bluuebunny/crossref_metadata_2025 huggingface dataset:
Caring for people in the ‘virtual ward’: the practical ramifications for acute nursing work
Authors: DEACON M.
Published: July 2003

This paper discusses an aspect of data analysis arising from an ethnographic study of acute mental health nursing conducted in an inner city psychiatric unit. The data were collected in the form of field notes. These were made during and following episodes of participant observation on one acute ward and on a psychiatric intensive care unit over a period of 20 months. Acute wards have been subject to continuing criticism as both sites of care and treatment for mentally ill people and of nursing work. However, the practical operations of this social world remain largely unexamined. It is argued that without this understanding, mundane work methods will always be regarded as an impediment to work that is regarded as ‘therapeutic’ and therefore, of importance. The focus will be on phenomena named the ‘virtual ward’. The research has demonstrated that the boundaries of nurses’ responsibility reach far beyond the ward's spatial environment, both physically and communicatively. The complexity of the work that takes place within the virtual ward and the skills required for its successful completion are taken for granted, but are also a practical source of frustration. Anonymized examples from the practice site studied are used to illuminate the discussion. Working within the virtual ward is part of the messy reality of nurses’ work. It is argued that understanding and respecting this reality is necessary if we are to be seriously and consistently ambitious about practice development.
Case management models and the care programme approach: how to make the CPA effective and credible
Authors: SIMPSON A. , MILLER C. , BOWERS L.
Published: July 2003

The care programme approach (CPA), a form of case management, is a key mental health policy in England. Yet after over 10 years, it remains poorly and unevenly implemented with few benefits for service users, carers or mental health staff. This paper reviews the wider literature on case management and identifies and considers the principal models that might have informed the development of the CPA. After discussing the evidence for each of the clinical, strengths, intensive and assertive case management models, the paper identifies the key components that appear to be central to effective case management across these models. These components are then considered in relation to the CPA. It is argued that the CPA has been undermined by a failure to incorporate and build on certain important features of the major models of case management. The paper concludes by suggesting the key developments required to make the CPA more effective and to underpin the policy with a unifying philosophy while endorsing it with much needed credibility among both clinicians and service users.
Mapping health on the Internet: strategies for learning in an information age
Authors: EDWARDS DEBORAH
Published: July 2003

Ross Scrivener

 Radcliffe Medical Press, Oxford

 2002, 160 pages, £19.95 (PB)

 ISBN: 1857755936.
Social policy: an introduction
Authors: BURNARD PHILIP
Published: July 2003

Ken Blakemore

 Open University Press, Buckingham

 2003, 2nd edn., 304 pages, £16.99 (PB), £55.00 (HB)

 ISBN: 0 335 20847 9
Single dose pharmacokinetics of piroxicam in cats
Authors: Heeb H. L. , Chun R. , Koch D. E. , Goatley M. A. , Hunter R. P.
Published: July 2003

Piroxicam (PIRO) is a nonsteroidal anti‐inflammatory drug (NSAID) recognized for its value as a chemopreventative and anti‐tumor agent. Eight cats were included in this study. PIRO was administered in a single oral (p.o.) and intravenous (i.v.) dose of 0.3 mg/kg. The study was designed as a randomized complete crossover with a 2‐week washout period. Serial blood samples were collected after each dose and plasma was analyzed for PIRO. Pharmacokinetic parameters of PIRO were determined using noncompartmental analysis. PIRO is well absorbed in the cat with a median bioavailability (F) of 80% (range 64–124%). The median i.v.t1/2was 12 h (range 8.6–14 h). The medianCmaxwas 519 ng/mL with a correspondingTmaxof 3 h. PIRO appears to be rapidly absorbed following p.o. administration in cats with a higherCmaxandAUCthan in dogs.
In vivo release of oxytetracycline from a biodegradable controlled‐release gel injected subcutaneously in Japanese quail (Coturnix coturnix japonica)
Authors: Tell L. A. , Sun Y. , Needham M. , Johnson J. R. , Shukla A.
Published: July 2003

