bluuebunny/crossref_metadata_2025 huggingface dataset:
The Nature of Gender: Work, Gender, and Environment
Authors: Nightingale Andrea
Published: March 2006

Gender has long been recognised as important within environmental issues, but there has been considerable debate over how to conceptualise the gender–environment nexus. As feminist theorising around women and gender has changed, so have conceptualisations about gender and environment, leading to a key debate within ecofeminism and related literatures about whether there is an essential or a contingent relationship between women and natural environments. Within geography, most political ecologists work with the assumption that the gender–environment nexus is a contingent relationship, and thus investigate how gender relations are salient in the symbolic and material construction of environmental issues. In this paper I seek to build from this work and again raise the issue of how gender is conceptualised in relation to environment. I begin by briefly reviewing some of the work that has been done on gender and environment and then draw from poststructural feminism to suggest that gender itself has been undertheorised in work on environment. Once gender is reconceptualised as a process, the dynamic relationship between gender, environment, and other aspects of social and cultural life can be brought into view. What emerges is the need for political ecologists to examine gender beyond the household and community and the need to reconceptualise the gender–environment nexus. A case study of community forestry in Nepal is used to illustrate the importance of interrogating the processes by which gender relations become salient and are reproduced symbolically and materially.
Groups as a Means or an End? Social Capital and the Promotion of Cooperation in Ghana
Authors: Porter Gina , Lyon Fergus
Published: March 2006

In the past two decades there has been a growing emphasis within the international development industry on promoting group activity. In this paper we chart how interpretation of the loose concept of social capital has shaped donor and NGO discourses on, and their preoccupation with, groups. Donors are using blueprints of group cooperation in an asocial and aspatial manner that ignores local specificities of place, space, and cultural context. An empirical case is examined that demonstrates how donor discourse is reinterpreted, translated, and even rejected by players at different spatial scales. The reasons for the continued donor preoccupation with groups in the face of local resistances are explored.
Processes influencing soil carbon storage following afforestation of pasture with Pinus radiata at different stocking densities in New Zealand
Authors: Scott Neal A. , Tate Kevin R. , Ross Des J. , Parshotam Aroon
Published: March 2006

Since 1992, afforestation with Pinus radiata D. Don in New Zealand has led to the establishment of over 600 000 ha of new plantation forests, about 85% of which are on fertile pastures used previously for grazing sheep and cattle. While this leads to rapid accumulation of carbon (C) in vegetation, the effects of afforestation on soil C are poorly understood. We examined key soil C cycling processes at the (former) Tikitere agroforestry experimental site near Rotorua, New Zealand. In 1973, replicated stands of P. radiata (100 and 400 stems/ha) were established on pastures, while replicated pasture plots were maintained throughout the first 26-year rotation. In 1996, soil C and microbial biomass C in 0–0.10 m depth soil, in situ soil respiration and net N mineralisation, and soil temperature were lower in the forest than in the pasture, and tended to decline with increasing tree-stocking density. In the 400 stems/ha stands, mineral soil C (0–0.50 m depth) was lower than in the pasture (104 and 126 Mg C/ha, respectively; P < 0.01). Carbon accumulation in the forest floor during the first rotation of these forest stands was 12 Mg C/ha. Using the Rothamsted soil C model (Roth-C), we examined how changes in plant C inputs following afforestation might lead to changes in soil C content to 0.30 m depth. Steady-state pasture inputs of 9.0 Mg C/ha.year were estimated using Roth-C; these C inputs were assumed to decrease linearly during the first 12 years following tree establishment (until canopy closure). Below-ground C inputs in the forest were estimated using steady-state relationships between litterfall and soil respiration; these inputs were assumed to increase linearly between years 1 and 12, after which they remained constant at 1.53 Mg C/ha.year until harvest. Measured changes in soil C (0−0.30 m) during the first rotation, in conjunction with the below-ground inputs, were used to estimate above-ground inputs (as a proportion of total litterfall [3.81 Mg C/ha.year]) to the soil. Our results suggest 10% of litterfall C over one rotation actually entered the mineral soil. Using these results and estimates of additional C inputs to the soil from harvest slash and weeds following harvest, we found mineral-soil C stocks would continue to decline during second and third rotations of P. radiata; the magnitude of this decline depended in part on how much slash enters the mineral soil matrix. We confirmed our modelling approach by simulating soil C changes to within 8% over 19 years following afforestation of pasture at another previously studied site, Purukohukohu. Whether afforestation leads to an increase or decrease in mineral-soil C may depend on previous pasture management; in highly productive pastures, high C inputs to the soil may maintain soil C at levels that cannot be sustained when trees are planted onto these grasslands.
The effect of organic mulch amendments on the physical and chemical properties and revegetation success of a saline-sodic minespoil from central Queensland, Australia
Authors: Grigg A. H. , Sheridan G. J. , Pearce A. B. , Mulligan D. R.
Published: March 2006

Saline-sodic clay minespoil materials excavated during open-cut coal mining in central Queensland, Australia, pose significant challenges for revegetation, particularly where suitable topsoil capping is not available. We examined the ability of sawdust or straw mulch amendments to ameliorate the adverse properties of these minespoils and improve the success of revegetation efforts. In laboratory studies, mulch application improved infiltration, increased soil moisture retention and reduced surface crust strength. In the field, mulches incorporated to a depth of 0.15 m at application rates of at least 20 t/ha straw or 80 t/ha sawdust were needed to mitigate against capillary rise of salts during drying cycles and support satisfactory vegetation cover. Further research is needed to determine whether improvements are maintained beyond the 4-year trial period reported here.
Influence of long-term irrigation on the distribution and availability of soil phosphorus under permanent pasture
Authors: Condron L. M. , Sinaj S. , McDowell R. W. , Dudler-Guela J. , Scott J. T. , Metherell A. K.
Published: March 2006

