bluuebunny/crossref_metadata_2025 huggingface dataset:
Age and Comorbidity in Acute Myocardial Infarction: A Report From the AMI‐Florence Italian Registry
Authors: Balzi Daniela , Barchielli Alessandro , Buiatti Eva , Franceschini Caterina , Mangani Irene , Del Bianco Laura , Monami Matteo , Valente Serafina , Franco Gensini Gian , Marchionni Niccolò ,   The AMI‐Florence Working Group
Published: January 2007

A total of 930 cases of ST‐segment elevation myocardial infarction were prospectively recorded in the Florence health district. Factors influencing survival or those associated with use of revascularization (percutaneous coronary intervention, 91%) were identified through multivariate analyses (Cox and logistic regression, respectively). The independent protective effect of coronary reperfusion therapy (CRT) was evident at 36 months (39% reduction in the risk of death). After adjusting for all multivariate predictors, CRT use was 63% less likely at age 85 years and older than at under 65 years (p<0.001). Since beyond advancing age, comorbidity appeared to be associated with a reduced chance of CRT, three chronic comorbidity score categories were calculated using information on past medical history. Increased 1‐year mortality in patients with higher comorbidity score categories derives, at least in part, from underutilization of CRT. Results confirm that although they might potentially benefit from CRT during ST‐segment elevation myocardial infarction, older and frail patients are excluded from CRT, even when eligible.
Cardiac Rehabilitation in the Elderly: Patient Selection and Outcomes
Authors: Ferrara Nicola , Corbi Graziamaria , Bosimini Enzo , Cobelli Franco , Furgi Giuseppe , Giannuzzi Pantaleo , Giordano Amerigo , Pedretti Roberto , Scrutinio Domenico , Rengo Franco
Published: January 2007

In Western countries, the aging and improving survival of patients with coronary heart disease are responsible for an increasing number of older adults (65 years of age and older) who are eligible for cardiac rehabilitation. The elderly with coronary heart disease represent a special population with changes induced by aging and lifestyle, comorbidity, cognitive dysfunction, and high risk of disability. Although the elderly account for the majority of cardiac admissions and procedures, studies on cardiac rehabilitation have traditionally focused on younger patients. In aged experimental animals, there is evidence that exercise training is able to improve hemodynamic parameters and biologic markers. Moreover, in older patients, exercise improves functional capacity and reduces myocardial work, similar to that seen in younger patients. As for younger patients, cardiac rehabilitation requires a multidisciplinary approach, including comprehensive assessment, treatment of risk factors and comorbidity, and psychosocial assessment. Cardiac rehabilitation is safe and helpful for elderly coronary patients. Physicians must be encouraged to prescribe cardiac rehabilitation programs for the elderly following major coronary events and coronary revascularization procedures.
Primary Percutaneous Coronary Intervention vs. Fibrinolytic Therapy for Acute ST‐Elevation Myocardial Infarction in the Elderly
Authors: Karha Juhana , Topol Eric J.
Published: January 2007

Advanced age is associated with worse prognosis among patients with acute ST‐elevation myocardial infarction. Many eligible elderly patients with acute ST‐elevation myocardial infarction, however, do not receive any reperfusion therapy at all. The risk of intracranial hemorrhage complicating fibrinolytic therapy increases with age. Furthermore, routine adjunctive stenting has made coronary angioplasty safer. In total, primary percutaneous coronary intervention is the preferred reperfusion strategy among elderly patients with acute ST‐elevation myocardial infarction, provided that it can be performed without excessive delay. The breakeven incremental delay with primary percutaneous coronary intervention compared with fibrinolytic therapy is not clear at this point and will need to be elucidated by future investigation.
The Great Escape: Junctional Escape—Capture Bigeminy
Authors: Henry Sonia A. , Makaryus Amgad N. , Loewinger Lee , Boal Bernard
Published: January 2007

Escape—capture bigeminy is a bigeminal rhythm in which each escape beat is followed by a captured beat. This dysrhythmia is very rare, because its manifestation requires the sinus interval to be longer than the escape interval. This is possible only with severe sinus nodal disease, where the intrinsic sinus rate is extremely low, or with a sinus rhythm associated with an accelerated junctional rhythm. The authors review the case of a 75‐year‐old man who presented with occasional dizziness and near‐syncopal episodes. He was diagnosed with escape—capture bigeminy and subsequently underwent pacemaker placement.
Dying on One's Own Terms
Authors: Basta Lofty L
Published: January 2007

Just as I select my ship as I am about to go on a voyage, so I shall choose my death when I am about to depart from life.—Lucius Annaeus Seneca, c. 4 BC—65 AD
Chagas' Disease and the Use of Implantable Cardioverter‐Defibrillators in Brazil
Authors: Rosa Ronaldo F. , Neto Argemiro S. , Franken Roberto A.
Published: January 2007

The authors discuss the epidemiology and pathogenesis of Chagas' disease in Brazil, including the use of treatment with a cardioverter‐defibrillator in patients with low ejection fraction. Select patients may benefit from resynchronization therapy associated with cardioverter‐defibrillator treatment. Electrophysiologic study is indicated in the assessment of the potential utility of an implantable cardioverter‐defibrillator.
Implantable Cardioverter‐Defibrillators: Expectations in the Brazilian Elderly
Authors: Gravina Claudia F. , Garcez‐Leme Luiz Eugěnio , Leme Mariana Deckers , Piegas Leopoldo
Published: January 2007

