bluuebunny/crossref_metadata_2025 huggingface dataset:
Streamlined Protocol for Antibody Staining of Large Numbers of Samples in Drosophila
Authors: Rebay Ilaria , Fehon Richard G.
Published: September 2008

INTRODUCTION

The most common reason for generating antibodies against a particular protein in Drosophila is to examine its pattern of endogenous expression during development. Thus, once an antibody is shown to stain transfected cells, the next step is to try staining Drosophila tissues, usually embryos (described here), imaginal discs, or ovaries. This streamlined protocol can be used for processing a large number of samples, e.g., screening monoclonal antibodies.
Human Lymphocyte Culture and Chromosome Analysis
Authors: Benn Peter , Delach Judith
Published: September 2008

INTRODUCTION

Phytohaemagglutinin (PHA), a lectin derived from the red kidney bean, is a powerful mitogen for human T-cells. When PHA is added in vitro to whole blood, mitotic cells can be found after 48 h, with a peak mitotic index at ~64-72 h. The convenience of peripheral blood as a source of human cells, the abundance of mitotic cells, and the simplicity of the cell culture technique make this the most convenient approach to study human chromosomes for both clinical and research purposes. This method of chromosome preparation provides metaphase cells that can be stained by a variety of methods or used for fluorescence in situ hybridization (FISH). The most common chromosome staining techniques involve exposing fixed preparations to a protease (e.g., trypsin), followed by an appropriate semipermanent stain. The characteristic banding patterns obtained reflect both structural and functional differences in different parts of the chromosomes. The staining procedure described here provides a Giemsa banding pattern using trypsin with Wright stain (i.e., GTW banding). This procedure is reliable and, with only minor modifications, suitable for preparing chromosomes from a variety of human tissues.
Preparation of Human Foreskin Fibroblasts for Human Embryonic Stem Cell Culture
Authors: Panula Sarita , Reijo Pera Renee A.
Published: September 2008

INTRODUCTION

In order to reduce the risk of immune rejection and zoonosis in human embryonic stem cell (hESC) transplantation recipients, animal-derived components, nonhuman sera, and animal feeder cells need to be removed from hESC culture. To limit the use of animal products, several different human feeder cell types have been tested for hESC culture. These include cells from fetal muscle and skin, as well as adult fallopian tube epithelial cells and adult muscle cells. Human foreskin fibroblasts (hFFs) have been used successfully as feeder cells to support the derivation and undifferentiated growth of hESCs and are commercially available through the American Type Culture Collection (ATCC). This protocol describes a method for preparing hFFs for hESC culture.
Culturing Human Embryonic Stem Cells in Feeder-Free Conditions: Figure 1.
Authors: McElroy Sohyun L. , Reijo Pera Renee A.
Published: September 2008

INTRODUCTION

Human embryonic stem cells (hESCs) have the potential to differentiate into all three germ layers and proliferate in long-term culture in vitro. hESCs can provide a cell source for the testing of novel therapies, drug screening, and functional genomics applications. Undifferentiated hESCs can be maintained and proliferated on mouse embryonic fibroblasts (MEFs) or human feeder cells. In this protocol, we describe the culture of hESCs in feeder-free conditions on Matrigel with MEF-conditioned medium. This protocol can be used for applications such as genetic modification of hESCs without feeder cell contamination.
Human Germ Cell Lineage Differentiation from Embryonic Stem Cells
Authors: Kee Kehkooi , Reijo Pera Renee A.
Published: September 2008

INTRODUCTION

Biological and ethical constraints hinder studies of human germ cell development despite its importance to reproductive health, including fertility and tumorigenesis. Thus, most of what we know of human germ cell development has been extrapolated from studies in model organisms. Human embryonic stem cells (hESCs) may provide an ideal system for probing the developmental genetics of germ cell formation and differentiation in vitro. The growth factors BMP (bone morphogenetic protein) 4, BMP7, and BMP8b are required for development of primordial germ cells (PGCs) in mice. It has been shown that these BMPs significantly increase germ cell differentiation from hESCs in vitro. This protocol describes a method to induce germ cell differentiation from hESCs by the addition of BMPs to hESC differentiation medium. The protocol can be used to study the basic mechanism of germ cell development in human cells.
Coimmunoprecipitation (co-IP) of Nuclear Proteins and Chromatin Immunoprecipitation (ChIP) from Arabidopsis
Authors: Fiil Berthe Katrine , Qiu Jin-Long , Petersen Klaus , Petersen Morten , Mundy John
Published: September 2008

INTRODUCTION

Transcriptional reprogramming occurs during development and in response to diverse stimuli and stresses. The isolation and characterization of nuclear proteins, particularly those binding to DNA and chromatin, are therefore important to understanding these processes. Two specific approaches to understanding the function of nuclear proteins involve the characterization of their protein-protein interactions, and of the transcriptional targets of specific transcription factors. Coimmunoprecipitation (co-IP) is a straightforward technique to study in vivo protein-protein interactions, and can identify interacting proteins or protein complexes present in cell extracts. Chromatin immunoprecipitation (ChIP) permits the identification of protein-DNA interactions in pull-down assays using specific antibodies against DNA-binding proteins, such as transcription factors or histone/chromatin-binding proteins. Here, we present detailed protocols for extraction of Arabidopsis seedlings, co-IP of nuclear proteins, and ChIP.
Is Histoplasma capsulatum a native inhabitant of Japan?
Authors: Kikuchi Ken , Sugita Takashi , Makimura Koichi , Urata Kensaku , Someya Takashi , Sasaki Takashi , Kamei Katsuhiko , Niimi Masakazu , Hiramatsu Keiichi , Uehara Yoshimasa
Published: September 2008

