bluuebunny/crossref_metadata_2025 huggingface dataset:
Extent of constitutive histone H2AX phosphorylation on Ser‐139 varies in cells with different TP53 status
Authors: Tanaka T. , Kurose A. , Huang X. , Traganos F. , Dai W. , Darzynkiewicz Z.
Published: July 2006

Abstract.  In response to DNA damage by genotoxic agents, histone H2AX is phosphorylated on Ser‐139. However, during the cell cycle, predominantly in S and G2M phase, histone H2AX is also phosphorylated in untreated normal and tumour cells. This constitutive H2AX phosphorylation is markedly reduced by exposure of cells to the reactive oxygen species scavenger N‐acetyl‐L‐cysteine. Therefore, it appears likely that constitutive H2AX phosphorylation reflects the ongoing oxidative DNA damage induced by the reactive oxygen species during progression through the cell cycle. Because the tumour suppressor p53 (tumour protein p53) is known to induce transcription of genes associated with cell response to oxidative stress, we have compared the intensity of constitutive H2AX phosphorylation, and the effect of N‐acetyl‐L‐cysteine on it, in cells with different tumour protein p53 status. These were human lymphoblastoid cell lines derived from WIL2 cells: TK6, a p53 wt line, NH32, a tumour protein p53 knock‐out derived from TK6, and WTK1, a WIL2‐derived line that expresses a homozygous mutant of tumour protein p53. Also tested were the tumour protein p53‐null promyelocytic HL‐60 cells. The degree of constitutive H2AX phosphorylation was distinctly lower in NH32, WTK1 and HL‐60 compared to TK6 cells in all phases of the cell cycle. Also, the degree of attenuation of constitutive H2AX phosphorylation by N‐acetyl‐L‐cysteine was less pronounced in NH32, WTK1, and HL‐60, compared to TK6 cells. However, the level of reactive oxygen species detected by the cells’ ability to oxidize carboxyl‐dichlorodihydrofluorescein diacetate was not significantly different in the cell lines studied, which would suggest that regardless of tumour protein p53 status, the level of oxidative DNA damage was similar. The observed higher level of constitutive H2AX phosphorylation in cells harbouring wt tumour protein p53 may thus indicate that tumour protein p53 plays a role in facilitating histone H2AX phosphorylation, an important step in the mobilization of the DNA repair machinery at the site of DNA double‐strand breaks.
Discourses of blame: deconstructing (hetero)sexuality, peer sexual violence and residential children’s homes
Authors: Barter Christine
Published: July 2006

Little research exists on sexuality and residential care, even less that position gender inequality as a central analytical component. This paper, based on a wider research project that focused on children’s and young people’s experiences of peer violence within residential care, seeks to help redress this disparity. The main aim of this paper is to explore how (hetero)sexuality, discourses of blame and male peer sexual violence are constructed and interrelated in residential child care. Young people’s and staff’s conceptualizations, evaluations and experiences are compared and contrasted to illuminate the range of discourses surrounding sexuality and sexual violence operating within residential settings. Findings relating to female culpability for male violence, normative male violence, female duplicity and challenges to traditional gender roles will be presented, and contextualized, in relation to previous studies. In addition, some broader implications of the research findings for residential practice are explored.
Effects of tixocortol pivalate on gastrointestinal disease in systemic mastocytosis: a preliminary study
Authors: FRIEDMAN B. S. , METCALFE D. D.
Published: July 2006

Tixocortol pivalate is a steroid reportedly without significant adrenal‐pituitary axis suppression when administered via the gastrointestinal tract. To determine whether this steroid would suppress the gastrointestinal manifestations of systemic mastocytosis, we performed an open clinical trial for safety and efficacy with tixocortal pivalate in four patients for periods of 8‐15 weeks. All patients showed a decrease in the symptoms of abdominal pain and frequency of stools. Laboratory parameters of malabsorption improved in parallel with symptom relief. Histopathologic abnormalities of the small bowel improved in one patient. There was no significant suppression of the pituitary adrenal axis. Two patients developed fluid retention while on tixocortol pivalate, which was attributed to a mineralocorticoid effect. One patient had a fall in am cortisol. In summary, this study strongly suggests that tixocortol pivalate, when administered orally, has gastrointestinal anti‐inflammatory activity comparable to conventional steroids, but may not be entirely without adrenal‐suppressive effect and may lead to fluid retention in some patients. Further studies are warranted to assess the value of tixocortol pivalate in the therapy of inflammatory diseases of the upper gastrointestinal tract.
Respiratory symptoms in Queensland schoolchildren: an association between month of birth and respiratory illness
Authors: DUFFY D. L. , MITCHELL C. A.
Published: July 2006

We have examined the relationship between month of birth and episodes of wheezing, productive cough, eczema and hayfever, and also standardized maximum midexpiratory flow (FEF25–75) in a representative sample of 4549 Australian primary schoolchildren (mean age 10.1 years). Children experiencing frequent wheezing (one or more episodes per month) were more likely to be born in spring and summer (odds ratio =1.6, 95% confidence interval = 1.1–2.4), but this was not seen for less severe disease. No effect of season or birth month was seen for the other illnesses examined or for lung function.
Effect of oral and inhaled cetirizine in allergen induced bronchoconstriction
Authors: RAFFERTY P. , GHOSH S. K. , DE VOS C. , PATEL K. R.
Published: July 2006

