bluuebunny/crossref_metadata_2025 huggingface dataset:
Research ethics committees: agents of research policy?
Authors: Hemminki Elina
Published: October 2005

The purpose of this commentary is to describe the unintended effects ethics committees may have on research and to analyse the regulatory and administrative problems of clinical trials.

Discussion

The Finnish law makes an arbitrary distinction between medical research and other health research, and the European Union's directive for good clinical trials further differentiates drug trials. The starting point of current rules is that clinical trials are lesser in the interest of patients and society than routine health care. However, commercial interests are not considered unethical. The contrasting procedures in research and normal health care may tempt physicians to continue introducing innovations into practice by relying on unsystematic and uncontrolled observations. Tedious and bureaucratic rules may lead to the disappearance of trials initiated by researchers. Trying to accommodate the special legislative requirements for new drug trials into more complex interventions may result in poor designs with unreliable results and increased costs. Meanwhile, current legal requirements may undermine the morale of ethics committee members.

Conclusion

The aims and the quality of the work of ethics committees should be evaluated, and a reformulation of the EU directive on good clinical trials is needed. Ethical judgement should consider the specific circumstance of each trial, and ethics committees should not foster poor research for legal reasons.
Cell type-specific characterization of nuclear DNA contents within complex tissues and organs
Authors: Zhang Changqing , Gong Fang Cheng , Lambert Georgina M , Galbraith David W
Published: October 2005

Background

Eukaryotic organisms are defined by the presence of a nucleus, which encloses the chromosomal DNA, and is characterized by its DNA content (C-value). Complex eukaryotic organisms contain organs and tissues that comprise interspersions of different cell types, within which polysomaty, endoreduplication, and cell cycle arrest is frequently observed. Little is known about the distribution of C-values across different cell types within these organs and tissues.

Results

We have developed, and describe here, a method to precisely define the C-value status within any specific cell type within complex organs and tissues of plants. We illustrate the application of this method to Arabidopsis thaliana, specifically focusing on the different cell types found within the root.

Conclusion

The method accurately and conveniently charts C-value within specific cell types, and provides novel insight into developmental processes. The method is, in principle, applicable to any transformable organism, including mammals, within which cell type specificity of regulation of endoreduplication, of polysomaty, and of cell cycle arrest is suspected.
Quantification of the response of circulating epithelial cells to neodadjuvant treatment for breast cancer: a new tool for therapy monitoring
Authors: Pachmann Katharina , Camara Oumar , Kavallaris Andreas , Schneider Uwe , Schünemann Stefanie , Höffken Klaus
Published: October 2005

Introduction

In adjuvant treatment for breast cancer there is no tool available with which to measure the efficacy of the therapy. In contrast, in neoadjuvant therapy reduction in tumour size is used as an indicator of the sensitivity of tumour cells to the agents applied. If circulating epithelial (tumour) cells can be shown to react to therapy in the same way as the primary tumour, then this response may be exploited to monitor the effect of therapy in the adjuvant setting.

Method

We used MAINTRAC® analysis to monitor the reduction in circulating epithelial cells during the first three to four cycles of neoadjuvant therapy in 30 breast cancer patients.

Results

MAINTRAC® analysis revealed a patient-specific response. Comparison of this response with the decline in size of the primary tumour showed that the reduction in number of circulating epithelial cells accurately predicted final tumour reduction at surgery if the entire neoadjuvant regimen consisted of chemotherapy. However, the response of the circulating tumour cells was unable to predict the response to additional antibody therapy.

Conclusion

The response of circulating epithelial cells faithfully reflects the response of the whole tumour to adjuvant therapy, indicating that these cells may be considered part of the tumour and can be used for therapy monitoring.
Increased CXCR4-dependent HIV-1 fusion in activated T cells: role of CD4/CXCR4 association
Authors: Zaitseva Marina , Romantseva Tatiana , Manischewitz Jody , Wang Jiun , Goucher David , Golding Hana
Published: October 2005

Activation of peripheral CD4+ T cells resulted in augmented fusion with X4 human immunodeficiency virus type 1 (HIV-1) envelope-expressing cells without parallel increases in the surface expression of CD4 or CXC chemokine receptor 4 (CXCR4). Our study used biochemical methods and biological assays to correlate the increased fusion potential of activated T cells with changes in CXCR4 isoforms and CD4-CXCR4 association. Western blot analyses of CXCR4, precipitated from resting T cells, identified several CXCR4 species with molecular weights of 47, 50, 62, and 98 kDa. After 24 h stimulation with phytohemagglutinin/interleukin-2, a marked reduction was seen in the 47-kDa, with a concomitant increase in the amounts of 50 and 62–64 kDa CXCR4. T cell activation also induced an increase in the coprecipitation of CXCR4 with CD4. The 62-kDa CXCR4 predominantly coprecipitated with CD4 and was shown to be ubiquitinated. Stripping of CD4 from the cell surface with pronase treatment prior to cell lysis only partially reduced coprecipitation of CD4 with the 62-kDa CXCR4, revealing a pool of intracellular CD4-CXCR4 complexes. Coprecipitation of CXCR4 with CD4 was reduced in activated cells treated with Brefeldin A and Monensin, suggesting that late endosomes play a role in intracellular association of CXCR4 with CD4. Confocal microscopy confirmed the colocalization of CD4 and CXCR4 within CD63+ endocytic compartments. These findings demonstrated a correlation between the enhanced susceptibility of activated T cells to HIV-1 fusion and accumulation of ubiquitinated 62–64 kDa CXCR4 species, which preferentially associated with CD4. The CD4-CXCR4 complexes may shuttle between late endosomes and the cell surface.
Decreased clearance ofPseudomonas aeruginosafrom airways of mice deficient in lysozyme M
Authors: Cole Alexander M , Thapa Dharma R , Gabayan Victoria , Liao Hsiang-I , Liu Lide , Ganz Tomas
Published: October 2005