Histoplasmosis is an infectious disease caused by inhaling spores of the fungal pathogen H. capsulatum and in Japan is considered an imported mycosis. However, some patients in Japan with histoplasmosis have no history of traveling overseas nor of risk of occupational exposure to Histoplasma. To investigate the possibility of native distribution of Histoplasma in Japan, 187 bat guano samples from 67 bat‐inhabited caves in 17 prefectures were collected. These were examined for H. capsulatum by culture and Histoplasma‐specific PCR in three independent laboratories. No H. capsulatum was detected by either method, therefore H. capsulatum is unlikely to be present in bat guano in Japanese caves.
Comparative evaluation ofTrichosporon asahiisusceptibility using ASTY colorimetric microdilution and CLSI M27‐A2 broth microdilution reference methods
Authors: Kalkanci Ayse , Mekha Nanthawan , Poonwan Natteewan , Makimura Koichi , Sugita Takashi
Published: September 2008

Thein vitroactivity of AMPH‐B, 5‐FC, FLCZ, MCZ, ITCZ, and VRCZ against 50 isolates ofT. asahiiwas determined using CLSI M27‐A2 microdilution and ASTY colorimetric methods. Observed agreement ranged from 96 to 100% according to the drug. Overall, the agreement between two methods was 97.7%. The ASTY colorimetric method was thus determined to be comparable to the CLSI reference method when testing the susceptibility ofT.asahiito a variety of antifungal agents.
Enhancing effects of the chemical adjuvant levamisole on the DNA vaccine pVIR‐P12A‐IL18‐3C
Authors: Huijun Lu , Xiaowei Huo , Ying Zhang , Min Zheng , Mingxiao Ma , Hongyong Zhang , Minglan Jin , Guoshun Shen , Leili Jia , Yue Ji , Xu Li , Kuoshi Jin , Ningyi Jin
Published: September 2008

DNA‐based vaccination is an attractive alternative for overcoming the disadvantages of inactivated virus vaccines; however, DNA vaccines alone often generate only weak immune responses. In this study, the efficacy of LMS as a chemical adjuvant on a DNA vaccine (pVIR‐P12A‐IL18‐3C) encoding the P1‐2A and 3C genes of the FMDV and swine IL‐18, which provides protection against FMDV challenge, was tested. All test pigs were administered booster vaccinations 28 days after the initial inoculation, and were challenged with 1000 ID50 FMDV O/NY00 20 days after the booster vaccination. Positive and negative control groups were inoculated with inactivated virus vaccine and PBS respectively. The DNA vaccine plus LMS induced greater humoral and cell‐mediated responses than the DNA vaccine alone, as evidenced by higher concentrations of neutralizing and specific anti‐FMDV antibodies, and by higher concentrations of T‐lymphocyte proliferation and IFN‐γ production, respectively. FMDV challenge revealed that the DNA vaccine plus LMS provided higher protection than the DNA vaccine alone. This study demonstrates that LMS may be useful as an adjuvant for improving the protective efficiency of DNA vaccination against FMDV in pigs.
Chlamydia trachomatis and Chlamydophila abortus induce the expression of secretory leukocyte protease inhibitor in cells of the human female reproductive tract
Authors: Wheelhouse Nick , Wattegedera Sean , Fleming Diana , Fitch Paul , Kelly Rodney , Entrican Gary
Published: September 2008

C. trachomatis and C. abortus are related Gram‐negative intracellular bacteria that cause reproductive failure due to infertility (C. trachomatis) or abortion (C. abortus). These organisms target epithelial cells in the reproductive tract and/or placenta, but the innate immune mechanisms that lead to protection or pathology and disease are poorly understood. SLPI is an innate immune molecule which protects mucosal surfaces from infection and injury. C. trachomatis and C. abortus were found to induce SLPI mRNA and peptide expression in HeLa (cervical epithelium) and JEG‐3 cells (trophoblast) respectively. Both cell lines constitutively expressed SLPI and, although infection enhanced this expression, killed organisms did not. These data demonstrate that Chlamydia/Chlamydophila grow in cells that express SLPI, suggesting that SLPI does not exert antimicrobial effects against these organisms. However, SLPI has multiple functions, and we speculate that it may play a role in controlling tissue inflammation and pathology.
Chlamydia pneumoniae induces interleukin‐6 and interleukin‐10 in human gingival fibroblasts
Authors: Rizzo Antonietta , Paolillo Rossella , Lanza Alfonso Galeota , Guida Luigi , Annunziata Marco , Carratelli Caterina Romano
Published: September 2008