This study examined the influence of irrigation on soil phosphorus (P) distribution and availability under permanent pasture in New Zealand. Soil samples (0–0.075, 0.075–0.15, 0.15–0.25 m) were taken from a long-term field experiment, which included a dryland and 2 irrigation treatments (irrigated at 10% and 20% soil moisture) that had received 25 kg P/ha annually as superphosphate for 52 years. Corresponding data for soil from an adjacent ‘wilderness’ site that had not been used for agriculture for 54 years were included for comparison. Analyses included total P, organic P, and inorganic P; isotopic exchange kinetics (IEK) was used to determine soil inorganic P pools of differing plant availability. Concentrations of total and inorganic P were greater in soil taken from the dryland treatment than the irrigated treatments at all depths. This was attributed to a combination of decreased pasture growth and P transfer in drainage and off-farm produce. Concentrations of organic P were greater in the irrigated treatments (e.g. 0–0.075 m: 672–709 mg P/kg) than in the dryland treatment (e.g. 0–0.075 m: 574 mg P/kg) as a consequence of increased pasture production and soil biological activity. Inorganic P availability (Cp and E1min) was also greater in the dryland treatment than the irrigated treatments. Furthermore, concentrations of inorganic P in the recalcitrant IEK pool (E>3m = E3m–1y + E>1y) in the 0–0.075 m soil from the dryland treatment (479 mg P/kg) were significantly greater than the 10% irrigated (346 mg P/kg) and 20% irrigated (159 mg P/kg) treatments, which was mainly attributed to physico-chemical reactions that decreased the exchangeability of accumulated inorganic P with time. Despite increased P retention capacity at depth (R/r1, 0.15–0.25 m: dryland 6.6, 10% irrigated 10.2, 20% irrigated 12.8), concentrations of total inorganic P in the 0.15–0.25 m soil layer were lower under irrigation (195–266 mg P/kg) than dryland (354 mg P/kg), which indicated that long-term flood irrigation increased P transfer by leaching. The findings of this study revealed that while irrigation improved the utilisation of applied fertiliser P it also resulted in increased P movement to depth in the soil profile.
Sulfur mineralisation in a coarse-textured soil after different sulfate fertilisation histories, and yield responses of wheat and lupin
Authors: Anderson G. C. , Fillery I. R. P. , Ripper F. H. , Leach B. J.
Published: March 2006

The effect of annual applications of gypsum at rates of 0, 5, 15, and 30 kg S/ha on grain yields of lupin, wheat, and canola were studied between 1992 and 1995, and the residual value of these sulfate (SO42–) inputs was examined in 1996 and 1997. The top 0.1 m of soil was collected in 1994, 1996, and 1997 to determine KCl-40 extractable S. Extensive soil sampling was undertaken in 1996 and 1997 to determine quantities of SO42– in soil layers to 0.5 m, and to measure rates of net SO42– mineralisation. Application of gypsum resulted in significantly higher grain yields in 2 out of 3 wheat crops, and in canola, but SO42– fertiliser use did not increase lupin yield. Wheat yields were also significantly higher in treatments with a prior history of gypsum use but lupin grain yields were not enhanced by previous gypsum applications. The quantities of SO42– in the top 0.2 m of soil decreased sharply in the first year after SO42– fertilisers were withheld with further declines in SO42– to low levels across all treatments in the second year gypsum was not applied. A buildup of SO42– was evident in soil between 0.3 and 0.5 m. However, S budgets for the top 0.5 m suggested that little SO42– leached below 0.5 m in the years studied. Previous sulfur fertilisation did not significantly affect the total growing season net SO42– mineralisation, which was 2.2–8.0 kg S/ha. Mineralisation of organic S over a summer fallow period (0–5.8 kg S/ha) and the total S net mineralised over 17 months (7–17 kg S/ha) was significantly affected by previous S fertiliser history. Net SO42– mineralisation was not correlated with the labile organic S fraction extracted using the KCl-40 soil test.
Capture of overland flow by a tree belt on a pastured hillslope in south-eastern Australia
Authors: Ellis T. W. , Leguédois S. , Hairsine P. B. , Tongway D. J.
Published: March 2006

We describe a rainfall simulator experiment designed to measure the capture, by a fenced tree belt, of excess water generated as Hortonian flow from a pasture slope. Three rainfall events (48, 49, and 75 mm/h for 13, 30, and 30 min, respectively) were applied, of which 15%, 29%, and 44%, respectively, ran off and drained onto the tree belt. The tree belt captured 100%, 32–68%, and 0–28% of the runoff from the 3 events, respectively. These captured runoff volumes represented 31–39%, 22–45%, and 0–29% increases in water supply to the trees, in addition to incident rainfall. Infiltration rates within the tree belt were up to 46% higher than in the pasture zone. This higher infiltration was mainly attributed to better soil surface conditions in the absence of stock and a 50-mm layer of tree litter. Overland flows within the tree belt formed tree litter into microterraces, which spread and slowed flows and allowed greater time for infiltration.
Two distinct cytoplasmic regions of the β2 integrin chain regulate RhoA function during phagocytosis
Authors: Wiedemann Agnès , Patel Jayesh C. , Lim Jenson , Tsun Andy , van Kooyk Yvette , Caron Emmanuelle
Published: March 2006

