bluuebunny/crossref_metadata_2025_split huggingface dataset:
Frequency Analysis of HBsAg‐specific B Lymphocytes in High‐responder Individuals to Recombinant Hepatitis B Vaccine: Comparison of LDA and ELISPOT Assays
Authors: Shokrgozar M. A. , Sam M. R. , Amirkhani A. , Shokri F. 
Published: October 2006

AbstractThe determination of the frequency of antigen‐specific lymphocytes may provide invaluable information for the evaluation of the immune response to different antigens and pathogens. Different methods are employed to determine the frequency of specific B lymphocytes in peripheral blood. In this study, the frequency of hepatitis B surface antigen (HBsAg)‐specific B lymphocytes was determined by a limiting dilution assay (LDA) and an enzyme‐linked immunospot assay (ELISPOT) in seven healthy adult high responders to recombinant HBsAg. Peripheral blood mononuclear cells isolated at different time intervals (1, 2, 4, 8 and 16 weeks) following administration of a booster dose were either transformed with Epstein–Barr virus (LDA) or stimulated with Pokeweed mitogen (ELISPOT). In an LDA, anti‐HBs positive wells were screened by a sandwich ELISA and the frequency of specific B lymphocytes was estimated based on the Poisson statistical analysis. In an ELISPOT, coloured spots representing specific B lymphocytes were finally enumerated by stereomicroscope. Our results showed a significant increase in the number of specific B lymphocytes in the first week by an ELISPOT compared with an LDA (1:190 versus 1:13,462) (P < 0.001). No significant differences were observed at other time intervals. A significant correlation was observed between the serum titer of anti‐HBs antibody and frequency of HBsAg‐specific B cells obtained by LDA and ELISPOT methods at different time intervals. The highest correlation was found at fourth week in LDA (r = 0.83, P < 0.01) and ELISPOT (r = 0.85, P < 0.01) assays. Furthermore, a significant correlation was observed between an LDA and ELISPOT at different time intervals (highest correlation in second week, r = 0.88, P < 0.008). These findings suggest that in addition to technical advantages, such as speed and simplicity, an ELISPOT is a more sensitive assay, compared with an LDA.
Regulatory Activity of Activated Murine Peripheral CD4+CD25− T Cells: A Possible Mechanism of Feedback Regulation on Adaptive Immunity
Authors: Lu Y. , Xu W. , Zhou Y. , Lv P. , Gao X.‐M. 
Published: October 2006

AbstractThis study was undertaken to investigate the possibility that peripheral CD4+CD25− T cells, once fully activated, transiently obtain suppressive function against other T cells. CD4+CD25− T cells, isolated from splenocytes of BALB/c mice, were stimulated with anti‐CD3 MoAb in the presence of feeder cells for 72 h. The activated CD4+ T cells (Tact) thus obtained were able to inhibit the activation and proliferation of bystander CD4+ T cells in a non‐MHC‐restricted manner. Tact‐mediated suppression was cell contact dependent, reversible by exogenous IL‐2 as well as anti‐GITR antibody. Furthermore, adoptive transfer of Tact cells significantly downregulated humoral response of BALB/c mice to s.c. immunization with ovalbumin. We argue that suppression mediated by activated CD4+CD25− T cells may play an important role in maintaining homeostasis of the immune system and preventing excessive T‐cell responses in vivo.
Changing indications for red cell transfusion from 2000 to 2004 in the North of England
Authors: Wallis J. P. , Wells A. W. , Chapman C. E. 
Published: October 2006

summary Falling donor numbers and the threat of transfusion‐transmitted variant Creutzfeldt‐Jakob disease may lead to shortages in the national blood supply. Knowledge of current patterns of transfusion and trends in usage will help predict future change in blood use. Our previous survey identified medical indications as the major reason for transfusion, but detailed information within this category was limited.We performed prospective surveys of indications for red cell transfusion for two 14‐day periods in 2004 in the North of England, concentrating on medical reasons for transfusion. Data were obtained for transfusion indications of 9003 units, which accounted for over 99% of red cell issues from the regional blood centre during the study. In 2004, medical patients received 62% (5558 units) of all transfused units, surgical patients 33% (3001 units) and Obstetric & Gynaecology patients 5% (444 units). These figures compare with 52, 41 and 6% for Medicine, Surgery, and Obstetrics & Gynaecology in 1999/2000.The three largest uses of blood within the medical category were for patients with primary haematological disorders (18·2% of all transfused blood), for management of gastrointestinal haemorrhage (13·8%) and for patients with nonhaematological malignancies (8·8%).There has been a significant reduction in use of blood for surgical indications over the last 5 years, but an absolute increase in use of blood for medical indications. Lower transfusion triggers, education, use of cell salvage, the increasing price of a unit of red cells and changing population demographics may all have contributed to the reduction in surgical blood use. Promotion of good transfusion practice and alternatives to allogeneic transfusion should now focus on medical and surgical use of blood transfusion.
ANOTHER LOOK AT THE PRESUMED‐VERSUS‐INFORMED CONSENT DICHOTOMY IN POSTMORTEM ORGAN PROCUREMENT
Authors: JACOB MARIE‐ANDRÉE 
Published: October 2006