A long‐acting, biodegradable, controlled‐release formulation of oxytetracycline (CR‐OTC) was evaluated in 18 adult Japanese quail (Coturnix coturnix japonica) following a single subcutaneous (s.c.) injection. Prior to characterizing the release of oxytetracycline (OTC) from the CR‐OTC, the pharmacokinetic parameters of intravenously (i.v.) administered OTC were determined. Concentrations of free OTC were measured using a bioassay. The plasma concentration–time profile of OTC after a single i.v. injection at 20 mg/kg was best fit to an open two‐compartmental model, with the following pharmacokinetic parameters: area under the curve (AUC) = 36.72 mg · h/L, terminal elimination half‐life = 2.34 h, clearance (Cl) = 0.545 L/kg/h. Plasma [OTC] was >1.0 μg/mL for at least 4 h following i.v. injection. The CR‐OTC gel was well tolerated at a dosage of 1500 mg/kg s.c. Plasma [OTC] rose to >1.0 μg/mL within 24 h; it remained >1.0 μg/mL for at least 10 days in all birds sampled at that time point (n = 9) and for at least 18 days in two of nine birds. Using a deconvolution technique, it was determined that approximately 54.8% of the administered OTC was released from the CR‐OTC over the 45‐day observation period. This long‐acting, biodegradable controlled‐release OTC formulation may have potential for the treatment of chlamydophila infections and other OTC‐sensitive bacteria in Japanese quail, however further studies are necessary to determine its safety and clinical application.
The effect of food or water deprivation on paracetamol pharmacokinetics in calves
Authors: Janus K. , Grochowina B. , Antoszek J. , Suszycki S. , Muszczynski Z.
Published: July 2003

This study investigated the effect of food or water deprivation on the pharmacokinetics of paracetamol in 30 Holstein‐Friesian preruminant calves (10 controls, 10 food withheld and 10 water‐deprived) aged 24–25 days. Control calves were given paracetamol at 24–25 days and again at 28–29 days of age. In the food withheld and water‐deprived calves paracetamol studies were performed before and after 4 days of food or water deprivation. In the control group there were no significant differences in pharmacokinetic parameters for paracetamol in 24–25 and 28–29‐day‐old calves. Witholding food for 4 days was associated with an increase in the mean residence time (MRT) of paracetamol (P < 0.01). When food was withheld total body clearance (ClB) of paracetamol was significantly decreased (P < 0.05). The volume of distribution (Vss) was not significantly altered. Similarly, water deprivation was associated with a significant increase in MRT and significant decrease in ClB of paracetamol (P < 0.01). The Vss was not significantly altered. Food or water deprivation also influenced the formation of major metabolites (glucuronide and sulphate) of paracetamol. It is concluded that food or water deprivation may impair the elimination drugs that undergo metabolism by UDP‐glucuronyltransferase and sulphotransferase in cattle.
Clinical efficacy of trimethoprim/sulfadiazine and procaine penicillin G in a Streptococcus equi subsp. zooepidemicus infection model in ponies
Authors: Ensink J. M. , Smit J. A. H , Van Duijkeren E.
Published: July 2003