Lysozyme is a ubiquitous and abundant, cationic, antimicrobial polypeptide of leukocytes and epithelia, but its biological function in host defense is largely unexplored. To ascertain the role of lysozyme during bacterial infection of murine airways, we exposed the airways of lysozyme M-deficient (lys M−/−) mice to the pulmonary pathogen Pseudomonas aeruginosa and examined the host's response to infection. Despite partial compensation as a result of the appearance of lysozyme P in the infected airways of lys M−/− mice, these lys M−/− mice showed decreased clearance of P. aeruginosa compared with their lys M+/− or lys M+/+ littermates. Lysozyme contributes to optimal clearance of P. aeruginosa from the murine airways.
Splenic macrophages and B cells mediate immunosuppression following abrupt withdrawal from morphine
Authors: Rahim Rahil T , Meissler Joseph J , Adler Martin W , Eisenstein Toby K
Published: October 2005

We have previously shown that abrupt withdrawal (AW) from morphine induces greater than 80% immunosuppression in murine spleen cells, as assessed by the capacity to mount an in vitro plaque-forming cell response to sheep red blood cells. Present studies about the mechanisms of immunosuppression following AW showed that addition of highly enriched (CD11b+) splenic macrophages (obtained by cell sorting or magnetic separation) from AW mice to cultures of normal, unfractionated spleen cells suppressed immune responses. Further, addition of highly enriched (CD19+) B cells (but not T cells) from AW mice to normal cells was also immunosuppressive. B cells from AW mice were also able to inhibit the proliferative response of normal spleen cells to concanavalin A but not to lipopolysaccharide. Overall, the data suggest that immunosuppression by AW spleen cells is a result of active suppression by macrophages and B cells.
Differential transcription of Eomes and T-bet during maturation of mouse uterine natural killer cells
Authors: Tayade Chandrakant , Fang Yuan , Black Gordon P , VA Paffaro , Erlebacher Adrian , Croy B Anne
Published: October 2005

During human and rodent uterine decidualization, transient but abundant numbers of uterine natural killer (uNK) cells appear, proliferate, and differentiate. uNK cells share features with peripheral NK cells but are specialized to promote interferon-γ (IFN-γ)-mediated, pregnancy-associated, structural changes in maternal placental arteries. In CD8+ T cells and NK cells, the transcription factors T-bet and eomesodermin (Eomes) regulate maturation and effector functions, including IFN-γ production. No studies are reported for uNK cells. Implantation sites in T-bet null mice, which have a defect in NK cell maturation, had uNK cells normal in morphology and number and normally modified spiral arteries. As Eomes null mice are not viable, real-time polymerase chain reaction comparisons between C57Bl/6J (B6) and alymphoid (Rag20/0γc0/0) mice were used to assess uNK cell expression of T-bet, Eomes, and the target genes IFN-γ, granzyme A, and perforin. Gestation dated (gd) uterine tissues (mixed cell composition) and 200 morphologically homogeneous, laser-capture, microdissected uNK cells of different maturation stages were used. In uterus, Eomes transcripts greatly outnumbered those of T-bet, whether donors were nonpregnant or pregnant, and increased to gd10. In uNK cells, transcripts for T-bet, Eomes, and IFN-γ were most abundant in mature stage cells, and transcripts for granzyme A and perforin were lower at this stage than in immature or senescent cells. Thus, Eomes dominance to T-bet discriminates regulation of the uNK cell subset from that observed for peripheral NK cells.
Local anesthetics inhibit priming of neutrophils by lipopolysaccharide for enhanced release of superoxide: suppression of cytochrome b558 expression by disparate mechanisms
Authors: Jinnouchi Akio , Aida Yoshitomi , Nozoe Kohji , Maeda Katsumasa , Pabst Michael J
Published: October 2005

Local anesthetics have anti-inflammatory effects in vivo and inhibit neutrophil functions in vitro, but how these agents act on neutrophils remains unclear. Phagocytosis and bactericidal activity of neutrophils are enhanced by exposure to bacterial components such as lipopolysaccharide (LPS); this process is termed priming, which for enhanced release of superoxide (O2−) causes mobilization of intracellular granules that contain cytochrome b558, a component of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. We studied whether local anesthetics affected LPS priming for enhanced release of O2− in response to triggering by the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP), and we investigated which element in the LPS signaling pathway might be the target of local anesthetics. Neutrophils were incubated with 10 ng/ml LPS and 1% plasma ± local anesthetics, washed, and triggered with fMLP. Local anesthetics all inhibited LPS priming, and 50% inhibition was at 0.1 mM tetracaine, 0.5 mM bupivacaine, 3.0 mM lidocaine, or 4.0 mM procaine. Local anesthetics inhibited LPS-induced mobilization of specific granules and secretory vesicles. Local anesthetics inhibited LPS-induced up-regulation of cytochrome b558 but not LPS-induced translocation of p47phox. Inhibition of priming by local anesthetics was reversed by washing and incubating for 5 min. Tetracaine alone, but not the other local anesthetics, inhibited LPS activation of p38 mitogen-activated protein kinase (MAPK) and MAPK kinase 3 (kinases in the LPS signaling pathway). The p38 MAPK inhibitors SB203580 and PD169316 also blocked LPS priming. Thus, tetracaine and the other local anesthetics inhibit by disparate mechanisms, but all the local anesthetics impaired up-regulation of cytochrome b558 and all impaired priming of NADPH oxidase by LPS.
Cannabinoid receptors in microglia of the central nervous system: immune functional relevance
Authors: Cabral G A , Marciano-Cabral F
Published: October 2005