Chlamydia pneumoniae is an obligate intracellular Gram‐negative bacterium with a unique biphasic developmental cycle that can cause persistent infections. In humans, Chlamydia causes airway infection and has been implicated in chronic inflammatory diseases, such as asthma and atherosclerosis. In addition, recent studies demonstrated that patients with severe periodontitis can harbor C. pneumoniae, which can increase the risk for a host inflammatory response with weighty clinical sequelae. Previous studies have established that periodontal pathogenic bacteria (i.e. Gram‐negative bacteria) can induce the synthesis and release of cytokines and other inflammatory mediators in human gingival fibroblasts. HGF are resident cells of the periodontium that respond to receptor stimulation by producing a variety of substances including cytokines and growth factors. Our results demonstrate that after 48 hr of incubation with viable C. pneumoniae HGF showed a proliferative response, as seen by both colorimetric MTT assay and direct cell count (30% and 35%, respectively). In addition, HGF incubated with viable or UV light‐inactivated C. pneumoniae organisms showed an increase in the levels of IL‐6 and IL‐10, but not IL‐4; on the contrary, HGF infected with heat‐killed bacteria did not show a significant production of any of the cytokines considered. In conclusion, the present study suggests that C. pneumoniae may modulate the expression of IL‐6 and IL‐10 by human gingival fibroblasts. Further studies are warranted to clarify the molecular mechanisms of C. pneumoniae in the regulation of cytokine expression by host cells and to elaborate the relevant clinical implications.
Impaired removal of DNA interstrand cross‐link in Nijmegen breakage syndrome and Fanconi anemia, but not in BRCA‐defective group
Authors: Tsuchida Ken , Komatsu Kenshi
Published: September 2008

Human diseases characterized by a high sensitivity to DNA interstrand cross‐links (ICL) and predisposition to malignance include Nijmegen breakage syndrome (NBS) and Fanconi anemia (FA), which is further classified to three groups: (1) FA core‐complex group; (2) FA‐ID complex group; and (3) breast cancer (BRCA)‐defective group. The relationships between these four groups and the basic defect in ICL repair remain unclear. To study the details of ICL repair in NBS and FA, a highly sensitive PPB (psoralen–polyethylene oxide–biotin) dot blot assay was developed to provide sensitive quantitative measurements of ICL during the removal process. Studies utilizing this assay demonstrated a decreased rate of ICL removal in cells belonging to the FA core‐complex group (e.g. groups A and G) and FA‐ID complex group (group D2), while ICL removal was restored to normal levels after these cells were complemented with wt‐FANCA, wt‐FANCG and wt‐FANCD2. Conversely, FA‐D1 cells with a defective BRCA2 protein displayed normal ICL removal, although they were compromised with respect to recombination. This normal ICL removal rate in recombination‐deficient cells was confirmed by using XRCC3‐defective Chinese hamster cells, which are similarly compromised with respect to recombination and are sensitive to mitomycin C. The present study also showed that cells from patients with Nijmegen breakage syndrome were defective in ICL removal, while they were impaired in the recombination. These results indicate an obvious defect of FA and NBS in the ICL repair process, except in the BRCA‐defective group, and a separate step of recombination‐mediated repair pathway between the BRCA group and NBS. (Cancer Sci 2008; 99: 2238–2243)
Associations of inflammatory factors with glycaemic status among middle‐aged and older Chinese people
Authors: Ye Xingwang , Franco Oscar H. , Yu Zhijie , Li Huaixing , Hu Frank B. , Liu Huaiyu , Wang Xiangdong , Tang Haiying , Liu Yong , Chen Yan , Lin Xu
Published: September 2008

Objective  Low‐grade chronic inflammation is associated with risk for type 2 diabetes. We investigated the associations between inflammatory factors and glycaemic status in a middle‐aged and older Chinese population.

Design  A population‐based cross‐sectional study of 3289 residents aged 50–70 years from Beijing and Shanghai.

Measurements  C‐reactive protein (CRP), interleukin‐6 (IL‐6) and soluble tumour necrosis factor α‐receptor two (sTNFR2) were assayed.

Results  Comparing the highest with the lowest quartile of CRP, the odds ratio (OR) of diabetes was significantly higher in women [3·66 (95% CI 2·23–6·03)] than in men [1·51 (0·95–2·41)] (P for interaction = 0·004), while the increased OR for impaired fasting glucose (IFG) was only observed in women [OR 2·03 (1·44–2·84)] (P for interaction = 0·022), after adjustment for age, geographic location, education, lifestyle factors, family history of diabetes, and use of antibiotics, aspirin and lipid‐lowering drugs. The multiple‐adjusted OR of IL‐6 for diabetes was also higher in women [2·95 (1·78–4·90)] than in men [2·23 (1·39–3·59)] (P for interaction = 0·045). sTNFR2 was not associated with diabetes but inversely associated with IFG in men [OR 0·59 (0·41–0·85)] and women [OR 0·78 (0·56–1·09)] (P for interaction = 0·13). In addition, CRP was significantly associated with increased HbA1c in both genders within euglycaemia after multiple adjustments.

Conclusions  Inflammatory markers are closely associated with diabetes and IFG in Chinese people. These associations appear to be stronger in women than in men. Furthermore, plasma CRP is positively associated with HbA1c even in euglycaemic individuals.
Elevated cerebrospinal fluid (CSF) leptin in idiopathic intracranial hypertension (IIH): evidence for hypothalamic leptin resistance?
Authors: Ball A. K. , Sinclair A. J. , Curnow S. J. , Tomlinson J. W. , Burdon M. A. , Walker E. A. , Stewart P. M. , Nightingale P. G. , Clarke C. E. , Rauz S.
Published: September 2008

Objective  The aetiology of idiopathic intracranial hypertension (IIH) is not known, but its association with obesity is well‐recognized. Recent studies have linked obesity with abnormalities in circulating inflammatory and adiposity related cytokines. The aim of this study was to characterize adipokine and inflammatory cytokine profiles in IIH.