Tissue chambers, implanted subcutaneously on both sides of the neck in eight ponies, were inoculated with Streptococcus equi subsp. zooepidemicus in order to compare the clinical efficacy of trimethoprim/sulfadiazine (TMP/SDZ) and penicillin G treatment in a purulent infection. The TMP/SDZ treatment consisted of one intravenous (i.v.) injection of 5 mg/kg TMP and 25 mg/kg SDZ and the same dose of TMP/SDZ per os (p.o.), both given 20 h after inoculation. The oral dose was then repeated every 12 h for 21 days. The penicillin treatment consisted of one i.v. injection of 20 000 IU/kg sodium penicillin G and intramuscular (i.m.) injection of 20 000 IU/kg procaine penicillin G, both given 20 h after infection. The i.m. dose was then repeated every 24 h for 21 days. Eight ponies, each with two tissue chambers, were used in a cross over design; in the first experiment the left tissue chamber (TC) was infected and in the second experiment the right. TMP/SDZ treatment resulted in a limited reduction of viable bacteria in the TC but did not eliminate the infection, resulting in abscessation in 10–42 days in all eight ponies. However, penicillin treatment eliminated the streptococci in seven of eight ponies, and only one pony suffered abscessation on day 10. This constitutes a significantly better efficacy of the penicillin treatment in this model. The most probable cause of the failure of TMP/SDZ to eliminate the streptococci is inhibition of the action of TMP/SDZ in the purulent TCF. Therefore, TMP/SDZ should not be used to treat purulent infections in secluded sites in horses.
Stereospecific biotransformation of albendazole in mouflon and rat‐isolated hepatocytes
Authors: Velík J. , Baliharová V. , Skálová L. , Szotáková B. , Wsól V. , Lamka J.
Published: July 2003

The anthelmintic albendazole (ABZ) undergoes a two‐step oxidation resulting first in the formation of chiral albendazole sulfoxide (ABZSO) followed by its transformation to albendazole sulfone (ABZSO2) in many farm and laboratory animal species. Although cloven‐hoofed game are also treated with ABZ, limited information concerning ABZ biotransformation in these species is available. The present study focused on in vitro ABZ sulfoxidation in hepatocytes from wild sheep‐mouflon (Ovis musimon) and comparison of ABZ sulfoxidation in mouflon and rat (Rattus norvergicus) hepatocytes. ABZ was used as a substrate for primary cultures of mouflon and rat hepatocytes. Time‐dependent stereospecific consumption of ABZSO and ABZSO2 formation has been investigated. The metabolites were determined by high‐performance liquid chromatography with both achiral and chiral stationary phases. Although total‐ABZSO formation did not significantly differ between mouflon and rat, after separation of the (+)‐ABZSO and (−)‐ABZSO enantiomers a significant difference between species was found. The enantiomeric ratio of (+)/(−)‐ABZSO in mouflon hepatocytes was 2.8–3.8, while rat hepatocytes biotransformed ABZ to almost racemic ABZSO, with an enantiomeric ratio of 1.0–1.1. The ratio were similar for two concentrations of substrate used and stable over several time intervals. The formation of ABZSO2 was more extensive in rat (approximately five times) than in mouflon hepatocytes.
Differential effects of phosphodiesterase inhibitors on platelet activating factor (PAF)‐ and adenosine diphosphate (ADP)‐induced equine platelet aggregation
Authors: Rickards K. J. , Andrews M. J. , Waterworth T. H. , Alexander G. B. C. , Cunningham F. M.
Published: July 2003

Compounds that activate adenylate cyclase, increasing intracellular cyclic adenosine monophosphate (cAMP), inhibit equine platelet aggregation. Cyclic AMP is broken down by phosphodiesterase (PDE) and, in the present study, the effects of theophylline, a nonselective PDE inhibitor, and selective inhibitors of PDE isoenzymes PDE3, PDE4 and PDE5, on equine platelet aggregation in response to platelet activating factor (PAF) and adenosine diphosphate (ADP) have been examined. Theophylline and the PDE3 inhibitors, trequinsin and quazinone, inhibited both PAF and ADP‐induced aggregation in a concentration dependent manner. The inhibition of PAF‐induced aggregation was, however, significantly greater than that of the response to ADP. The inhibitory effects of theophylline and the PDE3 inhibitors on ADP‐ but not PAF‐, induced aggregation were reversed by addition of the calcium ionophore, A23187. Rolipram and zaprinast, inhibitors of PDE4 and PDE5, respectively, had no effect on either PAF‐ or ADP‐induced aggregation. These results demonstrate that inhibition of aggregation caused by PAF or ADP can be achieved by selective inhibition of PDE3 but suggest that there may be agonist‐specific differences in the intracellular signalling pathways that regulate equine platelet aggregation.
Comparative effects of fluoroquinolones on subsets of T lymphocytes in normothermic and hyperthermic mice
Authors: Szczypka M. , Obmińska‐Mrukowicz B.
Published: July 2003