ABSTRACTIn this paper I problematise quite a simple assertion: that the two major frameworks used in assessing consent to post‐mortem organ donation, presumed consent and informed consent, are procedurally similar in that both are ‘default rules.’ Because of their procedural common characteristic, both rules do exclude marginalized groups from consent schemes. Yet this connection is often overlooked. Contract theory on default rules, better than bioethical arguments, can assist in choosing between these two rules. Applying contract theory to the question of post‐mortem organ donation suggests that the default rule should be one that goes against the wishes of the stronger party in consent decisions.
CLONING, PARENTHOOD, AND GENETIC RELATEDNESS
Authors: SPARROW ROBERT 
Published: October 2006

ABSTRACTIn this paper I examine what I take to be the best case for reproductive human cloning, as a medical procedure designed to overcome infertility, and argue that it founders on an irresolvable tension in the attitude towards the importance of being ‘genetically related’ to our children implied in the desire to clone. Except in the case where couples are cloning a child they have previously conceived naturally, cloning is unable to establish the right sort of genetic relation to make couples the parents of their cloned child. If anybody is the genetic parent of a cloned child it is the natural parent(s) of the DNA donor. Paradoxically, in order to resist the claims of the parents of the donor to the cloned child, the argument for human reproductive cloning must place more weight on the intention to parent a child, than we do in cases of ordinary reproduction. It must insist that the parental relation is established by the intentions of the couple who bring a clone into the world and not by their genetic relation to the child. The emphasis placed on intention as establishing the parental relationship works to undermine the justification for cloning in the first place. For cloning to play a useful role as a reproductive technology, it must allow couples to become parents who could do so no other way. However, to the extent that intention is sufficient to establish parenthood, adoption or surrogacy, which are existing alternatives to cloning, will serve equally well to allow couples to become parents.
THE CLAIM FROM ADOPTION REVISITED
Authors: RIVERA‐LÓPEZ EDUARDO 
Published: October 2006

ABSTRACTIn a recent paper published in this journal, Thomas S. Petersen makes a qualified defense of what he calls ‘the Claim from Adoption’, according to which, ‘instead of expending resources on bringing new children [in developed countries] into the world using reproductive technology and caring for these children, we ought to devote these resources to the adoption and care of existing destitute children’. My purpose in this paper is not to discuss Petersen’s argument in favor of that claim. Rather, I want to show that, even if the Claim from Adoption might be, other things being equal, true, it is not true all things considered. What is, all things considered, true is a much more complex set of statements, including a general prima facie obligation to adopt, as well as an obligation by the state of developed countries to discourage procreation and promote adoption of destitute children.
Immunohistochemical Staining and Enzyme Activity Measurements Show myo‐Inositol‐1‐Phosphate Synthase to Be Localized in the Vasculature of Brain
Authors: Wong Yun‐Hua H. , Kalmbach Sandra J. , Hartman Boyd K. , Sherman William R. 
Published: October 2006