Microglia, resident macrophages of the brain, function as immune effector and accessory cells. Paradoxically, they not only play a role in host defense and tissue repair but also have been implicated in a variety of neuropathological processes. Microglia, in addition to exhibiting phenotypic markers for macrophages, express CB1 and CB2 cannabinoid receptors. Recent studies suggest the existence of a third, yet-to-be cloned, non-CB1, non-CB2 cannabinoid receptor. These receptors appear to be functionally relevant within defined windows of microglial activation state and have been implicated as linked to cannabinoid modulation of chemokine and cytokine expression. The recognition that microglia express cannabinoid receptors and that their activation results in modulation of select cellular activities suggests that they may be amenable to therapeutic manipulation for ablating untoward inflammatory responses in the central nervous system.
S6 kinase 2 potentiates interleukin-3-driven cell proliferation
Authors: Cruz Rebecca , Hedden Lee , Boyer Derek , Kharas Michael G , Fruman David A , Lee-Fruman Kay K
Published: October 2005

Interleukin-3 (IL-3) mediates hematopoietic cell survival and proliferation via several signaling pathways such as the Janus kinase/signal transducer and activator of transcription pathway, mitogen-activated protein kinase (MAPK) pathway, and phosphoinositide-3 kinase (PI-3K) pathway. Mammalian target of rapamycin (mTOR) is one of the downstream targets of the PI-3K pathway, and it plays an important role in hematopoiesis and immune cell function. To better elucidate how mTOR mediates proliferation signals from IL-3, we assessed the role of S6 kinase 2 (S6K2), one of the downstream targets of mTOR, in IL-3 signaling. We show that S6K2 is activated by IL-3 in the IL-3-dependent Ba/F3 cell line and that this is mediated by mTOR and its upstream activator PI-3K but not by the MAPK kinase/extracellular signal-regulated kinase pathway. S6K2 is also activated in primary mouse bone marrow-derived mast cells upon IL-3 stimulation. Expression of a rapamycin-resistant form of S6K2, T388E, in Ba/F3 cells provides a proliferation advantage in the absence or presence of rapamycin, indicating that S6K2 can potentiate IL-3-mediated mitogenic signals. In cells expressing T388E, rapamycin still reduces proliferation at all doses of rapamycin, showing that mTOR targets other than S6K2 play an important role in IL-3-dependent proliferation. Cell-cycle analysis shows that T388E-expressing Ba/F3 cells enter S phase earlier than the control cells, indicating that the proliferation advantage may be mediated by a shortened G1 phase. This is the first indication that S6K2 plays a role in IL-3-dependent cell proliferation.
Cytokine responses and sudden infant death syndrome: genetic, developmental, and environmental risk factors
Authors: Blackwell C Caroline , Moscovis Sophia M , Gordon Ann E , Al Madani Osama M , Hall Sharron T , Gleeson Maree , Scott Rodney J , Roberts-Thomson June , Weir Donald M , Busuttil Anthony
Published: October 2005

Despite the success of the campaigns to reduce the risk of sudden infant death syndrome (SIDS), it still remains the major cause of postneonatal mortality. The incidence of SIDS is higher among ethnic groups in which there are also high incidences of serious infectious diseases. The risk factors for SIDS parallel those for susceptibility to infection, and recent data have provided evidence to support the mathematical model of the common bacterial toxin hypothesis. One current hypothesis for the etiology of SIDS is that the deaths are a result of overwhelming proinflammatory responses to bacterial toxins; as in inflammatory responses to sepsis, cytokines, induced by bacterial toxins, cause physiological changes leading to death. The genetic, developmental, and environmental risk factors for SIDS are reviewed in relation to colonization by potentially harmful bacteria and the inflammatory responses induced in the nonimmune infant to microorganisms or their products.
HMGN2: a novel antimicrobial effector molecule of human mononuclear leukocytes?
Authors: Feng Yun , Huang Ning , Wu Qi , Wang Boyao
Published: October 2005

Leukocytes are a central cellular element of innate-immune defense in mammals. In addition to the generation of toxic oxygen radicals and nitric oxide, leukocytes express and secrete a broad array of antimicrobial proteins and peptides. In the study, an antimicrobial polypeptide was isolated and purified from human peripheral blood mononuclear leukocytes in the presence of interleukin (IL)-2. Microsequencing provided that its N-terminal amino sequence was PKRKAEGDAK, which was identical to high mobility group nucleosomal-binding domain 2 (HMGN2). Mass spectrometric value and Western blot also indicated its individual character of HMGN2. The antimicrobial assays showed that the Escherichia coli-based production of HMGN2 had a potent antimicrobial activity against E. coli ML-35p, Pseudomonas aeruginosa ATCC 27853, and to some extent, against Candida albicans ATCC 10231. The HMGN2 α-helical domain had the same antimicrobial activity as HMGN2. The immunocytochemistry staining, enzyme-linked immunosorbent assay, and Western blot revealed that HMGN2 was present in the cytoplasm of mononuclear leukocytes and released to the extracellular environment when stimulated with IL-2. These results suggest that HMGN2 would be a novel antimicrobial effector molecule of human mononuclear leukocyte.
Splenic PGE2-releasing macrophages regulate Th1 and Th2 immune responses in mice treated with heat-killed BCG
Authors: Shibata Yoshimi , Henriksen Ruth Ann , Honda Ikuro , Nakamura Reiko M , Myrvik Quentin N
Published: October 2005