αMβ2 integrins mediate phagocytosis of opsonized particles in a process controlled by RhoA, Rho kinase, myosin II, Arp2/3, and actin polymerization. αMβ2, Rho, Arp2/3, and F-actin accumulate underneath bound particles; however, the mechanism regulating Rho function during αMβ2-mediated phagocytosis is poorly understood. We report that the binding of C3bi-opsonized sheep red blood cells (RBCs) to αMβ2 increases Rho-GTP, but not Rac-GTP, levels. Deletion of the cytoplasmic domain of β2, but not of αM, abolished Rho recruitment and activation, as well as phagocytic uptake. Interestingly, a 16–amino acid (aa) region in the membrane-proximal half of the β2 cytoplasmic domain was necessary for activating Rho. Three COOH-terminal residues (aa 758–760) were essential for β2-induced accumulation of Rho at complement receptor 3 (CR3) phagosomes. Activation of Rho was necessary, but not sufficient, for its stable recruitment underneath bound particles or for uptake. However, recruitment of active Rho was sufficient for phagocytosis. Our data shed light on the mechanism of outside-in signaling, from ligated integrins to the activation of Rho GTPase signaling.
A role for the RabA4b effector protein PI-4Kβ1 in polarized expansion of root hair cells in Arabidopsis thaliana
Authors: Preuss Mary L. , Schmitz Aaron J. , Thole Julie M. , Bonner Heather K.S. , Otegui Marisa S. , Nielsen Erik
Published: March 2006

The RabA4b GTPase labels a novel, trans-Golgi network compartment displaying a developmentally regulated polar distribution in growing Arabidopsis thaliana root hair cells. GTP bound RabA4b selectively recruits the plant phosphatidylinositol 4-OH kinase, PI-4Kβ1, but not members of other PI-4K families. PI-4Kβ1 colocalizes with RabA4b on tip-localized membranes in growing root hairs, and mutant plants in which both the PI-4Kβ1 and -4Kβ2 genes are disrupted display aberrant root hair morphologies. PI-4Kβ1 interacts with RabA4b through a novel homology domain, specific to eukaryotic type IIIβ PI-4Ks, and PI-4Kβ1 also interacts with a Ca2+ sensor, AtCBL1, through its NH2 terminus. We propose that RabA4b recruitment of PI-4Kβ1 results in Ca2+-dependent generation of PI-4P on this compartment, providing a link between Ca2+ and PI-4,5P2–dependent signals during the polarized secretion of cell wall components in tip-growing root hair cells.
Stu2p binds tubulin and undergoes an open-to-closed conformational change
Authors: Al-Bassam Jawdat , van Breugel Mark , Harrison Stephen C. , Hyman Anthony
Published: March 2006

Stu2p from budding yeast belongs to the conserved Dis1/XMAP215 family of microtubule-associated proteins (MAPs). The common feature of proteins in this family is the presence of HEAT repeat–containing TOG domains near the NH2 terminus. We have investigated the functions of the two TOG domains of Stu2p in vivo and in vitro. Our data suggest that Stu2p regulates microtubule dynamics through two separate activities. First, Stu2p binds to a single free tubulin heterodimer through its first TOG domain. A large conformational transition in homodimeric Stu2p from an open structure to a closed one accompanies the capture of a single free tubulin heterodimer. Second, Stu2p has the capacity to associate directly with microtubule ends, at least in part, through its second TOG domain. These two properties lead to the stabilization of microtubules in vivo, perhaps by the loading of tubulin dimers at microtubule ends. We suggest that this mechanism of microtubule regulation is a conserved feature of the Dis1/XMAP215 family of MAPs.
Vectorial insertion of apical and basolateral membrane proteins in polarized epithelial cells revealed by quantitative 3D live cell imaging
Authors: Hua Wei , Sheff David , Toomre Derek , Mellman Ira
Published: March 2006

Although epithelial cells are known to exhibit a polarized distribution of membrane components, the pathways responsible for delivering membrane proteins to their appropriate domains remain unclear. Using an optimized approach to three-dimensional live cell imaging, we have visualized the transport of newly synthesized apical and basolateral membrane proteins in fully polarized filter-grown Madin–Darby canine kidney cells. We performed a detailed quantitative kinetic analysis of trans-Golgi network (TGN) exit, passage through transport intermediates, and arrival at the plasma membrane using cyan/yellow fluorescent protein–tagged glycosylphosphatidylinositol-anchored protein and vesicular stomatitis virus glycoprotein as apical and basolateral reporters, respectively. For both pathways, exit from the TGN was rate limiting. Furthermore, apical and basolateral proteins were targeted directly to their respective membranes, resolving current confusion as to whether sorting occurs on the secretory pathway or only after endocytosis. However, a transcytotic protein did reach the apical surface after a prior appearance basolaterally. Finally, newly synthesized proteins appeared to be delivered to the entire lateral or apical surface, suggesting—contrary to expectations—that there is not a restricted site for vesicle docking or fusion adjacent to the junctional complex.
Controversy fuels trafficking of GPI-anchored proteins
Authors: Schuck Sebastian , Simons Kai
Published: March 2006

The model that glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) take a direct transport route to the apical membrane of epithelial cells has recently been challenged. In this issue, Paladino et al. (p. 1023) and Hua et al. (p. 1035) show that the original view nevertheless holds. This closes a chapter in the winding story of GPI-AP trafficking but opens another phase, as the controversy has stimulated the development of new methodology.
Physiological Features of the S- and M-cone Photoreceptors of Wild-type Mice from Single-cell Recordings
Authors: Nikonov Sergei S. , Kholodenko Roman , Lem Janis , Pugh Edward N.
Published: March 2006