The subsets of T lymphocytes in thymus, spleen and mesenteric lymph nodes were investigated in normothermic and hyperthermic mice treated with fluoroquinolones administered orally six times at 24 h intervals at doses of 15 or 75 mg/kg (flumequine, norfloxacin and ciprofloxacin) and 5 or 25 mg/kg (enrofloxacin). It has been found that fluoroquinolones can modulate CD3+, CD4+ and CD8+ marker expression on thymocytes, splenocytes and lymphocytes of mesenteric lymph nodes. Flumequine (15 mg/kg) decreased the percentage of immature CD4+CD8+ thymic cells and increased the percentage of mature CD4+ and CD8+. When the dose of flumequine was increased to 75 mg/kg a reduction in the maturation of thymocytes was observed. Administration of flumequine, norfloxacin and ciprofloxacin, irrespective of doses applied, increased the percentages of CD3+ splenocytes of CD4+ spleen cells. Exposure to enrofloxacin decreased the percentage of T helper‐inducer cells. Flumequine and ciprofloxacin augmented the percentage of CD3+ mesenteric lymph node cells and increased the percentage of CD8+ cells. In contrast, norfloxacin and enrofloxacin decreased the percentage of CD3+ mesenteric lymph node cells and the percentage of CD4+ cells.

Lipopolysaccharide (LPS) from E. coli (25 μg/mouse) increased the percentage of single‐positive CD4+ thymocytes, but did not affect the percentage of CD3+, CD4+ and CD8+ splenocytes and mesenteric lymph node cells. Flumequine and ciprofloxacin administered to mice pior to LPS potentiated its stimulant effect on the maturation of thymic cells ( increased percentage of mature CD4+ and CD8+ thymocytes). Pre‐treatment with norfloxacin or enrofloxacin either reduced or did not modify the stimulant effect of LPS on maturation of thymic cells. Flumequine, norfloxacin, enrofloxacin and ciprofloxacin administered prior to LPS decreased the percentage of CD8+ splenocytes and increased the percentage of CD4+ spleen cells. Norfloxacin and ciprofloxacin at a dose of 75 mg/kg reduced the percentage of CD8+ mesenteric lymph node cells in hyperthermic mice. Pretreatment with norfloxacin at a dose of 15 mg/kg augmented the percentage of mesenteric lymph node cells.

It was concluded that the modulating effects of fluoroquinolones depends on the chemial structure of drugs, dose administered as well as immunologic status.
Effect of alkalinization on the local analgesic efficacy of ketamine in the abaxial sesamoid nerve block in horses
Authors: López‐Sanromán J. , Cruz J. , Santos M. , Mazzini R. , Tabanera A. , Tendillo F. J.
Published: July 2003