By 2025, Brazil will have the sixth largest elderly population in the world. Thirty percent of the elderly currently have private health insurance and 70% have their health care paid for by the Unified Health System. The system, founded through a governmental entity, provides free health service to all citizens. In 2003, the government passed the Elderly Statement Law, claiming that the elderly have equal access to prevention, promotion, protection, and recovery of health. In spite of the high costs, the placement of implantable cardioverter‐defibrillators (ICDs) in the elderly is growing. There is a significant regional difference in the number of ICD placements, and there is also a significant sex bias: 73% of the ICDs were placed in elderly men. The majority of ICDs were placed in the elderly from ages 65 to 79 years. Nevertheless, 9.5% of ICDs were placed in octogenarians and nonagenarians, reflecting the valuation of biologic age rather than chronologic age.
The Slt2‐type MAP kinase Bmp3 of Botrytis cinerea is required for normal saprotrophic growth, conidiation, plant surface sensing and host tissue colonization
Authors: RUI OLIVER , HAHN MATTHIAS
Published: January 2007

Mitogen‐activated protein kinases (MAPKs) play important roles in signal transduction and regulation of various aspects of saprotrophic and pathogenic growth in plant pathogenic fungi. We have generated a Botrytis cinerea knock‐out mutant in the bmp3 gene encoding a homologue of the yeast Slt2 cell wall integrity MAPK. The Δbmp3 mutant showed reduced vegetative growth on various media, strongly impaired conidiation and loss of sclerotia formation. Growth retardation of the mutant was enhanced in media with low osmolarity, whereas nearly wild‐type growth rates were observed under high osmolarity conditions. The Δbmp3 mutant did not show increased susceptibility to cell wall damage induced by glucanase, Calcofluor White or Nikkomycin Z, but was more susceptible to the oxidizing agent paraquat and the phenylpyrrole fungicide fludioxonil. Δbmp3 conidia showed normal germination and germ tube growth on agar, but excessive germ tube elongation on hard surfaces and reduced penetration efficiency, indicating a defect in surface sensing. After penetration, development of necrotic lesions induced by the Δbmp3 mutant was retarded. All these defects were restored by genetic complementation of the mutant with the wild‐type bmp3 gene.
The mixed xylem sap proteome of Fusarium oxysporum‐infected tomato plants
Authors: HOUTERMAN PETRA M. , SPEIJER DAVE , DEKKER HENK L. , DE KOSTER CHRIS G. , CORNELISSEN BEN J. C. , REP MARTIJN
Published: January 2007

Secreted proteins are known to play decisive roles in plant–fungus interactions. To study the molecular details of the interaction between the xylem‐colonizing, plant‐pathogenic fungus Fusarium oxysporum and tomato, the composition of the xylem sap proteome of infected tomato plants was investigated and compared with that of healthy plants. Two‐dimensional gel separation and mass spectrometry yielded peptide masses and peptide sequences of 33 different proteins. Despite the absence of complete genome sequences of either tomato or F. oxysporum, 21 proteins were identified as tomato proteins and seven as fungal proteins. Thirteen of the tomato proteins were specific for infected plants. Sixteen tomato proteins were found in xylem sap for the first time, four of which were identified based on matches to expressed sequences only. Coding sequences for new proteins from F. oxysporum were identified through either direct matching to a database sequence, matching of peptide sequences to genome or expressed sequence tag databases of other Fusarium species, or PCR with degenerate primers on cDNA derived from infected plants followed by screening of a F. oxysporum BAC library. Together, these findings provide an excellent basis for further exploration of the interaction between xylem‐colonizing pathogens and their hosts.
The implications of US experiences with early childhood interventions for the UK Sure Start Programme
Authors: Gray R. , Francis E.
Published: January 2007

Background  The UK Government introduced a large‐scale early childhood intervention programme, Sure Start, in 1999. Sure Start is to be further expanded, to achieve national coverage. US experience is highly relevant for anticipating challenges that this expansion will raise.

Methods  This is a focused, narrative review. We examine the impact, funding, quality‐improvement and programme objectives of Head Start and Early Head Start programmes.

Results  (1) Early childhood interventions can make a significant difference to children’s life chances; (2) expansion without adequate funding threatens quality; (3) narrower objectives, which are easier to measure, can crowd out broader objectives, which are difficult to measure; (4) programmes must balance fidelity to the model and flexibility to local conditions; (5) multiple objectives may conflict; and (6) programmes may have differential impacts. We consider the implications of these findings for Sure Start, focusing on funding, quality control and parental involvement. We also consider that the potential Sure Start should offer for tackling health inequalities in early childhood and suggest ways in which this aspect of the Programme could be enhanced.

Conclusion  Head Start has been dogged by concerns about quality and effectiveness. Many of these problems stemmed from an over‐hasty expansion, which locked the Programme into inadequate funding and uneven project‐ and staff‐quality. These issues have been addressed through large funding increases and more rigorous performance measures. Nevertheless, concerns about the aims of the Programme and the extent of parental involvement in management remain. Current funding for Sure Start appears to be adequate, while systematic evaluation procedures have been built in from its inception. Concerns have been raised about the implications of expansion for funding, quality and for parental involvement in management of local programmes. US experience shows that these are centrally important issues and that, if they are not addressed early on, they can take many years to rectify.
A computer‐aided program for pattern‐matching of natural marks on the spotted raggedtooth shark Carcharias taurus
Authors: VAN TIENHOVEN A. M. , DEN HARTOG J. E. , REIJNS R. A. , PEDDEMORS V. M.
Published: January 2007

The ability to identify individual animals is a critical aid in wildlife and conservation studies requiring information on behaviour, distribution, habitat use, population and life‐history parameters. We present a computer‐aided photo‐identification technique that relies on natural marks to identify individuals of Carcharias taurus, a shark species that is critically endangered off the eastern Australian coast and considered globally vulnerable. The technique could potentially be applied to a range of species of similar form and bearing natural marks.

