < .01). The completion times did not differ significantly between the Internet and the paper mode. Except for perceived self‐efficacy to eat sufficient fruit (p < .05), no differences in the asthma and fruit scores were found when comparing between the 2 modes. All variables showed strong intraclass correlation coefficients (166 ≥ 0.64) between modes. Most items had good to very good agreement (kappa 0.61‐0.95). The percentages for global and exact agreement ranged per item from 61.3 to 100. Most children preferred the Internet mode on “general preference” and “ease of use” aspects but rated no preference on “understandability.” The majority rated the evaluation aspects of the Internet mode positively. Thus, Internet administration of a health and health behavior questionnaire is feasible at elementary schools, gives comparable responses, and is well accepted and preferred by children compared to the paper version. Therefore, it is recommended to use Internet as a tool in health and health behavior research among children. (J Sch Health. 2006;76(2):80‐86)
Comparisons Between Thai Adolescent Voices and Thai Adolescent Health Literature
Authors: Thongpriwan Vipavee , McElmurry Beverly J.
Published: February 2006

Abstract: Thai adolescents are hesitant to openly talk to adults; however, they are avid users of the Internet. In 2002, faculty of the Boromarajonani College of Nursing, Nopparat Vajira, Thailand, established a webboard to reach out to high school students for questions and answers on adolescent health. Adolescents pose health questions, which are answered by nursing faculty and students. A total of 106 questions were selected for content analysis. Thai adolescent studies for the years 1992 to 2004 were identified from searches of CINAHL, ERIC, MEDLINE, and PsycINFO databases. The selection criteria required that chosen articles have a Thai adolescent health focus, be written in English, and be retrievable. Of the 68 citations identified, 23 studies met inclusion criteria. Content of the Thai adolescent webboard was compared with a content analysis of the retrieved Thai adolescent research. Physiological development, sexuality, and risky behaviors were common literature themes, whereas Thai adolescents expressed concerns about love and dating relationships. Parenting and parent‐child relationships were discussed on the webboard but not in the literature. Analysis of the mental health revealed differences between the literature that covered psychosocial change, and the webboard questions concerned with body image, the need for emotional support, and satisfaction and conflicts of friendship. It is recommended that investigators consider incorporating adolescents as research team participants, particularly as they examine mental health promotion, adolescent and family relationships, and concerns of Thai adolescents. (J Sch Health. 2006;76(2):47‐51)
Performance of Antibodies against Tissue Transglutaminase for the Diagnosis of Celiac Disease: Meta-Analysis
Authors: Zintzaras Elias , Germenis Anastasios E.
Published: February 2006

A meta-analysis of studies investigating the diagnostic accuracy of enzyme-linked immunosorbent assays (ELISA) for antibodies against tissue transglutaminases (tTG) of various origins in celiac disease (CD) diagnosis was carried out. Twenty-one studies, with untreated CD patients and healthy/CD-free controls, were included in the meta-analysis. The diagnostic accuracy was estimated using a summary receiver operating characteristic (SROC) curve and pooled sensitivity (Se) and specificity (Sp). Multiple assays within a study were treated by considering all the assays within a study and by analyzing the most popular assay (i.e., the commercial anti-tTTG ELISA most frequently utilized in the papers in which multiple assays were included). The SROC curve indicated the absence of heterogeneity, and the superiority of recombinant human tTG (rh-tTG) and purified human tTG (ph-tTG) compared to guinea pig-tTG (gp-tTG). The sensitivities (most popular assay) for rh-tTG, ph-tTG, and gp-tTG were 94%, 90%, and 92%, respectively, and the specificities were 97%, 92%, and 96%, respectively. A sensitivity analysis (exclusion of studies with bias) altered the results of ph-tTG: Se, 95%; Sp, 98%. The sensitivities (all individual assays) for rh-tTG, ph-tTG, and gp-tTG were 94%, 94%, and 91%, respectively, and the specificities were 95%, 94%, and 89%, respectively. Human tTG ELISA is sensitive and specific, and it can be used for mass screening. Sensitivity analysis showed that ph-tTG might perform better.
Performance of a Time-Resolved Fluorescence Immunoassay for Measuring Varicella-Zoster Virus Immunoglobulin G Levels in Adults and Comparison with Commercial Enzyme Immunoassays and Merck Glycoprotein Enzyme Immunoassay
Authors: Maple P. A. C. , Gray J. , Breuer J. , Kafatos G. , Parker S. , Brown D.
Published: February 2006

