bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytokine profile of Plasmodium falciparum‐specific T cells in non‐immune malaria patients
Authors: Ramharter Michael , Willheim Martin , Winkler Heidi , Wahl Katharina , Lagler Heimo , Graninger Wolfgang , Winkler Stefan
Published: August 2003

CD3+ T cells are important sources of both pro‐ and anti‐inflammatory cytokines during Plasmodium falciparum malaria. We studied the frequency of interleukin‐2 (IL‐2), gamma interferon (IFN‐γ), tumour necrosis factor‐alpha (TNF‐α) and IL‐10 expressing CD3+ cells in 10 non‐immune malaria patients with uncomplicated malaria and in one patient with cerebral malaria after P. falciparum‐specific and non‐specific mitogenic stimulation. Analysis by fluorescence‐activated cell sorting was performed after drug‐induced clearance of parasites to allow previously sequestered T cells to be detected in peripheral blood. CD3+ cells of patients responded to P. falciparum infected erythrocytes with significant increases in the percentage of IL‐2, IFN‐γ, and TNF‐α, but also IL‐10, positive cells. CD3+ cells from malaria‐naïve donors were also responsive to specific stimulation albeit to a much lesser extent. Mitogenic stimulation of PBMC revealed no significant differences between cells of patients and controls. CD3+ cells of the patient with cerebral malaria were hyporesponsive both to the infecting parasite isolate as well as to our laboratory‐adapted P. falciparum isolate, whereas two patients with uncomplicated disease were more responsive to their infecting parasites than to the laboratory‐adapted isolate. The results indicate that the increased responsiveness of in vivo primed compared to malaria‐naïve CD3+ cells is Plasmodium‐specific and biased towards production of IFN‐γ and TNF‐α.
Evolving the multiple roles of ‘patients’ in health‐care research: reflections after involvement in a trial of shared decision‐making
Authors: Thornton Hazel , Edwards Adrian , Elwyn Glyn
Published: August 2003

Objective This paper offers ‘consumer‐led’ reflections by steering group members of a patient‐centred research study involving consumer advocates, patients’ associations and patients, throughout the whole study, from pre‐ to post‐study phases.

Original Study Design The study: ‘Shared decision making and risk communication in general practice’ incorporated systematic reviews, psychometric evaluation of outcome measures, and quantitative, qualitative and health economic analyses of a cluster randomized trial of professional skill development, all informed by consumer and patient engagement.

Setting and participants The work was produced by a wide collaboration led by researchers from the Department of General Practice, University of Wales College of Medicine, Cardiff, including a consumers’ advisory group and a patients’ association. The study participants were 20 general practitioners from Gwent, their practice staff, and almost 800 patients at these practices.

Discussion Consumers and patients contributed to several stages of the research from inception and design, securing of funding, implementation of the protocol, and interpretation and dissemination of the findings. ‘Patient involvement’ research initiatives that include an equally wide variety of ‘user’ participants as ‘health‐professional’ participants, accountable to a ‘Health in Partnership’ funded project, require a user‐led viewpoint to be presented and disseminated. This paper presents reflections on the processes of the research, the interpretations of study findings by the involved parties, and notes how this model is fundamental to effective research in the field of patient‐centred health care if future practice, policy and research are to change.
Exploring doctor and patient views about risk communication and shared decision‐making in the consultation
Authors: Davis Ruth E , Dolan Gina , Thomas Sue , Atwell Christine , Mead Donna , Nehammer Sarah , Moseley Laurie , Edwards Adrian , Elwyn Glyn
Published: August 2003

Background  There have been significant conceptual developments regarding shared decision‐making (SDM) and assessments of people's hypothetical preferences for involvement in treatment or care decisions. There are few data on the perceptions of patients and professionals about SDM in actual practice.

Objective To explore, from paired doctor–patient interviews, participants’ perceptions of SDM in the consultation and the level of consensus between the participants in the consultation process.

Design Qualitative analysis of semi‐structured interview data.

Setting and participants Twenty general practitioners received training packages in ‘risk communication’ (RC) and ‘SDM’ to use as tools within the consultation. Forty patients with one of four conditions, for which a range of treatment options is available, were selected. Patient/doctor pairs were interviewed separately following consultations at four stages –‘baseline’ [general practitioner's (GP) usual consultation style], SDM training, RC alone, and both RC and SDM training. Interviews were transcribed and analysed using NVivo software.

Results Risk communication interventions by doctors appeared to result in a greater perception of decisions being made in the consultation. High levels of satisfaction with consultations were evident before application of the interventions and did not change after the interventions. Doctors’ and patients’ perceptions of the consultations were highly congruent at all phases of the study.

Conclusion Shared decision‐making and RC approaches were helpful in selected consultations and showed no detrimental effects to patients. However, the use of RC and SDM made only small differences to decision‐making in consultations within the population studied. Increasing patient participation may be seen as more ethically justifiable than the traditional paternalistic approach but this needs to be set against the additional training costs incurred.
Clinical issues in shared decision‐making applied to breast cancer
Authors: Moumjid Nora , Carrère Marie‐Odile , Charavel Marie , Brémond Alain
Published: August 2003

Objectives To assess (1) the clinical issues addressed during the medical encounter; (2) the feasibility of the process of shared decision‐making in clinical practice and (3) patients’ desires concerning the question of ‘who should take the decision in breast cancer treatments?’

Design Qualitative pilot study based on clinical encounters using decision boards and information booklets.

Setting Centre Léon Bérard, a comprehensive cancer centre in the Rhône‐Alpes region of France.

Participants One breast cancer surgeon and 22 breast cancer patients.

