bluuebunny/crossref_metadata_2025 huggingface dataset:
A comparison of disease‐specific medical expenditures in Japan using the principal diagnosis method and the proportional distribution method
Authors: Tanihara Shinichi , Okamoto Etsuji , Une Hiroshi
Published: February 2011

Background  There are technical limitations to handling all the information mentioned in health insurance claims (HICs) in Japan. Therefore, conventional methods for estimating disease‐specific medical expenditures assume that all medical care expenditures in a given HIC are spent on only one principal diagnosis even when the HIC contains multiple diagnoses.

Objectives  To estimate the disease‐specific medical expenditures that consider all diagnosis on a given HIC.

Methods  Data were obtained from 169 622 outpatient HICs in May 2006 from health insurance provided by the employer. We compared the estimated disease‐specific medical expenditures of the conventional method with a proportional distribution method (PDM), which considers all diagnoses on the HICs.

Results  For diabetes mellitus and other diseases of the digestive system, the proportion of principal diagnoses among total diagnoses was 52.4% (4849/9251) and 19.6% (2614/13331), respectively. In addition, the ratio of the estimated disease‐specific medical expenditures between the conventional method and the PDM method was 1.49 for diabetes mellitus and 0.64 for other diseases of the digestive system. The estimation of disease‐specific medical expenditures using the conventional method may therefore have overestimated the expenditures on the disease category typically selected as the principal diagnosis and underestimated the expenditures on the disease category less likely to be selected as the principal diagnosis.

Conclusions  The conventional method for estimation of disease‐specific medical expenditures should be improved by utilizing all the diagnoses information on HICs.
Part 2 – primary health care workforce policy intricacies: multidisciplinary team1 case analysis
Authors: Félix‐Bortolotti Margot
Published: February 2011

Objective  To demonstrate the usefulness of theoretical framework in the analysis of complex policy issues related to primary health care (PHC) workforce for informed policy decisions and planning.

Approach  The study links complexity theories with political economy and connects them to theoretical work on structure and agency to guide the analysis of the PHC workforce issues and their related ramifications. This theoretical framework guides as well the literature review. With this approach, it is possible to ascertain and disturb the linear thinking which tends to surround questions or issues related to PHC human resources.

Conclusion  Workforce changes occur in a multifaceted structure of norms and values as well as within the policies, regulations and traditions, the ruling relations2 between professions, and between employers and employees. Similarly, changes in the population characteristics and in the political economy not only affect the demand for care but also have implications for the type of staff that can be hired.
The grey zones of birth and death
Authors: Glouberman Sholom
Published: February 2011

Rationale  The promise of science to find clear and distinct boundaries for birth and death seems to have failed. The more science advances the more blurred the boundaries become. An overview of the history and the current trajectory suggests that the more we advance the greater the blurring.

Method  The paper examines the history of scientific advance in the areas of birthing and dying and traces the path for the early promise for definitive answers to the new more uncertain and ambiguous positions we have come to.

Conclusions  The problems associated with birthing and dying increasingly require humanistic philosophical discussion rather than scientific experimentation.
EMG analysis of trapezius and masticatory muscles: experimental protocol and data reproducibility
Authors: SFORZA C. , ROSATI R. , De MENEZES M. , MUSTO F. , TOMA M.
Published: February 2011

Summary  We aimed to define a standardised protocol for the electromyographic evaluation of trapezius muscle in dentistry and to assess its within‐ and between‐session repeatability. Surface electromyography of trapezius, masseter and temporal muscles was performed in 40 healthy subjects aged 20–35 years during shoulder elevation, and maximum teeth clenching with and without cotton rolls. Two repetitions were made both within (same electrodes) and between sessions (different electrodes). Maximum voluntary clench on cotton rolls was used to standardise the potentials of the six analysed muscles with tooth contact; shoulder elevation was used to standardise the upper trapezius potentials. From the standardised electromyographic potentials, several indices (muscle symmetry; masticatory muscle torque and relative activity; total masticatory muscle activity; trapezius cervical load, percentage co‐contraction of trapezius during teeth clenching) were computed; random (technical error of measurement) and systematic (Student’s t‐test, Analysis of Variance) errors were assessed. For all indices, no systematic errors were found between the two separate data collection sessions. Within session, limited (lower than 8%) technical errors of measurement were found for temporalis and masseter symmetry, torque and activity indices, and the trapezius cervical load. Larger random errors were obtained for trapezius symmetry and total masticatory muscle activity (up to 20%). Between sessions, no significant differences were found for trapezius co‐contraction. In conclusion, a protocol for the standardisation of trapezius muscle that may be used within dental clinical applications was defined, and the repeatability of masseter, temporalis and trapezius electromyographic recordings for serial assessments was assessed in healthy subjects.
Retracted: Nitric oxide enhances plant ultraviolet‐B protection up‐regulating gene expression of the phenylpropanoid biosynthetic pathway
Authors: TOSSI VANESA , AMENTA MELINA , LAMATTINA LORENZO , CASSIA RAÚL
Published: February 2011