Hosts infected with low doses of mycobacteria develop T helper cell type 1 (Th1) immunity, but at relatively higher doses, a switch to Th2 immunity occurs. Prostaglandin E2 (PGE2) is a proposed mediator of the Th1-to-Th2 shift of immune responses, and mycobacterial products induce PGE2-releasing macrophages (PGE2-MØ) in the mouse spleen in a dose-dependent manner. Splenic PGE2-M Ø from Balb/c mice, given 0.01 or 1 mg heat-killed (HK) Mycobacterium bovis bacillus Calmette-Guerin (BCG) intraperitoneally (i.p.), were characterized by the ex vivo release of PGE2 (>10 ng/106 cells), cytokine production, and expression of PGG/H synthase (PGHS)-1, PGHS-2, cytosolic PGE synthase (PGES), and microsomal PGES-1. At Day 14 after the treatment, mice treated with 1 mg, but not 0.01 mg, BCG had increased levels of PGHS-2+ PGE2-MØ, total serum immunoglobulin E (IgE), and serum IgG1 antibodies (Th2 responses) against heat shock protein 65 and purified protein derivative. Cultures of spleen cells isolated from these mice expressed interleukin (IL)-4 and IL-10 in recall responses. Treatment of mice receiving 1 mg BCG with NS-398 (a PGHS-2 inhibitor, 10 mg/kg i.p., daily) resulted in enhanced interferon-γ (IFN-γ) production with reduced IL-4 and IL-10 production in recall responses. This treatment also resulted in decreased total serum IgE levels. Treatment of C57Bl/6 mice with HK-BCG (0.5 mg dose) also induced a mixture of Th1 and Th2 responses, although IFN-γ production was markedly increased, and IL-4 was decreased compared with Balb/c mice. Thus, our results indicate that by 14 days following treatment of mice with high doses of HK-BCG, splenic PGE2-MØ formation is associated with a PGHS-2-dependent shift from Th1-to-Th2 immune responses.
Cloning, expression, and functional characterization of cynomolgus monkey(Macaca fascicularis)CC chemokine receptor 1
Authors: Gupta Shipra , Schulz-Maronde Sandra , Kutzleb Christian , Richter Rudolf , Forssmann Wolf-Georg , Kapp Alexander , Forssmann Ulf , Elsner Jörn
Published: October 2005

The CC chemokine receptor 1 (CCR1) has emerged as a relevant factor contributing to inflammatory diseases such as allergic asthma. Commonly used animal models of allergic airway inflammation, especially murine models, have certain limitations. The elaborate, nonhuman, primate models of asthma display the highest comparability with the situation in humans. These models play an important role in the understanding of the pathogenesis of asthma. To improve the understanding in cynomolgus monkey models, we identified and characterized CCR1 in this nonhuman primate. Initially, we cloned the cynomolgus monkey CCR1 (cCCR1) gene, and the sequence analysis revealed high homology at the nucleotide (92%) and amino acid (88.4%) levels with its human counterpart. Human embryonic kidney 293 cells were stably transfected with cCCR1 and used in functional assays. Among those CCR1 ligands tested, CCL14(9-74) was most potent in the induction of intracellular Ca2+ fluxes as observed for human CCR1 (hCCR1). Complete cross-desensitization could be achieved between CCL14(9-74) and CCL15. However, CCL3 could not fully abrogate the response to the potent ligand CCL14(9-74). Competition-binding studies with radiolabeled CCL3 concordantly showed that CCL14(9-74) has a higher affinity to cCCR1 than hCCL3. Moreover, differential tissue-specific expression of cCCR1 was investigated by real-time quantitative polymerase chain reaction, displaying the highest levels in spleen. This study adds basic information needed for the evaluation of the role of CCR1 in the pathophysiology of asthma using the highly relevant cynomolgus monkey model and in addition, aids in the preclinical evaluation of potential novel drugs targeting CCR1.
Colon lamina propria dendritic cells induce a proinflammatory cytokine response in lamina propria T cells in the SCID mouse model of colitis
Authors: Drakes Maureen L , Blanchard Thomas G , Czinn Steven J
Published: October 2005

Intestinal immune responses are normally regulated to maintain a state of immune balance. Dendritic cells (DC) are antigen-presenting cells, which induce immune responses against microbes and other stimuli and are key players in the regulation of tolerance in the gut. These cells influence the differentiation of cytokine responses in T cells, and in the gut, in particular, such interactions may be critical to the course of inflammatory bowel disease (IBD). Using the CD45RBhi CD4+ T cell-reconstituted severe combined immunodeficient mouse model of colitis, we investigated the ability of isolated colon DC to stimulate immune responses in syngeneic and allogeneic spleen CD4+ T cells, as well as in colon T cells isolated from the same tissue as DC in IBD mice. We found that the frequency of DC in IBD mice colons and spleens was elevated in comparison with control mice, but colon and spleen DC exhibited different phenotypic and functional properties. Colon DC stimulated significantly higher levels of interferon-γ and interleukin-6 when cocultured with autologous colon T cells than in cocultures with syngeneic or allogeneic spleen T cells. These data suggest that in the IBD colon, DC-T cell interactions may create conditions with an abundance of proinflammatory cytokines, which favor the inflammatory state.
Interleukin-10 and the immune response against cancer: a counterpoint
Authors: Mocellin Simone , Marincola Francesco M , Young Howard A
Published: October 2005