Cone cells constitute only 3% of the photoreceptors of the wild-type (WT) mouse. While mouse rods have been thoroughly investigated with suction pipette recordings of their outer segment membrane currents, to date no recordings from WT cones have been published, likely because of the rarity of cones and the fragility of their outer segments. Recently, we characterized the photoreceptors of Nrl−/− mice, using suction pipette recordings from their “inner segments” (perinuclear region), and found them to be cones. Here we report the use of this same method to record for the first time the responses of single cones of WT mice, and of mice lacking the α-subunit of the G-protein transducin (Gtα−/−), a loss that renders them functionally rodless. Most cones were found to functionally co-express both S- (λmax = 360 nm) and M- (λmax = 508 nm) cone opsins and to be maximally sensitive at 360 nm (“S-cones”); nonetheless, all cones from the dorsal retina were found to be maximally sensitive at 508 nm (“M-cones”). The dim-flash response kinetics and absolute sensitivity of S- and M-cones were very similar and not dependent on which of the coexpressed cone opsins drove transduction; the time to peak of the dim-flash response was ∼70 ms, and ∼0.2% of the circulating current was suppressed per photoisomerization. Amplification in WT cones (A ∼4 s−2) was found to be about twofold lower than in rods (A ∼8 s−2). Mouse M-cones maintained their circulating current at very nearly the dark adapted level even when >90% of their M-opsin was bleached. S-cones were less tolerant to bleached S-opsin than M-cones to bleached M-opsin, but still far more tolerant than mouse rods to bleached rhodopsin, which exhibit persistent suppression of nearly 50% of their circulating current following a 20% bleach. Thus, the three types of mouse opsin appear distinctive in the degree to which their bleached, unregenerated opsins generate “dark light.”
Informational Properties of Anxiety
and Sadness, and Displaced Coping
Authors: Raghunathan Rajagopal , Pham Michel Tuan , Corfman Kim P.
Published: March 2006

Replicating Raghunathan and Pham (1999), results from two experiments confirm that while anxiety triggers a preference for options that are safer and provide a sense of control, sadness triggers a preference for options that are more rewarding and comforting. Results also indicate that these effects are driven by an affect-as-information process and are most pervasive when the source of anxiety or sadness is not salient. Finally, our results document a previously unrecognized phenomenon we term displaced coping, wherein affective states whose source is salient influence decisions that are seemingly—but not directly—related to the source of these affective states.
Cord blood transplantation for acute myelogenous leukemia using a conditioning regimen consisting of granulocyte colony‐stimulating factor‐combined high‐dose cytarabine, fludarabine, and total body irradiation
Authors: Tomonari Akira , Takahashi Satoshi , Ooi Jun , Nakaoka Takashi , Takasugi Kashiya , Uchiyama Michihiro , Tsukada Nobuhiro , Konuma Takaaki , Iseki Tohru , Tojo Arinobu , Asano Shigetaka
Published: March 2006

Abstract:  The cytotoxic effect of cytarabine (Ara‐C) on myeloid leukemic cells is enhanced by concomitant use of granulocyte colony‐stimulating factor (G‐CSF) in vitro. The feasibility of a conditioning regimen consisting of G‐CSF‐combined 24 g/m2 Ara‐C, 90 mg/m2 fludarabine, and 12 Gy total body irradiation was studied for five patients with acute myelogenous leukemia in cord blood transplantation (CBT). Graft vs. host disease (GVHD) prophylaxis consisted of cyclosporine and methotrexate. After the conditioning regimen, 2.48 × 107/kg (2.28–3.53) of cord blood nucleated cells was infused. Neutrophil counts consistently >0.5 × 109/L was achieved 24 d (22–32) after CBT. Grade I stomatitis and gastrointestinal toxicities occurred in all patients. Grades I and II acute GVHD occurred in one and four patients, respectively, which resolved without steroid therapy. Sepsis and aspergillosis occurred in two and one patients, respectively. All patients were alive without leukemia relapse at a follow up of 15 months (12–43) after CBT. This conditioning regimen could avoid the toxicities of high‐dose cyclophosphamide but might enhance the cytotoxic effect of Ara‐C. Large‐scale studies will be needed to determine the efficacy and safety of the conditioning regimen in CBT.
Long‐term and concentration‐dependent beneficial effect of efavirenz on HDL‐cholesterol in HIV‐infected patients
Authors: Pereira Sofia A. , Branco Teresa , Côrte‐Real Rita M. , Germano Isabel , Lampreia Fátima , Caixas Umbelina , Monteiro Emília C.
Published: March 2006

Aims To investigate the long‐term effects of efavirenz on cholesterol (TC), high‐density lipoprotein cholesterol (HDL‐C), low‐density lipoprotein (LDL‐C) and triglycerides (TG).

Methods Thirty‐four HIV‐infected patients who commenced efavirenz therapy were monitored for 36 months.

Results In patients with baseline HDL‐C < 40 mg · dL−1 an increase in HDL‐C from 31 ± 1 mg · dL−1 to 44 ± 2 mg · dL−1 (95% confidence interval 5.9, 21.9, P < 0.01) was observed and remained throughout the follow‐up period. Median efavirenz plasma concentration was 1.98 mg · L−1 and a direct correlation between percentage of HDL‐C variation or TC/HDL‐C ratio and efavirenz plasma concentrations was found.