Cetirizine is a potent, selective H1 histamine receptor antagonist. The effect of oral and inhaled eetirizine was assessed on the early bronchoconstrictor response to inhaled allergen in 10 mild atopie asthmatic patients in a double‐blind, randomized, plaeebo controlled trial. All were sensitive to Dermatophagoides pleronyssinus and this was used as the provoking allergen. The geometric mean PD20 FEV] values obtained at allergen challenge were measured as cumulative breath units (c.b.u.) and following oral cetirizine, inhaled cetirizine and placebo were 124–5, 75–7 and 76–7 c.b.u. respectively. These did not differ significantly. We conclude that neither oral nor inhaled cetirizine significantly attenuates the early response to inhaled allergen in atopie asthmatic subjects. However, the method of repeated allergen challenge is likely to be relatively insensitive. Clinical and Experimental Allergy, [o]. 23, pp. 528–531.
Codeine‐induced histamine release in intact human skin monitored by skin microdialysis technique: comparison ofintradermal injections with an atraumatic intraprobe drug delivery system
Authors: PETERSEN L.J. , NIELSEN H.J. , SKOV STAHL
Published: July 2006

Background
The skin microdiallysis technique makes it possible to measure histamine release in intact human skin in vivo directly. In this study we have used the microdialysis technique to characterize histamine release by codeine after intracutaneous injectioin and following skin challenge by a novel atraumatic delivery technique.

Objective
The purpose of the study was to compare histamine release in human skin by codeine. delivered by an intraprobe drug delivery system (IPD) and intracutaneous injections (ICT), with respect to dose‐response relations, kinetics of histamine appearance and decay, corelations between histamine release and skin respones, and reproducibility.

Methods
Hollow dialysis fibres were inserted intradermally in 12 healthy subjects. Twelve fibres were inserted in each subjects, six fibres in each arm. Each fibre was perfused at a rate of 3 μL/min, and samples were collected in 2 min fractions. By the IPD technique, codeine was administrered to the skin by adding codeine to the perfusion medium. Sequential IPD challenges were performed in one arm. and ICTs were done on the other arm.

Results
Sixfold serial dilutions of codeine (0.01‐3 mg/mL) caused a significant doserelated histamine release by ICT and IPD. Peak histamine release was found within the first 4 min after skin challenge by ICT and IPD, followed by a fast decline with a dialysate histamine half life of approximately 2‐3 min. Peak hisamine release was linearly correlates with cumulative release of the 20 min sampling period, and histamine release correlated with weal soze. The coefficient of variation on peak histamine releae was 18.9% and 4.8% for codeine ICT and IPD, respectively.

Conclusioin
We have described in detail codeine‐induced histamine release in intact human skin in vivo by the microdialysis technique. It was possible to administer codeine atraumaticallyl to the skin by intraprobe delivery. The skin microdialysis codeine atraumaticallly to the skin by intraprobe delivery. The skin microdialysis technique opens up possibilities for measurement of infllammatory mediators release in normal and diseases skin, and it will be possible to deliver immunopharmacologically active drugsto the skin by intraprobe delivery.
Selective restimulation of antigen or allergen preactivated T cells using OKT3 F(ab)2 results in the secretion of TH‐1 or TH‐2‐like cytokine patterns
Authors: JUTEL M. , WYSS‐CORAY T. , CARBALLIDO J. M. , BLASER K. , MULLER U. R. , PICHLER W. J.
Published: July 2006

Background
The synthesis of IgE is regulated by cytokines secreted from T‐helper cells. The studies on cytokine secretion by peripheral blood mononuclear cells (PBMC) upon stimulation with antigen or allergen are ditficult due to low levels of cytokines, especially of inlerieukin‐4 (IL‐4).

Objective
ln this study we tried lo establish a culture system, which could enable the measurement of the cytokine profiles in specifically activated cultures.

Methods
Three methods to potentiate cytokine secretion were evaluated: PBMC from bee venom or house dust mite (Dermatophagoides pteronyssinus) allergic patients as well as normal subjects were stimulated either with the major bee venom allergen phosphoiipase A2 (PLA) or with the major D. pteronyssinus allergen (Der p 1) or with the control antigens tetanus toxoid (TT) and purified protein derivate (PPD). After 7 days of culture the cells were restimulated either wilh pkistie bound 0KT3 F(ab)2 monoclonal antibodies (MoAbs), with the appropriate antigen + antigen presenting cells or with IL‐2. The secretion of cylokines (IL‐4, IFNγ) was measured after restimulation of the cultures (day 8).

Results
While OKT3 F(ab)2 was unable lo activate resting T cells, it could restimulate preactivaled cells. Restimulalion with OKT3 F(ab)2 induced higher IL‐4 and IFN7 secretion than restimulation with IL‐2 or antigen. TT and PLA stimulated a similar cytokine secretion prolile in normal and PLA allergic subjects with substantial levels of both lL‐4 and IFNγ. In contrast, PPD induced virtually only IFNγ secretion. Der p 1 stimulated mainly IL‐4 seerclion but also IFNγ production in some mite allergic palienis.

Conclusion
We have established a cell culture system, which combines antigen specificity with a strong cytokine inducing signial provided by anti‐CD3 MoAbs. TH‐1 and TH‐2 characteristic cytokine patterns can be observed in short‐term PBMC cultures already after 8 days of culture.
Aggravation of bronchial eosinophilia in mice by nasal and bronchial exposure to Staphylococcus aureus enterotoxin B
Authors: Hellings P. W. , Hens G. , Meyts I. , Bullens D. , Vanoirbeek J. , Gevaert P. , Jorissen M. , Ceuppens J. L. , Bachert C.
Published: July 2006

Background
The role of bacterial enterotoxins like Staphylococcus aureus enterotoxin B (SEB) in allergic asthma remains unknown. We used a mouse model of airway allergy to study the effects of nasal or bronchial contact with SEB on bronchial allergic inflammation.

