bluuebunny/crossref_metadata_2025 huggingface dataset:
Mucosal and systemic antibody formation in the rat after intranasal administration of three different antigens
Authors: HAMELEERS DONA M. H. , VAN DER VEN INEKE , BIEWENGA JEIKE , SMINIA TAEDE
Published: July 2007

In order to study the role of nasal‐associated lymphoid tissue (NALT) in the local nasal immune response, rats were immunized intra‐nasally with either of the following trinitrophenylated (TNP) antigens; the thymus‐dependent keyhole limpet haemocyanin (KLH), or the thymusindependent lipopolysaccharide (LPS), or with the particulatc (thymus‐dependent) sheep red blood cells (SRBC). Primary responses hardly occurred, while only TNP‐KLH elicited a considerable secondary response. The major responding organ was the posterior cervical lymph node. Specific antibody‐forming cells (AFC) occurred in the medulla and were mainly of the IgA or IgG isotype. Hardly any specific AFC were found in NALT or the surrounding mucosa. Intranasal immunization evoked no antibody response in the lung. Ample anti‐TNP antibodies could be detected in the sera of animals, primed and boosted with TNP‐KLH or TNP‐LPS. No specific serum antibodies occurred after immunization with TNP‐SRBC. The results are discussed in view of the immunoiogical defence in the upper respiratory tract.
Deoxyspergualin inhibits cytotoxic T lymphocytes but not NK or LAK cells
Authors: KERR P. G. , ATKINS R. C.
Published: July 2007

Deoxyspergualin (DOSP) is a new immunosuppressive agent which probably inhibits various functions of monocytes, B cells and T cells. We examined the effects of deoxyspergualin on cellular cytotoxicity, including cytotoxic T lymphocyte (CTL) mediated killer, natural killer (NK) cell and lymphokine activated killer (LAK) cell killing. Deoxyspergualin inhibited cellular cytotoxicity generated by 7 days allo‐antigenic challenge: it also inhibited cell killing if added on day 6 of this 7 day culture period. The drug did not significantly inhibit NK or LAK cell killing. The inhibitory effects of deoxyspergualin, however, were dependent on the serum used in the culture medium. Normal human serum (NHS) was associated with less inhibition than fetal calf serum (FCS). Finally, interleukin 2 (IL‐2) was able to prevent the inhibitory effects of deoxyspergualin on antigen‐specific cytotoxicity.
The effect of intrathymic injection of donor blood on the graft versus host reaction and cardiac allograft survival in the rat
Authors: HAMANO K. , FUJIKURA Y. , FUKUDA S. , ESATO K. , FUKUMOTO T.
Published: July 2007

Our previous paper reported that it was difficult to induce tolerance using donor specific transfusion (DST) in Lewis rats, while in DA and PVG rats it was possible to induce tolerance using DST. In this paper we investigated the effects of DST in a local graft versus host reaction (GVHR) model using these rat strains. The following results were obtained: (i) DST suppressed the GVHR in the DA rat but not in the Lewis rat; (ii) DST did not suppress GVHR in PVG rat strain, but lymphocytes, prepared from the PVG rats that were tolerant of Lewis heart grafts induced by DST, could suppress the the GVHR; and (iii) in Lewis rats, intrathymic injection of whole blood suppressed the GVHR, suggesting this route of antigen delivery might be effective in prolonging allograft survival. Cardiac allograft survival in this strain was prolonged in some cases after pretreatment with intrathymic donor whole blood. These results suggest the value of the GVHR in the assessment of new protocols to induce allograft tolerance in rats.
Suppressor cells and soluble factors from the spleens of UV‐irradiated mice
Authors: BRODIE M. , HALLIDAY W. J.
Published: July 2007

Several indices of splenic reactivity were measured daily after ultraviolet (UV) irradiation of mice. Spleen weight increased to a maximum after 5 days. Suppressor cell activity, manifested in various assays, peaked slightly earlier. The ability of passively transferred spleen cells to suppress the induction of contact hypersensitivity (CHS) was maximal after 4 days, as was the production of soluble suppressor factors in culture. The factors were detected by suppression of CHS in vivo and leucocyte adherence inhibition (LAI) in vitro. These assays appeared to detect different factors: an I‐J− factor active in CHS assays and an I‐J+ factor active in LAI assays. The latter factor required CD4−, CD8+ lymphocytes for its production.
Low affinity binding of mouse immunoglobulin to human CD5+ B cells
Authors: WESTON KATHRYN M. , RAISON ROBERT L.
Published: July 2007

Polyreactive immunoglobulin (Ig) secreted by CD5‐bearing B cells has the capacity to bind a broad range of self and foreign antigens. Flow cytometric analysis was used to detect low‐affinity binding of mouse Ig molecules to the surface of CD5‐bearing B cells from patients with chronic lymphocytic leukaemia (CLL). Mouse Ig isotypes G. A. and M. and IgG subclasses G1, G2a, and G3 bound to the B cell surface via a mechanism not involving the antigen‐binding site of the mouse Ig molecule. Fab and F(ab′)2 fragments of mouse Ig associated with the CLL cells in a similar manner to intact Ig. indicating that Fc receptor interactions were not involved. Dissociation of the mouse Ig from the B cell surface by three washing steps distinguished this lower‐affinity binding from high‐affinity binding which occurs through the antigen‐binding site of a mouse monoclonal antibody (MoAb) when it recognizes a specific ceil surface epitope. Blocking studies suggest that the low‐affinity binding occurred via surface IgM and surface IgD (sIgM/sIgD) on the CD5‐bearing B cells. The results are consistent with the expression of polyreactive Ig on the surface of CD5‐bearing B cells. The results are constituent with the expression of polyreactive Ig on the surface of CD5‐bearing B cells in CLL.
Molecular cloning, expression and characterization of ovine TNFα
Authors: NASH A. D. , BARCHAM G. J. , BRANDON M. R. , ANDREWS A. E.
Published: July 2007