Highly sensitive and specific, quantitative assays are needed to detect varicella-zoster virus (VZV) immunoglobulin G in human sera, particularly for determining immune status and response following vaccination. A time-resolved fluorescence immunoassay (TRFIA) has been developed, and its performance was compared to that of two commercial enzyme immunoassays (EIAs) and Merck glycoprotein EIA (gpEIA). The TRFIA had equivalent sensitivity (97.8%) and high specificity (93.5%) in relation to gpEIA. A commercial (Behring) EIA compared favorably with TRFIA in terms of sensitivity (98.4%) but had lower specificity (80.7%). Another commercial EIA (Diamedix) had high specificity (97.1%) but low sensitivity (76.4%) compared to TRFIA if equivocal test results were treated as negative for VZV antibody. A novel feature of the TRFIA was that the cutoff was generated using population mixture modeling and was expressed in mIU/ml, as the assay was calibrated using the British standard VZV antibody.
The Probiotic Bacterium
Lactobacillus casei
Induces Activation of the Gut Mucosal Immune System through Innate Immunity
Authors: Galdeano C. Maldonado , Perdigón G.
Published: February 2006

The mechanisms by which probiotic bacteria affect the immune system are unknown yet, but many of them are attributed to an increase in the innate or in the acquired immune response. To study the influence of the probiotic bacterium
Lactobacillus casei
in the expression of receptors involved in the innate immune response, this bacterium was orally administered to BALB/c mice. After, they were sacrificed; the small intestine and intestinal fluids were collected to measure secretory immunoglobulin A (IgA) specific for
L. casei
. Mononuclear cells from Peyer's patches were isolated to determine the CD-206 and TLR-2 receptors. In histological slices we determined the number of IgA
+
, CD4
+
, CD8
+
, and CD3
+
cells and two cytokines (interleulin-5 [IL-5] and IL-6). CD-206 and TLR-2 increased with respect to the untreated control. We did not observe an increase in the T population or in the IL-5-positive cells. IgA
+
cells and IL-6-producing cells increased after 7 days of
L. casei
administration. We did not find specific antibodies against
L. casei
. The main immune cells activated after oral
L. casei
administration were those of the innate immune response, with an increase in the specific markers of these cells (CD-206 and TLR-2), with no modification in the number of T cells.
Human Interleukin-15 Improves Engraftment of Human T Cells in NOD-SCID Mice
Authors: Sun Anyuan , Wei Haiming , Sun Rui , Xiao Weihua , Yang Yongguang , Tian Zhigang
Published: February 2006

Human nonobese diabetic-severe combined immune deficiency (NOD-SCID) mouse chimeras have been widely used as an in vivo model to assess human immune function. However, only a small fraction of transferred human T lymphocytes can be detected in human peripheral blood lymphocyte (huPBL)-NOD-SCID chimeras. To improve the reconstitution of human T lymphocytes in NOD-SCID mice, the use of recombinant human interleukin-15 (rhIL-15) as a stimulator of human lymphocytes was explored. Administration of rhIL-15 after transplantation of huPBLs into NOD-SCID mice increased reconstitution of human T lymphocytes in a dose-dependent manner, with an optimal dosage of 1 μg/mouse. The number of human T lymphocytes (HLA-ABC+CD3+) in the lymphoid organs or tissue of rhIL-15-treated huPBL-NOD-SCID mice increased 11- to 80-fold, and phytohemagglutinin-induced T-lymphocyte proliferation and cytokine production were significantly enhanced. Additionally, although mature human cells have not been thought to enter the murine thymus, human T lymphocytes were detected in the huPBL-NOD-SCID thymus after rhIL-15 treatment. Thus, rhIL-15 can be used to optimize long-term peripheral T-cell engraftment in these human-mouse chimeras and may also be useful in clinical treatment of T-cell deficiencies.
Performance of the TruGene Human Immunodeficiency Virus Type 1 Genotyping Kit and OpenGene DNA Sequencing System on Clinical Samples Diluted to Approximately 100 Copies per Milliliter
Authors: Gale Howard B. , Kan Virginia L. , Shinol Rebecca C.
Published: February 2006