Methods
The features of allergic asthma were induced in ovalbumin (OVA)‐sensitized mice (days 1–13) by repeated exposures to nebulized OVA (days 33–37). Nasal or bronchial application of SEB was performed on three occasions (days 33–35–37), and the effects on bronchial inflammation, IgE titres and expression levels of mRNA for T helper type 2 cytokines and other inflammatory mediators were evaluated.

Results
Both nasal and bronchial SEB enhanced the allergen‐induced bronchial inflammation, as reflected by more eosinophilic inflammation in the airway lumen and in bronchial tissue. Aggravation of experimental asthma correlated with higher expression of mRNA for IL‐5, IL‐4, IFN‐γ, IL‐12 p40, eotaxin‐1 and TGF‐β in bronchi. In addition, nasal SEB elevated concentrations of IL‐4, IL‐5 and IFN‐γ in serum and bronchial SEB increased titres of OVA‐specific and total IgE in serum.

Conclusion
Our data illustrate the potential of both nasal as well as bronchial SEB to aggravate several features of allergic asthma in a mouse model.
Allergen concentration in natural rubber latex
Authors: Yeang H.‐Y. , Hamilton R. G. , Bernstein D. I. , Arif S. A. M. , Chow K. ‐S. , Loke Y. ‐H. , Raulf‐Heimsoth M. , Wagner S. , Breiteneder H. , Biagini R. E.
Published: July 2006

Background
Hevea brasiliensis latex serum is commonly used as the in vivo and in vitro reference antigen for latex allergy diagnosis as it contains the full complement of latex allergens.

Objective
This study quantifies the concentrations of the significant allergens in latex serum and examines its suitability as an antigen source in latex allergy diagnosis and immunotherapy.

Methods
The serum phase was extracted from centrifuged latex that was repeatedly freeze‐thawed or glycerinated. Quantitation of latex allergens was performed by two‐site immunoenzymetric assays. The abundance of RNA transcripts of the latex allergens was estimated from the number of their clones in an Expressed Sequence Tags library.

Results
The latex allergens, Hev b 1, 2, 3, 4, 5, 6, 7 and 13, were detected in freeze‐thawed and glycerinated latex serum at levels ranging from 75 (Hev b 6) to 0.06 nmol/mg total proteins (Hev b 4). Hev b 6 content in the latex was up to a thousand times higher than the other seven latex allergens, depending on source and/or preparation procedure. Allergen concentration was reflected in the abundance of mRNA transcripts. When used as the antigen, latex serum may bias the outcome of latex allergy diagnostic tests towards sensitization to Hev b 6. Tests that make use of latex serum may fail to detect latex‐specific IgE reactivity in subjects who are sensitized only to allergens that are present at low concentrations.

Conclusion
Latex allergy diagnostics and immunotherapy that use whole latex serum as the antigen source may not be optimal because of the marked imbalance of its constituent allergens.
GH peak response to GHRH‐arginine: relationship to insulin resistance and other cardiovascular risk factors in a population of adults aged 50–90
Authors: Carmichael John D. , Danoff Ann , Milani Daniela , Roubenoff Ronenn , Lesser Martin L. , Livote Elayne , Reitz Richard E. , Ferris Steven , Kleinberg David L.
Published: July 2006

Objective  To assess the GH response to GHRH‐arginine in apparently healthy adults in relation to cardiovascular risk factors.

Design  Cross‐sectional.

Patients  Eighty‐six male and female volunteers aged 50–90.

Measurements  GH peak response to GHRH‐arginine and cardiovascular risk factors, including obesity, insulin resistance, low levels of high density lipoprotein (HDL) cholesterol, elevated triglycerides, and hypertension. The primary outcome measurement was GH response to GHRH‐arginine. The relationship between GH peak responses and cardiovascular risk factors was determined after data collection.

Results  GH peaks were highly variable, ranging from 2·3 to 185 µg/l (14% with GH peaks < 9 µg/l). An increasing number of cardiovascular risk factors were associated with a lower mean GH peak (P < 0·0001). By univariate analysis, fasting glucose, insulin, body mass index (BMI), HDL cholesterol and triglycerides were significantly associated with GH peak (all P < 0·0001). Multiple regression analysis revealed that fasting glucose, fasting insulin, BMI, triglycerides and sex accounted for 54% of GH peak variability. The role of abdominal fat as it relates to GH peak was explored in a subset of 45 subjects. Trunk fat and abdominal subregion fat measured by dual energy X‐ray absorptiometry (DXA) were inversely related to GH peak (P < 0·008 and 0·001, respectively). Analysis of this subgroup by multiple regression revealed that subregion abdominal fat became the significant obesity‐related determinant of GH peak, but still lagged behind fasting insulin and glucose.

Conclusions  GH response to secretagogues was highly variable in apparently healthy adults aged 50–90 years. Peak GH was significantly related to fasting glucose, insulin, BMI, HDL cholesterol, triglycerides, trunk fat and abdominal subregion fat, with fasting glucose ranking first by multiple regression analysis. There was a strong relationship between cardiovascular risk factors and low GH, with individual risk factors being additive. Although these data do not differentiate between low GH being a cause or an effect of these cardiovascular risk factors, they indicate that the relationship between low GH and increased cardiovascular risk may be physiologically important in the absence of pituitary disease.
Ultrasonography in detection of single and multiple abnormal parathyroid glands in primary hyperparathyroidism: comparison with radionuclide scintigraphy and surgery
Authors: Bhansali A. , Masoodi S. R. , Bhadada S. , Mittal B. R. , Behra A. , Singh P.
Published: July 2006

Background  Among the parathyroid imaging techniques, ultrasonography (USG) has the advantage of convenience, easy availability and low cost.