Tumour necrosis factor α (TNFα) is a cytokine with a wide range of effects on both lymphoid and non‐lymphoid cell types. By hybridization with a human TNFα cDNA probe the corresponding ovine cDNA was isolated from a lipopolysaccharide (LPS) stimulated alveolar macrophage cDNA library. The sequence of the cDNA clone showed that ovine TNFα encodes a polypeptide of 234 amino acids that, based on analysis of human TNFα is processed to a protein of 157 amino acids. The nucleotide and amino acid sequences showed a high degree of homology to the equivalent human and mouse molecules. In a mammalian COS cell expression system the ovine cDNA was found to encode a protein which was able to lyse actinomycin‐D treated WEHI‐164 cells and induce COS cells to produce and secrete interleukin 6 (IL‐6). Further experiments demonstrated the importance of sequences within the 3’untranslated region of the cDNA in determining the level of expression of ovine TNFα Northern blot analysis was used to analyse the kinetics of induction of ovine TNFα mRNA in alveolar macrophages stimulated with a variety of mitogens. Addition of LPS increased mRNA encoding TNFα at 1 hand 5 h but not 24 h post stimulation. In contrast, addition of phorbol myristic acid (PMA) led to increased TNFα mRNA at 5 h while the combination of PMA and ionomycin increased the level of specific mRNA detected at 1 h, 5 h and 24 h. From genomic analysis ovine TNFα appears to exist as a single copy.
Analysis of T cell receptor Vα and Vβ gene usage in synovia of patients with rheumatoid arthritis
Authors: OLIVE C. , GATENBY P. A. , SERJEANTSON S. W.
Published: July 2007

T cells are thought to play a fundamental role in the pathogenesis of rheumatoid arthritis (RA), Activated T cells expressing α/β T cell receptor (Tcr) infiltrate the rheumatoid synovium and could potentially initiate a local inflammatory response directed against joint constituents, A Tcr repertoire with restricted heterogeneity may reflect a selective expansion of T cells reactive to a few antigenic determinants within the synovium. To determine whether predominant Vα and/or Vβ gene usage of the expressed α/β Tcr repertoire is a feature of synovial T cells in patients with RA, the polymerase chain reaction (PCR) was used to amplify Tcr‐α and Tcr‐β chain transcripts. The peripheral blood and synovia of five patients with adult RA were examined and no evidence of preferential use of 19 Tcr Vα gene families was found. Similarly, most of the 18 Tcr Vβ gene families could be detected in RA synovia although there were quantitative differences in Tcr Vβ gene expression when compared to peripheral blood. This report shows that when the extremely sensitive assay of oligonucleotide hybridization of PCR amplified Tcr transcripts is used, permitting identification of specific V gene families, the α/β Tcr repertoire in the rheumatoid synovium is more diverse than was previously thought. Therefore, in patients with RA of long duration, the synovial T cell response is most likely to be polyclonal.
Susceptibility of multipotent haemopoietic stem cell deficient W/Wv mice to Plasmodiumberghei‐infection
Authors: ASAMI M. , OWHASHI M. , ABE T. , NAWA Y.
Published: July 2007

The susceptibility of haemopoietic stem cell deficient W/Wv mice to infection with Plasmodium herghei was examined. The mean survival time of W/Wv mice after the infection was shorter than that of the +/+ mice. Splenomegaly, a characteristic pathological change of the host after infection with malaria parasites was not observed in W/Wv mice. When haemopoietic activity of the infected mice was examined, a substantial increase in number of multipotent haemopoietic stem cells (CFU‐S) and the committed stem cells for granulocytes and maerophages (CFU‐GM) or for erythrocytes (CFU‐E) was observed in the bone marrow and spleen of +/+ but not of W/Wv mice. CFU‐S were not detected in W/Wv mice before or alter infection. The number of CFU‐GM and CFU‐E in bone marrow and spleen of W/Wv mice decreased after infection. Bone marrow grafting from +/+ to W/Wv mice 8 weeks before infection prolonged the mean survival time of the mice and effectively restored the number of CFU‐S in the spleen of W/Wv mice. These results indicate that multipotent haemopoietic stem cells play an important role in the host's defence mechanisms against P. berghei‐infection.
Beneficial effects of endotoxin treatment on metabolism in tumour‐bearing rats
Authors: ROFE A. M. , BOURGEOIS C. S. , COYLE P.
Published: July 2007

The effects of endotoxin treatment on host metabolism in tumour‐bearing investigated. Metabolism in control rats (non‐tumour‐bearing) was slightly altered by endotoxin treatment, whereas in tumour‐bearing rats a number of biochemical parameters that were intially perturbed by the presence of the tumour had returned to normal at 48 h post‐treatment. The beneficial effects included increased blood glucose and insulin concentrations, and decreased ketone body, triglyceride and lactate concentrations. Potentially non‐beneficial effects of endotoxin observed in both tumour‐bearing and control rats included decreased plasma cholesterol, and increased plasma phosphate, potassium and alkaline phosphatase levels.