The link between ultraviolet (UV)‐B, nitric oxide (NO) and phenylpropanoid biosynthetic pathway (PPBP) was studied in maize and Arabidopsis. The transcription factor (TF) ZmP regulates PPBP in maize. A genetic approach using P‐rr (ZmP+) and P‐ww (ZmP‐) maize lines demonstrate that: (1) NO protects P‐rr leaves but not P‐ww from UV‐B‐induced reactive oxygen species (ROS) and cell damage; (2) NO increases flavonoid and anthocyanin content and prevents chlorophyll loss in P‐rr but not in P‐ww and (3) the NO scavenger 2‐(4‐carboxyphenyl)‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl‐3‐oxide (cPTIO) blocks the UV‐B‐induced expression of ZmP and their targets CHS and CHI suggesting that NO plays a key role in the UV‐B‐regulated PPBP. Involvement of endogenous NO was studied in Arabidopsis nitric oxide dioxygenase (NOD) plants that express a NO dioxygenase gene under the control of a dexamethasone (DEX)‐inducible promoter. Expression of HY5 and MYB12, TFs involved in PPBP regulation, was induced by UV‐B, reduced by DEX in NOD plants and recovered by subsequent NO treatment. C4H regulates synapate esters synthesis and is UV‐B‐induced in a NO‐independent pathway. Data indicate that UV‐B perception increases NO concentration, which protects plant against UV‐B by two ways: (1) scavenging ROS; and (2) up‐regulating the expression of HY5, MYB12 and ZmP, resulting in the PPBP activation.
REDOX regulation of mitochondrial function in plants
Authors: GIRAUD ESTELLE , VAN AKEN OLIVIER , UGGALLA VINDYA , WHELAN JAMES
Published: February 2011

Mitochondrial components dynamically change in response to environmental and developmental cues. However, the regulatory pathways that underlie these changes are largely unknown. A global analysis of changes in mitochondrial components at the transcript, protein and metabolite levels was undertaken, to gain a greater insight into how mitochondrial functions are regulated and respond to various internal or external cues. At the transcript level, large‐scale changes in groups of genes suggest the presence of co‐regulatory mechanisms for these components. Furthermore, the pathways that regulate these changes appear to be integrated into regulatory pathways that alter a variety of functions in cells. However, the changes in transcripts are not always observed at the protein or the metabolite level. This is likely to be due to post‐transcriptional levels of regulation and also the fact that in‐depth profiles, which have been obtained for transcripts from a variety of studies, are currently not available for proteins and metabolites. Thus, while transcripts for genes give us a picture of what the cells are ‘thinking’ in relation to mitochondrial components, some of these responses may be lost in translation.
Histopathology provides a phenotype by which to characterize stripe rust resistance genes in wheat
Authors: Jagger L. J. , Newell C. , Berry S. T. , MacCormack R. , Boyd L. A.
Published: February 2011

The expression of the resistance phenotypes of QPst.jic‐2D and QPst.jic‐4B, two quantitative trait loci (QTL) for stripe rust resistance in wheat cv. Alcedo, were assessed relative to plant growth stage, while a histopathology analysis was undertaken to characterize the cellular interaction between Puccinia striiformis f. sp. tritici (the causal agent of stripe rust) and each QTL. QPst.jic‐2D expressed a partial resistant phenotype at seedling growth stages, with the level of resistance increasing as the wheat plant matured, conferring a disease‐free phenotype at heading. QPst.jic‐4B, however, did not express a resistant phenotype until booting (growth stage 41 on the Zadoks scale), displaying its full resistant phenotype at heading. Microscopic examination in flag leaves showed that infection sites formed in all genotypes tested, with full infection‐site establishment being observed by 36 h post‐inoculation (hpi). In lines carrying both QPst.jic‐2D and QPst.jic‐4B, as well as the parental cv. Alcedo, no microcolony formation, defined by the appearance of runner hyphae, was observed. Microcolony formation was observed in lines carrying only one, or neither QTL. Cell death associated with infection sites was observed for all genotypes, although the timing of first appearance and the extent of the cell death response varied considerably. In lines carrying both QPst.jic‐2D and QPst.jic‐4B cell death did not extend beyond one to three cells. In lines carrying only one QTL more extensive cell death was observed and cell death appeared later in lines with QPst.jic‐4B than QPst.jic‐2D. Cell death was also occasionally observed in lines without either QTL, although only at 264 hpi.
Direct evidence for pressure‐generated closure of plasmodesmata
Authors: Oparka K.J. , Prior D.A.M.
Published: February 2011

Pressure differentials were generated between adjacent leaf trichome cells of Nicotiana clevelandii using a modified micropressure probe/injection system. The location of the pressure differentials was altered using two main treatments: (a) puncturing one of the cells in the trichome to induce a large differential at its junction with the next basal cell, or (b) raising the pressure of the impaled cell and ‘clamping’ it at a higher value, thus creating a differential at its upper, as well as lower, wall. In both treatments basipetal intercellular transport of Lucifer yellow was impeded exactly at the sites predicted by the generated pressure differentials; at the base of punctured cells and at the top of clamped cells. In flaccid trichomes puncturing was without effect because the turgor of all the cells in the trichome was too low (<50 kPa) to generate a sufficiently large pressure differential between the punctured cell and its neighbours. Elevations of cell turgor (ΔP) in excess of 200 kPa or more were required to impede intercellular transport When cells were clamped at a high pressure and then subsequently allowed to lose their turgor by withdrawal of the probe the effects of the original pressure clamp were partially retained, indicating an inability to resume intercellular transport.
The adenine nucleotide translocator of higher plants is synthesized as a large precursor that is processed upon import into mitochondria
Authors: Winning Brenda M. , Sarah Caroline J. , Purdue P. Edward , Day Christopher D. , Leaver Christopher J.
Published: February 2011