Objective  To study the role of USG in localization (side and/or site) of abnormal parathyroid glands in primary hyperparathyroidism (PHPT) and to compare with radionuclide scintigraphy and the ‘gold standard’, surgery.

Methods  Forty‐six consecutive patients undergoing USG of the anterior neck for a diagnosis of PHPT in whom a nuclear scan (technetium‐99m sestamibi and/or thallium‐201/technetium‐99m pertechnetate scintigraphy) was also performed, were studied. The results of imaging were independently interpreted and correlated with reference to surgical findings.

Results  Forty‐six patients had 52 abnormal parathyroid glands on surgical exploration and surgery was successful in all but one. Forty‐one patients had a single adenoma, four had multigland disease and one had a paraganglioma. USG correctly localized the abnormal gland in 30 (73%) and scintigraphy was positive in 40 (98%) out of 41 patients with a single adenoma as confirmed on surgical exploration. Scintigraphy showed a positive concordant test in all 30 patients with a single abnormal gland detected on USG and picked up 10 out of 11 abnormal glands where USG was negative. The sensitivity and positive predictive value of USG for detecting a single abnormal gland was 73% and 100%, respectively, whereas the sensitivity and positive predictive value for scintigraphy was 98%. In patients with multigland disease, USG missed 3 (30%) out of 10 and scintigraphy missed 6 (60%) out of 10 abnormal parathyroid glands as confirmed on surgical exploration (P < 0·05). However, in two patients who had ectopic parathyroid gland, both the modalities localized the lesion in one (paraganglioma), whereas in the other neither test was helpful (left retro‐esophageal).

Conclusion  USG is a convenient, affordable and useful modality to localize abnormal enlarged parathyroid glands in the majority of patients with PHPT. However, when USG is negative, scintigraphy is complementary to it.
Profile, mean residence time of ACTH and cortisol responses after low and standard ACTH tests in healthy volunteers
Authors: Alía P. , Villabona C. , Giménez O. , Sospedra E. , Soler J. , Navarro M. A.
Published: July 2006

Objective  No consensus exists until now about the suitable dose of tetracosactin in the ACTH stimulation test for detecting adrenal insufficiency. Our aim was to characterize both the ACTH(1–24) and the cortisol profiles after standard high‐dose test (250 µg) (HDT) and low‐dose test (1 µg) (LDT) in healthy subjects in order to provide a deeper knowledge about the relationship between stimulus and response.

Design and patients  ACTH tests were performed in 10 healthy volunteers (five men, five women) with at least 1 week of difference.

Measurements  Plasma ACTH(1–24) and ACTH(1–39) and serum cortisol were measured before tetracosactin i.v. injection and at 5, 15, 30, 45, 60, 75 and 90 min after stimulus. Area under the curve (AUC) of ACTH(1–24) and cortisol, as well as mean residence time (MRT) for ACTH(1–24) were calculated in both tests.

Results  Elimination of ACTH(1–24) was faster in HDT than in LDT (MRTs of 0·14 vs 0·37, respectively, P = 0·008), but plasma concentrations were higher up to 60 min cortisol production in HDT reaching a higher maximum concentration (Cmax: 1144 vs 960 nmol/l) but delayed in time (75 vs 52·5 min). No significant relationship was observed between AUC or Cmax of ACTH(1–24) and AUC, Cmax and increment of cortisol in any of the tests. However, a negative correlation of basal cortisol values was observed with relative cortisol increment (HDT: r = 0·77 P = 0·009; LDT: r = 0·94 P < 0·0001), but not so with Cmax (HDT: r = 0·22 P = 0·55; LDT: r = 0·57 P = 0·09).

Conclusions  The elimination rate of ACTH in healthy volunteers was significantly lower in LDT than in HDT, but cortisol production rate appears to be identical in both tests, so that a maximum adrenal stimulation seems to exist. The use of LDT may be more adequate, although data from patients need studying.
Use of Tag single nucleotide polymorphisms (SNPs) to screen PTPN21: no association with Graves’ disease
Authors: Zeitlin A. A. , Heward J. M. , Brand O. J. , Newby P. R. , Franklyn J. A. , Gough S. C. L. , Simmonds M. J.
Published: July 2006

Objective  The protein‐tyrosine‐phosphate nonreceptor 22 gene (PTPN22) has recently been identified as a susceptibility locus for a number of autoimmune diseases including Graves’ disease (GD). PTPN21 is another member of the PTPN family and its gene PTPN21 maps to the first reported region of genetic linkage to GD, GD−1, on chromosome 14q31. The aim of this study was to determine whether PTPN21 is acting as a GD susceptibility locus in UK Caucasian subjects.

Design  A case control association study of seven Tag single nucleotide polymorphisms (SNPs) (rs1469602, rs8007288, rs1998670, rs11622270, rs2274736, rs2295136 and rs366476) selected to predict 51 un‐genotyped polymorphisms present within PTPN21.

Patients  Unrelated Caucasian patients of UK origin with GD and ethnically and gender matched control subjects with no family history of autoimmune disease were recruited. In total, DNA was obtained from 768 GD patients and 768 control subjects.