Although interleukin-10 (IL-10) is commonly regarded as an anti-inflammatory, immunosuppressive cytokine that favors tumor escape from immune surveillance, a wealth of evidence is accumulating that IL-10 also possesses some immunostimulating properties. In fact, IL-10 has the pleiotropic ability of influencing positively and negatively the function of innate and adaptive immunity in different experimental models, which makes it questionable to merely categorize this cytokine as a target of anti-immune escape therapeutic strategies or rather, as an immunological adjuvant in the fight against cancer. Here, we review available data about the immunostimulating anticancer properties of IL-10, and in particular, we focus on the hypothesis that in contrast to what occurs in secondary lymphoid organs, IL-10 overexpression within the tumor microenvironment may catalyze cancer immune rejection.
B lymphocytes mediate Fas-dependent cytotoxicity in MRL/lprmice
Authors: Bonardelle Danielle , Benihoud Karim , Kiger Nicole , Bobé Pierre
Published: October 2005

The Fas/Fas ligand (FasL) pathway is one of the two major effector mechanisms of T cell-mediated cytotoxicity. To prevent nonspecific killing by lymphoid cells, FasL expression on the cell surface of immune effector cells is strictly regulated. However, MRL/lpr autoimmune-prone mice massively overexpress FasL on their T lymphocytes, which render them able to kill Fas+ targets in vitro and in vivo. It is surprising that we show in the present work that B lymphocytes purified from MRL/lpr spleen cells express FasL to the same extent as T cells at the mRNA and protein level. These B cells are potent cytotoxic effectors against Fas+ but not Fas− targets. The B lymphocyte effectors were used ex vivo without any in vitro activation by B cell stimuli. Furthermore, we found that MRL/lpr B lymphocytes have the same cytotoxic potential as natural killer cells, which have been characterized as potent, Fas-mediated, cytotoxic effectors. The level of membrane-anchored FasL increases with the size of the B cell and cell-surface activation marker CD69 expression, indicating that the expression of FasL is up-regulated in parallel with the activation state of the B cell. The activated B cell population contained the major cytotoxic activity, and a minor part was associated with CD138/Syndecan-1+ plasma cells.
A Novel Mutation in Fibroblast Growth Factor 23 Gene as a Cause of Tumoral Calcinosis
Authors: Araya Kaori , Fukumoto Seiji , Backenroth Rebecca , Takeuchi Yasuhiro , Nakayama Kounosuke , Ito Nobuaki , Yoshii Nozomi , Yamazaki Yuji , Yamashita Takeyoshi , Silver Justin , Igarashi Takashi , Fujita Toshiro
Published: October 2005

Context: Tumoral calcinosis is a disease characterized by ectopic calcification and hyperphosphatemia due to enhanced renal tubular phosphate reabsorption. Fibroblast growth factor (FGF)23 was identified as a responsible factor in hypophosphatemic diseases caused by renal phosphate leak.

Objective: The objective of the study was to analyze the involvement of FGF23 in the development of tumoral calcinosis.

Design: Serum FGF23 level was evaluated in a patient with tumoral calcinosis by two kinds of ELISA: full-length assay that detects only full-length FGF23 with phosphate-lowering activity and C-terminal assay that measures full-length as well as C-terminal fragment of FGF23. FGF23 gene was analyzed by direct sequencing of PCR products, and mutant FGF23 was analyzed by Western blotting after expression in mammalian cells.

Patients: A family of tumoral calcinosis patients were studied.

Results: Serum FGF23 was extremely high when measured by C-terminal assay. In contrast, it was low normal by full-length assay. Analysis of FGF23 gene detected a serine to phenylalanine mutation in codon 129. No wild-type allele of this codon was found in the patient. The brother of the proband showed the same base change. When this mutant FGF23 was expressed in vitro, full-length and N-terminal fragments were barely detectable by Western blotting, whereas C-terminal fragment with the same molecular weight as that from wild-type FGF23 could be detected.

Conclusion: The production and serum level of C-terminal fragment of FGF23 are increased in this patient with tumoral calcinosis. Together with the recent similar report of FGF23 mutation, impaired action of full-length FGF23 seems to result in tumoral calcinosis.
Aging in Healthy Men Impairs Recombinant Human Luteinizing Hormone (LH)-Stimulated Testosterone Secretion Monitored under a Two-Day Intravenous Pulsatile LH Clamp
Authors: Liu Peter Y. , Takahashi Paul Y. , Roebuck Pamela D. , Iranmanesh Ali , Veldhuis Johannes D.
Published: October 2005

Context: Testosterone (Te) depletion in aging men in principle could reflect deficits in the hypothalamus, pituitary gland, or testis. Available pharmacological studies of possible failure of Leydig cell steroidogenesis remain inconclusive.

Objective: The objective of the study was to assess Te secretion in older and young men in response to near physiological LH stimulation.

Intervention: Pulsatile iv infusion of recombinant human LH was administered for 2 d to stimulate Te secretion during suppression of endogenous LH concentrations with a potent selective GnRH receptor antagonist (ganirelix).

Subjects/Context: Healthy older (aged 60–73 yr, n = 8) and young (19–30 yr, n = 13) men were studied in an academic setting.

Measures: Pulsatile LH and Te concentrations on the second day of exogenous LH stimulation were measured.