Conclusions There is evidence of a long‐term and concentration‐dependent beneficial effect of efavirenz on HDL‐C in HIV‐infected patients.
An evaluation of potential mechanism‐based inactivation of human drug metabolizing cytochromes P450 by monoamine oxidase inhibitors, including isoniazid
Authors: Polasek Thomas M. , Elliot David J. , Somogyi Andrew A. , Gillam Elizabeth M. J. , Lewis Benjamin C. , Miners John O.
Published: March 2006

AimsTo characterize potential mechanism‐based inactivation (MBI) of major human drug‐metabolizing cytochromes P450 (CYP) by monoamine oxidase (MAO) inhibitors, including the antitubercular drug isoniazid.

MethodsHuman liver microsomal CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A activities were investigated following co‐ and preincubation with MAO inhibitors. Inactivation kinetic constants (KIandkinact) were determined where a significant preincubation effect was observed. Spectral studies were conducted to elucidate the mechanisms of inactivation.

ResultsHydrazine MAO inhibitors generally exhibited greater inhibition of CYP following preincubation, whereas this was less frequent for the propargylamines, and tranylcypromine and moclobemide. Phenelzine and isoniazid inactivated all CYP but were most potent toward CYP3A and CYP2C19. Respective inactivation kinetic constants (KIandkinact) for isoniazid were 48.6 µmand 0.042 min−1and 79.3 µmand 0.039 min−1. Clorgyline was a selective inactivator of CYP1A2 (6.8 µmand 0.15 min−1). Inactivation of CYP was irreversible, consistent with metabolite‐intermediate complexation for isoniazid and clorgyline, and haeme destruction for phenelzine. With the exception of phenelzine‐mediated CYP3A inactivation, glutathione and superoxide dismutase failed to protect CYP from inactivation by isoniazid and phenelzine. Glutathione partially slowed (17%) the inactivation of CYP1A2 by clorgyline. Alternate substrates or inhibitors generally protected against CYP inactivation.

ConclusionsThese data are consistent with mechanism‐based inactivation of human drug‐metabolizing CYP enzymes and suggest that impaired metabolic clearance may contribute to clinical drug–drug interactions with some MAO inhibitors.
α‐Haemoglobin stabilising protein is a quantitative trait gene that modifies the phenotype of β‐thalassaemia
Authors: Lai Mei I. , Jiang Jie , Silver Nicholas , Best Steve , Menzel Stephan , Mijovic Aleksandar , Colella Stefano , Ragoussis Jiannis , Garner Chad , Weiss Mitchell J. , Thein Swee L.
Published: March 2006

It has been suggested that altered levels or function of α‐haemoglobin stabilising protein (AHSP), an erythroid‐specific protein that binds specifically to free α‐(haemo)globin, might account for some of the clinical variability in β‐thalassaemia. To assess the variation of AHSP expression, mRNA levels in circulating reticulocytes of 103 healthy individuals were measured by quantitative reverse transcription‐polymerase chain reaction. AHSP expression varied up to threefold, and did not correlate with age or sex. A systematic survey of the AHSP locus identified eight sequence variants, of which six were common. Four common variants, including the longer homopolymer (T18) in the putative promoter, are strongly associated with AHSP expression. Reporter assays in K562 cells showed that the activity of the shorter (T15) reporter was relatively lower than that of the T18 reporter. In a study of nine anaemic patients who were heterozygous for β‐thalassaemia and also heterozygous for the triplicated α‐globin gene (ααα/αα), frequency of the shorter homopolymer was higher than expected. AHSP expression is variable, with cis control accounting for some of its variance. In some families, the subtle altered levels in AHSP related to the AHSP genotype appears to be a relevant contributory factor in the haematological phenotype.
Role of interleukin (IL-10) in probiotic-mediated immune modulation: an assessment in wild-type and IL-10 knock-out mice
Authors: Sheil B , MacSharry J , O'Callaghan L , O'Riordan A , Waters A , Morgan J , Collins J K , O'Mahony L , Shanahan F
Published: March 2006

While the impact of Bifidobacterium infantis 35624 and other probiotics on cytokines has been shown in established colitis, the effects of B. infantis consumption in pre-inflammation of interleukin (IL)-10 knock-out (KO) mice and on the wild-type (WT) C57Bl/6 mice have not been well demonstrated. The objective of this study was to examine cytokine responses in mucosal and systemic lymphoid compartments of IL-10 KO mice early in disease and to compare with control WT mice. Mice were fed B. infantis or placebo for 5 weeks and culled prior to the onset of chronic intestinal inflammation (12–14 weeks). The spleen, Peyer’s patches and intestinal mucosa were removed and stimulated with various bacterial stimuli. Cytokine levels were measured by enzyme-linked immunosorbent assay. While basal intestinal and systemic cytokine profiles of WT and IL-10 KO mice were similar, transforming growth factor (TGF)-β was reduced in the spleen of IL-10 KO mice. Following probiotic consumption, interferon (IFN)-γ was reduced in the Peyer’s patch of both WT and IL-10 KO mice. Alterations in IFN-γ in the Peyer’s patches of WT mice (enhancement) versus IL-10 KO (reduction) were observed following in vitro stimulation with salmonella. Differential IL-12p40, CCL2 and CCL5 responses were also observed in IL-10 KO mice and WT mice. The cytokine profile of IL-10 KO mice in early disease was similar to that of WT mice. The most pronounced changes occurred in the Peyer’s patch of IL-10 KO mice, suggesting a probiotic mechanism of action independent of IL-10. This study provides a rationale for the use of B. infantis 35624 for the treatment of gastrointestinal inflammation.
Modulation of immune responses and suppression of experimental autoimmune myasthenia gravis by surgical denervation of the spleen
Authors: Bakhiet M , Yu L-Y , Özenci V , Khan A , Shi F-D
Published: March 2006