Abstract:Rabbit anti‐bovine myo‐inositol‐1‐phosphate synthase was used to examine the distribution of that enzyme in perfused and immersion‐fixed bovine brain and testis. In brain, intense and specific staining was found in the walls of all the vascular elements including cerebral capillaries. The remainder of brain parenchyma exhibited only low levels of background staining. In testis, an organ rich in the enzyme, blood vessels showed no specific staining. Instead, the enzyme was found in the seminiferous epithelium of the seminiferous tubules, perhaps localized in spermatozoa. To confirm the brain finding, the activity of myo‐inositol‐1‐phosphate synthase was measured in bovine brain microvessel preparations and brain pial vessels. In these preparations the activity of the enzyme was found on average to be 7 and 22 times enriched over that in whole brain, respectively. The activities of two other enzymes of inositol metabolism, myo‐inosose reductase and myo‐inositol‐1‐phosphatase, were also examined for their distribution in brain. Those enzymes were found to be generally distributed. The surprising finding of a vascular localization of myo‐inositol‐1‐phosphate synthase in brain raises new questions about the mechanism by which myo‐inositol is concentrated to such high cellular levels in the principal substance of that organ.
Involvement of Dihydropyridine‐Sensitive Ca2+ Channels in Adenosine‐Evoked Inhibition of Acetylcholine Release from Guinea Pig Ileal Preparation
Authors: Katsuragi Takeshi , Shirakabe Katsuya , Ogawa Shusuke , Soejima Osamu , Furukawa Tatsuo 
Published: October 2006

AbstractThe effects of adenosine and nifedipine on endogenous acetylcholine (ACh) release evoked by electrical stimulation from guinea pig ileal longitudinal muscle preparations exposed to physostigmine were evaluated using an HPLC with electrochemical detection (ECD) system. Resting ACh release, which was sensitive to tetrodotoxin (0.3 μM), was enhanced by Bay K. 8644 (0.5 μM; a Ca2+ antagonist) or 4‐aminopyridine (30 μM; a K+ channel blocker) but not by theophylline (100 μM; a P1 purinoceptor antagonist) or atropine (0.3 μM). The enhancement of the resting ACh release by Bay K 8644 was virtually unaffected by atropine. Electrically evoked ACh release was enhanced by around two‐ to fourfold in the presence of theophylline, atropine, Bay K 8644, 4‐aminopyridine, or atropine. On the other hand, the evoked ACh release was reduced by adenosine (10–30 μM), nifedipine (0.1–0.3 μM; a dihydropyridine Ca2+ channel antagonist), or bethanechol (1–3 μM) in a concentration‐related fashion. The reduction induced by adenosine or nifedipine was almost abolished by either theophylline or Bay K. 8644, whereas that induced by bethanechol was virtually unaffected by these drugs. The inhibition by adenosine of ACh release was not influenced in the presence of 4‐aminopyridine or atropine. However, this inhibition by adenosine was considerably enhanced by halving the Ca2+ concentration in the Krebs solution and was diminished by doubling the Ca2+ concentration. These findings suggest that adenosine produces a cholinergic neuromodulation presumably via modifying dihydropyridine‐sensitive Ca2+ channel activities in the cholinergic neurons, and thus L‐type Ca2+ channels may exist on the nerve terminals. It was also demonstrated that the HPLC‐ECD method used for detecting endogenous ACh was suitable for evaluating mechanisms of the neuromodulation.
Modulation of Calcium Fluxes Across Synaptosomal Plasma Membrane by Local Anesthetics
Authors: García‐Martín Elena , González‐Cabanillas Soledad , Gutiérrez‐Merino Carlos 
Published: October 2006

AbstractWe have studied the effects of local anesthetics (dibucaine, tetracaine, lidocaine, and procaine) on calcium fluxes through the plasma membrane of synaptosomes. All these local anesthetics inhibit the ATP‐dependent calcium uptake by inverted plasma membrane vesicles at concentrations close to those that promote an effective blockade of the action potential. The values obtained for the KC.5 of inhibition of calcium uptake are the following: 23 μM (dibucaine), 0.44 mM (lidocaine), 1.5 mM (procaine), and 0.8 mM (tetracaine). There is a good correlation between these K0.5 values and the concentrations of the local anesthetics that inhibit the Ca2+‐dependent Mg2+‐ATPase of these membranes. In addition, except for procaine, these local anesthetics stimulate severalfold the Ca2+ outflow via the Na+/Ca2+ exchange in these membranes. This effect, however, is observed at concentrations slightly higher than those that effectively inhibit the ATP‐dependent Ca2+ uptake, e.g., 80–700 μM dibucaine, 2–10 mM lidocaine, and 1–3 mM tetracaine. The results suggest that the Ca2+ buffering of neuronal cytosol is altered by these anesthetics at pharmacological concentrations.
Inhibition of Brain Type A Monoamine Oxidase and 5‐Hydroxytryptamine Uptake by Two Amphetamine Metabolites, p‐Hydroxyamphetamine and p‐Hydroxynorephedrine
Authors: Arai Yuichiro , Kim Se Kyung , Kinemuchi Hiroyasu , Tadano Takeshi , Satoh Shin‐Etsu , Satoh Nobunori , Kisara Kensuke 
Published: October 2006