Main outcome measures Analysis of patients’ reactions to a shared decision‐making process concerning surgery and chemotherapy, and analysis of its practical feasibility (i.e. duration of the consultations).

Results (1) Twenty‐one patients participated in the decision regarding surgery; all chose conservative treatment; 15 patients had their own say about chemotherapy (nine chose no chemotherapy, six chose chemotherapy). (2) Participating in treatment choice generated anxiety for a majority of patients. Some were dissatisfied because they had to make a choice and consequently to give up the other option. Finally, some were uncertain about making the right choice. Nevertheless, most were satisfied with the information given and the possibility of participating to the treatment decision‐making process. (3) The total duration of the entire process of shared decision‐making is consistent with the time spent with patients with such a severe disease.

Discussion/conclusion Most of the patients expressed their satisfaction regarding the possibility to participate in treatment decision‐making, knowing that offering treatment choice is very unusual in France. From this pilot study it appears that shared decision‐making is feasible in clinical practice. Nevertheless, a quantitative study based on a large sample of patients is necessary to: (1) confirm this hypothesis, (2) ensure that patients are willing to participate in their treatment decision‐making, and (3) measure the potential benefits related to this participation.
How is medical decision‐making shared? The case of haemophilia patients and doctors: the aftermath of the infected blood affair in France
Authors: Fillion Emmanuelle
Published: August 2003

Objective  This article looks at how users and doctors in France have rethought the question of shared decision‐making in the clinical field of haemophilia following a major crisis – that of the infected blood affair.

Design  We did a qualitative survey based on semi‐structured interviews in three regions of France.

Setting and participants  The interviews covered 31 clinical doctors of haemophilia and 31 users: 21 adult males with severe haemophilia (21/31), infected (14/21) or not (7/21) with HIV, the infected wife of one of the latter (1/31) and nine parents of young patients with severe haemophilia (9/31), either HIV positive (6/9) or negative (3/9).

Results and conclusions  The results show the infected blood affair to be a major individual and collective ordeal. It has caused users and doctors to rethink their roles within clinical relationships and to develop new ways of sharing medical decision‐making. Prior to the crisis, the dominant model was based upon a distinction between the medical aspect, governed by the doctors, and the psychosocial aspect, which involved the patients and their families. Since the crisis, medicoscientific knowledge has been shared between users and doctors. This general trend nevertheless permits the existence of different patient, family and doctor profiles which in turn correspond to different notions of what a clinical decision should be. Some users remain attached to the idea of complementarity between doctors and patients (new partnership model), whilst others put doctors and patients on an equal footing (negotiation model). On the doctors’ side, whilst some still prefer the initial model for therapeutic decision‐making, the majority have reassessed their perceptions and viewpoints. A certain number believe that decisions should be made by both doctor and patient in accordance with scientific procedures (decision‐making controlled by scientific standards) or regulatory procedures (decision‐making controlled by legal standards). Yet others feel that multiple points of view are acceptable within the decision‐making process (decision‐making model as interactivity).
The problematic of decision‐sharing: deconstructing ‘cholesterol’ in a clinical encounter
Authors: Gwyn Richard , Elwyn Glyn , Edwards Adrian , Mooney Annabelle
Published: August 2003

Shared decision‐making is increasingly advocated as a means of interacting with patients but there is also a widely accepted view that many factors will militate against this ideal. While some patients may not wish to take on the responsibility of decision‐making, it is also evident that many find it difficult to assimilate probabilities about future events and overestimate the likelihood of some outcomes, especially when terms such as ‘stroke’, ‘bleeding’ and ‘heart attack’ are used in consultation and bring with them emotional connotations and reactions. Under such circumstances, should clinicians portray risks as best they can, in the hope that even a marginally improved understanding will be an improvement on unilateral professional decision‐making? Or, conversely, should they ‘guide’ the decision process, acting in a way that is known as ‘professional agency'? Developing some perspectives put forward in recent work by the authors and applying it to a distinct clinical context, this paper will provide (i) a discourse analytic exploration of a single extended example from clinical practice employing aspects of Bakhtin's theory of dialogism, and (ii) a discussion and summary of what we can learn from this analysis in the context of shared decision‐making and risk communication.
Effects of Thermotherapy Treatments on Systemic Agrobacterium vitis in Dormant Grape Cuttings
Authors: Mahmoodzadeh H. , Nazemieh A. , Majidi I. , Paygami I. , Khalighi A.
Published: August 2003

Agrobacterium tumefaciens biovar 3 (A. vitis) was eradicated or reduced below the level of detection in dormant cuttings of grape [Vitis vinifera cultivar (cv.) Thompson seedless] and rootstock NAZ3 (V. vinifera × V. rupestris) by hot‐water treatment (exposure to 50°C for 30 min). Strains of A. vitis varied in their sensitivity to heat, but were generally more sensitive than strains of A. tumefaciens biovar 1 or biovar 2. Populations of about 103 CFU/ml in broth were killed by a 30‐min treatment. Biovar 1 strains were apparently unaffected by 50°C, even when exposed for 30 min. Non‐tumorigenic biovar 1 strains were recovered from hot‐water‐treated cuttings. An assessment was also made of the effect of treatment on growth parameters of cuttings in a field nursery after 9 months. The effect of hot‐water treatment on the vitality and growth of vines varied with different cultivars or rootstocks. The number and length of canes, root dry weight and length and diameter of trunks, increased in most instances. Treatment and time usually did affect bud survival and, in most cases, increased the level of callus formation at the base of the cuttings. Hot‐water treatment may offer a simple, effective, economical and environmentally safe means of eradicating A. vitis from dormant grape cuttings.
Inhibited Long‐Distance Movement of Potato Leafroll Virus to Tubers in Potato Genotypes Expressing Combined Resistance to Infection, Virus Multiplication and Accumulation
Authors: Syller J.
Published: August 2003