Two maize genes and cDNAs encoding the mitochondrial adenine nucleotide translocator (ANT), a nuclear‐encoded inner mitochondrial membrane carrier protein, have previously been isolated in this laboratory. Sequence analysis revealed the existence of much longer open reading frames than the corresponding fungal and mammalian ANT genes. Potato ANT cDNAs have subsequently been isolated and sequenced and alignment of the deduced plant amino acid sequences with the equivalent fungal and mammalian polypeptides indicated that the plant proteins contain N‐terminal extensions. When the plant cDNA clones are expressed in vitro they direct the synthesis of precursor proteins that are specifically processed at the N‐terminus upon import into isolated mitochondria. N‐terminal amino acid sequence data obtained from the native proteins purified from both maize and potato mitochondria has allowed identification of the putative processing sites. Further import analysis has shown that two distinct regions of the maize precursor protein contain targeting information, the 97 amino acids at the N‐terminus and the 267 C‐terminal amino acids. This is the first report that provides experimental evidence that the adenine nucleotide translocator of higher plants is synthesized as a large precursor protein that is specifically cleaved upon import into mitochondria. Import of ANT into higher plant mitochondria therefore appears to be different to the corresponding process in fungal and mammalian systems where targeting of ANT to mitochondria is mediated by internal signals and there is no N‐terminal processing.
Mutagenesis of a race‐specific rust resistance gene in Antirrhinum majus using a transposon‐tagging protocol
Authors: Aitken E.A.B. , Callow J.A. , Newbury H.J.
Published: February 2011

In a transposon tagging experiment, lines of Antirrhinum majus exhibiting both race‐specific resistance (homozygous for the dominant R gene) to the rust pathogen Puccinia antirrhini and a high frequency of transposition were crossed with homozygous susceptible lines. From a screen of 11153 F1 progeny, 15 individuals were detected showing susceptibility to rust race α. Six of these exhibited a susceptibility phenotype (classified as type B) not previously observed. A control experiment involving the same tagging strategy but employing lines that do not exhibit high rates of transposition did not yield any susceptible mutants from a screen of 6243 progeny. In experiments on the heritability and stability of the mutation, the six plants exhibiting susceptibility phenotype B produced progeny in which the R locus had reverted to an active form (i.e. some of the progeny were resistant), a classic characteristic of transposon‐tagged plant genes. Reversion was shown to occur somatically, and its rate was temperature dependent. Inheritance studies showed that the mutations in two of the susceptible plants from the tagging protocol map at, or very close to, the race α‐specific resistance gene. The results are consistent with the transposon tagging of a race‐specific gene for rust resistance.
Co‐immunolocalization of topoisomerase II and chloroplast DNA in developing, dividing and mature wheat chloroplasts
Authors: Marrison Joanne L. , Leech Rachel M.
Published: February 2011

This paper describes the first localization of immunofluorescence of topoisomerase II in developing chloroplasts. In order to investigate the relationship between topoisomerase II and chloroplast DNA (ctDNA) replication during chloroplast development the 7‐day‐old wheat leaf was used. Topoisomerase II was immunolabelled and fluorescein tagged and the ctDNA simultaneously stained with 4,6‐diamidino‐2‐phenylindole (DAPI) in the same sections. Topoisomerase II was detected at every stage of chloroplast development and maximal levels of topoisomerase II were found in chloroplasts at the time of ctDNA replication. Topoisomerase II was localized around the plastid periphery, exactly mirroring the position of the ctDNA. After chloroplast division both topoisomerase II and ctDNA are seen to be restricted to small discrete areas within the plastid, but at different sites. These findings strongly suggest a role for topoisomerase II in ctDNA decatenation prior to chloroplast division.
Efficient initiation of translation at non‐AUG triplets in plant cells
Authors: Gordon Karl , Fütterer Johannes , Hohr Thomas
Published: February 2011

The efficiency of translation initiation at triplets differing at one residue from AUG was tested by transient expression in protoplasts from two different plant species. All possible alternative codons were tested. Some triplets showed significant CAT activity, with CUG (30% of the AUG activity) being most active. Most others had between 5 and 15% of the activity obtained from AUG, whereas UUG and AUC yielded about 2% and the two composed only of purines, AAG and AGG, had no significant activity. Translation initiation from AUC, especially, responded to leader sequences outside the immediate context which did not affect translation initiation from AUG.
A pathogen‐induced gene of barley encodes a protein showing high similarity to a protein kinase regulator
Authors: Brandt Jakob , Thordal‐Christensen Hans , Vad Knud , Gregersen Per L. , Collinge David B.
Published: February 2011

A full length cDNA of a barley leaf messenger, found to increase in amount during infection attempts by the powdery mildew fungus (Erysiphe graminis), is characterized. The messenger encodes a polypeptide of 261 amino acid residues with a calculated mass of 29.2 kDa and a pl of 4.6. Sequence comparisons as well as serological studies demonstrate that the encoded protein is closely related to a family of mammalian proteins believed to have functions associated with the multifunctional Ca2+‐dependent protein kinases.
Fatty acid elongase is required for shoot development in rice
Authors: Ito Yukihiro , Kimura Fumiko , Hirakata Kazuma , Tsuda Katsutoshi , Takasugi Tomoaki , Eiguchi Mitsugu , Nakagawa Kiyotaka , Kurata Nori
Published: February 2011