Critical interactions between the nervous system and the immune system during experimental autoimmune myasthenia gravis (EAMG) were examined in an animal model for human MG after immunization of adult female Lewis rats with Torpedo acetylcholine receptor (AChR) and complete Freund’s adjuvant. Immunized rats depicted marked clinical severity of the disease. Using enzyme-linked immunospot (ELISPOT) assay and in situ hybridization techniques, immune responses in these animals were examined and showed elevated numbers of anti-AChR IgG secreting B cells and AChR reactive interferon (IFN)-γ-secreting cells, enhanced mRNA expression of the proinflammatory cytokines IFN-γ and tumour necrosis factor (TNF)-α as Th1 subset and the anti-inflammatory cytokines interleukin (IL)-4 and IL-10 as a Th2 subset, and transforming growth factor (TGF)-β as a Th3 cytokine. Corticosterone and prostaglandin E2 (PGE2) levels were measured by radioimmunoassay and illustrated increased production after immunization. Surgical denervation of the spleen reduced significantly the clinical severity of the disease, suppressed the numbers of IgG and IFN-γ-secreting cells, down-regulated the mRNA expression for cytokines and reduced corticosterone and PGE2 production. As controls, sham-operated rats were used and showed results as the EAMG non-denervated control rats. The data present herein, and for the first time, substantial effects of the nervous system on immune responses that may influence the outcome of EAMG. These effects were not dependent on cytokine inhibitory mediators such as prostaglandins or stress hormones. IL-10 and TGF-β, the two potent immunosuppressive cytokines, were also suppressed, indicating a general suppression by splenic denervation. More investigations are initiated at our laboratories to understand the evident neural control over the immune system during challenges leading to the break of tolerance and development of autoimmunity, which may assist in innovative therapeutic approaches.
The presence of antibodies to oxidative modified proteins in serum from polycystic ovary syndrome patients
Authors: Palacio J R , Iborra A , Ulcova-Gallova Z , Badia R , Martínez P
Published: March 2006

Polycystic ovary syndrome (PCOS) affects 5–10% of women of reproductive age. Free radicals, as a product of oxidative stress, impair cells and tissue properties related to human fertility. These free radicals, together with the oxidized molecules, may have a cytotoxic or deleterious effects on sperm and oocytes, on early embryo development or on the endometrium. Aldehyde-modified proteins are highly immunogenic and circulating autoantibodies to new epitopes, such as malondialdehyde (MDA), may affect the reproductive system. Autoantibodies or elevated reactive oxygen species (ROS) in serum are often associated with inflammatory response. The purpose of this work is to investigate whether PCOS women show increased levels of oxidized proteins (protein–MDA) and anti-endometrial antibodies (AEA) in their sera, compared with control patients, and to determine whether AEA specificity is related to oxidized protein derivatives. Sera from 31 women [10 patients with PCOS (PCOS group) and 21 women with male factor of infertility (control group)] were chosen from patients attending for infertility. Anti-endometrial antibodies were determined by enzyme-linked immunosorbent assay (ELISA) with an endometrial cell line (RL-95). Antibodies against MDA modified human serum albumin (HSA–MDA) were also determined by ELISA. Oxidized proteins (protein–MDA) in serum were determined by a colorimetric assay. Patients with PCOS have significantly higher levels of AEA and anti-HSA–MDA, as well as oxidized proteins (protein–MDA) in serum than control patients. For the first time, we describe an autoimmune response in PCOS patients, in terms of AEA. The evidence of protein–MDA in the serum of these patients, together with the increased antibody reactivity to MDA-modified proteins (HSA–MDA) in vitro, supports the conclusion that oxidative stress may be one of the important causes for abnormal endometrial environment with poor embryo receptivity in PCOS patients.
Ischaemia is linked to inflammation and induction of angiogenesis in pancreatic islets
Authors: Linn T , Schmitz J , Hauck-Schmalenberger I , Lai Y , Bretzel R G , Brandhorst H , Brandhorst D
Published: March 2006

β-cell replacement is the only way to restore euglycaemia in patients with type-1 diabetes. Pancreatic tissue, processed for subsequent clinical islet transplantation, is exposed to ischaemia causing injury and death in a large number of islets before and after transplantation. In this review we summarize what is known on the sources of environmental stress for pancreatic islets, such as insufficient oxygen supply during pancreas procurement and in culture prior to intraportal transplantation, nutritional and oxygen deprivation during the isolation process, and the consequences of hyperglycaemia. An increasingly recognized role in the modulation of β-cell function and these environmental stress factors plays the vascular network of the pancreatic islets. Islet revascularization by angiogenesis is relevant for the survival of the graft subsequent to transplantation. Potential strategies offered by therapeutic induction of revascularization to ameliorate the detrimental impact of these factors on the quality of islet transplants are discussed.
Imatinib mesylate inhibits proliferation of rheumatoid synovial fibroblast-like cells and phosphorylation of Gab adapter proteins activated by platelet-derived growth factor
Authors: Kameda H , Ishigami H , Suzuki M , Abe T , Takeuchi T
Published: March 2006