Plants of two potato clones which, in preliminary greenhouse assessments, showed resistance to multiplication and accumulation of potato leafroll virus (PLRV) were graft or aphid inoculated with the virus and grown in the greenhouse; plants of a moderately susceptible cultivar were used for comparison in all experiments. A high concentration of aphid‐borne inoculum was used to ensure strong infection pressure. Clone M62759 appeared to be highly resistant to PLRV infection, whereas clone PS1706 was more susceptible. Both clones expressed a high level of resistance to virus multiplication, when primary or secondary infection was assayed by enzyme‐linked immunosorbent assay. Moreover, PLRV was detected in only few or none of the progeny plants of clone M62759, which thus strongly inhibited virus transport to tubers. The study on PLRV translocation from aphid‐inoculated shoots to uninoculated shoots sprouted from the same tubers showed that no specific mechanisms are likely to impair PLRV movement through the tubers of the resistant genotypes. These results indicate that three valuable components of the resistance to PLRV are probably closely linked in the genotype, a combination that seems to occur rather rarely in potato clones. Nevertheless, selecting potato genotypes for the complex resistance to PLRV may prove to be a worthwhile part of breeding programmes, provided that the genetic mechanisms governing particular types of resistance are better recognized.
Interaction of Xylella fastidiosa with Different Cultivars of Nicotiana tabacum: a Comparison of Colonization Patterns
Authors: Alves E. , Kitajima E. W. , Leite B.
Published: August 2003

The number of colony forming units (CFU) inside xylem vessels of leaf petioles were evaluated in tobacco by scanning electron microscopy followed by an image analysis. Symptom expression of Nicotiana tabacum, cultivars TNN, Havana and RP1, were correlated with the presence of bacteria inside leaf petiole vessels. The symptom expression were evaluated in terms of colony forming units per gram of colonized tissue and percentage of colonized vessels. No statistical differences were found among the varieties tested. Havana expressed the most intense symptoms, best indicating a better experimental host. We also observed that leaf symptoms could be reversed with the application of ammonium sulphate along with pruning. This routine was effective in delaying symptom development. In summary, colonization efficiency was similar in tobacco varieties. Fertilization may affect Xylella fastidiosa symptom expression and pruning may be used as an aid to diminish X. fastidiosa advance within the plant.
In vitro and In vivo Effects of Some Fungicides against the Chickpea Blight Pathogen, Ascochyta rabiei
Authors: Demirci F. , Bayraktar H. , Babalioǧullu I. , Dolar F. S. , Maden S.
Published: August 2003

The effects of various fungicides on mycelial growth and spore germination of Ascochyta rabiei were determined by incorporating them into potato dextrose agar and measuring colony diameter and observing colony growth and spore germination at 20 ± 2°C. Eight fungicides prevented spore germination of the pathogen at concentrations of 0.125–2 μg/ml, three hindered mycelial growth at 2–4 μg/ml and seven failed to inhibit mycelial growth even at 128 μg/ml. The reference fungicide for the pathogen, chlorothalonil, stopped conidial germination at low rates but did not prevent mycelial growth at 128 μg/ml. Thirteen fungicides were tested against seed infections of the pathogen, and benomyl + thiram, carbendazim and carbendazim + chlorothalonil seed treatments gave more than 85% inhibition on both vacuum‐infiltrated and naturally infected seeds. Coating the seeds with polymers did not increase the effectiveness of fungicides. Three fungicides; (azoxystrobin, chlorothalonil and mancozeb), gave the highest protection in the field but protection decreased with increased inoculum pressure. Addition of humic acid to fungicide suspensions did not affect their performance.
PCR‐based Specific Detection of Ustilaginoidea virens and Ephelis japonica
Authors: Zhou Y.‐L. , Izumitsu K. , Sonoda R. , Nakazaki T. , Tanaka E. , Tsuda M. , Tanaka C.
Published: August 2003

A PCR‐based technique for detection of clavicipitaceous pathogens in rice and related grasses was developed. The target pathogens were Ustilaginoidea virens, which causes rice false smut, and Ephelis japonica, which causes rice udbatta disease and black choke in grasses. To design specific primers, a comparison was made on genetic diversity on the rDNA internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene of U. virens, Ephelis japonica, as well as some other clavicipitaceous fungi. Each fungus was successfully detected by using a specific primer set with high sensitivity. Species‐specific primers designed here were capable of detecting these pathogens in plant tissues. The PCR detection was consistent with conventional histological observation. This nested PCR assay was sensitive and reliable for the detection of U. virens and E. japonica, and thus can be a used to study disease cycles and early prediction of false smut and udbatta‐disease incidence in fields.
Biocontrol of Fusarium and Verticillium Wilt of Tomato by Penicillium oxalicum under Greenhouse and Field Conditions
Authors: Larena I. , Sabuquillo P. , Melgarejo P. , De Cal A.
Published: August 2003