Organisms are covered extracellularly with cuticular waxes that consist of various fatty acids. In higher plants, extracellular waxes act as indispensable barriers to protect the plants from physical and biological stresses such as drought and pathogen attacks. However, the effect of fatty acid composition on plant development under normal growth conditions is not well understood. Here we show that the ONION1 (ONI1) gene, which encodes a fatty acid elongase (β‐ketoacyl CoA synthase) involved in the synthesis of very‐long‐chain fatty acids, is required for correct fatty acid composition and normal shoot development in rice. oni1 mutants containing a reduced amount of very‐long‐chain fatty acids produced very small shoots, with an aberrant outermost epidermal cell layer, and ceased to grow soon after germination. These mutants also showed abnormal expression of a KNOX family homeobox gene. ONI1 was specifically expressed in the outermost cell layer of the shoot apical meristem and developing lateral organs. These results show that fatty acid elongase is required for formation of the outermost cell layer, and this layer is indispensable for entire shoot development in rice.
Root hair‐specific expansins modulate root hair elongation in rice
Authors: ZhiMing Yu , Bo Kang , XiaoWei He , ShaoLei Lv , YouHuang Bai , WoNa Ding , Ming Chen , Hyung‐Taeg Cho , Ping Wu
Published: February 2011

Root hair growth requires intensive cell‐wall modification. This study demonstrates that root hair‐specific expansin As, a sub‐clade of the cell wall‐loosening expansin proteins, are required for root hair elongation in rice (Oryza sativaL.). We identified a gene encodingEXPA17(OsEXPA17) from a rice mutant with short root hairs. Promoter::reporter transgenic lines exhibited exclusiveOsEXPA17expression in root hair cells. The OsEXPA17 mutant protein (OsexpA17) contained a point mutation, causing a change in the amino acid sequence (Gly104→Arg). This amino acid alteration is predicted to disrupt a highly conserved disulfide bond in the mutant. Suppression ofOsEXPA17by RNA interference further confirmed requirement for the gene in root hair elongation. Complementation of theOsEXPA17mutant with other root hair EXPAs (OsEXPA30 and Arabidopsis EXPA7) can restore root hair elongation, indicating functional conservation of these root hair EXPAs in monocots and dicots. These results demonstrate that members of the root hair EXPA sub‐clade play a crucial role in root hair cell elongation in Graminaceae.
Arabidopsis chloroplast lipid transport protein TGD2 disrupts membranes and is part of a large complex
Authors: Roston Rebecca , Gao Jinpeng , Xu Changcheng , Benning Christoph
Published: February 2011

In most plants the assembly of the photosynthetic thylakoid membrane requires lipid precursors synthesized at the endoplasmic reticulum (ER). Thus, the transport of lipids from the ER to the chloroplast is essential for biogenesis of the thylakoids. TGD2 is one of four proteins in Arabidopsis required for lipid import into the chloroplast, and was found to bind phosphatidic acid in vitro. However, the significance of phosphatidic acid binding for the function of TGD2 in vivo and TGD2 interaction with membranes remained unclear. Developing three functional assays probing how TGD2 affects lipid bilayers in vitro, we show that it perturbs membranes to the point of fusion, causes liposome leakage and redistributes lipids in the bilayer. By identifying and characterizing five new mutant alleles, we demonstrate that these functions are impaired in specific mutants with lipid phenotypes in vivo. At the structural level, we show that TGD2 is part of a protein complex larger than 500 kDa, the formation of which is disrupted in two mutant alleles, indicative of the biological relevance of this TGD2‐containing complex. Based on the data presented, we propose that TGD2, as part of a larger complex, forms a lipid transport conduit between the inner and outer chloroplast envelope membranes, with its N terminus anchored in the inner membrane and its C terminus binding phosphatidic acid in the outer membrane.
MR spectroscopy in opiate maintenance therapy: association of glutamate with the number of previous withdrawals in the anterior cingulate cortex
Authors: Hermann Derik , Frischknecht Ulrich , Heinrich Milena , Hoerst Mareen , Vollmert Christian , Vollstädt‐Klein Sabine , Tunc‐Skarka Nuran , Kiefer Falk , Mann Karl , Ende Gabriele
Published: February 2011

Pre‐clinical research indicates that opioids reduce extracellular glutamate in acute opioid treatment, whereas during withdrawal, glutamatergic neurotransmission is increased and withdrawal symptoms can be blocked by glutamate receptor antagonists. The glutamate hypothesis of addiction suggests that withdrawal‐associated hyperglutamatergic states destabilize the glutamatergic system chronically and contribute to relapse. magnetic resonance spectroscopy at three tesla optimized for glutamate assessment (TE 80 ms) was performed in the anterior cingulate gyrus (ACC) and frontal white matter (fWM) of 17 opiate‐dependent patients during opiate maintenance therapy and 20 healthy controls. Controlling for age and gray matter content, glutamate in the ACC was positively associated with the number of previous withdrawals. For glutamate + glutamine (Glx), a significant group–age interaction was found. Whereas Glx declines with age in healthy controls, Glx increases with age in opiate‐dependent patients. The number of previous withdrawals did not correlate with age. In fWM spectra, increased Cho concentrations were observed in opiate‐dependent patients. Both new findings, the positive correlation of glutamate and previous withdrawals and increasing Glx with age in contrast to an age‐dependent Glx decrease in controls indicate a destabilization of the glutamate system in opiate‐dependent patients and support the glutamate hypothesis of addiction. Increased Cho concentrations in fWM corroborate findings of WM abnormalities in opioid‐dependent subjects.
Evidence that vasopressin V1b receptors mediate the transition to excessive drinking in ethanol‐dependent rats
Authors: Edwards Scott , Guerrero Miguel , Ghoneim Ola M. , Roberts Edward , Koob George F.
Published: February 2011