Results: Serum ganirelix concentrations and infused LH pulse increments were similar by age. In contrast, older subjects manifested: 1) reduced mean Te concentrations (P = 0.016), Te peak heights (P = 0.014), increments (P = 0.010), summed areas (P < 0.013), and interpeak Te concentrations (P = 0.023); 2) decreased Te to LH concentration ratios (P = 0.002); 3) diminished LH-Te feed-forward synchrony (P = 0.020); and 4) a blunted amplitude (P = 0.036) and advanced phase (P = 0.013) of diurnal Te rhythms.

Conclusion: A novel regimen of pulsatile LH stimulation for 48 h during GnRH receptor blockade unmasks deficits in pulsatile, basal, synchronous, and nyctohemeral Te secretion in healthy older men. These findings do not exclude concomitant defects in GnRH outflow and/or Te-negative feedback in the aging male.
Comparison of Apolipoprotein B100 Metabolism between Continuous Subcutaneous and Intraperitoneal Insulin Therapy in Type 1 Diabetes
Authors: Duvillard Laurence , Florentin Emmanuel , Baillot-Rudoni Sabine , Lalanne-Mistrich Marie-Laure , Brun-Pacaud Agnès , Petit Jean-Michel , Brun Jean-Marcel , Gambert Philippe , Vergès Bruno
Published: October 2005

Objective: In type 1 diabetic patients, the replacement of sc insulin infusion with ip insulin infusion restores the normal physiological gradient between the portal vein and the peripheral circulation, which is likely to modify lipoprotein metabolism.

Design: To check this hypothesis, we performed two apolipoprotein (apo) B100 kinetic studies in seven type 1 diabetic patients, first under sc insulin infusion and then 3 months after the beginning of ip insulin infusion.

Results: Glycemic control was similar under sc insulin infusion and ip insulin infusion, as assessed by glycated hemoglobin A1c and the capillary glycemic curve determined during the kinetic study. Very low-density and intermediate-density lipoprotein apoB100 pool size, production rate, and fractional catabolic rate (FCR) were similar under sc insulin infusion and ip insulin infusion. The low-density lipoprotein apoB100 FCR tended to decrease under ip insulin (0.45 ± 0.06 vs. 0.55 ± 0.11 pool/d), but the difference did not reach statistical significance (95% confidence interval for the difference, −0.33, 0.11). The low-density lipoprotein apoB100 pool size and production rate remained unchanged under ip insulin infusion compared with sc insulin infusion.

Conclusion: In type 1 diabetic patients, the replacement of sc insulin infusion with ip insulin infusion does not induce profound modifications of apoB100-containing lipoprotein production and FCRs.
A Novel Electrode for Ozone Generation
Authors: Kaneda Kazuhiro , Ikematsu Mineo , Iseki Masahiro , Takaoka Daizo , Higuchi Tohru , Hattori Takeshi , Tsukamoto Takeyo , Yasuda Masashi
Published: October 2005

A new type of electrode for ozone generation was fabricated by radio-frequency (RF) sputtering. High current efficiency (8%) of ozone generation is achieved at a very low current density (ca. 10 mA/cm2) with the electrode.
Are the Naranjo Criteria Reliable and Valid for Determination of Adverse Drug Reactions in the Intensive Care Unit?
Authors: Kane-Gill Sandra L , Kirisci Levent , Pathak Dev S
Published: October 2005

BACKGROUND

The Naranjo criteria are frequently used for determination of causality for suspected adverse drug reactions (ADRs); however, the psychometric properties have not been studied in the critically ill.

OBJECTIVE

To evaluate the reliability and validity of the Naranjo criteria for ADR determination in the intensive care unit (ICU).

METHODS

All patients admitted to a surgical ICU during a 3-month period were enrolled. Four raters independently reviewed 142 suspected ADRs using the Naranjo criteria (review 1). Raters evaluated the 142 suspected ADRs 3–4 weeks later, again using the Naranjo criteria (review 2). Inter-rater reliability was tested using the kappa statistic. The weighted kappa statistic was calculated between reviews 1 and 2 for the intra-rater reliability of each rater. Cronbach alpha was computed to assess the inter-item consistency correlation. The Naranjo criteria were compared with expert opinion for criterion validity for each rater and reported as a Spearman rank (rs) coefficient.

RESULTS

The kappa statistic ranged from 0.14 to 0.33, reflecting poor inter-rater agreement. The weighted kappa within raters was 0.5402–0.9371. The Cronbach alpha ranged from 0.443 to 0.660, which is considered moderate to good. The rs coefficient range was 0.385–0.545; all rs coefficients were statistically significant (p < 0.05).

CONCLUSIONS

Inter-rater reliability is marginal; however, within-rater evaluation appears to be consistent. The inter-item correlation is expected to be higher since all questions pertain to ADRs. Overall, the Naranjo criteria need modification for use in the ICU to improve reliability, validity, and clinical usefulness.
Levetiracetam for Mood Stabilization and Maintenance of Seizure Control Following Multiple Treatment Failures
Authors: Stoner Steven C , Lea Jessica W , Wolf Angel L , Berges Arnaldo A
Published: October 2005

OBJECTIVE

To report the case of a patient who experienced adverse events in succession to antiepileptic medications being used for both antiepileptic and mood-stabilization benefit.

CASE SUMMARY

A 46-year-old white woman developed hyponatremia with carbamazepine, hyperammonemia with divalproex, cognitive impairment with topiramate, and hyponatremia with oxcarbazepine. The patient was stabilized physically and psychiatrically on levetiracetam without any noted adverse events.