Treatments with conidia of Penicillium oxalicum produced in a solid‐state fermentation system were applied at similar densities (6 × 106 spores/g seedbed substrate) to tomato seedbeds in water suspensions (T1: 5 days before sowing, or T2: 7 days before transplanting; 15 days after sowing), or in mixture with the production substrate (T3: 7 days before transplanting; 15 days after sowing). Treatments T2 and T3 significantly (P = 0.05) reduced fusarium wilt of tomato in both greenhouse (artificial inoculation) (33 and 28%, respectively) and field conditions (naturally infested soils) (51 and 72%, respectively), while treatment T1 was efficient only in greenhouse (52%). Verticillium wilt disease reduction was obtained with T3 in two field experiments (56 and 46%, respectively), while T1 and T2 reduced disease only in one field experiment (52% for both T1 and T2). Treatment with conidia of P. oxalicum plus fermentation substrate (T3) resulted in better establishment of a stable and effective population of P. oxalicum in seedbed soil and rhizosphere providing populations of approx. 107 CFU/g soil before transplanting. Results indicate that it will be necessary to apply P. oxalicum at a rate of approx. 106–107 CFU/g in seedbed substrate and rhizosphere before transplanting for effective control of fusarium and verticillium wilt of tomato, and that formulation of P. oxalicum has a substantial influence on its efficacy.
Role of birds in transmission of classical swine fever virus
Authors: Kaden V. , Lange E. , Steyer H. , Bruer W. , Langner CH.
Published: August 2003

Active transmission of classical swine fever virus (CSFV) was studied in six birds (five ravens, one hooded crow) and two laying hens. Cloacal swabs, blood and organs of birds and hens as well as blood and organ samples of pigs which had been fed with faeces derived from CSFV infected birds or which had come in contact with faeces of infected hens were negative for CSFV. None of the animals seroconverted during the study. This result demonstrates that active virus transmission by these animals is unlikely. Dissemination of CSFV from wild boar to domestic pigs is discussed.
Bacteriological Status of Canine Milk and Septicaemia in Neonatal Puppies – a Retrospective Study
Authors: Schäfer‐Somi S. , Spergser J. , Breitenfellner J. , Aurich J. E.
Published: August 2003

Results of the bacteriological examination of milk samples from 46 bitches were evaluated retrospectively, and correlated with findings of the bacteriological examination of organs from dead, septicaemic puppies in their litters (n = 33). The aim of this study was to investigate, in how many cases of clinical and subclinical mastitis of the bitches, the same bacteria can be detected in their septicaemic neonates. One group of lactating bitches was clinically healthy (group I, n = 38), whereas in eight bitches different puerperal disorders were found (group II). Twenty‐five septicaemic puppies were from group I, eight from group II. Out of a broad spectrum of bacteria isolated from the milk of clinically healthy and diseased bitches, only Escherichia coli, Klebsiella pneumoniae and/or ß‐haemolytic (haem.) Streptococcus sp. could be isolated from organs of their septicaemic puppies. This was the case in three bitches with mastitis and in one clinically healthy bitch only. Staphylococcus intermedius, although frequently isolated from canine milk, does not seem to be a cause of septicaemia in neonates. It is assumed that in most cases of neonate septicaemia, bacteria from the bitches’ milk are not the primary cause.
Isolation of Mycoplasmas from Nasal Swabs of Calves affected with Respiratory Diseases and Antimicrobial Susceptibility of their Isolates
Authors: Hirose K. , Kobayashi H. , Ito N. , Kawasaki Y. , Zako M. , Kotani K. , Ogawa H. , Sato H.
Published: August 2003

Nasal swabs of 293 calves were examined for Mycoplasma. The samples were collected from calves affected with respiratory diseases on 71 farms in various parts of Japan between 1996 and 1997. Mycoplasma bovirhinis was isolated from 47 of 293 calves (16.0%). Mycoplasma alkalescens, M. bovis, M. arginini, M. bovigenitalium and Acholeplasma spp. were isolated from 19 (6.5%), seven (2.4%), four (1.4%), four (1.4%) and 18 (6.1%) calves, respectively. Pasteurella multocida and P. haemolytica were isolated from 60% of Mycoplasma‐positive calves. However, other bacteria were not isolated from calves. To evaluate the antimicrobial susceptibility of their isolates, 68 M. bovirhinis, 21 M. alkalescens and 10 M. bovis strains were examined for 12 antimicrobial agents. All isolates showed higher susceptibility to tiamulin than to the other drugs used in the study. However, erythromycin had no effect on any of the Mycoplasma strains studied. The field isolates were less susceptible than the type strains to some drugs, such as spiramycin, oxytetracycline and tylosin.
Spatial Relative Risk Mapping of Pseudorabies‐Seropositive Pig Herds in an Animal‐Dense Region
Authors: Berke O. , Grosse Beilage E.
Published: August 2003

This paper describes a new approach for spatial relative risk mapping as a tool for geographical risk assessment. The spatial epidemiological analysis is based on geographically referenced data about pseudorabies (Aujeszky's disease) virus infections at farm level in a region of high animal density in Germany at the beginning of the national eradication project. On the basis of serological findings 186 farms were classified as positive out of a total of 482 investigated farms listed in veterinary administrative registers. Geographical cluster analysis was used to identify two areas of high risk (RR = 2.4 and 3.3). Non‐parametric density estimation was used to estimate the proportion of infected farms per square kilometre. Furthermore, the spatial relative risk function was approximated through the prevalence ratio defined by the ratio of the local prevalence and the overall prevalence of the farms outside the cluster regions. The corresponding approximated relative risk map indicates and quantifies a clear spatial pattern of disease frequency.
Diagnosis of Porcine Proliferative Enteropathy: Detection of Lawsonia intracellularis by Pathological Examinations, Polymerase Chain Reaction and Cell Culture Inoculation
Authors: Dittmar M. , Hoelzle L. E. , Hoelzle K. , Sydler T. , Corboz L. , Miserez R. , Wittenbrink M. M.
Published: August 2003