The TruGene human immunodeficiency virus type 1 (HIV-1) genotyping kit/OpenGene DNA sequencing system (Bayer HealthCare, Tarrytown, NY) reliably produced clinically acceptable resistance profiles for reverse transcriptase and protease inhibitors on patient samples diluted to ∼100 copies/ml following extraction with the QIAamp viral RNA minikit (QIAGEN Inc., Valencia, CA). One modification of the standard protocol was made to guarantee PCR amplification: a centrifugation step to concentrate virus was added before RNA extraction. For genotypic antiretroviral resistance testing, no significant differences in the identification and sensitivity of detection for codon mutations, base mutations, and multibase sites were found between the original and diluted samples.
Modulation of Immunity-Related Gene Expression in Small Intestines of Mice by Oral Administration of Lactoferrin
Authors: Wakabayashi Hiroyuki , Takakura Natsuko , Yamauchi Koji , Tamura Yoshitaka
Published: February 2006

Oral administration of lactoferrin (LF), an antimicrobial and immunomodulatory protein, shows a protective effect against infectious diseases, possibly via immunomodulation of the host. Initially, we confirmed an immunomodulatory effect of LF by observing changes in the number of cells in the leukocyte subsets in the peripheral blood and spleens of mice 1 day after oral administration of LF. Then we developed a quantitative reverse transcription-PCR method for 20 immunity-related genes of antimicrobial proteins, pattern recognition receptors, cytokines, and lymphocyte mobilization-related proteins, and we assessed the expression of these genes in the small intestines of mice 2 h after administration of water, bovine serum albumin (BSA), or LF. Expression of the LF gene was lower in mice administered LF than in mice administered water or BSA, implying a negative-feedback control. Expression of gamma interferon (IFN-γ) and interleukin-10 (IL-10) was lower in both BSA- and LF-administered mice than in water administered mice, suggesting a nonspecific effect of protein ingestion. Expression of NOD2, IFN-β, and IL-12p40 was higher with LF administration than with water or BSA administration. The expression levels of these three genes were correlated. This study indicated that oral administration of LF modulates the small intestinal expression of genes closely related to the host defense in a specific or a nonspecific manner.
Gamma Interferon Production in Response toMycobacterium bovisBCG andMycobacterium tuberculosisAntigens in Infants Born to Human Immunodeficiency Virus-Infected Mothers
Authors: Van Rie Annelies , Madhi Shabir A. , Heera Jayvant R. , Meddows-Taylor Stephen , Wendelboe Aaron M. , Anthony Fiona , Violari Avy , Tiemessen Caroline T.
Published: February 2006

In utero sensitization to infectious pathogens can establish immunological memory and may influence the immune response to unrelated antigens. Little is known about the influence of intrauterine human immunodeficiency virus (HIV) exposure on the cellular immune response to mycobacterial antigens. Whole-blood culture gamma interferon (IFN-γ) production in response to mycobacterial antigens was measured at birth and 6 weeks of age to determine the characteristics of the IFN-γ response in HIV-exposed infants toMycobacterium bovisBCG and mycobacterial antigens. At birth, we observed an increased immune activation in response to phytohemagglutinin among HIV-exposed, uninfected infants. In a proportion of these infants, we also observed an increased immune activation in response to purified protein derivative, BCG, and early secreted antigen target 6. Increases in the IFN-γ response to the four antigens between birth and 6 weeks of age, observed in all HIV-unexposed infants, was absent in a substantial proportion of HIV-exposed, uninfected infants. The immunological differences persisted at 6 weeks of age, suggesting a sustained impact of in utero immune priming by HIV. Intrauterine exposure to HIV affects the infants' cellular immune response to mycobacterial antigens, either specifically or as a consequence of nonspecific, broadly reactive immune activation. Further studies will be important to help determine optimal vaccination and disease prevention strategies for this vulnerable population group.
Evaluation of a Tetraplex Microsphere Assay for
Bordetella pertussis
Antibodies
Authors: Prince Harry E. , Lapé-Nixon Mary , Matud Jose
Published: February 2006