Design  Paired serum and cerebrospinal fluid (CSF) specimens were collected from 26 patients with IIH and compared to 62 control subjects. Samples were analysed for leptin, resistin, adiponectin, insulin, IL‐1β, IL‐6, IL‐8 (CXCL8), TNFα, MCP‐1 (CCL2), hepatocyte growth factor, nerve growth factor and PAI‐1 using multiplex bead immunoassays.

Results  CSF leptin was significantly higher in patients with IIH (P = 0·001) compared to controls after correction for age, gender and body mass index (BMI). In the control population, BMI correlated with serum leptin (r = 0·34; P = 0·007) and CSF leptin (r = 0·51; P < 0·0001), but this was not the case for the IIH population. Profiles of other inflammatory cytokines and adipokines did not differ between IIH patients and controls once anthropometric factors had been accounted for.

Conclusions  IIH was characterized by significantly elevated CSF leptin levels which did not correlate with BMI. We suggest that CSF leptin may be important in the pathophysiology of IIH and that obesity in IIH may occur as a result of hypothalamic leptin resistance.
Characterization of macroprolactin and assessment of markers of autoimmunity in macroprolactinaemic patients
Authors: Kavanagh‐Wright Lucille , Smith Thomas P. , Gibney James , McKenna T. Joseph
Published: September 2008

Objective  It has been reported that macroprolactin is a complex of PRL and an immunoglobulin G (IgG). This study further characterizes macroprolactin and evaluates for other markers of autoimmunity using a cohort of macroprolactinaemic sera.

Patients and normal subjects  Following treatment of hyperprolactinaemic sera (n = 58) with polyethylene glycol (PEG), PRL values fell from 524–13 546 mU/l (Range) to 452–8455 mU/l, while in macroprolactinaemic sera (n = 41), PRL concentration fell from 525–5747 to 98–378 mU/l (PEG treated normoprolactinaemic reference range, 68–230 mU/l in males, 70–390 mU/l in females).

Design  PRL was measured in sera prior to and following gel filtration chromatography, ultrafiltration, treatment with protein A‐sepharose, protein G‐sepharose, antihuman IgG‐agarose and sodium thiocyanate (NaSCN). The binding of radio‐labelled PRL in macroprolactinaemic sera was also measured. Sera were assayed for antithyroid and antinuclear antibodies. C‐reactive protein (CRP) and CD5 positive B cells were also measured. Comparisons were made between values obtained in normal, hyperprolactinaemic and macroprolactinaemic sera.

Results  Macroprolactinaemic sera indicated the presence of an IgG molecule and/or IgG fragments with one or more molecules of PRL. In 97% of the sera macroprolactin had a molecular weight of 204 kDa. Treatment of macroprolactinaemic sera with NaSCN caused dissociation of macroprolactin, releasing monomeric PRL. Macroprolactinaemic sera did not yield evidence of an increase in markers of autoimmunity when compared with hyperprolactinaemic or normal sera.

Conclusions  Comprehensive analysis of macroprolactin confirmed its composition as an IgG molecule or fragment with a PRL molecule. The occurrence of macroprolactin does not appear to be associated with autoimmunity.
Vitamin D deficiency and supplementation during pregnancy
Authors: Yu C. K. H. , Sykes L. , Sethi M. , Teoh T. G. , Robinson S.
Published: September 2008

Objective  Vitamin D is essential for skeletal health and prolonged deficiency results in infantile rickets and adult osteomalacia. The aim of this study is to determine the vitamin D status in pregnancy and to evaluate the effects of daily and of single‐dose vitamin D supplementation.

Design  A prospective randomized study at St Mary's Hospital London.

Patients A total of 180 women (Indian Asian, Middle Eastern, Black and Caucasian) were recruited at 27 weeks gestation and randomized into three treatment groups: a single oral dose of 200 000 IU vitamin D, a daily supplement of 800 IU vitamin D from 27 weeks until delivery and a no treatment group.

Measurements  Vitamin D (25‐hydroxyvitamin D), PTH and corrected calcium levels in mothers at 27 weeks and at delivery and cord 25‐hydroxyvitamin D and corrected calcium levels.

Results  The final maternal 25‐hydroxyvitamin D levels were significantly higher in the supplemented group [daily dose (median) 42 (IQR 31–76) nmol/l, stat dose (median) 34 (IQR 30–46) nmol/l vs. median 27 (IQR 27–39) nmol/l in the no treatment; P < 0·0001] and significantly fewer women with secondary hyperparathyroidism in the supplemented group (10% in daily dose vs. 12% in stat dose vs. 27% in the no treatment; P < 0·05). Cord 25‐hydroxyvitamin D levels were significantly higher with supplementation [daily dose median 26 (IQR 17–45) nmol/l, stat dose median 25 (IQR 18–34) nmol/l vs. median 17 (IQR 14–22) nmol/l in no treatment; P = 0·001].

Conclusion  Single or daily dose improved 25‐hydroxyvitamin D levels significantly. However, even with supplementation, only a small percentage of women and babies were vitamin D sufficient. Further research is required to determine the optimal timing and dosing of vitamin D in pregnancy.
Small, dense low‐density lipoproteins (LDL) are predictors of cardio‐ and cerebro‐vascular events in subjects with the metabolic syndrome
Authors: Rizzo Manfredi , Pernice Vincenzo , Frasheri Arian , Di Lorenzo Gabriele , Rini Giovam Battista , Spinas Giatgen A. , Berneis Kaspar
Published: September 2008

Objective  Small, dense low‐density lipoproteins (LDL) are a feature of the metabolic syndrome (MS) but their predictive role still remains to be established. We performed a 2‐year follow‐up study in 124 subjects with MS (63 ± 6 years), as defined by the American Heart Association/National Heart, Lung and Blood Institute guidelines, to assess clinical and biochemical predictors of cerebro‐ and cardio‐vascular events.