A total of 21 pigs aged 7–17 weeks with clinical symptoms suggestive for Porcine Proliferative Enteropathy were examined for Lawsonia intracellularis by analysing the following parameters: (i) intestinal gross and histological lesions, (ii) presence of comma‐shaped bacteria in enterocytes by Warthin–Starry and a modified Ziehl–Neelsen stain, (iii) PCR amplification of L. intracellularis DNA from intestinal mucosa by using two oligonucleotide primer pairs targeting a 255‐bp DNA fragment of the 16S rDNA‐gene and a 319‐bp DNA fragment of the L. intracellularis chromosome. Specificity of PCR reactions was confirmed by using DNA extracted from the L. intracellularis reference strain N343 (ATCC 55672) as well as by DNA sequence comparisons of PCR amplification products with data bank entries. Intestinal gross lesion indicative for PPE were observed in 20 pigs (95.2%). For all 21 pigs, the L. intracellularis aetiology was confirmed by histological as well as bacterioscopical examinations. Specific PCR amplification products were obtained from 20 pigs (95.2%). Taking PCR positivity as the definite criterion, L. intracellularis was diagnosed in 20 pigs from 11 herds in seven Swiss cantons (Argovia, Berne, Fribourg, Grisons, Lucerne, Schwyz, Thurgovia). To grow L. intracellularisin vitro, the cell culture method of Lawson et al. (J. Clin. Microbiol. 1993: 31, 1136–1142) was adopted. Inocula prepared from heavily infected fresh and frozen ileal mucosa of 15 pigs were cultured in rat enterocytic IEC‐18 cells (ATCC CRL 1589). Six cell culture passages of 10 days each were completed. The reference strain N343 was examined for cultivability, accordingly. Except for occasional specific PCR amplifications from cell cultures up to the second passage, any indications for growth of L. intracellularis in IEC‐18 cells were not found.
Osmosensitive properties of rapid and slow delayed rectifier K+ currents in guinea‐pig heart cells
Authors: Ogura Toshitsugu , Matsuda Hiroyuki , Shibamoto Toshishige , Imanishi Sunao
Published: August 2003

1. Changes in cell volume affect a variety of sarcolemmal transport processes in the heart. To study whether osmotically induced cell volume shrinkage has functional consequences for K+ channel activity, guinea‐pig cardiac preparations were superfused with hyperosmotic Tyrode's solution (1.2–2‐fold normal osmolality). Membrane currents and cell surface dimensions were measured from whole‐cell patch‐clamped ventricular myocytes and membrane potentials were recorded from isolated ventricular muscles and non‐patched myocytes.

2. Hyperosmotic treatment of myocytes quickly (< 3 min to steady state) shrank cell volume (approximately 20% reduction in 1.5‐fold hyperosmotic solution) and depressed the delayed rectifier K+ current (IK). Analysis using different activation protocols and a selective inhibitor (5 µmol/L E4031) indicated that the IK inhibition was due to osmolality and cell volume‐dependent changes in the two subtypes of the classical cardiac IK (rapidly activating IKr and slowly activating IKs); 1.5‐fold hyperosmotic treatment depressed the amplitudes of IKr and IKs by approximately 30 and 50%, respectively.

3. Superfusion of muscles and myocytes with 1.5‐fold hyperosmotic solution lengthened the action potentials by 14–17%. Hyperosmotic treatment also caused 6–7 mV hyperpolarization that is most likely due to a concentrating of intracellular K+.

4. The inhibition of IK helps explain the lengthening of action potentials observed in osmotically stressed heart cells. These results, together with the reported IK stimulation by hyposmotic cell swelling, provide further support for cell volume‐sensitive properties of cardiac electrical activity.
Diethylenetriamine NITRIC oxide adduct relaxes precontracted mouse tracheal smooth muscle*
Authors: Lam Chen Fuh , Lan Rommel S , Van Heerden Peter V , Ilett Kenneth F , Henry Peter J
Published: August 2003

1. Inhaled diethylenetriamine nitric oxide adduct (DETA/NO) has been shown to be a selective pulmonary vasodilator in animal and human studies. The aims of the present study were to investigate the effect of DETA/NO on mouse precontracted isolated tracheal smooth muscle preparations and to determine the active component of this compound.

2. Mouse isolated tracheal smooth muscle rings were precontracted with carbachol (10−7 mol/L). Changes in isometric tension were recorded after cumulative addition of DETA (30–300 µmol/L; n = 6), DETA/NO (30–300 µmol/L; n = 9) or diluent control (n = 3). In addition, some preparations (n = 5) were pretreated with the soluble guanylyl cyclase (sGC) inhibitor 1H‐[1,2,4]‐oxadiazolo[4,3,‐a]quinoxalin‐1‐one (ODQ; 30 µmol/L) before precontraction and exposure to DETA/NO.

3. Addition of DETA/NO caused a concentration‐dependent relaxation of tracheal smooth muscle at 100 and 300 µmol/L, with an EC25%R of 109 µmol/L (95% confidence interval 72.6–164 µmol/L). The nucleophile amine carrier DETA had no effect on isometric tension. However, the relaxant effect of DETA/NO was completely abolished by pretreatment with ODQ.

4. We conclude that DETA/NO induces a concentration‐dependent relaxation of mouse carbachol‐contracted isolated tracheal smooth muscle that is mediated by NO released from DETA/NO via the activation of sGC.
Intermittent hypobaric hypoxia increases the ability of neutrophils to generate superoxide anion in humans
Authors: Hitomi Y , Miyamura M , Mori S , Suzuki K , Kizaki T , Itoh C , Murakami K , Haga S , Ohno H
Published: August 2003

1. We investigated the effect of intermittent exposure to hypobaric hypoxia on the ability of neutrophils to generate ·O2–.