Alcoholism is a devastating condition that represents a progression from initial alcohol use to dependence. Although most individuals are capable of consuming alcohol in a limited fashion, the development of alcohol dependence in a subset of individuals is often associated with negative emotional states (including anxiety and depression). Since the alleviation of this negative motivational state via excessive alcohol consumption often becomes a central goal of alcoholics, the transition from initial use to dependence is postulated to be associated with a transition from positive to negative reinforcement mechanisms. Vasopressin is a neuropeptide known to potentiate the effects of CRF on the HPA axis, and emerging evidence also suggests a role for centrally located vasopressin acting on V1b receptors in the regulation of stress‐ and anxiety‐like behaviors in rodents. The present study determined state‐dependent alterations in vasopressin/V1bR signaling in an animal model of ethanol dependence. The V1bR antagonist SSR149415 dose‐dependently reduced excessive levels of ethanol self‐administration observed in dependent animals without affecting the limited levels of ethanol drinking in non‐dependent animals. Ethanol self‐administration reduced V1b receptor levels in the basolateral amygdala of non‐dependent animals, a neuroadaptation that could theoretically facilitate the positive reinforcing effects of alcohol. In contrast, V1bR levels were seemingly restored in ethanol‐dependent rats, a switch that may in part underlie a transition from positive to negative reinforcement mechanisms with dependence. Together, our data suggest a key role for vasopressin/V1bR signaling in the transition to ethanol dependence.
Alpha‐synuclein and heroin craving in opiate‐dependent patients on injectable heroin maintenance
Authors: Dürsteler‐MacFarland Kenneth M. , Brugger Isabelle , Bönsch Dominikus , Schmid Otto , Kornhuber Johannes , Bleich Stefan , Wiesbeck Gerhard A.
Published: February 2011

Research suggests that alpha‐synuclein (SNCA) and NACP‐Rep1, a polymorphic complex microsatellite repeat ~10 kb upstream of the SNCA gene translational start, may be involved in substance‐use behaviors and craving. This study was the first to examine the effects of diacetylmorphine (DAM) on peripheral SNCA protein expression along with craving in opiate‐dependent patients and to compare their NACP‐Rep1 allele lengths with those of healthy controls. Using an experimental design, opiate‐dependent patients on injectable heroin maintenance were investigated at four time points, twice pre‐ and post‐injection of DAM. SNCA protein levels of 30 DAM‐maintained patients were measured using enzyme‐linked immunosorbent assay. Participant‐rated effects were assessed in 42 patients by Tiffany's Heroin Craving Questionnaire (HCQ), Gossop's Short Opiate Withdrawal Scale and Visual Analogs. NACP‐Rep1 alleles of 42 patients and 101 controls were analyzed. One‐way repeated‐measures ANOVAs provided significant overall effects for SNCA protein content (P = 0.028), craving (P < 0.001), withdrawal symptomatology (P < 0.001) and mood (P < 0.001), indicating that DAM injections may not only reduce craving but also SNCA protein expression. However, there was no association between protein expression and craving. Relative to controls, patients had significantly longer NACP‐Rep1 alleles (P < 0.001). NACP‐Rep1 allele lengths correlated positively with HCQ total scores averaged across all time points (r = 0.420; P = 0.006) as well as with post‐DAM HCQ total scores in the morning (r = 0.488, P = 0.001) and afternoon (r = 0.423, P = 0.005). The findings provide evidence of a contributory role of SNCA and NACP‐Rep1 for opiate dependence.
Role of ghrelin in food reward: impact of ghrelin on sucrose self‐administration and mesolimbic dopamine and acetylcholine receptor gene expression
Authors: Skibicka Karolina P. , Hansson Caroline , Egecioglu Emil , Dickson Suzanne L.
Published: February 2011