Methods and results  Beyond traditional cardiovascular risk factors, we measured LDL size and subclasses by gradient gel electrophoresis. Clinical events were registered in the 25% of subjects. At univariate analysis subjects with events had increased prevalence of elevated fasting glucose (P = 0·0117), smoking (P = 0·0015), family history of coronary artery disease (P = 0·0033) and higher levels of total‐ and LDL‐cholesterol (P = 0·0027 and P = 0·0023, respectively); LDL size was lower (P < 0·0001), due to reduced larger subclasses and increased small, dense LDL (all P < 0·0001). At multivariate analysis the following were independent predictors of events (univariate odd ratios were calculated): low HDL‐cholesterol (OR 15·4, P = 0·0238), elevated fasting glucose (OR 12·1, P = 0·0102), elevated small, dense LDL (OR 11·7, P = 0·0004), elevated blood pressure (OR 9·2, P = 0·0392), smoking (OR 4·8, P = 0·0054).

Conclusions  This is the first study that assessed the predictive role of small, dense LDL beyond traditional cardiovascular risk factors in subjects with MS.
Sediment Fe:PO4 ratio as a diagnostic and prognostic tool for the restoration of macrophyte biodiversity in fen waters
Authors: GEURTS JEROEN J. M. , SMOLDERS ALFONS J. P. , VERHOEVEN JOS T. A. , ROELOFS JAN G. M. , LAMERS LEON P. M.
Published: September 2008

1. Globally, freshwater wetlands, including fen waters, are suffering from biodiversity loss due to eutrophication, water shortage and toxic substances, and to mitigate these pressures numerous restoration projects have been launched. Water quality data are generally used to evaluate the chances of reestablishment of aquatic vegetation in fen waters and shallow peat lakes. Here we investigated whether sediment characteristics, which are less prone to fluctuate in time, would result in more reliable predictions.

2. To test if sediment characteristics can indeed be used not only for an easy and early diagnosis of nutrient availability and water quality changes in fen waters, but also for the prognosis of biodiversity response, we recorded the aquatic vegetation and collected surface water, sediment pore water and sediment samples in 145 fen waters in the Netherlands, Ireland and Poland.

3. Endangered macrophyte species were more closely related to surface water chemistry than common species in terms of occurrence and abundance. Sites featuring endangered species appeared to have significantly lower turbidity and pH, and lower concentrations of SO4, PO4, total phosphorus (TP) and NH4 than other sites.

4. PO4 and TP concentrations in the water layer increased markedly at PO4 concentrations above 5–10 μmol L−1 in the sediment pore water. High surface water PO4 and TP concentrations appeared to be SO4‐induced and only occurred below certain threshold values for pore water Fe:PO4 (3.5 mol mol−1) and total sediment Fe:P (10 mol mol−1).

5. Interestingly, the occurrence of endangered species also correlated strongly with sediment and sediment pore water ratios; the number of endangered species increased markedly at pore water Fe:PO4 ratios above 1 mol mol−1, whereas their actual abundance had the greatest increase at ratios above 10 mol mol−1. Additionally, endangered species seemed to be more sensitive to accumulation of potentially toxic substances such as sulphide and ammonium than non‐endangered species.

6. As an indicator of both biogeochemical processes and biodiversity, pore water Fe:PO4 ratios could be a valuable diagnostic and prognostic tool for the restoration of water quality and biodiversity in fen waters, e.g. for selecting the most promising sites for restoration and for optimization of restoration measures.
The disappearing act of factor VIII
Authors: LENTING P. J. , CHRISTOPHE O. D. , GUÉGUEN P.
Published: September 2008

Summary.  Factor VIII (FVIII) is a plasma protein critical to the haemostatic system. This notion is illustrated by the severe bleeding disorder that is associated with its functional absence, known as haemophilia A. In addition, several epidemiological studies have revealed an association between the presence of elevated levels of FVIII and thrombotic complications. In view of its relation to thrombotic and haemorrhagic disorders, it is not surprising that FVIII has gained wide attention from the research community in the previous decades. This research has led to a better understanding of not only the structural, functional and physiological aspects of this intriguing protein, but also of the pathogenesis of haemostatic defects associated with FVIII. In the present review, focus will be on the interaction between FVIII and surface receptors that are able to capture FVIII. These interactions are of importance for FVIII, as they may affect both function and survival of FVIII.
Signalling mechanisms regulating the activation of human eosinophils by mast‐cell‐derived chymase: implications for mast cell–eosinophil interaction in allergic inflammation
Authors: Wong Chun K. , Ng Sinnie S. M. , Lun Samantha W. M. , Cao Ju , Lam Christopher W. K.
Published: September 2008