2. Seven male volunteers were exposed intermittently to hypobaric hypoxia, equivalent to an altitude of 4500 m, for 7 successive days. Peripheral blood samples were collected before and after the 2 h course of hypobaric hypoxia on days 1 and 7 and neutrophils were subjected to a chemiluminescence assay for ·O2– production.

3. On day 1, 2 h exposure to hypobaric hypoxia induced granulocytosis (P < 0.01), but the ability of neutrophils to generate ·O2– was unchanged.

4. On day 7, such granulocytosis was not observed, suggesting acclimatization to hypobaric hypoxia.

5. The ability of neutrophils to generate ·O2– was significantly increased on day 7 (P < 0.01), although there was no definite change in the mRNA expression of NADPH oxidase subunits in the cells.

6. The results suggest that the ability of neutrophils to generate ·O2– may be gradually potentiated by intermittent exposure to hypobaric hypoxia, even after the number of neutrophils in peripheral blood stabilizes.
Approaches for introducing peptides into intact and functional arteriolar smooth muscle: manipulation of protein kinase‐based signalling‡
Authors:   Kay Richards,, Davis Michael J , Potocnik Simon J , Murphy Timothy V , Bishara Nour B , Rajanayagam MA Sharmini , Darby Ian A , Hill Michael A
Published: August 2003

1. An exact understanding of signal transduction pathways within intact and functional arteriolar smooth muscle is made difficult by limited access to the intracellular environment due to the cell membrane. The aim of the present studies was to determine the feasibility of using polycationic lipids and reverse permeabilization for the introduction of peptide inhibitors into smooth muscle cells of the intact arteriolar wall.

2. Isolated cannulated arterioles were exposed to polycationic lipid preparations together with varying concentrations of the protein β‐galactosidase (30–90 µg/mL). Similar experiments were also performed using cultured smooth muscle cells. Staining for the chromogenic substrate of β‐galactosidase (5‐bromo‐4‐chloro‐3‐indolyl‐β‐d‐galactosidase; X‐gal) demonstrated incorporation of the protein into cultured cells but not intact arteriolar smooth muscle. Similarly, polycationic lipid treatment did not enable loading of arteriolar smooth muscle (as assessed by cAMP‐mediated vasodilation) with the protein kinase (PK) A inhibitory peptide PKI.

3. In contrast, reverse permeabilization, using high ATP concentrations in the presence of EGTA enabled introduction of PKI and inhibition of forskolin‐mediated vasodilatation. Furthermore, arterioles maintained full viability following reverse permeabilization, as demonstrated by an ability to develop spontaneous myogenic tone.

4. Reverse permeabilization provides a method for introducing peptide inhibitors into functional arteriolar smooth muscle and manipulating signal transduction. Protein transfection using polycationic lipids appears to be limited by the barrier provided by the adventitia or inherent differences between cells under cultured conditions compared within the intact arteriole.
P2Y receptor‐mediated enhancement of permeation requires Ca2+ signalling in vascular endothelial cells
Authors: Tanaka Naoko , Kawasaki Kumiko , Nejime Namie , Kubota Yoko , Takahashi Koichi , Hashimoto Michio , Kunitomo Masaru , Shinozuka Kazumasa
Published: August 2003

1. We investigated the effects of 2‐methylthioATP (2meS‐ATP; a P2Y receptor agonist) on the permeation of fluorescein isothiocyanate (FITC)‐labelled dextran, transendothelial electrical resistance (TEER) and intracellular calcium levels ([Ca2+]i) in cultured endothelial cells isolated from the rat caudal artery.

2. The cellular transport of FITC‐labelled dextran was enhanced and TEER of the endothelial monolayer was reduced by 2meS‐ATP. Both these effects were prevented by pyridoxalphosphate‐6‐azophenyl‐2′,4′‐disulphonic acid, a P2Y receptor antagonist, which also inhibited the increase in [Ca2+]i induced by 2meS‐ATP in endothelial cells.

3. The increase in [Ca2+]i induced by 2meS‐ATP was inhibited by thapsigargin (a Ca2+ pump inhibitor) and by U‐73122 (a phospholipase C inhibitor).

4. These findings suggested that activation of the P2Y receptor enhances the passage of material in the endothelium, which is associated with Ca2+ signalling in endothelial cells.
Oxidized low‐density lipoprotein inhibits vascular endothelial growth factor‐induced endothelial progenitor cell differentiation
Authors: Imanishi Toshio , Hano Takuzo , Matsuo Yoshiki , Nishio Ichiro
Published: August 2003

1. Bone marrow‐derived endothelial progenitor cells (EPC) in the peripheral blood of adult animals and humans have been shown to be incorporated into neovascularization. In contrast, hypercholesterolaemia impairs angiogenesis and collateral vessel formation in response to regional tissue ischaemia. We investigated whether oxidized LDL (oxLDL) affected human EPC differentiation.

2. When isolated human mononuclear cells (MNC) were incubated with vascular endothelial growth factor (VEGF), the number of differentiated, adherent EPC, as assessed by an in vitro culture assay, was increased in a dose‐dependent manner (P < 0.01). When MNC were incubated with oxLDL at 1, 5 and 10µg/mL in the presence of 100 ng/mL VEGF for 24 h, oxLDL dose‐dependently reduced the number of differentiated, adherent EPC.

3. Vascular endothelial growth factor‐induced EPC differentiation was significantly inhibited by pharmacological phosphatidylinositol 3‐kinase blockers (either 10 nmol/L wortmannin or 10 µmol/L LY294002). Interestingly, immunoblotting analysis revealed that oxLDL dose‐dependently led to dephosphorylation and, thus, deactivation of Akt in the presence of VEGF. Finally, these inhibitory effects induced by oxLDL were abolished by pretreatment with 1 µmol/L atorvastatin (P < 0.01).