AbstractTwo amphetamine metabolites, p‐hydroxyam‐phetamine (p‐OHA) and p‐hydroxynorephedrine (p‐OHN), selectively inhibited the A form of monoamine oxidase (MAO) in rat and mouse forebrain homogenates. Of these two metabolites, p‐OHA inhibited MAO‐A more strongly than p‐OHN. This MAO‐A‐selective inhibition by p‐OHA or p‐OHN was found to be competitive with respect to deamination of its substrate, 5‐hydroxytryptamine (5‐HT). The degree of MAO‐A inhibition was not changed by 90 min of preincubation of the enzyme preparations with either metabolite, and the activity inhibited by p‐OHA after the preincubation recovered completely to the control level after repeated washing. Uptake of 5‐HT or dopamine into mouse forebrain synaptosomes was highly reduced by both p‐OHA and p‐OHN. Both metabolites were more potent in reducing dopamine uptake than in reducing 5‐HT uptake. In reduction of 5‐HT and of dopamine uptake, p‐OHA was more potent than p‐OHN. These results indicate that p‐OHA is a more selective inhibitor of brain MAO‐A activity and 5‐HT uptake than its subsequent metabolite, p‐OHN. These two actions of p‐OHA might, together with possible 5‐HT efflux into the synaptic cleft, greatly contribute to head twitch, a brain 5‐HT‐mediated animal behavior induced by p‐OHA.
Increased γ‐Aminobutyric Acid Receptor Function in the Cerebral Cortex of Myoclonic Calves with an Hereditary Deficit in Glycine/Strychnine Receptors
Authors: Lummis Sarah C. R. , Gundlach Andrew L. , Johnston Graham A. R. , Harper Peter A. W. , Dodd Peter R. 
Published: October 2006

AbstractInherited congenital myoclonus (ICM) of Poll Hereford cattle is a neurological disease in which there are severe alterations in spinal cord glycine‐mediated neurotransmission. There is a specific and marked decrease, or defect, in glycine receptors and a significant increase in neuronal (synaptosomal) glycine uptake. Here we have examined the characteristics of the cerebral γ‐aminobutyric acid (GABA) receptor complex, and demonstrate that the malfunction of the spinal cord inhibitory system is accompanied by a change in the major inhibitory system in the cerebral cortex. In synaptic membrane preparations from ICM calves, both high‐ and low‐affinity binding sites for the GABA agonist [3H]muscimol were found (KD= 9.3 ± 1.5 and 227 ± 41 nM, respectively), whereas only the high‐affinity site was detectable in controls (KD= 14.0 ± 3.1 nM). The density and affinity of benzodiazepine agonist binding sites labelled by [3H]diazepam were unchanged, but there was an increase in GABA‐stimulated benzodiazepine binding. The affinity for t‐[3H]butylbicyclo‐o‐benzoate, a ligand that binds to the GABA‐activated chloride channel, was significantly increased in ICM brain membranes (KD= 148 ± 14 nM) compared with controls (KD= 245 ± 33 nM). Muscimol‐stimulated 36Cl‐ uptake was 12% greater in microsacs prepared from ICM calf cerebral cortex, and the uptake was more sensitive to block by the GABA antagonist picrotoxin. The results show that the characteristics of the GABA receptor complex in ICM calf cortex differ from those in cortex from unaffected calves, a difference that is particularly apparent for the low‐affinity, physiologically relevant GABA receptors. Study of the GABA receptor complex in ICM calves may provide a greater insight into the interactions of the various binding sites on the GABA‐benzodiazepine receptor protein molecule.
Gangliosides Stimulate Calcium Flux in Neuro‐2A Cells and Require Exogenous Calcium for Neuritogenesis
Authors: Wu Gusheng , Vaswani Kuldeep K. , Lu Zi‐Hua , Ledeen Robert W. 
Published: October 2006