Allergic diseases such as asthma and allergic dermatitis are associated with the degranulation of mast cells. Chymase, a mast‐cell‐specific protease, is the major component in mast cell granules that can induce eosinophil infiltration into inflammatory sites. We examined the immunopathological mechanisms for the activation of eosinophils by chymase in allergic inflammation. Cytokines were measured by cytometric bead array Flex Sets multiplex assay using flow cytometry and enzyme‐linked immunosorbent assay. Adhesion molecules, migration and intracellular signalling pathways were assessed by flow cytometry, Boyden chamber assay and Western blot, respectively. Chymase suppressed the apoptosis of eosinophils and induce the release of the cytokine interleukin‐6 (IL‐6) and chemokines CXCL8, CCL2 and CXCL1 by eosinophils dose‐dependently. It also up‐regulated the surface expression of adhesion molecule CD18 and stimulated the chemokinetic migration of eosinophils. The expressions of adhesion molecules, cytokines and chemokines, and chemokinetic migration were differentially regulated by the activation of extracellular signal‐regulated kinase, p38 mitogen‐activated protein kinase, Akt, Janus‐activated kinase and nuclear factor‐κB pathways. Chymase therefore plays a pivotal immunological role in the interaction between mast cells and eosinophils in allergic diseases such as allergic dermatitis by inducing adhesion molecule‐mediated chemokinetic migration and inflammatory cytokines and chemokines of eosinophils, through multiple intracellular signalling molecules and transcription factor. Our results therefore provide a further biochemical basis for the pathogenesis of allergic inflammation consequent on the interaction between mast cells and eosinophils, and give insight for the development of new therapies.
Applications of simulation technology in psychiatric mental health nursing education
Authors: BROWN J. F.
Published: September 2008

The purpose of this paper is to review the use of simulation in education across the health professionals, to describe the development and implementation of innovative simulation techniques for an undergraduate psychiatric mental‐health nursing course, and to identify lessons learned and future directions for successful simulation experiences in psychiatric nursing.
People with mental disabilities negotiating life in the risk society: a theoretical approach
Authors: ERIKSSON B. G. , HUMMELVOLL J. K.
Published: September 2008

Risk assessment and risk management owing to the potential threat from people with serious mental disorders living in the local community have been given much attention in psychiatric and mental healthcare research. Research connected to risks associated with living in the community for people with mental disabilities has also been published. This paper focuses on the situation of people with mental health problems and disabilities from a quite different perspective. In the light of the societal change towards what has been named the ‘risk society’, the consequences for vulnerable groups, especially people with mental disabilities, are discussed. Furthermore, the purpose is to shed light on how structural factors, like the process of individualization and ‘de‐population’ of societal institutions, are affecting the living conditions for people with mental disabilities – positively as well as negatively. The importance of trust in the risk society is highlighted, but also the important role that the local community might play in relation to people with mental disabilities. Finally, some potentialities and new roles that this development towards a risk society and increased individualization paradoxically can open up for people with mental disabilities are outlined.
Nurse staffing, bed numbers and the cost of acute psychiatric inpatient care in England
Authors: BOWERS L. , FLOOD C.
Published: September 2008

The aim of this analysis was to describe the composition, variability and factors associated with nurse staffing costs in acute psychiatric inpatient care. Numbers of acute inpatient beds in England have fallen, creating an occupancy crisis. Numbers of acute inpatient nursing staff are linked to quality of care. Variance in staffing and beds has considerable resource implications, but little is known about how these costs are structured. The sample comprised survey data from 136 wards in 26 NHS Trusts, matched with nationally available data on service levels, population and outcomes. The cost of providing acute inpatient care varied fivefold between different Trusts. This variation comprised of numbers of beds/population, numbers of nurses/beds and the proportion of nurses qualified. These variations were not fully accounted for by differing levels of social deprivation. Although service provision levels in London were higher, wide variation in costs existed in every region. Associations between nursing cost per bed and performance indicators were found. As investment in acute inpatient care varies widely, we need to know much more about the relationship of inputs to outputs, so that empirically based standard service levels can be defined.
Reflections on the notion of post‐natal depression following examination of the scoring pattern of women on the EPDS during pregnancy and in the post‐natal period
Authors: JOMEEN J. , MARTIN C. R.
Published: September 2008

The scoring pattern of case‐positive women on the Edinburgh Postnatal Depression Scale (EPDS) was explored over the antenatal and post‐natal period. Individual analysis of EPDS scores over the antenatal and post‐natal period revealed a picture of depressive symptom reporting inconsistent with the notion of a distinct and separate entity of post‐natal depression. A fundamental assumption that post‐natal depression has a typical pattern of comparatively higher post‐natal depressive symptomology may be illusory when individual symptom profiles are explored.
Mental illness: diagnostic title or derogatory term? (Attitudes towards mental illness) Developing a learning resource for use within a clinical call centre. A systematic literature review on attitudes towards mental illness
Authors: PUTMAN S.
Published: September 2008

With one in three people likely to experience mental health problems during their lifetime, it is paradoxical that stigma and negative attitudes towards mental illness are so prevalent in the UK today. This systematic literature review was completed to investigate what the most common negative attitudes towards mental illness are, and the most common recommendations made to address them. The findings were used to inform teaching resources used in an National Health Service Direct call centre. Guidelines for undertaking a systematic review, produced by the Centre for Reviews and Dissemination, were used. Terms were set and a search of electronic databases and peer‐reviewed academic journals was completed, from which 16 primary research papers (from the UK) were obtained and used. These were assessed, using evidence‐based critical appraisal tools, to obtain data pertinent to the original question. This paper describes the process, including a detailed account of the methodologies employed to gather and analyse relevant data. Put into context, alongside key drivers (e.g. government papers), the findings are presented and discussed, along with underlying theories, where appropriate. Recommendations for professional practice are then presented.
Promoting positive risk management: evaluation of a risk management panel
Authors: KALINIECKA H. , SHAWE‐TAYLOR M.
Published: September 2008