The use of natural marks for photo‐identification is a non‐invasive technique for identifying individual animals. As photo‐identification databases grow larger, and their implementation spans several years, the historically used visual‐matching processes lose accuracy and speed. A computerized pattern‐matching system that requires initial user interaction to select the key features aids researchers by considerably reducing the time needed for identification of individuals.

Our method uses a two‐dimensional affine transformation to compare two individuals in a commonly defined reference space. The methodology was developed using a database of 221 individually identifiable sharks that were photographically marked and rephotographed over 9 years, demonstrating both the efficacy of the technique and that the natural pigment marks of C. taurus are a reliable means of tracking individuals over several years.

Synthesis and applications. The identification of individual animals that are naturally marked with spots or similar patterns is achieved with an interactive pattern‐matching system that uses an affine transformation to compare selected points in a single‐user computer‐aided interface. Our technique has been used successfully on C. taurus and we believe the methodology can be applied to other species of a similar form that have natural marks or patterns. The identification of individuals allows accurate tracking of their movements and distribution, and contributes to better population estimates for improved wildlife management and conservation planning.
Coexistence of Gonadotrophin‐Releasing Hormone and Galanin: Immunohisto‐chemical and Functional Studies
Authors: Coen C. W. , Montagnese C. , Opacka‐Juffry J.
Published: January 2007

Our observation of galanin‐immunoreactive perikarya with morphological characteristics strikingly similar to those containing gorvadotrophin‐releasmg hormone (GnRH) led us to undertake double‐label immunohistochemical studies on the distribution of these peptides. A considerable proportion of GnRH‐containing perikarya in the rat preoptic area were found to be immunoreactive for galanin. In order to establish whether this coexistence contributes to the control of luteinizing hormone (LH) release, the two peptides have been perfused alone and in combination through anterior pituitary cell columns derived from female rats in various endocrine conditions. Galanin at 0.1 μ M had a small stimulatory effect on LH release from cells obtained on the day of pro‐oestrus; this effect was less than that obtained with GnRH at 4 nM. Cells from oestrous rats showed no LH release in response to galanin. The studies undertaken thusfar have not indicated that galanin has a priming effect on the release of LH induced by subsequent treatment with galanin or a potentiating effect when administered together with GnRH. Delta sleep‐inducing peptide, another neuropeptide recently reported to be coexistent with GnRH, was also found to be capable of inducing a small discharge of LH from anterior pituitary cells obtained on the day of pro‐oestrus. Further studies will be required to establish the full consequences of the coexistence of these peptides and GnRH.
Computer‐Aided Mapping of Vasopressin Neurons in the Hypothalamus of the Male Golden Hamster: Evidence of Magnocellular Neurons that do not Project to the Neurohypophysis
Authors: Mahoney Paul D. , Koh Edward T. , Irvin Robert W. , Ferris Craig F.
Published: January 2007

Vasopressin‐sensitive neurons in the region of the anterior hypothalamus are necessary for the mediation of flank marking behavior in the Golden hamster. The precise nature of the vasopressinergic innervation to the anterior hypothalamus is unknown. In this study we seek to examine the potential sources of this innervation by mapping and counting the vasopressin‐immunoreactive neurons that contribute to the hypothalamo‐neurohypophysial system, and those that do not.

Vasopressin‐immunoreactive neurons in the hypothalamus were visualized by immunocytochemistry. Sections were mapped with a computer‐aided microscope system, and labeled neurons counted. Two‐dimensional maps were stacked into a three‐dimensional wireframe model which could be manipulated for further examination. The average number of vasopressin neurons was 3,135, with over 60% of all perikarya localized to the lateral supraoptic nucleus.

In a double‐labeling study, neurons contributing to the hypothalamo‐neurohypophysial system were retrogradely labeled by the injection of horseradish peroxidase into the neurohypophysis. The enzyme reaction product was visualized by treatment with tetramethylbenzidine followed by nickel‐conjugated diaminobenzidine. Sections were subsequently stained for vasopressin by immunocytochemistry. Single‐ and double‐stained neurons from serial sections were mapped and counted. Wireframe and contoured three‐dimensional representations were generated. The average number of neurons projecting to the neurohypophysis was 5,619. However, an average of 981 neurons was immunoreactive to vasopressin but devoid of horseradish peroxidase. The greatest number of these non‐projecting perikarya were found in and around the anterior hypothalamus, localized primarily in the lateral and medial aspect of the supraoptic nuclei, the ventral area of the paraventricular nucleus, and the nucleus circularis.