4. Our data indicate that oxLDL inhibits VEGF‐induced EPC differentiation through the dephosphorylation of Akt.
Effect of ATP on Ca2+ uptake in the presence of high glucose in renal proximal tubule cells
Authors: Han Ho Jae , Park Soo Hyun , Lee Yun Jung , Lee Jeong Chi , Lee Jang Hern
Published: August 2003

1. Calcium regulation has been reported to be associated with the development of diabetic nephropathy. Thus, changes in Ca2+ uptake induced by ATP, an important regulator of Ca2+ uptake, in the diabetic condition and related signal pathways were examined in primary cultures of rabbit renal proximal tubule cells (PTC).

2. Under low (5 mmol/L) glucose conditions, 10−4 mol/L ATP inhibited Ca2+ uptake early on (< 30 min), whereas Ca2+ uptake was stimulated at later time points (> 2 h). However, under high (25 mmol/L) glucose conditions, ATP stimulated both the early and late uptake of Ca2+.

3. The adenylate cyclase inhibitor SQ 22536, the protein kinase (PK) A inhibitor PKI amide 14–22, Rp‐cAMP, staurosporine, bisindolylmaleimide I and H‐7 (PKC inhibitors) blocked the change in ATP effect on Ca2+ uptake in the presence of 25 mmol/L glucose. However, none one of these drugs blocked the effect of ATP on Ca2+ uptake in the presence of 5 mmol/L.

4. At 25 mmol/L, glucose increased cAMP content and PKC activity, whereas ATP had no effect on either parameter.

5. In conclusion, high glucose levels alter ATP‐induced Ca2+ uptake via cAMP and PKC pathways in the PTC.
Oestrogen improves glucose metabolism and insulin signal transduction in HepG2 cells
Authors: Xie Ping , Liu Mei‐Lian , Gu Ya‐Peng , Lu Jin , Xu Xia , Zeng Wei‐Min , Song Hui‐Ping
Published: August 2003

1. Many clinical studies have suggested a relationship between oestrogen and insulin sensitivity. In the present study, HepG2 cells were divided into four groups: (i) control, incubated with 1 nmol/L insulin; (ii) the HI group, which was incubated with 100 nmol/L insulin to induce insulin resistance; (iii) the E2 group, in which control cells were incubated with 1 nmol/L insulin plus 1 nmol/L oestradiol; and (iv) the HI + E2 group, in which insulin‐resistant cells were incubated with 100 nmol/L insulin + 1 nmol/L oestradiol.

2. A high concentration of insulin decreased the activity of phosphofructo‐1‐kinase (PFK), pyruvate dehydrogenase (PDH) and glycogen synthase (GS), as well as decreasing the expression of insulin receptor (IR) and insulin receptor substrate‐2 (IRS‐2). High insulin had no effect on glucose transport or the expression of insulin receptor‐1 (IRS‐1).

3. The addition of oestradiol to control cells increased glucose transport, the activity of PFK, PDH and GS and the expression of IRS‐1 and IRS‐2, but had no effect on the expression of IR.

4. Treatment of insulin‐resistant HepG2 cells with oestradiol attenuated HI‐induced decreases, except for IR, and the expression of IRS‐1 was significantly higher than control, attaining levels seen in group 3. The expression of IRS‐2 was significant higher than in insulin‐resistant cells, but did not reach control levels. Changes in the activity of PFK, PDH and GS were the same as the changes seen in the expression of IRS‐2.

5. These results suggest that high concentrations of insulin induce insulin resistance in HepG2 cells, whereas oestradiol improves glucose metabolism and insulin signal transduction of cells by enhancing the activity of key enzymes involved in glucose metabolism and the expression of IRS‐1 and IRS‐2.
The unbearable whiteness of being (in nursing)
Authors: Puzan Elayne
Published: August 2003

The unbearable whiteness of being (in nursing)

My purpose in writing this paper is to uncover some of the ways in which nursing participates in, reproduces, and resists the detrimental practices associated with white cultural privilege and to share some instances of its personal and social costs. It draws upon the body of scholarship which interrogates racism as it is enacted through whiteness in North America. Whiteness is depicted not as a preordained biological property, but as a socially constructed category of race, wherein non‐white people are racially designated, while whites escape such designation and occupy positions which allow them to carry on as if what they say is neutral, rather than historically and ideologically situated. While the concept of whiteness may not have much resonance in nursing, it offers another way to talk about racism, one that does not stop with the scrutiny of the racialized Other. The presumed neutrality of whiteness has been institutionalized so that its authority to define knowledge, membership, and language, as well as its ability to stipulate and enforce the rules and regulations of everyday concourse and discourse within nursing is concealed.
Increased fatty acid oxidation in transgenic mice overexpressing UCP3 in skeletal muscle
Authors: Wang S. , Subramaniam A. , Cawthorne M. A. , Clapham J. C.
Published: August 2003

Aim:  To determine the rates of substrate oxidation by skeletal muscle in vitro as well as tissue‐specific glucose uptake in vivo in transgenic mice overexpressing uncoupling protein‐3 (UCP3) in skeletal muscle.

Methods:  Soleus muscle was isolated from transgenic mice overexpressing UCP3 in skeletal muscle and wild‐type mice. Rates of [1‐14C]‐palmitate oxidation and [2‐14C]‐pyruvate oxidation were determined by in vitro incubation of the soleus muscle. Tissue glucose uptake rates were characterized during a glucose tolerance test using 2‐deoxy‐[1‐3H]‐glucose as a tracer.