Results  No association of any of the seven Tag SNPs was detected with GD. Preliminary evidence of association of rs2274736 was found with younger age of GD onset (0–30 years) (OR = 1. 48 [95% CI = 1·11–1·97]). No other correlations with clinical phenotype or previously established susceptibility loci were detected.

Conclusions  Using a Tag SNP approach we screened PTPN21 as a susceptibility locus for GD and found no evidence for association with disease. Preliminary evidence for association of rs2274736 with younger age of GD onset requires replication in similar sized data sets to exclude a false positive result. Methods such as the Tag SNP approach significantly reduce the amount of genotyping required when screening candidate loci, including those within regions of chromosomal linkage.
PPAR‐γ expression in pituitary tumours and the functional activity of the glitazones: evidence that any anti‐proliferative effect of the glitazones is independent of the PPAR‐γ receptor
Authors: Emery Michelle N. , Leontiou Chrysanthia , Bonner Sarah E. , Merulli Chiara , Nanzer Alexandra M. , Musat Madalina , Galloway Malcolm , Powell Michael , Nikookam Khash , Korbonits Márta , Grossman Ashley B.
Published: July 2006

Objective  It has been reported that both normal pituitary and pituitary tumours express PPAR‐γ, a nuclear hormone receptor, the expression being more abundant in pituitary tumours, and that this is the basis for the reported antiproliferative effects of the thiazolidinedione, rosiglitazone, in animal models. However, the mechanisms for the responsivity to rosiglitazone have remained unclear.

Design and measurements To investigate this further, ‘real‐time’ PCR was used to assess PPAR‐γ mRNA expression, and Western blotting and immunohistochemistry to study its protein expression, in 46 human pituitary tumours and normal pituitary tissue. Cell proliferation of the GH3 pituitary cell line was assessed by [3H]‐thymidine‐incorporation after 48 h rosiglitazone and pioglitazone (10−4 m– 10−10 m) treatment alone, or rosiglitazone in combination with the PPAR‐γ antagonist GW9662.

Results  PPAR‐γ mRNA and protein was found to be expressed in normal pituitary and was variably expressed in pituitary tumours, but were increased specifically in nonfunctioning pituitary adenomas. However, very little staining was observed with immunohistochemistry, with only occasional cell nuclei stained, and no difference was detectable between controls and tumours. Rosiglitazone at 10−4 mand 10−5 mconcentrations inhibited cell proliferation (10−4 m14·0% ± 1·5% and 10−5 m67% ± 4%[mean ± SEM]vsControl 100% ± 3%,P < 0·0001) while lower concentrations showed no significant effect. Following withdrawal of rosiglitazone 10−5 m, the cells fully recovered at a further 48 h, while lower doses showed a ‘rebound’ of stimulation. Pioglitazone was of similar potency to rosiglitazone in inhibiting proliferation. The PPAR‐γ antagonist did not show a significant reversal of the antiproliferative effect of rosiglitazone, and indeed suppressed proliferation on its own.

Conclusions  Our data suggest that the antiproliferative action of rosiglitazone is probably not via PPAR‐γ.
53BP1 contributes to survival of cells irradiated with X‐ray during G1 without Ku70 or Artemis
Authors: Iwabuchi Kuniyoshi , Hashimoto Mitsumasa , Matsui Tadashi , Kurihara Takayuki , Shimizu Hiroko , Adachi Noritaka , Ishiai Masamichi , Yamamoto Ken‐ichi , Tauchi Hiroshi , Takata Minoru , Koyama Hideki , Date Takayasu
Published: July 2006

Ionizing radiation (IR) induces a variety of DNA lesions. The most significant lesion is a DNA double‐strand break (DSB), which is repaired by homologous recombination or nonhomologous end joining (NHEJ) pathway. Since we previously demonstrated that IR‐responsive protein 53BP1 specifically enhances activity of DNA ligase IV, a DNA ligase required for NHEJ, we investigated responses of 53BP1‐deficient chicken DT40 cells to IR. 53BP1‐deficient cells showed increased sensitivity to X‐rays during G1 phase. Although intra‐S and G2/M checkpoints were intact, the frequency of isochromatid‐type chromosomal aberrations was elevated after irradiation in 53BP1‐deficient cells. Furthermore, the disappearance of X‐ray‐induced γ‐H2AX foci, a marker of DNA DSBs, was prolonged in 53BP1‐deficient cells. Thus, the elevated X‐ray sensitivity in G1 phase cells was attributable to repair defect for IR‐induced DNA‐damage. Epistasis analysis revealed that 53BP1 plays a role in a pathway distinct from the Ku‐dependent and Artemis‐dependent NHEJ pathways, but requires DNA ligase IV. Strikingly, disruption of the 53BP1 gene together with inhibition of phosphatidylinositol 3‐kinase family by wortmannin completely abolished colony formation by cells irradiated during G1 phase. These results demonstrate that the 53BP1‐dependent repair pathway is important for survival of cells irradiated with IR during the G1 phase of the cell cycle.
Crystal structure of human Rad GTPase of the RGK‐family
Authors: Yanuar Arry , Sakurai Shigeru , Kitano Ken , Hakoshima Toshio
Published: July 2006