Endotoxin caused haemorrhaging in the encapsulated tumour, and this was associated with histological evidence of endorhelial damage, red cell infiltration into surrounding tumour tissue and a marked decrease in cell viability. The in vivo uptake of glucose by the tumour, measured by 2‐deoxy [U‐14C]glucose uptake, was decreased by 96% following endotoxin treatment, and this was associated with a two‐fold increase in glucose uptake by muscle. It is concluded that endotoxin treatment has major effects on cell viability and the integrity of vasculature in the tumour, which limits glucose uptake by the tumour and thereby decreases the energy and substrate requirements of the tumour, thus benefiting the host. It is suggested that tumour cytotoxicity and intra‐tumour haemorrhage are the result of endotoxin stimulating cytokine release from macrophages that are already activated by the presence of the tumour. While the effects of endotoxin on metabolism in tumour‐bearing rats appeared to be primarily the result of anti‐tumour activity, other findings, particularly the raised plasma alkaline phosphatase activity that was observed in endotoxin‐treated control and tumourbearing rats, indicate possible side effects that may need to be considered in any treatment strategy.
Relocation of antigens to the cell surface membrane can enhance immune stimulation and protection
Authors: BOTH G. W. , ANDREW M. E. , BOYLE D. B. , COUPAR B. E. H. , BELLAMY A. R.
Published: July 2007

The major outer capsid glycoprotein of rotaviruses, VP7, is normally synthesized and directed to the ER, where it is required for virus assembly. By substituting a foreign signal sequence for the VP7 signal peptide, a secreted form of VP7 with an authentic amino terminus was produced. Secreted VP7 was further modified by the addition of a transmembrane anchor and cytoplasmic domain to its C‐terminus. When the novel chimeric protein was expressed in transfected cells it became anchored in the cell surface membrane. The antigenicity of the chimeric protein was compared with that of the intracellular form of VP7 using recombinant vaccinia viruses to deliver the antigens in vivo. The novel antigen produced enhanced stimulation of both B and T lymphocytes of the immune system, and in mice it was able to induce protection against rotavirus‐induced diarrhoeal disease. Other secreted and intracellular antigens show a similar improved level of antigenicity as a result of their relocation to the cell surface. Surface localization may therefore have general utility in the development of recombinant subunit vaccines.
Effects of PMA, cytokines and dexamethasone on the expression of cell surface Fc receptors and mRNA in U937 cells
Authors: GRATTAGE LYNN P. , McKENZIE IAN F. C. , HOGARTH P. MARK
Published: July 2007

Fc receptors (FcR) are of importance in immune and inflammatory reactions. FcR expression as mRNA and surface protein was therefore examined in the myelomonocytic cell line, U937, after stimulation with phorbol ester (PMA), in the presence of seven different cytokines (interferon‐γ [IFNγ], IFNα, granulocyte‐macrophage colony‐stimulating factor [GM–CSF], tumour necrosis factor‐α [TNFα], TNFβ, interleukin‐β [IL‐1β], IL‐2) or dexamethasone. HLA class I and CD lib expression were also examined. Cell surface expression of FcRI and II was measured by flow cytometry using monoclonal antibodies, and the mRNA of FcRII was measured with cDNA or oligonucleotide probes. The major findings were: PMA increased cell surface FcRI, FcRII and CD11b, but decreased HLA; PMA caused a fivefold increase in all three FcRII RNA transcripts (2.5, 1.5 and 0.9 kb) in Northern analysis; IFNγ, IFNα and GM‐CSF increased the expression of FcRI and II, and there was no effect with IL‐1β, IL‐2, TNFα or TNFβ (only GM‐CSF increased the expression of CD11b); all cytokines further increased FcRI and FcRII expression in the presence of PMA; HLA expression was also increased in the presence of PMA, IFNα and IFNγ; dexamethasone reduced the levels of FcRI and II in cells stimulated with PMA with or without cytokines. Thus stimulatory agents and cytokines can alter the expression of surface FcγR and mRNA encoding FcRI or II, providing potential control mechanisms for the modulation of these receptors in inflammatory responses.
Retention of tissue‐specific phenotype in a panel of colon carcinoma cell lines: Relationship to correlates
Authors: WHITEHEAD R. H. , ZHANG H. H. , HAYWARD I. P.
Published: July 2007

A panel of eight cell lines has been derived from colon carcinomas. These cell lines have both been characterized according to standard criteria of growth rate, response to mitogens (epidermal growth factor and basic fibroblast growth factor), xenograft growth and growth in soft agar, and according to the ability of the cells to express epitopes known to be expressed by cells in the normal intestinal mucosa. The expression of epitopes present in columnar (absorptive) cells has been assessed using a panel of monoclonal antibodies to brush border peptidases and disaccharidases, villin and brush border‐specific peptides. Goblet cell epitopes have been determined by monoclonal antibodies to mucin and carcinoembryonic antigen. An antibody to chromogranin was used to identify endocrine cells. Using these antibodies we found that all the cell lines reacted with at least one of the antibodies to columnar cells. Similarly, varying proportions of cells in six of the eight cell lines stained with antibodies to mucin. None of the cells expressed chromogranin. Expression of a differentiated colonic phenotype, as measured from antibody staining, did not correlate with measurements of malignancy, such as the ability of the cells to grow in soft agar or as xenografts. Similarly, there was no correlation between retention of a colonic phenotype and the initial pathological stage of the tumour from which the cell lines were derived.
Serological responses and immunity produced in salmonids by vaccination with Australian strains of Vibrio anguillarum
Authors: MUNDAY B. L. , CARSON J. , WHITTINGTON R. , ALEXANDER J.
Published: July 2007