By comparing the number of non‐projecting neurons found by double‐staining to the total cell count of the entire vasopressin system, it was estimated that approximately 30% of all vasopressin neurons in and around the anterior hypothalamus did not project to the neurohypophysis. Based on the distribution and localization of the non‐projecting perikarya, it is speculated that these neurons may provide neurotransmitter for vasopressin‐dependent flank marking in the male Golden hamster.
Kainic Acid Lesion of Vasopressinergic Neurons in the Hypothalamus Disrupts Flank Marking Behavior in Golden Hamsters
Authors: Ferris Craig F. , Irvin Robert W. , Potegal Michael , Axelson John F.
Published: January 2007

It is well established that flank marking behavior in the Golden hamster is controlled by vasopressin‐sensitive neurons localized to the anterior hypothalamus; however, the source(s) of vasopressinergic innervation to this area is unknown. Previous analysis by immunocytochemistry showed distinct populations of vasopressinergic magnocellular neurons localized to the supraoptic nucleus, paraventricular nucleus and the nucleus circularis that did not project to the neurohypophysis. In the present study, these same hypothalamic nuclei were lesioned by microinjection of kainic acid to determine which, if any, of these populations of vasopressin neurons are involved in the control of flank marking. Unilateral lesions in the areas of the nucleus circularis and supraoptic nucleus at the rostro‐caudal plane of the anterior hypothalamus abolished odor‐induced flank marking behavior. Lesions in the paraventricular nucleus at the level of the anterior hypothalamus did not consistently inhibit flank marking, while lesions of magnocellular neurons rostral or caudal to the anterior hypothalamus were ineffective. The microinjection of vasopressin into the anterior hypothalamus following lesion of the paraventricular and supraoptic nuclei stimulated flank marking, evidence that treatment with kainic acid did not damage the efferent component of this behavior. However, animals with lesions in the nucleus circularis did not respond to the microinjection of vasopressin; hence, it is uncertain whether lesions in this area disrupt vasopressinergic innervation to the anterior hypothalamus or simply destroy the motor neurons controlling flank marking. In summary, the data clearly demonstrate that vasopressin neurons localized primarily to the medial aspect of the supraoptic nucleus are necessary for sensory integration of odor‐induced flank marking, and as such, may be one possible source of neurotransmitter controlling this behavior.
Effects of Estrogen and Ectopic Pituitary Transplants on the Responsiveness of Pituitary Prolactin Release to Luteinizing Hormone‐Releasing Hormone and Dopamine
Authors: Esquifino A. I. , Steger R. W. , Fernandez‐Ruiz J. , Bartkes A. , Amador A. G. , Chandrashekar V.
Published: January 2007

The responsiveness of prolactin release to regulatory inputs depends on the functional state of the lactotrophs. In the present study, we have examined the effects of luteinizing hormone‐releasing hormone (LHRH; 10–7 or 10–6 M) on the release of prolactin in vitro from hyperplastic pituitaries of estrogen‐treated male Fischer rats, ectopic pituitary transplants and in situ pituitaries of grafted and control rats. The effects of dopamine (10–8 or 10–7 M) in this system were also examined. The extent of inhibition of prolactin release by dopamine was not related to the amounts of prolactin secreted under basal conditions or to plasma prolactin levels. LHRH significantly suppressed prolactin secretion in all groups but its effect was most pronounced in the ectopic pituitary transplants and in the hyperplastic pituitaries of animals after chronic exposure to estrogen followed by a period of recovery. Thus, the effects of LHRH on prolactin release appear to be related to the secretory activity and/or to the absolute or relative number of the lactotrophs.
Alterations in Cyclic AMP Concentration and Adenylate Cyclase Activity in Specific Brain Areas of Rats with Inherited Hypothalamic Diabetes Insipidus (Brattleboro Rats)
Authors: Bahner Udo , Palkovits Miklós , Geiger Helmut , Schmid Gerhard , Heidland August
Published: January 2007

The concentration of cyclic AMP (cAMP) and the activity of sodium‐fluoride‐stimulated adenylate cyclase was measured in 29 microdissected brain areas of homozygous Brattleboro rats and their Long‐Evans control rats. In ten of the investigated brain areas a decreased cAMP level was measured in Brattleboro rats. It was particularly decreased in the supraoptic nucleus, cingulate and parietal cortex, hippocampus, habenula and organum vasculosum laminae terminalis. Significantly lower cAMP levels were also found in the periventricular nucleus, bed nucleus of the stria terminalis, area postrema and locus coeruleus. An increased cAMP concentration was detected only in the subcommissural organ of Brattleboro rats. In most brain areas, where cAMP was decreased, sodium fluoride‐stimulated adenylate cyclase activity was significantly increased (supraoptic nucleus, parietal cortex, periventricular nucleus, bed nucleus of the stria terminalis, locus coeruleus) or unchanged (hippocampus, habenula, organum vasculosum laminae terminalis). The coincidence of alterations in cAMP concentration and adenylate cyclase activity in brain areas of Brattleboro rats with relatively dense vasopressinergic innervation and/or vasopressin receptor population in control rats, suggests an influence of brain vasopressin on the cAMP‐adenylate cyclase second messenger system.
Circadian and Estral Changes in the Hypothalamic Prostaglandin E2 Content and [3H]Prostaglandin E2 Binding in Female Rats
Authors: Bommelaer‐Bayet M. C. , Wisner A. , Renard C. A. , Levi F. A. , Dray F.
Published: January 2007

Prostaglandin E2, (PGE2) is involved in the luteinizing hormone‐releasing hormone‐stimulated luteinizing hormone surge in female rats and may act via specific membrane receptors. The following studies were performed to determine whether there were any changes in the hypothalamic PGE2 binding and/or PGE2 content which were specific to proestrus and not to the rest of the estrous cycle. Groups of female Wistar rats were sacrificed at 3‐h intervals throughout the estrous cycle to determine both the circadian and circaestral changes in the hypothalamic PGE2 content and [3H]PGE2 binding. The hypothalamic PGE2 content was maximal at 1700 h on each of the 4 consecutive days of the estrous cycle but was independent of the stage of the cycle. [3H]PGE2 binding also displayed a circadian rhythm; the lowest binding occurred near the circadian peak of PGE2, suggesting that the PGE2 binding sites were occupied by endogenous PGE2. Since such circadian rhythms were not observed in the hypothalamus of male rats, they may be under the control of ovarian steroids. Also, since PGE2 binding and the PGE2 content both exhibit a diurnal pattern independent of the day of the cycle, there may be changes in the PGE2 receptor‐mediated process coupled to an adenylyl cyclase which could explain the luteinizing hormone surge in proestrus.
Differential Localization of Estrogen Receptors in Various Vasopressin Synthesizing Nuclei of the Rat Brain
Authors: Axelson John F. , van Leeuwen Fred W.
Published: January 2007