Rad (Ras associated with diabetes) is an RGK‐family small GTPase that is over‐expressed in the skeletal muscle of humans with type II diabetes. Unlike other small GTPases, RGK family members including Rad lack several conserved residues in the GTPase domain. Here, we report the crystal structure of the GTPase domain of human Rad in the GDP‐bound form at 1.8 Å resolution. The structure revealed unexpected disordered structures of both switches I and II. We showed that the conformational flexibility of both switches is caused by non‐conservative substitutions in the G2 and G3 motifs forming the switch cores together with other substitutions in the structural elements interacting with the switches. Glycine‐rich sequences of the switches would also contribute to the flexibility. Switch I lacks the conserved phenylalanine that makes non‐polar interactions with the guanine base in H‐Ras. Instead, water‐mediated hydrogen bonding interactions were observed in Rad. The GDP molecule is located at the same position as in H‐Ras and adopts a similar conformation as that bound in H‐Ras. This similarity seems to be endowed by the conserved hydrogen bonding interactions with the guanine base‐recognition loops and the magnesium ion that has a typical octahedral coordination shell identical to that in H‐Ras.
From climate change to population change: the need to consider annual life cycles
Authors: ÅDAHL EMMA , LUNDBERG PER , JONZÉN NICLAS
Published: July 2006

Detailed studies of organisms' life cycles are important for understanding population response to climate change. However, in general one cannot make strong inference about the overall population response from such studies, unless the full annual cycle of the species in question is covered. Here, we present a theoretical framework for the understanding of population response to climate change. Owing to the combined effects of demography, intraspecific feedback, and a possible use of environmental cues, environmentally induced changes in survival and/or reproduction do not necessarily lead to a straightforward change in population size. This framework can guide our thinking about how abiotic conditions work their way to the population level. More specifically, it can help us to identify mechanisms that need to be examined when predicting population change in response to expected climate change.
Coral bleaching, reef fish community phase shifts and the resilience of coral reefs
Authors: BELLWOOD DAVID R. , HOEY ANDREW S. , ACKERMAN JOHN L. , DEPCZYNSKI MARTIAL
Published: July 2006

The 1998 global coral bleaching event was the largest recorded historical disturbance of coral reefs and resulted in extensive habitat loss. Annual censuses of reef fish community structure over a 12‐year period spanning the bleaching event revealed a marked phase shift from a prebleach to postbleach assemblage. Surprisingly, we found that the bleaching event had no detectable effect on the abundance, diversity or species richness of a local cryptobenthic reef fish community. Furthermore, there is no evidence of regeneration even after 5–35 generations of these short‐lived species. These results have significant implications for our understanding of the response of coral reef ecosystems to global warming and highlight the importance of selecting appropriate criteria for evaluating reef resilience.
The optimization of microwave‐assisted extraction of decursin from Angelica gigas Nakai root
Authors: Lee Gee‐Dong , Lee Seon‐Young , Kim Kwang‐Soo , Kwon Joong‐Ho
Published: July 2006

Response surface methodology was used to display the characteristics of microwave‐assisted extraction (MAE) of soluble solids and decursin from Angelica gigas Nakai root, thereby allowing MAE conditions to be optimized. These included ethanol concentration, microwave power and extraction time. The maximal solid yield was predicted to be 44.2% under conditions of 44.9% ethanol concentration, 115.6 W microwave power and 5.8 min extraction time. Decursin content of the extract was estimated to be maximal at 2.52% using 97.7% ethanol, 90.2 W microwave power, and 6.1 min extraction time. The estimated optimal MAE conditions (67% ethanol, 100 W and 6 min) to maximize both components were verified by comparing estimated values with experimental values (42.2% soluble solids, 2.2% decursin), which were higher than those (35.8% solids, 2.0% decursin) obtained by conventional reflux extraction (60% ethanol, 95 °C and 6 h).
Thermal, microstructural and textural characterisation of gelatinised corn, cassava and yam starch blends
Authors: Karam Laura B. , Ferrero Cristina , Martino Miriam N. , Zaritzky Noemi E. , Grossmann María Victoria E.
Published: July 2006

The functional properties of mixtures of maize, cassava and yam starches and their relationships with microstructural characteristics were investigated. Experiments were performed following the simplex‐centroid design with internal points and restrictions (upper limit) for yam starch proportion. The statistical model used (Scheffé canonical equation) was a powerful tool to predict the pastes behaviour within the limits of the experimental area. Polynomials with second level interactions were applied to obtain the surface response. Viscoelastic attributes of mixtures differed from those of individual starches. As shown by differential scanning calorimetry and microscopical observations, physical properties of the mixtures depended on the type of networks obtained. These networks varied upon yam, corn and cassava starch proportions as each one has a particular characteristic: gelatinisation temperature, granule size, swelling capacity and amylose/amylopectin ratio among others.
Prevalence and characterisation of Bacillus cereus in vacuum packed potato puree
Authors: Rajkovic Andreja , Uyttendaele Mieke , Courtens Tine , Heyndrickx Mark , Debevere Johan
Published: July 2006

Refrigerated processed foods of extended durability (REPFED) potato puree was analysed for Bacillus cereus contamination along the production line and during the product shelf‐life. Isolated B. cereus strains were tested for their psychrotrophic character and the ability to produce enterotoxins. Bacillus cereus contamination during four subsequent productions was in the range of 2.3–4.0 log cfu g−1. Productions five and six were significantly less contaminated with B. cereus (≤1 log cfu g −1). All B. cereus isolates from the first four productions were able to grow at 7 ° and 10 °C, whereas the majority of the isolates from productions five and six did not. No B. cereus isolates grew at 4 °C. randomly amplified polymorphic DNA (RAPD) fingerprinting showed that the most of B. cereus contamination originated from one source. In total, 30.4% of isolates expressed enterotoxic character. The present study points out the necessity to prevent an ‘in house’ colonisation and contamination during food processing in order to accomplish the safety of REPFED throughout the shelf‐life. It also indicates the most critical steps in the production line of ready‐to‐eat potato puree and impact of failures regarding the food safety. The data provided can be used for risk assessment studies regarding B. cereus in REPFED.
Development of a dipstick enzyme‐linked immunosorbent assay for the organophosphorus insecticide parathion‐methyl
Authors: Kim Min Jung , Shim Jee Youn , Lee Yong Tae , Lee Hye‐Sung
Published: July 2006