The purpose of this study was to evaluate whether a National Health Service Trust risk management panel (RMP) service was effective in promoting positive risk management by supporting mental health clinicians/teams in their management of people with mental health problems who presented with high risk of harm to themselves or to others. Fifteen mental health workers completed a questionnaire regarding their experience of the RMP. Content analysis revealed several themes. The majority of participants found the RMP helpful because of the support provided by shared case discussion and collective responsibility for risk management offered by the RMP. Medical records of the 23 individuals referred into the RMP were also examined. Trends in diagnoses, risks and RMP recommendations were identified. This study points to the importance of the organization's role alongside the individual clinician's in positive risk management and in supporting people with mental health problems using the recovery model. Improvements in developing a RMP are suggested.
An evaluation of a Mental Health Act educational resource
Authors: KETTLES A. M. , KIGER A. M. , PHILIP K. , CALDOW J.
Published: September 2008

The Mental Health (Care and Treatment) (Scotland) Act 2003 was developed with the participation of patients and carers. This Act is fundamentally different from all previous versions of the Mental Health Acts in the UK because it is based on Millan's 10 principles. Its objective is to ensure that people with mental disorder receive effective care and treatment. It is crucial that all staff who deal with persons with mental disorder are familiar with the Act and incorporate it into practice. This depends on the effectiveness of educational resources in supporting staff to develop their practice. The project presented here was commissioned by the National Health Service Education for Scotland (NES) to evaluate the accessibility and use of the educational resources produced for all frontline staff in Scotland. The aim was to evaluate the educational resources produced by NES to prepare the frontline staff in relation to the Mental Health (Care and Treatment) (Scotland) Act 2003. Evaluation was conducted through a questionnaire survey, focus groups and individual interviews. The NES educational resource was effective in assisting professionals in implementing the complex and progressive requirements of the new Mental Health Act. The three formats reached a wide audience, and are an effective model for future initiatives.
Introduction of a child and adolescent mental health link worker: education and health staff focus group findings
Authors: HUNTER A. , PLAYLE J. , SANCHEZ P. , CAHILL J. , McGOWAN L.
Published: September 2008

Policy guidance suggests that outcomes for adolescents with mental health problems can be improved by secondary education services (SES) and child and adolescent mental health services (CAMHS) working more closely. This study reports on staff experiences of the introduction of a mental health link worker (MHLW). The findings of two focus groups are presented, conducted with staff from CAMHS and SES. These focus groups formed part of the overall wider evaluation of the MHLW role. The groups explored staff perceptions and experiences following the introduction of the MHLW, and elicited their views on the effectiveness of this innovative role. Qualitative methods were employed, and analysis was conducted using the principles of grounded theory and the constant comparative method. The findings revealed that the MHLW was well received by both groups, despite the identification of potential barriers. A number of key themes emerged, which included the ability of the link worker to improve communication and to encourage mutual understanding between services. The issues raised by these themes are discussed and recommendations are made for future practice and research.
Social constraint and an absence of sex‐biased dispersal drive fine‐scale genetic structure in white‐winged choughs
Authors: BECK N. R. , PEAKALL R. , HEINSOHN R.
Published: September 2008

This study used eight polymorphic microsatellite loci to examine the relative effects of social organization and dispersal on fine‐scale genetic structure in an obligately cooperative breeding bird, the white‐winged chough (Corcorax melanorhamphos). Using both individual‐level and population‐level analyses, it was found that the majority of chough groups consisted of close relatives and there was significant differentiation among groups (FST = 0.124). However, spatial autocorrelation analysis revealed strong spatial genetic structure among groups up to 2 km apart, indicating above average relatedness among neighbours. Multiple analyses showed a unique lack of sex‐biased dispersal. As such, choughs may offer a model species for the study of the evolution of sex‐biased dispersal in cooperatively breeding birds. These findings suggest that genetic structure in white‐winged choughs reflects the interplay between social barriers to dispersal resulting in large family groups that can remain stable over long periods of times, and short dispersal distances which lead to above average relatedness among neighbouring groups.
High connectivity on a global scale in the pelagic wahoo, Acanthocybium solandri (tuna family Scombridae)
Authors: THEISEN T. C. , BOWEN B. W. , LANIER W. , BALDWIN J. D.
Published: September 2008

The population genetic structure and phylogeography of wahoo, Acanthocybium solandri, were investigated on a global scale with intron six of lactate dehydrogenase‐A (ldhA6, 8 locations, N = 213) and mtDNA cytochrome b (Cytb, 10 locations, N = 322). Results show extensive sharing of haplotypes across the wahoo's entire global range, and analyses were unable to detect significant structure (nuclear FST = 0.0125, P = 0.106; mtDNA ΦST < 0.0001, P = 0.634). Power analyses indicated 95% confidence in detecting nuclear FST ≥ 0.0389 and mtDNA ΦST ≥ 0.0148. These findings appear unique, as most other tunas, billfishes, and oceanic sharks exhibit significant population structure on the scale of East–West Atlantic, Atlantic vs. Indian‐Pacific, or East–West Pacific. Overall nuclear heterozygosity (H = 0.714) and mtDNA haplotype diversity (h = 0.918) are both high in wahoo, while overall mtDNA nucleotide diversity (π= 0.006) and nuclear nucleotide diversity (π=0.004) are uniformly low, indicating a recent increase in population size. Coalescence analyses yield an estimate of effective female population size (NeF) at ~816 000, and a population bottleneck ~690 000 years ago. However, conclusions about population history from our Cytb data set are not concordant with a control region survey, a finding that will require further investigation. This is the first example of a vertebrate with a single globally distributed population, a finding we attribute to extensive dispersal at all life stages. The indications of a worldwide stock for wahoo reinforce the mandate for international cooperation on fisheries issues.
Hitching a lift on the road to speciation
Authors: SMADJA CAROLE , GALINDO JUAN , BUTLIN ROGER
Published: September 2008