The objective of this study was to determine the effects of the alkalinization on the local analgesic efficacy of 1% ketamine in the abaxial sesamoid nerve block in horses. Thirty‐six mature healthy horses were randomly assigned to four groups for the following treatments; an abaxial sesamoid block with 5 mL of saline solution (control saline); an abaxial sesamoid block with 5 mL of a solution containing 1% ketamine (KETs 1%); an abaxial sesamoid block with 5 mL of a solution containing saline solution and 0.5 mEq of sodium bicarbonate (control bicarbonate); and an abaxial sesamoid block with 5 mL of a solution containing 1% ketamine and 0.5 mEq of sodium bicarbonate (KETb 1%). All blocks were performed in one randomly selected front leg. To determine analgesia, hoof withdrawal from thermal stimulus from radiant heat lamp was assessed. Before each block, the hoof withdrawal reflex latency (HWRL) (time between lamp illumination and withdrawal of the hoof) was determined; after the block, local analgesic effects were determined using the heat lamp at 2 and 5 min after the injection and then every 5 min for 1 h. In KETs 1% group, there were significant increases in HWRL between basal values and values from 2 to 10 min after an abaxial sesamoid block. In KETb 1% group, significant increases in HWRL was collected between the basal value and values from 2 to 25 min following an abaxial sesamoid block. In KETs 1% group, of the nine horses, four had an abaxial sesamoid block that was unsuccessful. However, in KETb 1% group, only one of the nine horses had an abaxial sesamoid nerve block that was unsuccessful. The alkalinization of a 1% ketamine solution produced a more consistent and persistent local analgesia in horses when compared with 1% ketamine solution alone.
Pharmacokinetics and sedative effects of intramuscular medetomidine in domestic sheep
Authors: Kästner S. B. R. , Wapf P. , Feige K. , Demuth D. , Bettschart‐ Wolfensberger R. , Akens M. K. , Huhtinen M.
Published: July 2003

Intramuscular (i.m.) administration of medetomidine (MED) may avoid the severe pressor effects caused by peripheral actions of MED associated with intravenous (i.v.) dosing. The purpose of this study was to determine the pharmacokinetics, the time course of sedation and occurence of hypoxaemia after i.m. administration of MED in domestic sheep. The MED was injected i.m. at a dose of 30 μg/kg in nine domestic sheep. Blood was sampled at 0, 5, 10, 20, 30, 40, 60, 120, 180, 240, 360 and 600 min after MED. Sedation was assessed and arterial blood samples were taken before and 35 min after MED application. Mean (SD) pharmacokinetic parameters of i.m. MED were: absorption half‐life: 13.2 (7.5) min; terminal half‐life: 32.7 (14.9) min; time to peak concentration: 29.2 (8.9) min; peak concentration: 4.98 (1.89) ng/mL; volume of distribution: 3.9 (2.4)  l/kg; total body clearance: 81.0 (21.5) mL/(min kg). Peak sedation occurred between 30 and 40 min after injection of MED. The degree of sedation correlated with individual plasma concentrations (rS: 0.926). One animal became hypoxaemic (PaO2 = 54.1 mmHg).
RENAL AND CARDIOVASCULAR DRUGS. Pharmacologic identification of putative D1dopamine receptors in feline kidneys
Authors: Flournoy W. S. , Wohl J. S. , Albrecht‐Schmitt T. J. , Schwartz D. D.
Published: July 2003

The presence of dopamine (DA) receptors in feline kidneys is a matter of contention. Radioligand binding and Western blotting studies were employed to determine whether DA receptors are present in feline kidneys. The pharmacologic profile of the selective D1‐receptor antagonist [3H]‐SCH 23390 was studied in renal cortical membrane preparations from cats by conducting saturation binding isotherm and competitive binding experiments. [3H]‐SCH 23390 bound to feline renal cortical membranes in a manner consistent with labeling of a D1‐like receptor. The binding profile revealed a single site D1‐like or D1receptor in the feline renal cortex with aKd = 7.8 ± 1.0 nmol/L andBmax = 76.5 ± 19.5 fmol/mg. Competitive binding studies for [3H]‐SCH 23390 against unlabeled agonists yielded the followingKivalues and rank order of competition: SKF38393 (Ki = 0.47 ± 0.26 μm) > fenoldopam (Ki = 3.12 ± 1.1 μm) > DA (Ki = 933.1 ± 1.6 μm). Competitive binding studies for [3H]‐SCH‐23390 against unlabeled antagonists yielded the following rank order of competition: SCH 23390 (Ki = 1.97 ± 0.81 μm) > spiperone (Ki = 3.79 ± 0.79) > metoclopramide (Ki = 4.26 ± 2.4 μm). Western blot analysis with anti‐DA D1receptor antibodies detected a single band withMrof 74 kDa corresponding to a D1DA receptor. These results suggest that a putative D1‐like or D1receptor exists in feline kidneys different from those previously identified in rat, dog or human kidneys.
Reduced rates of axonal and dendritic growth in embryonic hippocampal neurones cultured from a mouse model of Sandhoff disease
Authors: Pelled D. , Riebeling C. , Van Echten‐Deckert G. , Sandhoff K. , Futerman A. H.
Published: July 2003