DISCUSSION

The adverse events in this report have been associated with the medications in question. The patient's presentation is unique, as she developed adverse events in succession to antiepileptic drugs being used to treat both a seizure disorder and symptoms of mood instability. The Naranjo rankings for the reported adverse events indicated the associations were probable (carbamazepine, divalproex, oxcarbazepine) and possible (topiramate). After repeated incidences of intolerability to these drugs, levetiracetam was initiated and provided both seizure control and mood-stabilizing benefits, which eventually led to hospital discharge.

CONCLUSIONS

Levetiracetam may provide mood-stabilizing qualities through a mechanism that is unique from that of other antiepileptic agents used for their mood-stabilizing properties. There are potential advantages with levetiracetam, as no specific therapeutic drug monitoring parameters need to be followed after its introduction. Additionally, this case emphasizes the importance of therapeutic drug monitoring and frequent assessments to prevent physical and psychiatric adverse reactions.
Methylphenidate May Treat Apathy Independent of Depression
Authors: Padala Prasad R , Petty Frederick , Bhatia Subhash C
Published: October 2005

OBJECTIVE

To report a case of apathy treated with methylphenidate in which improvement in apathy was independent of improvement of depression.

CASE SUMMARY

A 47-year-old woman with a 20-year history of recurrent major depression was diagnosed as having significant apathy with lack of initiative and motivation. Over the course of a 4-week treatment regimen with methylphenidate, her apathy, as measured by the Apathy Evaluation Scale, improved, with her score decreasing from 57 to 31. During this period, her depression, as assessed by the 21-item Hamilton Rating Scale for Depression, remained unchanged.

DISCUSSION

Our report of improvement of apathy with methylphenidate is consistent with other reports in the literature, although previous studies have not specifically used the rating scales to assess apathy. Even though this patient had experienced apathy for a long time, it had not been detected due to lack of direct questioning. In this case, as noted, the improvement of apathy was independent of improvement in depression.

CONCLUSIONS

A high degree of suspicion and specific inquiry is required for identification of apathy. Once detected, methylphenidate may be beneficial in its treatment, a strategy that may work independently of augmentation of antidepressants.
National campaign effects on secondary pupils' bullying and violence
Authors: Mooij Ton
Published: October 2005

Background. Research on pupils' bullying (1991) and violence (1993) motivated the Dutch Ministry of Education, Culture and Science to initiate a national campaign on school safety. The government campaign was undertaken from 1995 to 2000.

Aim. To test for differences in secondary pupils' bullying and violence before and after the campaign while controlling for different contextual variables.

Samples. In 1991, a representative survey on bullying was conducted in 36 secondary schools with 1,055 pupils from Year 2 and 4 classes (age range 13–16 years). In 1993, a survey on violent behaviour took place in 71 secondary schools with 1,998 pupils from Year 3 and 4 classes (age range 14–16 years). In 2000, a survey on bullying and violence was conducted in 60 secondary schools with 9,948 pupils from Year 1 to 6 classes (age range 12–18 years).

Methods. The data from pupils in identical school years were compared with respect to bullying (1991–2000) and violence (1993–2000). The statistical relations were analysed in two stepwise multiple regression analyses. Year of investigation (1991–2000, 1993–2000) was the respective dependent variable. The independent variables were pupils' bullying or violence scores, sex, school year, contextual lesson, school, and community variables.

Results. In both regression analyses, the contextual lesson and school variables discriminated between the measurement years. Also, compared with 1991, the pupils in 2000 scored lower for being a bully and higher for being bullied directly. Compared with 1993, the pupils in 2000 scored lower for being a victim of intentional damage to property or emotional violence, lower for being a perpetrator of disruptive behaviour in school, and higher for being a perpetrator of intentional damage to property.

Conclusions. The differences between the contextual variables measured before and after the campaign reflect changes in educational and instructional situations. Independent of these differences, the national campaign appears to have helped improve the awareness of pupils' social behaviour and elicit, in particular, a decrease in pupils' violent behaviour. However, more specific pedagogical and preventative support for pupils socially at risk appears to be needed to have a more prosocial impact on the behaviour of secondary school pupils.
I used to like it but now I don't: The effect of the transfer test in Northern Ireland on pupils' intrinsic motivation
Authors: Remedios Richard , Ritchie Kathryn , Lieberman David A.
Published: October 2005

Background. Research has suggested that the pressure of exams could undermine pupils' interest in their subjects, but almost all of this research has been conducted in laboratory settings. The Transfer Test in Northern Ireland provides an unusual opportunity to assess the effects of exam pressure in real life because some 10‐ and 11‐year‐olds sit a Transfer Test to be admitted to grammar school while others are not tested until they are 14.

Aim. To assess the effect of exams on pupils' interest in their subjects both during the period before the exam and after the results are known.

Sample. The sample comprised 66 pupils preparing to sit the Transfer Test and 55 not preparing for the test.

Method. Pupils' interest in their school subjects was assessed by questionnaires administered 2 weeks before the Transfer Test and then again 2 weeks after the results were announced.

Results. Surprisingly, prior to sitting the test, there was no significant difference in motivation between the test and no‐test pupils. However, after sitting the test, the motivation of the test pupils decreased significantly relative to their no‐test counterparts, despite the fact that most achieved the grades they needed for admission to grammar school.

Conclusions. Exams provide a valuable tool for assessing academic progress, but under some circumstances they can reduce pupils' interest in the subjects they are studying.
Mental health, coping and family‐functioning in parents of young people with obsessive‐compulsive disorder and with anxiety disorders
Authors: Derisley Jo , Libby Sarah , Clark Sarah , Reynolds Shirley
Published: October 2005

Objective. To compare mental health, coping and family‐functioning in parents of young people with obsessive‐compulsive disorder (OCD), anxiety disorders, and no known mental health problems.