The decision to eat is strongly influenced by non‐homeostatic factors such as food palatability. Indeed, the rewarding and motivational value of food can override homeostatic signals, leading to increased consumption and hence, obesity. Ghrelin, a gut‐derived orexigenic hormone, has a prominent role in homeostatic feeding. Recently, however, it has emerged as a potent modulator of the mesolimbic dopaminergic reward pathway, suggesting a role for ghrelin in food reward. Here, we sought to determine whether ghrelin and its receptors are important for reinforcing motivation for natural sugar reward by examining the role of ghrelin receptor (GHS‐R1A) stimulation and blockade for sucrose progressive ratio operant conditioning, a procedure used to measure motivational drive to obtain a reward. Peripherally and centrally administered ghrelin significantly increased operant responding and therefore, incentive motivation for sucrose. Utilizing the GHS‐R1A antagonist JMV2959, we demonstrated that blockade of GHS‐R1A signaling significantly decreased operant responding for sucrose. We further investigated ghrelin's effects on key mesolimbic reward nodes, the ventral tegmental area (VTA) and nucleus accumbens (NAcc), by evaluating the effects of chronic central ghrelin treatment on the expression of genes encoding major reward neurotransmitter receptors, namely dopamine and acetylcholine. Ghrelin treatment was associated with an increased dopamine receptor D5 and acetylcholine receptor nAChRβ2 gene expression in the VTA and decreased expression of D1, D3, D5 and nAChRα3 in the NAcc. Our data indicate that ghrelin plays an important role in motivation and reinforcement for sucrose and impacts on the expression of dopamine and acetylcholine encoding genes in the mesolimbic reward circuitry. These findings suggest that ghrelin antagonists have therapeutic potential for the treatment of obesity and to suppress the overconsumption of sweet food.
Short‐term development of behavioral sensitization to amphetamine requires N‐methyl‐D‐aspartate‐ and nicotinic‐dependent mechanisms in the nucleus accumbens
Authors: Degoulet Mickaël , Rostain Jean‐Claude , Abraini Jacques H. , David Hélène N.
Published: February 2011

Repeated administration of psychostimulant drugs, such as amphetamine, induces an enhanced behavioral response to subsequent drug challenge. This behavioral sensitization is proposed to model the increased drug craving observed in human psychostimulant abusers. Current thinking is that the ventral tegmental area, but not the nucleus accumbens, plays a critical role in the development of behavioral sensitization. Here, we report that the concomitant blockade of glutamatergic and nicotinic ionotropic receptors in the core of the nucleus accumbens blocks the development of behavioral sensitization to amphetamine and further abolishes the increase in extracellular dopamine release induced by amphetamine in the nucleus accumbens. These findings demonstrate that the development of behavioral sensitization to amphetamine depends, in addition to the well‐known role of the ventral tegmental area, on glutamatergic and nicotinic‐dependent mechanisms in the core of the nucleus accumbens and further indicate that the dopaminergic mesolimbic pathway must be viewed as a single coordinated system of critical importance in the development of behavioral sensitization to psychostimulant drugs.
Collaboration and co‐ownership in research: dynamics and dialogues between patient research partners and professional researchers in a research team
Authors: Nierse Christi J. , Schipper Karen , van Zadelhoff Ezra , van de Griendt Joos , Abma Tineke A.
Published: February 2011

Background  Different strategies by which patients can be involved in research include consultation, control and collaboration. This article focuses on collaboration within research teams and considers this with reference to a research project about setting a social–scientific agenda for health research from the perspectives of patients with a chronic kidney disease (CKD).

Objective  To examine the dynamics and dialogues in a collaboration between patient research partners and professional researchers.

Design  A responsive methodology was used in the research project. Two patient research partners participated in the research team. Twenty‐seven patients with CKD and parents of young children with CKD were interviewed, seven focus groups were organized and observations were held at a dialysis centre.

Results  During the collaboration, the research partners and professional researchers engaged in a mutual learning process in all stages of the project. The professional researchers gave the research partners a sense of ownership in the research process. The research partners could relate to the lives of patients by using their experiential knowledge. In the context of collaborative working, this helped shape an agenda for research.

Conclusion and discussion  The active involvement of patients as research partners can add value to a research strategy, especially when research partners and professional researchers engage in a dialogue that is open, inclusive and deliberative. Issues for discussion include the possibility of ‘over‐involvement’, the research profile and training of research partners and whether participation of patients is restricted to certain types of research.
Outcomes in pediatric solid‐organ transplantation
Authors: LaRosa Christopher , Jorge Baluarte H. , Meyers Kevin E. C.
Published: February 2011

LaRosa C, Jorge Baluarte H, Meyers KEC. Outcomes in pediatric solid‐organ transplantation.  Pediatr Transplantation 2011: 15:128–141. © 2010 John Wiley & Sons A/S.

Abstract:  Pediatric solid‐organ transplantation is an increasingly successful treatment for organ failure. Five‐ and 10‐yr patient survival rates have dramatically improved over the last couple of decades, and currently, over 80% of pediatric patients survive into adolescence and young adulthood. Waiting list mortality has been a concern for liver, heart, and intestinal transplantation, illustrating the importance of transplant as a life‐saving therapy. Unfortunately, the success of pediatric transplantation comes at the cost of long‐term or late complications that arise as a result of allograft rejection or injury, immunosuppression‐related morbidity, or both. As transplant recipients enter adolescence treatment, non‐adherence becomes a significant issue, and the medical and psychosocial impacts transition to adulthood not only with regard to healthcare but also in terms of functional outcomes, economic potential, and overall QoL. This review addresses the clinical and psychosocial challenges encountered by pediatric transplant recipients in the current era. A better understanding of pediatric transplant outcomes and adult morbidity and mortality requires further ongoing assessment.
KIR/HLA‐I mismatching and risk of relapse in paediatric patients undergoing non‐haploidentical allogeneic haematopoietic stem cell transplantation
Authors: Scquizzato Elisa , Zambello Renato , Teramo Antonella , Baesso Ilenia , Varotto Stefania , Albergoni Maria Paola , Boscaro Elisa , Cesaro Simone , Pillon Marta , Calore Elisabetta , Gazzola Maria Vittoria , Semenzato Gianpietro , Messina Chiara , Trentin Livio
Published: February 2011

Scquizzato E, Zambello R, Teramo A, Baesso I, Varotto S, Albergoni MP, Boscaro E, Cesaro S, Pillon M, Calore E, Gazzola MV, Semenzato G, Messina C, Trentin L. KIR/HLA‐I mismatching and risk of relapse in paediatric patients undergoing non‐haploidentical allogeneic haematopoietic stem cell transplantation. Pediatr Transplantation 2011: 15:198–204. © 2010 John Wiley & Sons A/S.