The Australian populations of rainbow trout (Onchorhynchm mykiss) and Atlantic salmon (Salmo salar) were found to have similar immunological responses to local strains of Vibrio anguillarum as those reported for the more genetically diverse populations of these fish and strains of V. anguillarum found in the Northern Hemisphere. In addition, our studies more precisely defined the respective responses of rainbow trout and Atlantic salmon to immersion vaccination by the bath and dip methods.
Desiccation-tolerance specific gene expression in leaf tissue of the resurrection plant Sporobolus stapfianus
Authors: Le Tuan Ngoc , Blomstedt Cecilia K. , Kuang Jianbo , Tenlen Jennifer , Gaff Donald F. , Hamill John D. , Neale Alan D.
Published: July 2007

The desiccation tolerant grass Sporobolus stapfianus Gandoger can modulate cellular processes to prevent the imposition of irreversible damage to cellular components by water deficit. The cellular processes conferring this ability are rapidly attenuated by increased water availability. This resurrection plant can quickly restore normal metabolism. Even after loss of more than 95% of its total water content, full rehydration and growth resumption can occur within 24 h. To study the molecular mechanisms of desiccation tolerance in S. stapfianus, a cDNA library constructed from dehydration-stressed leaf tissue, was differentially screened in a manner designed to identify genes with an adaptive role in desiccation tolerance. Further characterisation of four of the genes isolated revealed they are strongly up-regulated by severe dehydration stress and only in desiccation-tolerant tissue, with three of these genes not being expressed at detectable levels in hydrated or dehydrating desiccation-sensitive tissue. The nature of the putative proteins encoded by these genes are suggestive of molecular processes associated with protecting the plant against damage caused by desiccation and include a novel LEA-like protein, and a pore-like protein that may play an important role in peroxisome function during drought stress. A third gene product has similarity to a nuclear-localised protein implicated in chromatin remodelling. In addition, a UDPglucose glucosyltransferase gene has been identified that may play a role in controlling the bioactivity of plant hormones or secondary metabolites during drought stress.
Plant natriuretic peptide active site determination and effects on cGMP and cell volume regulation
Authors: Wang Yu Hua , Gehring Chris , Cahill David M. , Irving Helen R.
Published: July 2007

Natriuretic peptides (NP) were first identified in animals where they play a role in the regulation of salt and water balance. This regulation is partly mediated by intracellular changes in cyclic GMP (cGMP). NP immunoanalogues occur in many plants and have been isolated, with two NP encoding genes characterised in Arabidopsis thaliana L. (AtPNP-A and AtPNP-B). Part of AtPNP-A contains the region with homology to human atrial (A)NP. We report here on the effects of recombinant AtPNP-A and smaller synthetic peptides within the ANP-homologous region with a view to identifying the biologically active domain of the molecule. Furthermore, we investigated interactions between AtPNP-A and the hormone, abscisic acid (ABA). ABA does not significantly affect Arabidopsis mesophyll protoplast volume regulation, whereas AtPNP-A and synthetic peptides promote water uptake into the protoplasts causing swelling. This effect is promoted by the membrane permeable cGMP analogue, 8-Br-cGMP, and inhibited by guanylate cyclase inhibitors indicating that increases in cGMP are an essential component of the plant natriuretic peptides (PNP) signalling cascade. ABA does not induce cGMP transients and does not affect AtPNP-A dependent cGMP increases, hence the two regulators differ in their second messenger signatures. Interestingly, AtPNP-A significantly delays and reduces the extent of ABA stimulated stomatal closure that is also based on cell volume regulation. We conclude that a complex interplay between observed PNP effects (stomatal opening and protoplast swelling) and ABA is likely to be cell type specific.
Cell wall disassembly events in boysenberry (Rubus idaeus L. × Rubus ursinus Cham. & Schldl.) fruit development
Authors: Vicente Ariel Roberto , Powell Ann , Greve L. Carl , Labavitch John M.
Published: July 2007

Boysenberry fruit was harvested at five developmental stages, from green to purple, and changes in pectin and hemicellulose solubilisation and depolymerisation, polymer neutral sugar contents, and the activities of cell wall degrading enzymes were analysed. The high xylose to glucose ratio in the 4% KOH-soluble hemicellulose fraction suggests that xylans are abundant in the boysenberry cell wall. Although the cell wall changes associated with fruit development do not proceed in discrete stages and the cell wall disassembly is a consequence of highly regulated changes occurring in a continuum, the results suggest that the temporal changes in cell wall degradation in boysenberry account for at least three stages: an early stage (green to 75% red colour), associated with metabolism of cellulose and cross-linking glycans; an intermediate period (75 to 100% red colour), characterised by substantial pectin solubilisation without depolymerisation in which α-arabinofuranosidase increases markedly and 50% of the wall arabinose is lost; and a final stage (100% red colour to purple), characterised mainly by a reduction of pectic galactose content and a dramatic increase in pectin depolymerisation associated with higher polygalacturonase, pectin methylesterase, acetyl esterase and β-galactosidase activities. From a biotechnological perspective enzymes involved in pectin matrix disassembly seem to be the better candidates to affect boysenberry fruit late-softening by genetic intervention. A model for cell wall disassembly in boysenberry fruit is proposed.
PIP5KIγ is required for cardiovascular and neuronal development
Authors: Wang Yanfeng , Lian Lurong , Golden Jeffrey A. , Morrisey Edward E. , Abrams Charles S.
Published: July 2007