Vasopressin (VP) cells in the bed nucleus of the stria terminalis, medial amygdaloid nucleus and supraoptic and paraventricular nuclei are influenced by gonadal steroids. The present paper examined whether VP cells in the bed nucleus of the stria terminalis, medial amygdaloid nucleus, and supraoptic and paraventricular nuclei contain estrogen receptors. Brains from adult short‐term castrated, colchicine‐treated male rats were fixed with 4% paraformaldehyde and 0.5% glutaraldehyde. In the immunocytochemical double‐staining procedure Vibratome sections were first incubated with an estrogen receptor antibody (#H222) and stained with diaminobenzidine‐Ni+. Following methanol‐hydrogen peroxide washes, sections were incubated with anti‐neurophysin and stained with diaminobenzidine. Parvocellular cells in the bed nucleus of the stria terminalis and medial amygdaloid nucleus were double‐stained with a blue‐black nucleus (indicating the estrogen receptors) surrounded by brown cytoplasm (resulting from VP‐neurophysin‐immunoreactivity). Our results provide the first direct anatomical evidence supporting the hypothesis that gonadal steroids' influence of parvocellular VP cells in the bed nucleus of the stria terminalis and medial amygdaloid nucleus is mediated directly via estrogen receptors localized in nuclei of VP neurons. We were unable to co‐localize any estrogen receptors in VP and oxytocin cells of magnocellular size in the supraoptic, paraventricular and anterior commissural nuclei, suggesting that estrogen indirectly affects these magnocellular hypothalamic cells.
Quantification of S100A12 (EN‐RAGE) in Blood Varies with Sampling Method, Calcium and Heparin
Authors: Larsen A. , Bronstein I. B. , Dahl O. , Wentzel‐Larsen T. , Kristoffersen E. K. , Fagerhol M. K.
Published: January 2007

S100A12 is a calcium‐binding protein predominantly found in neutrophil granulocytes and monocytes. Its usefulness in monitoring inflammatory disease states depends on documentation that assay results are reliable. This study aimed at defining guidelines for blood sampling, selection of optimal material handling and reference intervals in healthy controls while taking into account the basic features of S100A12. An enzyme linked immunosorbent assay was developed based upon antibodies induced in rabbits by injection of recombinant S100A12. Our studies confirm that oligomers of S100A12 are generated in the presence of calcium. Structural changes in S100A12 mediated by calcium influence the interaction with antibody. This is proposed as the background for our very low readings of S100A12 in Ethylene Diamine Tetraacetic Acid (EDTA) plasma. Individual S100A12 levels did not change substantially over a 5‐week sampling period. Based upon testing of 150 blood donors we suggest reference intervals of S100A12 in serum to be 49–1340 μg/l for women and 27–1750 μg/l for men. The estimated mean concentrations were 234 μg/l in serum samples (range 12–15791), 114 μg/l (range 3–17282) in re‐calcified EDTA plasma and 48 μg/l (range 2–14843) in heparin plasma. Without adding calcium to EDTA plasma before running the assay, concentrations were around 2 μg/l (16 persons). S100A12 quantification is assumed to become relevant for diagnostic use in many disease states. The importance of the handling and analysing conditions for a reliable result was examined. We recommend serum collected in gel‐containing tubes as the preferred sample material and have suggested reference intervals for healthy individuals.
CD25‐expressing B‐lymphocytes in Rheumatic Diseases
Authors: Amu S. , Strömberg K. , Bokarewa M. , Tarkowski A. , Brisslert M.
Published: January 2007

B cells play an important role in the development of autoimmune diseases due to their production of autoantibodies, antigen‐presenting capacity and production of pro‐inflammatory cytokines. The purpose of the present study was to analyse B cells from rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) patients, with respect to their expression of the IL‐2 receptor (IL‐2R) subunit CD25. Using flow cytometry, we found that CD25+ B cells from RA patients expressed significantly higher frequencies of CD122 and CD132 than CD25+ B cells from control subjects, indicating a fully functional IL‐2R. These CD25+ B cells also expressed higher frequencies of the co‐stimulatory molecule CD80, whereas IgM and IgA expression was decreased compared with CD25+ B cells from healthy controls. In addition B cells from SLE patients co‐expressed CD25 together with CD80, CD122, and CD132, but to a lower degree IgD and IgM, when compared with healthy controls. Taken together, our results indicate that CD25+ B cells from RA and SLE patients are in a highly activated state, display a more mature phenotype and suggest that this B cell subset may be involved in the pathogenesis of RA and SLE.
The Major Histocompatibility Complex Haplotypes Dictate and the Background Genes Fine‐tune the Dominant versus the Cryptic Response Profile of a T‐cell Determinant within a Native Antigen: Relevance to Disease Susceptibility and Vaccination
Authors: Sinha P. , Snyder J. A. , Kim E. Y. , Moudgil K. D.
Published: January 2007