Abstract:  In HSCT setting, KIR‐driven alloreactivity might be better predicted if the donor KIR genotype is considered in addition to the recipient HLA genotype. The prediction of NK cell alloreactivity relies on the missing ligand in the recipient, a scenario that can be found in HLA‐identical and non‐identical allotransplants. The aim of this study was to investigate at genetic level the prognostic impact of recipient HLA‐I lacking for donor KIR on allotransplanted patients outcome. We analysed donors KIR genotype and HLA genotype of 60 paediatric patients who received related (n = 15) or unrelated (n = 45) transplantation. When patients were grouped based on the KIR gene type involved in the KIR/HLA‐I mismatch, we did not observe any relapse in the group of patients characterized by mismatches involving only inhibitory KIR. On the contrary, all relapses were observed in patients showing at least one activating gene involved in the mismatch (p < 0.05). Although the biological mechanism accounting for this putative genetic rule is still to be clarified, we suggest that a careful survey of KIR/HLA‐I mismatching should be taken into account in the selection of donor in related and unrelated HSCT.
DNA Analysis to Find Rare Blood Donors when Antisera is not Available
Authors: Reid Marion E.
Published: February 2011

In order to screen for antigen‐negative blood donors, it is necessary to have appropriate, potent antisera in sufficient volume. Anti‐Doa and anti‐Dob are notoriously weakly reactive antibodies, available only in small volumes, usually in sera containing other alloantibodies, and often deteriorate on storage. Thus, it has not been possible to test large numbers of blood samples to find Do (a‐) or Do (b‐) blood donors. At the NYBC, we now type selected donors for DOA and DOB by DNA analysis. Initially, we tested DNA Prepared from donors who had been typed by hemagglutination for one or both antigens. We found that four donors, whose RBCs previously typed as Do (a+b‐), had both DOA and DOB alleles, and when retested, the RBCs were Do (a+b+w). We have now tested over 300 donors for DO by PCR‐RFLP using either Eam1105 I or BseRI restriction enzymes. Blood from DOA/DOA donors has survived better than “crossmatch compatible” blood for Patients with anti‐Dob and such results suggest that anti‐Dob is a more frequent cause of transfusion reactions than reported. Furthermore, we have demonstrated that PCR‐RFLP can be used to screen for antigen‐negative donors in other blood group systems when appropriate antisera are not available. When interpreting the results, it is important to remember that the genotype may not reflect the phenotype. Our strategy has been to perform DNA analysis for the DO alleles on those donors who have been shown by hemagglutination to lack antigens corresponding to multiple alloantibodies in patients’ plasma. In this way, we have been able to supply rare blood to numerous patients, whose serum contained at least 5 additional alloantibodies of clinical significance.
Rare Blood Donors: The Past and the Future
Authors: Woodfield Graeme
Published: February 2011

For many years, there has been a clinical requirement for the provision of units of rare blood for patients with either single or multiple blood group antibodies present. To co‐ordinate this need, the Rare Blood Donor Working Party of the International Society of Blood Transfusion was established in 1985. Since then, the Working Party has been active in promoting information on the procedures and methods for testing, packaging, transport and delivery of rare blood internationally. The Working Party has a close relationship with the Blood Group Reference Laboratory in the United Kingdom. The availability of the World Health Organization International Donor Panel permits the rapid and accurate locating of rare blood units.

The Rare Blood Donor Working Party conducts surveys, attends to technical queries and is active in education. This can assist countries that are upgrading their blood services and give them a greater understanding of the international help available when confronted with a difficult‐to‐transfuse patient.

In the future, it is anticipated that a wider range of countries will contribute to the International Donor Panel. It is likely that with the many major population movements throughout the world and the potential consequent development of rare antibodies or mixtures of antibodies, the need for the availability of rare blood donations will remain for the foreseeable future.
Molecular Characterization of the Immune Response to Factor VIII
Authors: Lollar Pete
Published: February 2011

Inhibitory antibodies to factor VIII arise from an alloimmune response in patients with hemophilia A infused with factor VIII and as an autoimmune response in a variety of settings. The immune response to factor VIII is T‐cell dependent. Helper T cells recognize numerous epitopes in the factor VIII molecule. B cell epitopes in both the alloimmune and autoimmune responses are much more restricted, usually involving two major epitopes in the A2 and C2 domains and apparently minor epitopes in the light chain activation peptide (ap) region and the A3 domain. Anti‐C2 antibodies inhibit the binding of factor VIII to phospholipid and may also interfere with the binding of factor VIII to von Willebrand factor. Anti‐A2 and anti‐A3 antibodies block the binding of factor VIII to factor X and factor IXa, respectively, in the intrinsic pathway factor X activation complex. The mechanism of inhibition of anti‐ap antibodies is unknown. A murine hemophilia A model has been developed to study the immunogenicity of factor VIII. This model may lead to improved approaches to prevent development of inhibitory antibodies and to reverse the immune response if it develops.
Characterisation and natural variation of a dehydrin gene in Quercus petraea (Matt.) Liebl.
Authors: Vornam B. , Gailing O. , Derory J. , Plomion C. , Kremer A. , Finkeldey R.
Published: February 2011