All eukaryotic cells contain the phospholipid phosphatidylinositol 4, 5-bisphosphate (PIP
2
) that serves multiple roles in signal transduction cascades. Type I phosphatidylinositol-4-phosphate 5-kinase (PIP5KI) catalyzes the synthesis of PIP
2
by phosphorylating phosphatidylinositol 4 phosphate. Although the classical isoforms of PIP5KI (designated as α, β, and γ) all generate the same phospholipid product, they have significantly dissimilar primary structures and expression levels in different tissues, and they appear to localize within different compartments within the cell. Therefore, it appears likely that PIP5KI isoforms have overlapping, but not identical, functions. Here we show that targeted disruption of PIP5KIγ causes widespread developmental and cellular defects. PIP5KIγ-null embryos have myocardial developmental defects associated with impaired intracellular junctions that lead to heart failure and extensive prenatal lethality at embryonic day 11.5 of development. Loss of PIP5KIγ also results in neural tube closure defects that were associated with impaired PIP
2
production, adhesion junction formation, and neuronal cell migration. These data, along with those of other PIP5KI isoforms, indicate that individual PIP5KI isoenzymes fulfill specific roles in embryonic development.
Probing genetic overlap among complex human phenotypes
Authors: Rzhetsky Andrey , Wajngurt David , Park Naeun , Zheng Tian
Published: July 2007

Geneticists and epidemiologists often observe that certain hereditary disorders cooccur in individual patients significantly more (or significantly less) frequently than expected, suggesting there is a genetic variation that predisposes its bearer to multiple disorders, or that protects against some disorders while predisposing to others. We suggest that, by using a large number of phenotypic observations about multiple disorders and an appropriate statistical model, we can infer genetic overlaps between phenotypes. Our proof-of-concept analysis of 1.5 million patient records and 161 disorders indicates that disease phenotypes form a highly connected network of strong pairwise correlations. Our modeling approach, under appropriate assumptions, allows us to estimate from these correlations the size of putative genetic overlaps. For example, we suggest that autism, bipolar disorder, and schizophrenia share significant genetic overlaps. Our disease network hypothesis can be immediately exploited in the design of genetic mapping approaches that involve joint linkage or association analyses of multiple seemingly disparate phenotypes.
Lipid-lowering effects of anti-angiopoietin-like 4 antibody recapitulate the lipid phenotype found in angiopoietin-like 4 knockout mice
Authors: Desai Urvi , Lee E-Chiang , Chung Kyu , Gao Cuihua , Gay Jason , Key Billie , Hansen Gwenn , Machajewski Dennis , Platt Kenneth A. , Sands Arthur T. , Schneider Matthias , Van Sligtenhorst Isaac , Suwanichkul Adisak , Vogel Peter , Wilganowski Nat , Wingert June , Zambrowicz Brian P. , Landes Greg , Powell David R.
Published: July 2007

We used gene knockout mice to explore the role of Angiopoietin-like-4 (Angptl4) in lipid metabolism as well as to generate anti-Angptl4 mAbs with pharmacological activity. Angptl4 −/− mice had lower triglyceride (TG) levels resulting both from increased very low-density lipoprotein (VLDL) clearance and decreased VLDL production and had modestly lower cholesterol levels. Also, both Angptl4 −/− suckling mice and adult mice fed a high-fat diet showed reduced viability associated with lipogranulomatous lesions of the intestines and their draining lymphatics and mesenteric lymph nodes. Treating C57BL/6J, ApoE −/−, LDLr −/−, and db/db mice with the anti-Angptl4 mAb 14D12 recapitulated the lipid and histopathologic phenotypes noted in Angptl4 −/− mice. This demonstrates that the knockout phenotype reflects not only the physiologic function of the Angptl4 gene but also predicts the pharmacologic consequences of Angptl4 protein inhibition with a neutralizing antibody in relevant models of human disease.
Detecting genetic variation in microarray expression data
Authors: Greenhall Jennifer A. , Zapala Matthew A. , Cáceres Mario , Libiger Ondrej , Barlow Carrolee , Schork Nicholas J. , Lockhart David J.
Published: July 2007

The use of high-density oligonucleotide arrays to measure the expression levels of thousands of genes in parallel has become commonplace. To take further advantage of the growing body of data, we developed a method, termed “GeSNP,” to mine the detailed hybridization patterns in oligonucleotide array expression data for evidence of genetic variation. To demonstrate the performance of the algorithm, the hybridization patterns in data obtained previously from SAMP8/Ta, SAMP10/Ta, and SAMR1/Ta inbred mice and from humans and chimpanzees were analyzed. Genes with consistent strain-specific and species-specific hybridization pattern differences were identified, and ∼90% of the candidate genes were independently confirmed to harbor sequence differences. Importantly, the quality of gene expression data was also improved by masking the probes of regions with putative sequence differences between species and strains. To illustrate the application to human disease groups, data from an inflammatory bowel disease study were analyzed. GeSNP identified sequence differences in candidate genes previously discovered in independent association and linkage studies and uncovered many promising new candidates. This approach enables the opportunistic extraction of genetic variation information from new or pre-existing gene expression data obtained with high-density oligonucleotide arrays.
Rethinking the paradigms that inform behavioral treatment research for substance use disorders
Authors: Morgenstern Jon , McKay James R.
Published: July 2007

Aims  Over the last three decades, the randomized controlled trial or ‘psychotherapy technology’ approach has been the dominant model of inquiry in research on addiction treatment. This period has yielded impressive discoveries, but recent failures to confirm core research hypotheses such as occurred in Project MATCH and the Cocaine Collaborative Treatment Study have raised questions about future research directions. The paper identifies several testable assumptions of the psychotherapy technology model.