Understanding how speciation can take place in the presence of homogenizing gene flow remains a major challenge in evolutionary biology. In the early stages of ecological speciation, reproductive isolation between populations occupying different habitats is expected to be concentrated around genes for local adaptation. These genomic regions will show high divergence while gene exchange in other regions of the genome should continue relatively unimpaired, resulting in low levels of differentiation. The problem is to explain how speciation progresses from this point towards complete reproductive isolation, allowing genome‐wide divergence. A new study by Via and West (2008) on speciation between host races of the pea aphid, Acyrthosiphon pisum, introduces the mechanism of ‘divergence hitchhiking’ which can generate large ‘islands of differentiation’ and facilitate the build‐up of linkage disequilibrium, favouring increased reproductive isolation. This idea potentially removes a major stumbling block to speciation under continuous gene flow.
Alteration in vaginal microflora, douching prior to pregnancy, and preterm birth
Authors: Thorp John M. , Dole Nancy , Herring Amy H. , McDonald Thaddeus L. , Eucker Barbara , Savitz David A. , Kaczor Diane
Published: September 2008

We aimed to determine the relationship of douching prior to pregnancy and bacterial vaginosis (BV) during pregnancy on preterm birth, addressing individual and joint effects. We used a prospective cohort study and assessed vaginal microflora using gram stains and Nugent's criteria. Douching behaviour was based on self‐report about the 12 months prior to pregnancy. Preterm births were categorised as spontaneous or medically indicated. A total of 2561 women provided vaginal specimens and 1492 provided self‐reports on douching behaviour.

Bacterial vaginosis assessed at 24–28 weeks' gestation in the absence of douching prior to pregnancy was associated with spontaneous preterm birth (odds ratio = 2.74 [95% confidence interval 1.13, 6.66]) as was douching in the absence of BV (OR = 2.20 [1.29, 3.75]). The combination of BV and douching was unrelated to spontaneous or medically indicated preterm birth. We concluded that acute alterations in vaginal microflora at mid‐pregnancy or douching prior to pregnancy were associated with an increased risk of preterm birth, but the combination did not appear to increase the risk further than would be expected.
Essential role of MYB transcription factor: PvPHR1 and microRNA: PvmiR399 in phosphorus‐deficiency signalling in common bean roots
Authors: VALDÉS‐LÓPEZ OSWALDO , ARENAS‐HUERTERO CATALINA , RAMÍREZ MARIO , GIRARD LOURDES , SÁNCHEZ FEDERICO , VANCE CARROLL P. , LUIS REYES JOSÉ , HERNÁNDEZ GEORGINA
Published: September 2008

Phosphorus (P), an essential element for plants, is one of the most limiting nutrients for plant growth. A few transcription factor (TF) genes involved in P‐starvation signalling have been characterized for Arabidopsis thaliana and rice. Crop production of common bean (Phaseolus vulgaris L.), the most important legume for human consumption, is often limited by low P in the soil. Despite its agronomic importance, nothing is known about transcriptional regulation in P‐deficient bean plants. We functionally characterized the P‐deficiency‐induced MYB TF TC3604 (Dana Farber Cancer Institute, Common Bean Gene Index v.2.0), ortholog to AtPHR1 (PvPHR1). For its study, we applied RNAi technology in bean composite plants. PvPHR1 is a positive regulator of genes implicated in P transport, remobilization and homeostasis. Although there are no reports on the regulatory roles of microRNAs (miRNA) in bean, we demonstrated that PvmiR399 is an essential component of the PvPHR1 signalling pathway. The analysis of DICER‐like1 (PvDCL1) silenced bean composite plants suppressed for accumulation of PvmiR399 and other miRNAs suggested that miR399 is a negative regulator of the ubiquitin E2 conjugase: PvPHO2 expression. Our results set the basis for understanding the signalling for P‐starvation responses in common bean and may contribute to crop improvement.
The rapid light response of leaf hydraulic conductance: new evidence from two experimental methods
Authors: SCOFFONI CHRISTINE , POU ALÍCIA , AASAMAA KRÕÕT , SACK LAWREN
Published: September 2008

Previous studies have shown a rapid enhancement in leaf hydraulic conductance (Kleaf) from low to high irradiance (from <10 to >1000 µmol photons m−2 s−1), using the high‐pressure flow meter (HPFM), for 7 of 14 tested woody species. However, theoretical suggestions have been made that this response might arise as an artifact of the HPFM. We tested the Kleaf light response for six evergreen species using refined versions of the rehydration kinetics method (RKM) and the evaporative flux method (EFM). We found new evidence for a rapid, 60% to 100% increase in Kleaf from low to high irradiance for three species. In the RKM, the leaf rehydration time constant declined by up to 70% under high irradiance relative to darkness. In the EFM, under higher irradiance, the flow rate increased disproportionately to the water potential gradient. Combining our data with those of previous studies, we found that heterobaric species, i.e. those with bundle sheath extensions (BSEs) showed a twofold greater Kleaf light response on average than homobaric species, i.e. those without BSEs. We suggest further research to characterize this substantial dynamic at the nexus of plant light‐ and water‐relations.