To increase testing efficiency, a microsphere-based multianalyte immune detection (MAID) system was developed to measure serum immunoglobulin G (IgG) and IgA recognizing two
Bordetella pertussis
antigens, pertussis toxin (PT) and filamentous hemagglutinin antigen (FHA). The assay was performed as two separate duplexes. One duplex measured IgG to PT and FHA, and the other measured IgA to PT and FHA. The two duplexes were then combined and analyzed as a tetraplex. The MAID system and an in-house enzyme-linked immunosorbent assay (ELISA) system were used to evaluate 100 sera from blood donors and 220 consecutive sera submitted for
B. pertussis
antibody testing. For both the MAID and ELISA systems, antibody levels were defined as increased if greater than the blood donor group 95th percentile value. The qualitative concordance rates between MAID and ELISA results for the 220 consecutively submitted sera were as follows: PT IgG, 99%; PT IgA, 94%; FHA IgG, 93%; FHA IgA, 94%. The overall concordance rate was 95% (836 of 880 result sets). For 29 of 44 (66%) discordant result sets, the discordant MAID result was supported by the MAID and ELISA results for other
B. pertussis
antibodies. The MAID and in-house ELISA systems were also used to evaluate 20 sera previously tested for pertussis antibodies at a pertussis vaccine research laboratory; MAID results for all four analytes did not significantly differ from results obtained by the research laboratory. These findings show that antibodies to
B. pertussis
antigens can be measured easily and accurately using a tetraplex microsphere system.
Kinetics and Diagnostic and Prognostic Potential of Quantitative Western Blot Analysis and Antigen-Specific Enzyme-Linked Immunosorbent Assay in Experimental Canine Leishmaniasis
Authors: Talmi-Frank D. , Strauss-Ayali D. , Jaffe C. L. , Baneth G.
Published: February 2006

Quantitative computerized Western blot analysis of antibody responses during experimental canine
Leishmania infantum
infection distinguished between immunodominant and nonimmunodominant protein bands. Six infected beagles, positive by both PCR and parasite culture, were monitored over 75 weeks postinfection and during a 12-week allopurinol treatment course. All dogs were symptomatic at the time of treatment. Of 12 antigenic bands examined, the immunodominant bands (12, 14, 24, 29, 48, and 68 kDa) showed significantly increased intensities (
P
< 0.01) and higher frequencies of recognition than the nonimmunodominant bands at all time points. Detection of the former bands at 6 weeks postinfection preceded seroconversion by enzyme-linked immunosorbent assay (ELISA) both on crude
Leishmania
antigen or the recombinant proteins rK39 and HSP70. Reactivity with the 14-, 48-, and 68-kDa bands signified early infection, whereas increased reactivity with the 14-, 24-, and 29-kDa bands was associated with posttreatment parasite persistence and potential unfavorable prognosis. Total lane intensity (TLI) emerged as a sensitive marker for early infection and increased as early as 4 weeks postinfection. TLI had a significantly higher (
P
< 0.01) relative increase rate than crude
Leishmania
antigen or HSP70 or rK39 ELISA at all time points. These immunodominant antigens and TLI, as determined by quantitative Western blotting, will be valuable for early detection and treatment evaluation of canine leishmaniasis.
Detection of κ and λ Light Chain Monoclonal Proteins in Human Serum: Automated Immunoassay versus Immunofixation Electrophoresis
Authors: Jaskowski Troy D. , Litwin Christine M. , Hill Harry R.
Published: February 2006

Recently, turbidimetric immunoassays for detecting and quantifying κ and λ free light chains (FLC) have become available and are promoted as being more sensitive than immunofixation electrophoresis (IFE) in detecting FLC monoclonal proteins. In this study, we assessed the ability of these turbidimetric assays to detect serum monoclonal proteins involving both free and heavy-chain-bound κ and λ light chains compared to standard immunofixation electrophoresis. Sera demonstrating a restricted band of protein migration (other than a definite M spike) by serum protein electrophoresis (SPE), which may represent early monoclonal proteins, were also examined. When compared to IFE, percent agreement, sensitivity, and specificity for the κ-FLC and λ-FLC were 94.6, 72.9, and 99.5% and 98.5, 91.4, and 99.7%, respectively, in detecting monoclonal proteins involving free and heavy-chain-bound light chains. The majority of sera (73.7%) demonstrating a restricted band of protein migration on SPE demonstrated abnormal IFE patterns suggestive of multiple myeloma or monoclonal gammopathy of unknown significance, but gave normal κ/λ FLC ratios using the turbidimetric immunoassays. In conclusion, the κ and λ FLC assays are significantly less sensitive (72.9 to 91.4%) than IFE, but specific in detecting serum monoclonal proteins. Moreover, the κ/λ ratio has little value in routine screening since the majority of sera with abnormal IFE patterns had normal κ/λ FLC ratios.