Methods  A review is conducted on four substance use disorder behavioral interventions—motivational interviewing, cognitive–behavioral treatment, behavioral couples treatment and 12‐Step‐oriented treatment—to determine whether these assumptions are supported by research findings.

Results  Overall, the review suggests weak support for the technology model of psychotherapy research. Lack of support is interpreted as indicating flaws in several model assumptions about how to conceptualize and measure patient responsivity and the interaction of non‐specific and specific therapeutic factors. The paper offers alternative strategies for addressing these issues drawn from the general psychotherapy process literature and provides illustrative examples of how these could be used to spur innovation in addiction treatment research.

Conclusions  The addiction treatment research field is coming up against the limitations of the psychotherapy technology model as the dominant paradigm guiding treatment research. It is important for addiction treatment researchers to explore alternative conceptualizations and methodologies in order to understand more clearly how treatment works.
Atomoxetine pharmacokinetics in healthy Chinese subjects and effect of the CYP2D6*10 allele
Authors: Cui Yi M. , Teng Choo H. , Pan Alan X. , Yuen Eunice , Yeo Kwee P. , Zhou Ying , Zhao Xia , Long Amanda J. , Bangs Mark E. , Wise Stephen D.
Published: July 2007

What is already known about this subject

• Atomoxetine is a cytochrome P450 2D6 (CYP2D6) substrate and its pharmacokinetics has been characterized in a predominantly White population during clinical development.

• There are scant East Asian pharmacokinetic data available.

• The CYP2D6*10 allele is particularly prevalent in East Asian populations and may contribute to the known ethnic differences in CYP2D6 metabolic capacity.

What this study adds

• The pharmacokinetics of multiple‐dose 80 mg daily atomoxetine observed in Chinese healthy subjects appears comparable to previous data in other ethnic populations.

• Homozygous CYP2D6*10 subjects appear to have higher exposures, but are not a clearly distinct group compared with other CYP2D6 extensive metabolizers.

Aims

To characterize atomoxetine pharmacokinetics, explore the effect of the homozygous CYP2D6*10 genotype on atomoxetine pharmacokinetics and evaluate the tolerability of atomoxetine, in healthy Chinese subjects.

Methods

Twenty‐four subjects, all CYP2D6 extensive metabolizers (EM), were randomized to receive atomoxetine (40 mg qd for 3 days, then 80 mg qd for 7 days) or matching placebo (2 : 1 ratio) in a double‐blind fashion. Atomoxetine serum concentrations were measured following single (40 mg) and multiple (80 mg) doses. Adverse events, clinical safety laboratory data and vital signs were assessed during the study.

Results

Atomoxetine was rapidly absorbed with median time to maximum serum concentrations of approximately 1.5 h after single and multiple doses. Atomoxetine concentrations appeared to decrease monoexponentially with a mean apparent terminal half‐life (t1/2) of approximately 4 h. The apparent clearance, apparent volume of distribution and t1/2 following single and multiple doses were similar, suggesting linear pharmacokinetics with respect to time. Homozygous CYP2D6*10 subjects had 50% lower clearances compared with other EM subjects, resulting in twofold higher mean exposures. No clinically significant changes or abnormalities were noted in laboratory data and vital signs.

Conclusions

The pharmacokinetics of atomoxetine in healthy Chinese subjects appears comparable to other ethnic populations. Multiple dosing of 80 mg qd atomoxetine was well tolerated in this study.
Myocardial iron loading by magnetic resonance imaging T2* in good prognostic myelodysplastic syndrome patients on long‐term blood transfusions
Authors: Chacko Joseph , Pennell Dudley J. , Tanner Mark A. , Hamblin Terry J. , Wonke Beatrix , Levy Terry , Thomas Peter W. , Killick Sally B.
Published: July 2007

Magnetic resonance imaging (MRI) was used to quantify myocardial iron loading by T2* in 11 transfusion‐dependent good prognostic myelodysplastic syndrome (MDS) patients. Myocardial T2*, left ventricular function and hepatic T2* were measured simultaneously. Patients had been on transfusion therapy for 13–123 months and had serum ferritin levels of 1109–6148 μg/l at the time of study. Five patients had not commenced iron chelation and had been transfused with a median of 63 red cell units and had a median serum ferritin level of 1490 μg/l. Six patients were on iron chelation and had been transfused with a median of 112 red cell units and had a median serum ferritin level of 4809 μg/l. Hepatic iron overload was mild in two, moderate in seven and severe in two patients. The median liver iron concentration was 5·9 mg/g dry weight in chelated patients and 9·5 mg/g in non‐chelated patients (P = 0·17; not significant). Myocardial T2* indicated absent iron loading in 10/11 patients (91%; 95% confidence interval 62–98%) and borderline‐normal in one patient. Left ventricular function was normal in all patients. No correlation was observed between increasing serum ferritin levels, hepatic iron overload and myocardial T2*. A long latent period relative to hepatic iron loading appears to predate the development of myocardial iron loading in transfusion‐dependent MDS patients.
Parenting education for young fathers in prison
Authors: Meek Rosie
Published: July 2007