In our previous study, polyclonal antibodies against the organophosphorus insecticide parathion‐methyl were obtained and competitive enzyme‐linked immunosorbent assays (ELISAs) for this pesticide were developed. In this study, using these antibodies and an enzyme tracer, a direct competitive ELISA for parathion‐methyl in dipstick format was developed. The dipstick ELISA using Immunodyne membrane as a solid support, allowed the quick visual detection of parathion‐methyl in concentrations above 10 μg L−1. The IC50 value of the dipstick ELISA using reflectance detection was 8.8 μg L−1 with a detection limit of 0.3 μg L−1. The recoveries of parathion‐methyl from spiked lettuce and rice samples using the dipstick ELISA ranged from 89% to 106%.
Characterisation of ice cream containing flaxseed oil
Authors: Goh Kelvin K.T. , Ye Aiqian , Dale Nicola
Published: July 2006

Five batches of a 12% (w/w) fat ice cream formulation were prepared using different amounts of flaxseed oil (0%, 3%, 6%, 9% and 12% w/w) to replace milkfat. Meltdown rates, texture of ice cream, particle size of fat globules and microstructure of the ice cream were determined. Increasing the proportion of flaxseed oil in ice cream resulted in an increase in the meltdown rate and a decrease in the ice cream hardness. These were attributed to the low melting temperature of flaxseed oil and the varying extent of fat flocculation. In ice cream containing a high proportion of milkfat, the fat globules formed large particles largely due to flocculation rather than partial‐coalescence. The extent of fat flocculation decreased as the proportion of flaxseed oil was increased. Evidence of fat coalescence was observed in ice cream containing 12% flaxseed oil. The fat flocculates stabilised the air cells and strengthened the foam structure of the ice cream.
Changes in the volatile flavour components of Jinhua ham during the traditional ageing process
Authors: Zhang Jianhao , Wang Li , Liu Yuan , Zhu Jianhui , Zhou Guanghong
Published: July 2006

Changes in volatile flavour compounds of Jinhua ham were studied during the traditional ageing process of sun‐drying, loft‐ageing and post‐ageing. About seventy‐eight volatile compounds were tentatively identified, including aldehydes (ten to fifteen), carboxylic acids (eight to ten), alcohols (thirteen to fourteen), ketones (eight), esters (eight to ten), aromatic hydrocarbons (five), sulphur‐containing compounds (two to three) and pyrazines (zero to four) which were the main components. The percentage content of aldehydes and ketones reduced over time as the temperature rose with each stage of the process, whereas alcohols, carboxylic acids, esters, pyrazines and sulphur‐containing compounds increased gradually. Jinhua ham's high‐temperature maturating process (40 days at 30–37 °C) made the percentage content of carboxylic acids arising from Maillard reactions markedly increase (from sun‐dried ham 8.45% to fully aged ham 19.23%). This was the most obvious difference between Jinhua ham and the Mediterranean dry‐cured hams, and was the main factor leading to the unique flavour of Jinhua ham.
A phenomenological study of fatigue in patients with primary biliary cirrhosis
Authors: Jorgensen Roberta
Published: July 2006

Aim. This paper reports the findings of research on the lived experience of fatigue in patients with primary biliary cirrhosis – a type of chronic liver disease.

Background. Fatigue is a universal phenomenon, commonly associated with both acute and chronic illness, but also seen in healthy individuals. However, it is not often addressed in healthcare encounters despite its disabling nature, perhaps because of the multitude of contributing factors and the lack of effective treatments. It is a common and debilitating symptom in patients with primary biliary cirrhosis. Despite the prevalence of fatigue in this condition, patients’ experiences of this symptom have not been researched.

Method. An interpretive‐phenomenological approach developed by van Manen was chosen and this combines features of descriptive and interpretive phenomenology. Eight participants were interviewed during 2005 about their fatigue experience. Thematic analysis was conducted.

Findings. Five themes were identified: an unreliable body, fatigue as always present yet insidious, planning a life to conserve energy, struggling to maintain normality and emotional consequences. These themes reflected the unrelenting, intrusive nature of fatigue into the lives of those affected.

Conclusion. This research will provide empathic insight into the fatigue experience in this condition and generate communication about coping strategies. It will add to the body of research on fatigue in chronic conditions and may generate ideas for intervention research.
How and what university students learn through online and face‐to‐face discussion: conceptions, intentions and approaches
Authors: Ellis R. A. , Goodyear P. , Prosser M. , O'Hara A.
Published: July 2006

Abstract
This paper reports a phenomenographic investigation into students' experiences of learning through discussion – both online and face to face (F2F). The study context was a second‐year undergraduate course in psychology for social work in which the teacher had designed discussion tasks to begin in F2F mode and to continue online. A combination of open‐ended questionnaires and semi‐structured interviews was used to investigate students' conceptions of what they were learning, their intentions and their approaches to learning through discussion. Analysis of the interview and open‐ended questionnaire data identified a number of qualitatively different conceptions, intentions and approaches to learning through discussion. Associations were found between what students thought they were learning through discussions, their approaches to learning through discussion and their course grade. Students with a cohesive conception and students adopting a deep approach (to learning through online discussion) got better course grades. There was no significant difference between deep and surface approaches to F2F discussion and course grade. The outcomes of this study have implications for the design of online and F2F discussion tasks and in particular for helping students adopt richer conceptions of what they stand to gain through discussion.
Development of real‐time visual feedback assistance in singing training: a review
Authors: Hoppe D. , Sadakata M. , Desain P.
Published: July 2006