The immune system of a healthy individual responds vigorously to foreign microbial antigens. However, all potentially immunogenic regions (determinants) within an antigen are not functionally of equal relevance in mediating host immunity against the pathogen. Moreover, some of these antigenic determinants are well processed and presented (immunodominant), while others are not revealed (cryptic) from the native antigen. Nevertheless, cryptic determinants are good immunogens in the pre‐processed peptide form. Defining the factors influencing the dominance versus the crypticity of antigenic determinants is critical to advancing our understanding of the individual variations in host immunity to infection, autoantigens and vaccination. In this study based on a model antigen, hen eggwhite lysozyme (HEL), we describe that the major histocompatibility complex (MHC) haplotypes imprint and the non‐MHC genes modify the dominance versus the crypticity of a specific antigenic determinant. Both the H‐2q‐ and the H‐2d‐bearing mice raised potent response to native HEL, but responded differently to its determinant region 57–78, which was dominant in the H‐2q but cryptic in the H‐2d mice. The H‐2q‐ but not the H‐2d‐bearing mice of three different genetic backgrounds yielded patterns of graded reactivity to epitope 57–78 showing the fine‐tuning effect of the non‐MHC genes. Interestingly, the F1 (H‐2q × H‐2d) mice retained the dominant response profile of the H‐2q parent regardless of the contributing gender, and also responded to a new sub‐determinant 61–75. These results highlight the genetic factors influencing the dominance/crypticity of a specific antigenic determinant.
The PEP‐carboxylase kinase gene family in Glycine max (GmPpcK1–4): an in‐depth molecular analysis with nodulated, non‐transgenic and transgenic plants
Authors: Xu Wenxin , Sato Shirley J. , Clemente Thomas E. , Chollet Raymond
Published: January 2007

Phosphoenolpyruvate carboxylase (PEPC) is a widely distributed metabolic enzyme among plant and prokaryotic species. In vascular plants, the typical PEPC is regulated post‐translationally by a complex interplay between opposing metabolite effectors and reversible protein phosphorylation. This phosphorylation event is controlled primarily by the up‐/down‐regulation of PEPC‐kinase (PpcK), an approximately 31‐kDa Ser/Thr‐kinase. As a sequel to earlier investigations related to PEPC phosphorylation in N2‐fixing nodules of Glycine max, we now present a detailed molecular analysis of the PpcK multigene family in nodulated soybeans. Although the GmPpcK1–4 transcripts are all expressed throughout nodule development, only the nearly identical GmPpcK2/3 homologs are nodule‐enhanced and up‐/down‐regulated in vivo by photosynthate supply from the shoots. In contrast, GmPpcK1 is a ‘housekeeping’ gene, and GmPpcK4 is a highly divergent member, distantly removed from the legume PpcK subfamily. Real‐time qRT–PCR analysis indicates that GmPpcK2/3 are overwhelmingly the dominant PpcKs expressed and up‐/down‐regulated throughout nodule development, mirroring the expression properties of nodule‐enhanced PEPC (GmPpc7). In situ RT–PCR investigation of the spatial localization of the GmPpcK1–4 and GmPpc7 transcripts in mature nodules is entirely consistent with this view. Complementary histochemical and related RNA gel‐blot findings with nodulated, GmPpcK1/3 promoter::GUS‐expressing T2 plants provide direct experimental evidence that (i) PpcK gene expression is controlled primarily at the transcriptional level; and (ii) the contrasting expression properties of GmPpcK1/3 are conferred largely by regulatory element(s) within the approximately 1.4‐kb 5′‐upstream region. As a result of our multifaceted analyses of GmPpcK1–4, GmPpc7 and PEPC‐phosphorylation in the soybean nodule, it is proposed that the GmPpcK2/3 homologs and GmPpc7 together comprise the key molecular ‘downstream players’ in this regulatory phosphorylation system within the mature nodule's central zone.
Spontaneous mutations caused by a Helitron transposon, Hel‐It1, in morning glory, Ipomoea tricolor
Authors: Choi Jeong‐Doo , Hoshino Atsushi , Park Kyeung‐Il , Park In‐Sook , Iida Shigeru
Published: January 2007

Helitrons are newcomers among eukaryotic DNA transposons and have originally been identified by computational analysis in the genomes of Arabidopsis, rice and nematode. They are distinguished from other transposons in their structural features, and their proposed transposition mechanisms are involved in rolling circle replication. Computer‐predicted autonomous Helitrons with conserved terminal sequences 5′‐TC and CTRR‐3′ are presumed to encode a putative transposase, Rep/Hel‐TPase, which contains a characteristic nuclease/ligase domain for the replication‐initiation protein (Rep) and a DNA helicase domain (Hel). Plant Helitrons are thought to encode an additional transposase, RPA‐TPase, which is related to the largest subunit of the replication protein A (RPA70). Although Helitrons are found in diverse genomes, neither an autonomous element nor a transposition event has been reported. Here we show that a spontaneous pearly‐s mutant of Ipomoea tricolor cv. Pearly Gates, exhibiting white flowers and isolated in approximately 1940, has an 11.5‐kbp novel Helitron, named Hel‐It1, integrated into the DFR‐B gene for anthocyanin pigmentation. Hel‐It1 shows the predicted plant Helitron structure for an autonomous element with the conserved termini and carrying the two putative transposase genes, Rep/Hel‐TPase and RPA‐TPase, which contain a nonsense and a frameshift mutation, respectively. Hel‐It1‐related elements are scattered in the Ipomoea genome, and only a fraction of the pearly‐s plants were found to carry Hel‐It1 at another insertion site. The pearly‐s mutant appears to bear an autonomous element and to express the wild‐type RPA‐TPase transcripts. The structures of a putative autonomous element and its transposase genes are discussed.
Analysis of the sucrose synthase gene family in Arabidopsis
Authors: Bieniawska Zuzanna , Paul Barratt D. H. , Garlick Andrew P. , Thole Vera , Kruger Nicholas J. , Martin Cathie , Zrenner Rita , Smith Alison M.
Published: January 2007