This paper explores the effectiveness of delivering an intensive parenting class to groups of young fathers in prison. Evaluation was based on course feedback from a total of 75 participants. Results demonstrate that even within a group of young prisoners of similar ages there were diverse parenting educational requirements, indicating a need for courses to remain flexible and participant‐led. In considering parenting support needs, three‐quarters of all course participants highlighted the importance of better visiting procedures to allow them to keep in contact with their children whilst in prison. Participants expressed reluctance in accessing parenting support services after release, with the majority of the young fathers indicating that they did not need or want to access formal post‐release provision. Results suggest that further efforts need to be made to support young fathers in custody and after release, with implications for preventing reoffending and engaging young men in parenting education.
Stepfather involvement in social interventions made by youth protection services in stepfamilies
Authors: Parent Claudine , Saint‐Jacques Marie‐Christine , Beaudry Madeleine , Robitaille Caroline
Published: July 2007

Despite efforts made by management and caseworkers to promote active parental participation in the protective context, fathers or other male figures are often brushed aside from intervention. This paper presents the results of qualitative research on methods used by youth protection caseworkers (n = 22) working with stepfather families. The main objective is to identify items that encourage or discourage stepfather involvement in psychosocial interventions. Results showed that certain items do not apply solely to stepfathers, but influence youth protection caseworker decision‐making from a broader perspective. Particular characteristics associated with being a stepfather significantly influence involvement practices espoused by caseworkers, notably the absence of legal status and biological connection with the mother's children.
My life in care: experiences of non‐participation in decision‐making processes
Authors: Leeson Caroline
Published: July 2007

The extent to which young people are involved in legal decision‐making depends on assumptions and perceptions about their ability to participate in decision‐making in general. This paper draws on research with four young people, looking at their experiences of involvement in a variety of decision‐making processes whilst in the care of the local authority. Through narratives, games and other activities, the thoughts and emotions of the four young people are explored, identifying the development of feelings of helplessness, low self‐esteem and poor confidence that have followed the lack of opportunities made available to them to make decisions about their own lives. The efficacy and tension of corporate parenting is also explored with suggestions from the participants on how the care system could be constructed differently to facilitate their voice and that of much younger children than themselves. Thus, the debate becomes one of adult ability and preparedness to involve young people in decisions about their own lives, rather than whether they are able to participate effectively.
Mineral metabolism disturbances in patients with chronic kidney disease
Authors: Kestenbaum B. , Belozeroff V.
Published: July 2007

Background  Kidney disease, especially chronic kidney disease (CKD), is a worldwide public health problem with serious adverse health consequences for affected individuals. Secondary hyperparathyroidism, a disorder characterized by elevated serum parathyroid hormone levels, and alteration of calcium and phosphorus homeostasis are common metabolic complications of CKD that may impact cardiovascular health.

Materials and methods  Here, we systematically review published reports from recent observational studies and clinical trials that examine markers of altered mineral metabolism and clinical outcomes in patients with CKD.

Results  Mineral metabolism disturbances begin early during the course of chronic kidney disease, and are associated with cardiovascular disease and mortality in observational studies. Vascular calcification is one plausible mechanism connecting renal‐related mineral metabolism with cardiovascular risk. Individual therapies to correct mineral metabolism disturbances have been associated with clinical benefit in some observational studies; clinical trials directed at more comprehensive control of this problem are warranted.

Conclusions  There exists a potential to improve outcomes for patients with CKD through increased awareness of the Bone Metabolism and Disease guidelines set forth by the National Kidney Foundation–Kidney Disease Outcomes Quality Initiative. Future studies may include more aggressive therapy with a combination of agents that address vitamin D deficiency, parathyroid hormone and phosphorus excess, as well as novel agents that modulate circulating promoters and inhibitors of calcification.
Leptin, leptin soluble receptor and coronary atherosclerosis
Authors: Hoefle G. , Saely C. H. , Risch L. , Rein P. , Koch L. , Schmid F. , Aczél S. , Marte T. , Langer P. , Drexel H.
Published: July 2007

Background  The adipose tissue‐related hormone leptin plays an important role in the regulation of body weight. The associations of leptin and leptin soluble receptor (sOb‐R) with coronary artery disease (CAD) are not clear.

Design  We measured leptin and sOb‐R in 543 consecutive patients (379 men, 164 women) referred for coronary angiography for the evaluation of CAD. Coronary artery stenoses with lumen narrowing ≥ 50% were considered significant.

Results  Serum leptin correlated significantly with body mass index (rs = 0·443), with insulin resistance as assessed by the homeostasis model for the assessment of insulin resistance (rs = 0·339), with serum triglycerides (rs = 0·181), with systolic as well as diastolic blood pressure (rs = 0·170 and rs = 0·133, respectively) and, inversely, with sOb‐R (rs = –0·346; P < 0·01 for all correlations). Coronary angiography revealed significant coronary artery stenoses in 331 (61%) of our patients. Serum leptin was significantly lower in patients with significant coronary artery stenoses than in patients without such lesions (8·5 ± 7·8 vs. 13·2 ± 12·2 ng mL−1; P < 0·001). Multivariate logistic regression analysis proved serum leptin inversely and independently associated with the presence of significant coronary artery stenoses (standardized adjusted odds ratio 0·746, 95% confidence interval 0·566–0·983, P = 0·038). In contrast to serum concentrations of leptin, serum concentrations of sOb‐R did not significantly differ between patients with significant stenoses and those without such lesions (22·4 ± 8·3 vs. 23·1 ± 12·1 ng mL−1; P = 0·655).