Receptors for platelet-derived growth factor (PDGF) are abundantly expressed on synovial fibroblast-like (SFL) cells from patients with rheumatoid arthritis (RA), and stimulation with PDGF enhances both the anchorage-dependent and -independent growth of RA–SFL cells. To elucidate the molecular mechanisms responsible for the excessive growth of RA–SFL cells and to seek a novel molecular-targeting therapy for RA, we examined the expression of adapter proteins and the effect of the specific inhibition of PDGF receptor activation by imatinib mesylate. Cultured SFL cells were used in the present study after 2–5 passages. The anchorage-dependent and -independent growth patterns of the SFL cells were evaluated using a tetrazolium-based assay and colony formation in 0·3% agar, respectively. Adapter proteins Gab1 and Gab2 were expressed in RA–SFL cells, and both proteins were rapidly (< 1 min) tyrosine-phosphorylated after the stimulation of RA–SFL cells with 10 ng/ml of PDGF and, to a lesser extent, after stimulation with 100 ng/ml of epidermal growth factor (EGF). The inhibition of PDGF receptor tyrosine kinase activation by 1 µM or less of imatinib mesylate specifically suppressed the PDGF-dependent, but not EGF-dependent, tyrosine phosphorylation of various proteins. Moreover, imatinib mesylate abolished both the anchorage-dependent and -independent proliferation of RA–SFL cells induced by PDGF stimulation. These results suggest that Gab adapter proteins are expressed and likely to be involved in the growth signalling of rheumatoid synovial cells and that imatinib mesylate, a key drug in the treatment of chronic myeloid leukaemia, may also be effective for the treatment of RA.
Influence of major surgery on the mannan-binding lectin pathway of innate immunity
Authors: Ytting H , Christensen I J , Basse L , Lykke J , Thiel S , Jensenius J C , Nielsen H J
Published: March 2006

The mannan-binding lectin (MBL) pathway of complement activation is important in host defence against pathogens and possibly against cancer. We investigated the effect of major surgery on two central components of the MBL pathway; MBL and the MBL-associated serine protease MASP-2, and for comparison also measured the interleukin (IL)-6 and C-reactive protein (CRP) levels. Serial blood samples were obtained from patients belonging to two different cohorts. Cohort 1 comprised 60 patients undergoing open or laparoscopic colectomy for benign disease (n = 12) or colon cancer (n = 48). Cohort 2 comprised 27 patients undergoing elective, open surgery for colorectal cancer, and was included in order to cover blood sampling between days 2 and 6. As expected, the surgical stress induced a marked acute phase response, as evidenced by a large increase in IL-6 (18-fold) and CRP (13-fold) levels with maximum at 12 h and 2 days, respectively. However, in both cohorts the levels of MBL and MBL-associated serine protease 2 (MASP-2) were largely unaffected, except for a minor but significant increase around day 8 in cohort 1. The preoperative levels of IL-6 and CRP were correlated significantly in both cohorts (r = 0·71, P < 0·0001 and r = 0·65, P = 0·005, respectively). Preoperative MASP-2 correlated with preoperative CRP (r = 0·59, P = 0·001) and IL-6 (r = 0·55, P = 0·02) in cohort 2 only. In contrast to the marked effects on the levels of IL-6 and CRP, the surgery influenced only marginally the two proteins of the MBL pathway.
The C-terminus of complement regulator Factor H mediates target recognition: evidence for a compact conformation of the native protein
Authors: Oppermann M , Manuelian T , Józsi M , Brandt E , Jokiranta T S , Heinen S , Meri S , Skerka C , Götze O , Zipfel P F
Published: March 2006

The complement inhibitor Factor H has three distinct binding sites for C3b and for heparin, but in solution uses specifically the most C-terminal domain, i.e. short consensus repeats (SCR) 20 for ligand interaction. Two novel monoclonal antibodies (mABs C14 and C18) that bind to the most C-terminal domain SCR 20 completely blocked interaction of Factor H with the ligands C3b, C3d, heparin and binding to endothelial cells. In contrast, several mAbs that bind to the N-terminus and to the middle regions of the molecule showed no or minor inhibitory effects when assayed by enzyme-linked immunosorbent assay (ELISA) and ligand interaction assays. This paradox between a single functional binding site identified for native Factor H versus multiple interaction sites reported for deletion constructs is explained by a compact conformation of the fluid phase protein with one accessible binding site. On zymosan particles mAbs C14 and C18 blocked alternative pathway activation completely. Thus demonstrating that native Factor H makes the first and initial contact with the C terminus, which is followed by N terminally mediated complement regulation. These results are explained by a conformational hypothetical model: the native Factor H protein has a compact structure and only one binding site accessible. Upon the first contact the protein unfolds and exposes the additional binding sites. This model does explain how Factor H mediates recognition functions during complement control and the clustering of disease associated mutations in patients with haemolytic uraemic syndrome that have been reported in the C-terminal recognition domain of Factor H.
Modulation of endothelial monolayer permeability induced by plasma obtained from lipopolysaccharide-stimulated whole blood
Authors: Nooteboom A , Bleichrodt R P , Hendriks T
Published: March 2006