The properties and expression patterns of the six isoforms of sucrose synthase in Arabidopsis are described, and their functions are explored through analysis of T‐DNA insertion mutants. The isoforms have generally similar kinetic properties. Although there is variation in sensitivity to substrate inhibition by fructose this is unlikely to be of major physiological significance. No two isoforms have the same spatial and temporal expression patterns. Some are highly expressed in specific locations, whereas others are more generally expressed. More than one isoform is expressed in all organs examined. Mutant plants lacking individual isoforms have no obvious growth phenotypes, and are not significantly different from wild‐type plants in starch, sugar and cellulose content, seed weight or seed composition under the growth conditions employed. Double mutants lacking the pairs of similar isoforms sus2 and sus3, and sus5 and sus6, are also not significantly different in these respects from wild‐type plants. These results are surprising in the light of the marked phenotypes observed when individual isoforms are eliminated in crop plants including pea, maize, potato and cotton. A sus1/sus4 double mutant grows normally in well‐aerated conditions, but shows marked growth retardation and accumulation of sugars when roots are subjected to hypoxia. The sucrose synthase activity in roots of this mutant is 3% or less of wild‐type activity. Thus under well‐aerated conditions sucrose mobilization in the root can proceed almost entirely via invertases without obvious detriment to the plant, but under hypoxia there is a specific requirement for sucrose synthase activity.
Cuticular defects lead to full immunity to a major plant pathogen
Authors: Chassot Céline , Nawrath Christiane , Métraux Jean‐Pierre
Published: January 2007

In addition to its role as a barrier, the cuticle is also a source of signals perceived by invading fungi. Cuticular breakdown products have been shown previously to be potent inducers of cutinase or developmental processes in fungal pathogens. Here the question was addressed as to whether plants themselves can perceive modifications of the cuticle. This was studied using Arabidopsis thaliana plants with altered cuticular structure. The expression of a cell wall‐targeted fungal cutinase in A. thaliana was found to provide total immunity to Botrytis cinerea. The response observed in such cutinase‐expressing plants is independent of signal transduction pathways involving salicylic acid, ethylene or jasmonic acid. It is accompanied by the release of a fungitoxic activity and increased expression of members of the lipid transfer protein, peroxidase and protein inhibitor gene families that provide resistance when overexpressed in wild‐type plants. The same experiments were made in the bodyguard (bdg) mutant of A. thaliana. This mutant exhibits cuticular defects and remained free of symptoms after inoculation with B. cinerea. The expression of resistance was accompanied by the release of a fungitoxic activity and increased expression of the same genes as observed in cutinase‐expressing plants. Structural defects of the cuticle can thus be converted into an effective multi‐factorial defence, and reveal a hitherto hidden aspect of the innate immune response of plants.
Change in pancreatic B‐cell function (HOMA‐B) varies in different populations with similar genetic backgrounds but different environments
Authors: Heald A. H. , Anderson S. G. , Patel J. , Rudenski A. , Vyas A. , Yates A. , Hughes E. , Prabharakan D. , Reddy S. , Durrington P. , Gibson J. M. , Bhatnagar D. , Cruickshank J. K. , Laing I.
Published: January 2007

Objective  To determine whether pancreatic B‐cell function varies in different populations with similar genetic backgrounds but different environments.

Research design/methods  We compared a specific migrant Gujarati community in the UK (n = 205) with people still resident in the same villages of origin in Gujarat, India (n = 246). Pancreatic B‐cell function (HOMA‐B) was determined and the influence of age, migration and other factors was explored.

Results  As anticipated, there was an age‐related decline in log(HOMA‐B) in both groups. However, the age‐related fall in log(HOMA‐B) was more pronounced in the UK than in Gujarat (normalized β−0.29 vs. −0.14, P for difference = 0.03). The decline of HOMA‐B with age persisted after adjustment for body mass index (UK β = −0.31; Gujarat β = −0.16, P = 0.015, P < 0.001). There was no significant change in insulin sensitivity (HOMA‐S) with age at either site, although insulin sensitivity was lower in the UK. Fasting non‐estrified fatty acid (NEFA) levels rose with age in the UK but not in Gujarat (P = 0.003 for difference in gradients). In multiple linear regression analysis, lower log(HOMA‐B) was independently associated with higher fasting log(NEFA) levels; normalized β = −0.24, P < 0.001, age; β = −0.16, P = 0.005, higher log(insulin‐like growth factor binding protein‐1); β = −0.19, P = 0.007 and lower body mass index; β = 0.26, P = 0.001. This model accounted for 25% of the variability in HOMA‐B.