AbstractThe neuritogenic effect of exogenous ganglioside has been documented with a variety of neuronal and neuroblastoma systems, but the mechanism is not understood. Involvement of Ca2+ is suggested by this study which demonstrates that treatment of Neuro‐2A cells with bovine brain gangliosides (BBG) in Ca2+‐depleted medium failed to produce neurite outgrowth. This was in contrast to treatment with retinoic acid or dibutyryl cyclic AMP which induced differentiation under the same conditions. Addition of BBG to Neuro‐2A cells caused small, but significant, increases in both influx and efflux of Ca2+. It thus appears that although neuritogenesis can proceed by more than one mechanism, that induced by BBG requires exogenous Ca2+ and involves stimulation of Ca2+ flux.
Exogenous Gangliosides Stimulate Breakdown of Neuro‐2A Phosphoinositides in a Manner Unrelated to Neurite Outgrowth
Authors: Vaswani Kuldeep K. , Wu Gusheng , Ledeen Robert W. 
Published: October 2006

AbstractGangliosides administered exogenously are wellknown effectors of differentiation in many neuroblastoma lines and primary neuronal cultures. Previous studies suggested the phosphoinositide signaling mechanism could be a contributing factor. We have found that treatment of Neuro‐2A cells with bovine brain ganglioside mixture (BBG) causes breakdown of phosphoinositides, as measured by increased levels of inositol phosphates. The effect was optimal at 60 min and required a minimal BBG concentration of 25 μM. However, addition of neomycin, which blocked phosphoinositide breakdown, had no observable effect on gangliosidestimulated neurite outgrowth. A similar result was obtained with ψ‐tectorigenin, which also inhibited phosphoinositide hydrolysis. When cells were treated with maitotoxin, an agent that promotes phosphoinositide breakdown, there was no enhancement of neurite outgrowth. These findings indicate that although exogenous gangliosides elevate inositol phosphate formation over a prolonged period in neuro‐2A cells, this reaction is not integral to the differentiation of these cells. The possibility of secondary effects influencing neurite type and structure cannot be excluded.
Neurotoxins Distinguish Between Different Neuronal Nicotinic Acetylcholine Receptor Subunit Combinations
Authors: Luetje Charles W. , Wada Keiji , Rogers Scott , Abramson Stewart N. , Tsuji Kuniro , Heinemann Steve , Patrick Jim 
Published: October 2006

AbstractNeuronal and muscle nicotinic acetylcholine receptor subunit combinations expressed in Xenopus oocytes were tested for sensitivity to various neurotoxins. Extensive blockade of the α3β2 neuronal subunit combination was achieved by 10 nM neuronal bungarotoxin. Partial blockade of the α4β2 neuronal and α1β1γδ muscle subunit combinations was caused by 1,000 nM neuronal bungarotoxin. The α2β2 neuronal subunit combination was insensitive to 1,000 nM neuronal bungarotoxin. Nearly complete blockade of all neuronal subunit combinations resulted from incubation with 2 nM neosurugatoxin, whereas 200 nM neosurugatoxin was required for partial blockade of the α1β1γδ muscle subunit combination. The α2β2 and α3β2 neuronal subunit combinations were partially blocked by 10,000 nM lophotoxin analog‐1, whereas complete blockade of the α4β2 neuronal and α1β1γδ muscle subunit combinations resulted from incubation with this concentration of lophotoxin analog‐1. The α1β1γδ muscle subunit combination was blocked by the α‐conotoxins G1A and M1 at concentrations of 100 nM. All of the neuronal subunit combinations were insensitive to 10,000 nM of both α‐conotoxins. Thus, neosurugatoxin and the α‐conotoxins distinguish between muscle and neuronal subunit combinations, whereas neuronal bungarotoxin and lophotoxin analog‐1 distinguish between different neuronal subunit combinations on the basis of differing α subunits.
Liver, Brain, and Heart Metallothionein Induction by Stress
Authors: Hidalgo Juan , Borrás Marta , Garvey Justine S. , Armario Antonio 
Published: October 2006

AbstractTo date, stress has been reported to induce metallothionein (MT) synthesis in the liver only. In the present experiment, the effects of food and water deprivation alone or of immobilization stress plus food and water deprivation on liver, brain, and heart MT have been studied in adult male rats. Liver and brain MT levels were increased by immobilization stress as soon as 6 h after the onset of stress. Eighteen hours of immobilization, which is accompanied by food and water deprivation, further increased liver and brain MT levels and significantly increased heart MT content. A specific effect of immobilization was evident in all three tissues, because the effect of food and water deprivation alone was significantly lower than that of immobilization plus starvation. Changes in MT apparently were not related to changes in cytosolic Zn.