The aim of this study was to elucidate the time course of the permeability response of endothelial monolayers after exposure to plasma obtained from lipopolysaccharide (LPS)-treated human whole blood; to investigate the role of apoptosis in monolayer permeability, and to inhibit the permeability increase, particularly after addition of the plasma stimulus. Human umbilical vein endothelial cells (HUVEC) were cultured on semiporous membranes and the permeability for albumin was measured after exposure, according to different schedules, to LPS-conditioned plasma. Apoptotic HUVEC were measured by both flow cytometry and ELISA. A variety of agents, including antibodies against cytokines, inhibitors of NF-κB, and a caspase inhibitor, were added to HUVEC, either prior to or after the stimulus. A maximum increase of the permeability was achieved after 4–6 h of exposure to LPS-conditioned plasma. This response was not accompanied by an increase in the number of apoptotic HUVEC. Administration of antibodies against both Tumour Necrosis Factor-α (TNF-α) and Interleukin-1β (IL-1β) to HUVEC within 1 h after stimulation significantly reduced the permeability increase. Similarly, pyrollidine di-thiocarbamate (PDTC), but not N-acetylcysteine, could prevent the permeability response, and was still effective when added within 2 h after LPS-conditioned plasma. The TNF-α/IL-1β signal present in LPS-conditioned plasma appears to increase endothelial permeability through intracellular pathways that very likely involve the activation of NF-κB. Although poststimulatory inhibition of the permeability response proves to be possible with agents such as PDTC, the window of opportunity appears very small if placed in a clinical perspective.
A modified chromogenic assay for the measurement of very low levels of factor VIII activity (FVIII:C)
Authors: YATUV R. , DAYAN I. , BARU M.
Published: March 2006

Summary. A precise and sensitive chromogenic assay for the measurement of very low levels of factor VIII (FVIII) in plasma has been developed. The assay is based on modifications of a commercially available chromogenic assay. The modifications include reduction of sample final dilution factor and prolongation of the development period. The modified assay allows accurate and precise measurement of FVIII in the range of 0.001–0.02 IU mL−1. The detection limit is 0.0005 IU mL−1and the quantitation limit is 0.0015 IU mL−1. This assay can be used in research and to study the clinical efficacy of low circulating levels of FVIII in haemophilia A patients.
Von Willebrand's disease and angiodysplasia treated with thalidomide
Authors: HIRRI H. M. , GREEN P. J. , LINDSAY J.
Published: March 2006

Summary.  A 54‐year‐old man with type IIB VWD and severe angiodysplasia had such a large blood loss from the bowel that it was difficult to keep up with transfusion requirements. Treatment with factor eight concentrate barely slowed the loss. D.D.A.V.P., Octreotide, and recombinant activated Factor VII, tried separately, were ineffective. The use of Thalidomide at a dose of 150 mg daily has rendered him free from blood loss for the last six months and we suggest would be worth a trial in similar cases.
A phase II randomized controlled trial of a novel male contraception, an intra‐vas device
Authors: Song L. , Gu Y. , Lu W. , Liang X. , Chen Z.
Published: March 2006

The aim of this study was to introduce a novel male contraceptive method, an intra‐vas device (IVD), and to assess the efficacy, safety and satisfaction of recipients compared with no‐scalpel vasectomy (NSV). A phase II randomized controlled trial was conducted in China in 2003. Two hundred and eighty‐eight male subjects seeking vasectomy were randomly assigned to the IVD or NSV group. Follow up included a telephone questionnaire on the 14th postoperative day and visits at the third and 12th postoperative months. The follow‐up rates at the three time points were 100%, 100% and 96.5% respectively. There was no technical failure in any subject. The surgical conditions were similar in both groups, but the IVD group experienced an additional 5 min of operative time (p < 0.001). The IVD group recovered normal activity and sexual intercourse more rapidly (both p < 0.05). The azoospermia rate was lower in the IVD group than in the NSV group at the third and 12th postoperative months. The rate of contraceptive success based on semen analyses was similar in both groups, especially at the 12th postoperative month (94.3% in the IVD group vs. 98.6% in the NSV group; p = 0.054). The IVD group had less risk of complications (i.e. pain, congestive epididymitis and sperm granuloma). More subjects reported satisfaction with IVD sterilization than with NSV. The two procedures were similar in terms of surgical complications. The IVD was slightly less effective, but had a lower risk of later adverse events than the NSV technique. The IVD group also reported a higher level of satisfaction.
Effect of storage in short‐ and long‐term commercial semen extenders on the motility, plasma membrane and chromatin integrity of boar spermatozoa
Authors: De Ambrogi Marco , Ballester Juan , Saravia Fernando , Caballero Ignacio , Johannisson Anders , Wallgren Margareta , Andersson Magnus , Rodriguez‐Martinez Heriberto
Published: March 2006

For artificial insemination (AI) in pigs, preservation of liquid boar semen at 16–20 °C is still common practice as sperm cryopreservation remains suboptimal in this species. To meet the different needs of the swine industry, several extenders have been developed to preserve semen in liquid form for short‐ and long‐term storage. In the present study, three different commercial extenders devised for short‐term (BTS+) or long‐term preservation (MR‐A and X‐Cell), were used to test whether storage of semen from four mature, fertile boars at 17 °C for 96 h would affect sperm characteristics relevant for fertility, such as motility, membrane integrity and chromatin stability. Computer‐assisted sperm analysis, and stainings with the acylated membrane dye SYBR‐14/propidium iodide, and acridine orange in connection with flow cytometry were used to evaluate these variables. Percentages of total motile spermatozoa decreased slightly, but significantly, after 72–96 h. While membrane integrity values varied during the period of study, no significant changes in either membrane integrity or chromatin stability were, however, registered. This suggests a customary 96‐day storage at 17 °C in these extenders was too short an interval to cause losses of integrity in nuclear DNA in the boar population studied.