Conclusions  HOMA‐B as a measure of B‐cell function declines more rapidly with age in the migrant UK group than in Gujarat. This may be a direct consequence of chronically higher NEFA exposure in the UK group.
Prevalence of autoimmune diseases in islet transplant candidates with severe hypoglycaemia and glycaemic lability: previously undiagnosed coeliac and autoimmune thyroid disease is identified by screening
Authors: Walter M. , McDonald C. G. , Paty B. W. , Shapiro A. M. J. , Ryan E. A. , Senior P. A.
Published: January 2007

Aims  Autoimmune diseases such as Addison's or coeliac disease can contribute to hypoglycaemia or malabsorption and are more common in Type 1 diabetes (T1DM). This brief report describes the prevalence of known and newly detected autoimmune disease in clinical islet transplant candidates with longstanding T1DM and severe hypoglycaemia and/or glycaemic lability who are routinely screened for coexisting autoimmune disease.

Methods  One hundred and twenty‐four C‐peptide negative T1DM subjects [77 (62%) female, mean age 44 ± 9 years, diabetes duration 28 ± 11 years, body mass index 24.9 ± 3.5 kg/m2] with indications for clinical islet transplantation at the University of Alberta were screened for autoimmune disease by history and measurement of anti‐transglutaminase antibodies (positive > 10 U/ml), 09.00 h cortisol (followed by adrenocorticotrophic hormone‐stimulation if < 495 nmol/l) and thyroid‐stimulating hormone to determine the prevalence of coeliac disease, Addison's disease and autoimmune thyroid disease, respectively.

Results  Forty per cent of subjects had one or more coexisting autoimmune disease. The prevalence of autoimmune disease was 35%, coeliac disease 8% and Addison's disease 1.6%. In 11 individuals (9%), one or more autoimmune disease were newly detected (seven coeliac disease and five thyroid disease). Seven of 10 cases of coeliac disease were newly detected. A gluten‐free diet in individuals with newly diagnosed coeliac disease reduced gastrointestinal symptoms, but indications for clinical islet cell transplantation persisted.

Conclusions  Coexisting autoimmune disease is common in candidates for clinical islet cell transplantation. Screening in this group identified a substantial number of previously unrecognized cases. Clinicians should consider the presence of autoimmune disease even in the absence of classical symptoms.
Prognostic role of B‐type natriuretic peptide in patients with diabetes and acute decompensated heart failure
Authors: Aspromonte N. , Feola M. , Milli M. , Scardovi A. B. , Coletta C. , Carbonieri E. , Giovinazzo P. , Di Giacomo T. , Barro S. , Rosso G. L. , Ceci V. , Milani L. , Valle R.
Published: January 2007

Background  Several studies have reported the prognostic value of natriuretic peptides, but their predictive value in patients with diabetes mellitus is unknown. The aim of the study was to test the hypothesis that measurement of brain natriuretic peptide (BNP) levels in ambulatory patients with congestive heart failure (CHF) and diabetes can predict the occurrence of cardiovascular events at 6‐month follow‐up.

Methods  We enrolled 145 consecutive patient with diabetes [age 72 ± 9 years, hypertension (21%), ischaemic heart disease (52%), atrial fibrillation (22%), preserved left ventricular function (29%)] seen in the outpatient heart failure clinic after an acute episode of cardiac failure.

Results  The median (25th/75th interquartile range) BNP concentrations at discharge were 186 (75–348) pg/ml. At 6‐month clinical follow‐up 10/145 (7%) subjects had died and 31/145 (21%) had been readmitted because of cardiac decompensation. BNP values of 200 and 500 pg/ml were found to have the best compromise between sensitivity (88 and 46%, respectively) and specificity (71 and 89%, respectively) for predicting events at 6 months. Multivariate Cox regression analysis identified only two parameters as predictors of events: serum creatinine [hazard ratio (HR) = 3.3; P = 0.02], and BNP plasma level BNP cut‐off values (HR = 3.8; P = 0.03 for 201–499 pg/ml and HR = 7.7; P = 0.001 for ≥ 500 pg/ml).

Conclusion  These results suggest that BNP and serum creatinine are strong predictors of clinical events in patients with diabetes and CHF. In these patients, clinical outcome might be stratified by plasma BNP levels.
Mechanisms of protective effects induced by blockade of the renin–angiotensin system: novel role of the pancreatic islet angiotensin‐generating system in Type 2 diabetes
Authors: Leung P. S.
Published: January 2007

Large clinical trials have shown that inhibition of the renin–angiotensin system (RAS) can delay and/or prevent the onset of Type 2 diabetes mellitus (T2DM) in high‐risk individuals, such as those with hypertension or chronic heart failure. Moreover, a meta‐analysis of these randomized clinical studies concluded that the mean weighted relative risk of development of T2DM was reduced by 25% in those patients treated with angiotensin II receptor blockers or angiotensin‐converting enzyme inhibitors. In spite of these firm clinical data, the mechanistic pathways mediating the protective activity of RAS blockade have yet to be resolved. Of particular interest is the recently identified local pancreatic RAS and, perhaps more importantly, the finding that it is up‐regulated in animal models of T2DM. This putative local RAS may regulate pancreatic islet blood flow, oxygen tension, and islet (pro)insulin biosynthesis. It might also mediate the generation of reactive oxygen species, thereby causing oxidative stress‐induced pancreatic β‐cell apoptosis and fibrosis. Moreover, findings that RAS blockade improved β‐cell secretory function and cell mass in experimental animal models of Type 2 diabetes indicate that inhibition of RAS activation may play a pivotal role in protecting islet cell function, and furthermore may prevent the development of overt T2DM. Such data supporting the involvement of the local pancreatic RAS in islet function, as well as a causal relationship between RAS activation and T2DM, and RAS induced β‐cell dysfunction, mandate further investigation into the role of RAS in the pathogenesis of the progressive islet impairment observed in patients with T2DM.