Results:  Oxidation of [1‐14C]‐palmitate to CO2 by isolated soleus muscle was increased in UCP3 transgenic mice (0.45 ± 0.03 vs. 0.24 ± 0.02 µmol/h/g). [2‐14C]‐pyruvate oxidation, which is a measure of the activity of pyruvate carboxylase in introducing pyruvate carbon into the tricarboxylic acid cycle, was increased 1.4‐fold in the presence of fatty acid in the UCP3 transgenic mice (3.84 ± 0.28 vs. 5.36 ± 0.29 µmol/h/g). The plasma glucose concentration after an overnight fast was significantly lower in the UCP3 transgenic mice (3.56 ± 0.37 vs. 5.11 ± 0.33 m/mol). Only brown adipose tissue from the UCP3 transgenic mice showed increased tissue glucose uptake rates compared with the wild‐type mice. Skeletal muscle uptake rates of 2‐deoxyglucose were either unchanged (soleus and gastrocnemius) or reduced (diaphragm) in the UCP3 transgenic mice.

Conclusions:  The improved glucose tolerance in the UCP3 transgenic mice does not appear to be the result of increased uptake into peripheral tissues. The increased fatty acid oxidation in skeletal muscle of UCP3 transgenic mice supports the proposed role of UCP3 in the export of fatty acid anions from mitochondria during fatty acid oxidation.
Plasma adiponectin in overweight, nondiabetic individuals with or without insulin resistance
Authors: Shand B. I. , Scott R. S. , Elder P. A. , George P. M.
Published: August 2003

Aim:  Adiponectin is a protein produced exclusively by adipocytes with putative insulin‐sensitizing and anti‐atherogenic properties. This cross‐sectional study investigated the relationship between plasma adiponectin and a range of anthropometric, glycaemic, lipid and inflammatory parameters in overweight and obese subjects expressing characteristics of the metabolic syndrome.

Methods:  Subjects were selected for the study from a clinical database, if they were non‐diabetic, overweight [body mass index (BMI) > 25] and had features of the metabolic syndrome. The subjects were grouped according to BMI (25–30, 31–35 and >35 kg/m2) and then stratified for insulin resistance [homeostasis model assessment (HOMA) %S]. One hundred and ninety‐seven patients (109 males and 88 females) were selected for the study by taking an equal number with the highest and lowest HOMA indices from each of the three BMI groups. Plasma adiponectin concentration was measured in duplicate by radioimmunoassay, and the relationship between these levels and the other parameters was investigated using correlation and multiple linear regression analyses.

Results:  Plasma adiponectin concentration was higher in females than males (median 10.3 vs. 7.1 µg/ml, p < 0.001) despite being matched for BMI. In both genders, adiponectin levels were inversely related to BMI, waist circumference, percentage body fat, insulin resistance and the fasting plasma concentration of leptin. A direct correlation in both sexes was found between adiponectin levels and high‐density lipoprotein (HDL)‐cholesterol, apolipoprotein A1 and age. Multiple linear regression analyses showed that the independent determinants of low plasma adiponectin concentrations were gender, age, BMI, insulin resistance and HDL‐cholesterol. An association between reduced adiponectin and increased high‐sensitivity plasma C‐reactive protein concentration was observed only in female subjects and was independent of anthropometric variables. Our observation that adiponectin levels increase with age differs from the majority of other studies and may simply reflect the demographics of the population studied.

Conclusions:  This study shows that adiponectin is an important molecular link between obesity, insulin resistance and atherogenic lipoproteins. It is possible that plasma adiponectin concentration may be a convenient marker for identifying subjects with the metabolic syndrome who may progress to impaired glucose tolerance. Longitudinal studies are required in order to verify this clinical application of adiponectin.
Sibutramine enhances insulin sensitivity ameliorating metabolic parameters in a double‐blind, randomized, placebo‐controlled trial
Authors: Tambascia M. A. , Geloneze B. , Repetto E. M. , Geloneze S. R. , Picolo M , Magro D. O.
Published: August 2003

Aim:  To assess the effect of sibutramine‐assisted weight reduction program on insulin sensitivity and metabolic parameters in obese normal glucose tolerant individuals over a period of 24 weeks.

Research Design and Methods:  A double‐blind, placebo‐controlled, parallel, prospective clinical trial was carried out at our medical centre. Forty female normal glucose tolerant patients, body mass index: 34.3 ± 2.9 kg/m2 and age: 41.1 ± 9.9 (range: 19–58 years), were randomized to placebo or sibutramine, 10 mg once daily.

Results:  Seventeen patients from sibutramine group and 14 placebo had completed the study protocol. Significant weight change was seen in sibutramine (p < 0.01) (−5.6 kg or −6.1% vs. +0.9 kg or +1.1% in placebo). Insulin sensitivity enhanced in sibutramine group (Kitt: from 4.03 ± 1.97 to 5.09 ± 2.48%/min; p < 0.05). Homeostasis model assessment‐IR (HOMA‐IR) decreased from 7.8 ± 6.9 to 5.6 ± 4.5 (p < 0.05). HOMA‐β also decreased from 508 ± 381 to 374 ± 256 (p < 0.05). No changes were observed in the placebo control group regarding insulin sensitivity or secretion. Concomitant reductions were observed in the sibutramine group in lipid parameters (triglycerides and high‐density lipoprotein‐cholesterol), uric acid and gamma‐glutamyl transferase (p < 0.05).

Conclusions:  Sibutramine has demonstrated efficacy in reducing weight in non‐diabetic women along with amelioration in insulin sensitivity and additional improvement in metabolic parameters.