For the first time in sessile oak [Quercus petraea (Matt.) Liebl.], the isolation and characterisation of a full‐length dehydrin gene and its promoter region, as well as its allelic variation in natural populations, is reported. Dehydrins (Dhn) are stress‐related genes important for the survival of perennial plants in a seasonal climate. A full‐length dehydrin gene (Dhn3) was characterised at the nucleotide level and the protein structure was modelled. Additionally, the allelic variation was analysed in five natural populations of Quercus petraea (Matt.) Liebl. sampled along an altitudinal gradient in the French Pyrenees. The analysed sequences contain typical domains of the Kn class of dehydrins in the coding region. Also, the 5′untranslated region (promoter) of the gene was amplified, which shows typical motifs essential for drought‐ and cold‐responsive gene expression. Single nucleotide substitutions and indels (insertions/deletions) within the coding region determine large biochemical differences at the protein level. However, only low levels of genetic differentiation between populations from different altitudes were detectable.
Seed longevity and fire: germination responses of an exotic perennial herb in NW Patagonian grasslands (Argentina)
Authors: Franzese J. , Ghermandi L.
Published: February 2011

Fire affects grassland composition by selectively influencing recruitment. Some exotic species can increase their abundance as a consequence of fire‐stimulated seed germination, but response may depend on seed age. Rumex acetosella L. (Polygonaceae, sheep’s sorrel) is a cosmopolitan herb that has invaded NW Patagonia’s grasslands. This species forms persistent soil seed banks and increases after disturbances, particularly fire. We studied how fire and seed longevity influence R. acetosella germination. In 2008, we conducted laboratory experiments where we exposed different‐aged seeds (up to 19 years old) to heat, smoke, charcoal, ash and control treatments. Total percentage germination and mean germination time depended on both seed age and fire treatment. Germination of younger seeds decreased with increasing temperature. There was no general pattern in germination responses of different‐aged seeds to smoke, charcoal and ash. While smoke improved the germination of fresh seeds, charcoal decreased germination. Germination of untreated seeds was negatively correlated with seed age, and mean germination time increased with seed age. In most treatments, fresh seeds had lower germination than 1–5‐year‐old seeds, indicating an after‐ripening requirement. Smoke stimulates R. acetosella germination, causing successful recruitment during post‐fire conditions. Fresh seeds are particularly responsive to fire factors, possibly because they have not experienced physical degradation and are more receptive to environmental stimuli. Knowing the colonisation potential from the soil seed bank of this species during post‐fire conditions will allow us to predict their impact on native communities.
Potentiality of evenly distributed hypervariable microsatellite markers in marker‐assisted breeding of rice
Authors: Narshimulu Gonela , Jamaloddin Mohammed , Vemireddy Lakshminarayana R. , Anuradha Ghnata , Siddiq Eea
Published: February 2011

With 4 figures and 1 table

To provide ready‐to‐use markers for background selection in marker‐assisted breeding of rice, we have extracted 840 hypervariable microsatellite markers (hvRMs) evenly distributed in the genome from 18 828 Class I simple sequence repeats. These hvRMs were grouped into bins of 5‐Mb intervals with 5–8 bins depending on the chromosome length. In each bin, 10 hvRMs were selected based on their microsatellite repeat number and type. Of 840, a representative of 36 hvRMs were selected and its polymorphic information content (PIC) was estimated by amplifying 24 diverse rice cultivars. The findings revealed that hvRMs show high PIC values and more alleles when compared to the 25 randomly chosen microsatellite markers. The informative hvRMs developed for background selection across cultivars in this study along with the already established molecular markers tightly linked to the genes of agronomic value could be of value for the development of a ‘MAS kit’ for speeding up the efforts of plant breeders for targeted crop improvement.
Phenotypic characterization of sweet potato [Ipomoea batatas (L.) Lam.] genotypes in relation to prediction of chemical quality constituents by NIRS equations
Authors: Lebot Vincent , Ndiaye André , Malapa Roger
Published: February 2011

With 5 figures and 3 tables

This study evaluates the potential of NIRS (near‐infrared reflectance spectroscopy) to predict the major constituents (starch, sugars, cellulose, proteins and minerals) of the sweet potato root. Overall, 240 accessions were morphologically described, chemically analysed and their NIR spectra recorded. No correlations were observed between aerial and underground traits, and between morphological traits and major constituents. Calibration equations, developed on 190 accessions, showed high explained variances in cross‐validation (r2cv) for starch (0.82), sugars (0.91), proteins (0.89) and minerals (0.74) but no response for cellulose (0.21). The predictions were tested on an independent set of 50 randomly selected accessions. The r2pred values for starch, sugars and proteins were, respectively, of 0.71, 0.82 and 0.87 with ratios of performance to deviation (RPD) of 2.11, 2.29 and 2.93. New calibration equations developed on 240 accessions showed improved RPD values for starch (2.65), sugars (2.75), cellulose (1.58), proteins (3.47) and minerals (2.34), indicating that larger sets could improve prediction. NIRS could be used in sweet potato breeding programmes to predict starch, sugars and proteins contents in the roots.