Method. Parents of young people with OCD (N=28), other anxiety disorders (N=28), and no known mental health problems (N=62) completed the Brief Symptom Inventory (Derogatis, 1993), the Coping Responses Inventory (Moos, 1990), and the McMaster family assessment device (Epstein, Baldwin, & Bishop, 1983).

Results. Parents of children with OCD and anxiety disorders had poorer mental health and used more avoidant coping than parents of non‐clinical children. There were no group differences in family‐functioning.

Conclusion. The similarities across the parents of clinically referred children suggest that there is a case for encouraging active parental involvement in the treatment of OCD in young people.
Factors associated with the efficacy of a group intervention for anger in people with intellectual disabilities
Authors: Rose John , Loftus Michael , Flint Beverley , Carey Louise
Published: October 2005

Objectives. There is a growing literature that suggests cognitive behavioural interventions aimed at reducing inappropriately expressed anger by people with intellectual disabilities are effective. However, interventions provide little information about which aspects of the individual or their treatment may be contributing to the overall efficacy of the approach.

Design. A group intervention is compared with a waiting list control. A cross‐sectional regressional method was used to explore the relationship between change in a provocation inventory score over the course of an intervention and a number of other variables.

Methods. Data was collected from 50 participants who attended a series of groups with the aim of reducing aggressive behaviour. This was compared with a waiting list control of 36. Outcome was measured by a provocation inventory, which was administered pre‐, post‐group, and at follow‐up. Other variables were also measured including, receptive vocabulary, age, gender, experience of primary therapist and whether staff accompanied participants to the group or not.

Results. A 2 × 2 split plot ANOVA identified a statistically significant treatment effect. However, analysis examining clinical significance was more equivocal with only 11 of the individuals in the intervention group showing reliable change as defined by Jacobson and Truax (1991). Regression analysis indicated that 17.5% of the variance in change of provocation inventory scores could be accounted for by the independent variables. Immediately on completion of the group, being accompanied by a member of staff and increased receptive language scores contributed significantly to the variance.

Conclusions. A reduction in provocation inventory scores immediately after the group was more likely to occur if the participant was accompanied by a member of staff who knew them and if they had a higher score on the test of receptive vocabulary. This pattern changed with time. The implications of the results will be discussed and the relatively large amount of variance that is not accounted for will also be considered.
Knowledge of post‐concussional syndrome in naïve lay‐people, general practitioners and people with minor traumatic brain injury
Authors: Mackenzie J. A. , McMillan T. M.
Published: October 2005

Background. Expectations about effects of minor traumatic brain injury (MTBI) may play a role in maintaining symptoms, the ability to simulate post‐concussion syndrome (PCS) and the extent to which professionals might associate patient complaints with MTBI. In turn, expectations will be related to knowledge about MTBI. This study investigates knowledge of PCS in people with MTBI compared with general practitioners (GPs) and lay‐people, using recall (a vignette) and symptom guessing or recognition (checklists).

Methods. Thirty GPs, 30 people with MTBI, and 30 lay‐people generated symptoms from a vignette describing a MTBI. They then completed a PCS checklist or a checklist containing PCS and ‘bogus’ items pertaining to the vignette.

Results. Only 1 of 90 participants reported enough PCS symptoms to indicate ‘caseness’ from the vignette alone. Using checklists, more GPs (60%) than controls (20%) ticked enough symptoms to attain PCS ‘caseness’. Few people with MTBI who did not themselves achieve PCS ‘caseness’ by self‐report using checklists achieved ‘caseness’ using the vignette and checklists. The frequency of non‐physical symptom reporting was higher than for physical symptom reporting. The addition of ‘bogus’ items to checklists did not affect responding.

Conclusions. It is unlikely that pre‐injury expectations about MTBI maintain PCS symptoms. PCS was difficult to simulate without prompting, even by GPs and PCS ‘cases’. The use of ‘caseness’, rather than simple symptom scores, and further development of checklists with ‘bogus’ items may provide a more valid assessment of MTBI effects. GPs may benefit from further information about PCS and available treatments.
Smoking and peer groups: Results from a longitudinal qualitative study of young people in Northern Ireland
Authors: Stewart‐Knox Barbara J. , Sittlington Julie , Rugkåsa Jorun , Harrisson Sheila , Treacy Margaret , Abaunza Pilar Santos
Published: October 2005

Previous research has indicated that young people are under considerable social pressure to take up smoking. This study has therefore sought to explore and better understand the mechanisms through which peer‐related social factors operate to encourage young people to smoke. Individual qualitative interviews were held with adolescent children aged 11–12 years (N=102) within youth clubs based in economically deprived areas of Northern Ireland, and then followed up on two occasions during the subsequent 3 years (N=51/39). The data implied that, although peers influence smoking uptake, this seldom happens through direct persuasion, but rather as the result of the young person striving to conform to the normative behaviour of the peer group with which they identify. The findings are consistent with social identity theory and self‐categorization theory in that for both smoking and non‐smoking 14‐year‐olds smoking activity appears to provide a means through which to define social groups, to accentuate similarity within groups and differences between groups. In‐group favouritism was expressed in the sharing of cigarettes within the in‐group and in the negative stereotyping of out‐group members. There was some evidence that group affiliation may be negotiated differently for boys and girls. These findings imply that successful intervention needs to reconsider the normative processes that encourage young people to smoke.