Abstract
Four real‐time visual feedback computer tools for singing lessons (Singad, Albert, Sing & See, and WinSINGAD), and the research carried out to evaluate the usefulness of these systems are reviewed in this article. We report on the development of user‐functions and the usability of these computer‐assisted learning tools. Both quantitative and qualitative studies confirm the efficiency of real‐time visual feedback in improving singing abilities. Having addressed these findings, we suggest further quantitative investigations of (1) the detailed effect of visual feedback on performance accuracy and on the learning process, and (2) the interactions between improvement of musical performance and the type of visual feedback and the amount of information it presents, the skill level of the user and the teacher's role.
The Patient Satisfaction Scale – an empirical investigation into the Finnish adaptation
Authors: Suhonen Riitta , Leino‐Kilpi Helena , Välimäki Maritta , Kim Hesook Suzie
Published: July 2006

Rationale and aims  The aim of this paper is to empirically investigate the performance characteristics of the Finnish adaptation PSS‐Fin of the Patient Satisfaction Scale (PSS) intended to measure patient satisfaction with nursing care. The PSS‐Fin includes three sub‐scales: technical‐scientific, informational and interaction/support care‐needs.

Methods  The PSS‐Fin was used in a cross‐sectional survey measuring the satisfaction of Finnish surgical patients (n = 454) with the nursing care they received. The feasibility, internal consistency, stability, equivalence and construct validity of the measure were investigated. Evaluation was based on statistical methods.

Results  The PSS is brief and easy to use, and it produced low missing data. Cronbach’s alpha coefficient ranged from 0.79 to 0.89 for the sub‐scales. The items correlated strongly with the sub‐scales and the sub‐scales with the total PSS as well. Test–retest reliability of 0.7 showed reasonable stability over time. Three factor analytic procedures supported for the three‐factor solution with a technical‐scientific, informational and interaction/support factors, explaining approximately 77% of the variance. The PSS had some equivalence with another satisfaction instrument, but also exhibited the ability to discriminate between each other. In multiple regression analysis the informational care‐needs sub‐scale was the most significant factor explaining patient satisfaction.

Conclusion  The PSS‐Fin demonstrated good psychometric properties and conceptual rigour and is thus reliable tool for examining patient satisfaction with nursing care.
Infectious pancreatic necrosis virus establishes an asymptomatic carrier state in kidney leucocytes of juvenile Atlantic cod, Gadus morhua L.
Authors: Garcia J , Urquhart K , Ellis A E
Published: July 2006

Juvenile Atlantic cod (10 g) were infected with infectious pancreatic necrosis virus (IPNV) by intraperitoneal injection and cohabitation. Fish showed no signs of disease but IPNV could be re‐isolated from kidney tissue for up to 12 weeks. On weeks 2, 5, 8, 10, 11 and 12 following infection, kidney leucocytes were fractionated on Percoll gradients, and cells separated into plastic adherent and non‐adherent cell populations after overnight incubation. IPNV was detectable in lysates of both cell populations and in supernatants by culture in CHSE‐214 cells. Wells containing 105−106 macrophages had an IPNV TCID50 of about 103/well and in serially diluted macrophages the minimum number of cells required to detect virus ranged from 101 to 104. These data indicate that about one in 104 macrophages were infected and the mean number of virus/infected cell was about 10. Replication of IPNV in the macrophages was low as the titre of the virus in macrophage lysates did not increase between days 1 and 3 of culturing the macrophages, but virus was released into the supernatant over this time.
Effects of anterior cruciate ligament (ACL) injury on postural control and muscle activity of head, neck and trunk muscles
Authors: TECCO S. , SALINI V. , CALVISI V. , COLUCCI C. , ORSO C. A. , FESTA F. , D'ATTILIO M.
Published: July 2006

summary The aim of the study was to evaluate the effects that an anterior cruciate ligament injury of the knee has on postural control and activity of neck, head and trunk muscles to investigate the existence of connections between the masticatory system and body posture. Surface electromyographic activity of the muscles at mandibular rest position, and during maximal voluntary clenching and posturometric and stabilometric measurements of 25 adult patients having pathology on the left knee were compared with a control non‐pathological group. At rest, the patients showed a higher muscular activity of anterior temporalis, masseter, sternocleidomastoid and lower trapezius, compared with the control subjects (P < 0·05). At maximal voluntary clenching, the patients showed a lower muscular activity of the right anterior temporalis and masseter and a higher muscular activity of the lower trapezius, compared with the control subjects. For the stabilometric measurements, all the subjects showed a significant reduction in the postural centre of pressure path length during the test with eyes open and cotton rolls, compared with the test with eyes closed and mandibular rest position (P < 0·05). In addition, the patients showed a significant displacement of the postural centre of pressure in a forward direction (P < 0·05) and into the right side (P < 0·05), compared with the control subjects. Anterior cruciate ligament injury appears to be associated to a change in the activity of head, neck and trunk muscles and to a change in the position of the postural centre of pressure. Cotton rolls seem to improve the stability of the subject.