Conclusions  Serum leptin but not sOb‐R is significantly lower in patients with angiographically determined CAD. Despite its association with cardiovascular risk factors, leptin should not be simply regarded as a promoter of atherosclerosis.
Fatty acid incorporation in endothelial cells and effects on endothelial nitric oxide synthase
Authors: Couloubaly S. , Deloménie C. , Rousseau D. , Paul J. L. , Grynberg A. , Pourci M. L.
Published: July 2007

Background  The nature of fatty acids provided by the diet as well as plasma lipid metabolism can modify the composition and properties of plasma membrane and thus the activity of membrane proteins. In humans, as well as in experimental models, diabetes is associated with both an alteration in serum lipid profile and a documented endothelial dysfunction. This in vitro study investigated on an immortalized human endothelial cell line (EA.hy 926) the specific effects of several free fatty acids (FFAs) on the composition of cellular membranes and the regulation of endothelial nitric oxide synthase (eNOS).

Materials and methods  0·1% of lipid deprived serum was added to the incubation medium with 25 mm glucose in order to study the effects of individual fatty acids: myristic acid, palmitic acid, stearic acid, oleic acid or linoleic acid at 100 µm bound with albumin. The effects of the FFAs on the endothelial nitric oxide synthase were investigated on mRNA level by quantitative PCR, on protein level and Ser1177 phosphorylation by Western blot and on enzymatic activity on living cells using radiolabelled arginine.

Results  Free linoleic acid increased the membrane content in n‐6 fatty acids (mainly C18: n‐6 and its metabolites) with a decrease in saturated and monounsaturated fatty acids. These conditions decreased the basal eNOS activity and reduced the phosphorylation of eNOS‐Ser1177 due to activation by histamine. Free palmitic acid enriched the membranes with 16 : 0 with a slight decrease in monounsaturated fatty acids. These conditions increased eNOS activation without increasing Ser1177 phosphorylation upon histamine activation. The addition of the other FFAs also resulted in modifications of membrane composition, which did not to affect eNOS‐Ser1177 phosphorylation.

Conclusion  Among the fatty acids used, only modification of the membrane composition due to linoleic acid supply disturbed the basal enzymatic activity and Ser1177 phosphorylation of eNOS in a way that limited the role of histamine activation. Linoleic acid might involve the dysfunction of both eNOS basal activity and its phosphorylation status and may then contribute to an impaired vasodilatation in vivo.
Alpha interferon for hepatitis C virus infection in haemophilic patients
Authors: Telfer Paul , Devereux Helen , Colvin Brian , Hayden Sheila , Dusheiko Geoffrey , Lee Christine
Published: July 2007

We have conducted a controlled trial in 20 haemophilic patients in which intravenous recombinant interferon alpha‐2a, 3 mega units thrice weekly, was used to treat chronic hepatitis C infection. The study endpoints included complete and paritial normalization of serum ALT, and loss of serum HCV RNA as determined by polymerase chain reaction (PCR). Interferon treatment was effective, and resulted in improvement in ALT in nine (45%) and a loss of HCV RNA in five patients (26%), but a sustained normalization of ALT has been seen in only one case. Responses were poor in those with HIV coinfection or with HCV genotype 1 (Simmonds classification). Troublesome side‐effects were reported in 80%. The occurrence of a factor VIIIc inhibitor during the study was possibly an autoimmune complication of interferon. In conclusion, we have shown lower response rates than those seen in post‐transfusion hepatitis C infection and suspect that current interferon regimes are unlikely to influence the natural history of HCV infection in haemophilia. Consideration should be given to trials of higher dosage interferon, and of long‐term maintenance therapy for those who relapse.
Methylation of proteins involved in translation
Authors: Polevoda Bogdan , Sherman Fred
Published: July 2007

Methylation is one of the most common protein modifications. Many different prokaryotic and eukaryotic proteins are methylated, including proteins involved in translation, including ribosomal proteins (RPs) and translation factors (TFs). Positions of the methylated residues in six Escherichia coli RPs and two Saccharomyces cerevisiae RPs have been determined. At least two RPs, L3 and L12, are methylated in both organisms. Both prokaryotic and eukaryotic elongation TFs (EF1A) are methylated at lysine residues, while both release factors are methylated at glutamine residues. The enzymes catalysing methylation reactions, protein methyltransferases (MTases), generally use S‐adenosylmethionine as the methyl donor to add one to three methyl groups that, in case of arginine, can be asymetrically positioned. The biological significance of RP and TF methylation is poorly understood, and deletions of the MTase genes usually do not cause major phenotypes. Apparently methylation modulates intra‐ or intermolecular interactions of the target proteins or affects their affinity for RNA, and, thus, influences various cell processes, including transcriptional regulation, RNA processing, ribosome assembly, translation accuracy, protein nuclear trafficking and metabolism, and cellular signalling. Differential methylation of specific RPs and TFs in a number of organisms at different physiological states indicates that this modification may play a regulatory role.