Sandhoff disease is a lysosomal storage disease in which ganglioside GM2 accumulates because of a defective β‐subunit of β‐hexosaminidase. This disease is characterized by neurological manifestations, although the pathogenic mechanisms leading from GM2 accumulation to neuropathology are largely unknown. We now examine the viability, development and rates of neurite growth of embryonic hippocampal neurones cultured from a mouse model of Sandhoff disease, the Hexb–/– mouse. GM2 was detected by metabolic labelling at low levels in wild type (Hexb+/+) neurones, and increased by approximately three‐fold in Hexb–/– neurones. Hexb–/– hippocampal neurones were as viable as their wild type counterparts and, moreover, their developmental programme was unaltered because the formation of axons and of the minor processes which eventually become dendrites was similar in Hexb–/– and Hexb+/+ neurones. In contrast, once formed, a striking difference in the rate of axonal and minor process growth was observed, with changes becoming apparent after 3 days in culture and highly significant after 5 days in culture. Analysis of various parameters of axonal growth suggested that a key reason for the decreased rate of axonal growth was because of a decrease in the formation of collateral axonal branches, the major mechanism by which hippocampal axons elongate in culture. Thus, although the developmental programme with respect to axon and minor process formation and the viability of hippocampal neurones are unaltered, a significant decrease occurs in the rate of axonal and minor process growth in Hexb–/– neurones. These results appear to be in contrast to dorsal root ganglion neurones cultured from 1‐month‐old Sandhoff mice, in which cell survival is impaired but normal outgrowth of neurones occurs. The possible reasons for these differences are discussed.
Apoptotic mitochondrial pathway in neurones and astrocytes after neonatal hypoxia‐ischaemia in the rat brain
Authors: Benjelloun N. , Joly L.‐M. , Palmier B. , Plotkine M. , Charriaut‐Marlangue C.
Published: July 2003

Neuronal apoptosis plays an essential role in early brain development and contributes to secondary neuronal loss after acute ischaemia. Recent studies have provided evidence that caspase‐3 is an important downstream event after hypoxia‐ischaemia in the immature brain, but a minor event in the adult brain. Our investigations have focused on cell populations that expressed apoptotic effectors in the enzymatic death pathway including cytochrome c, caspase‐9 and caspase‐3. Expression, activation and cellular localization of these proteins were studied using cleavage of fluorogenic substrate and immunohistochemistry in neonatal rat brain after unilateral focal ischaemia. Caspase‐3 enzyme activity was elevated in brain homogenate between 6 and 48 h after reperfusion. This activation was preceded by that of caspase‐9, between 3 and 24 h. Apoptotic cell death was  finally accomplished by poly‐ADP‐ribose polymerase cleavage, an endogenous caspase‐3 substrate. In addition, immunodetection demonstrated that cytochrome c and activated caspase‐9 and caspase‐3 were expressed not only in the neurones, the primarily affected cells, but also within the astrocytes, which constituted a dense network delineating the infarct. These results suggested that glial injury may promote the formation of cystic lesions such as those observed clinically in the newborn brain.
Expression of Tie‐2 in human peripheral and autonomic nervous system
Authors: Poncet S. , Gasc J.‐M. , Janzer R. C. , Meyer S. , Juillerat‐Jeanneret L.
Published: July 2003

Tie‐2, a tyrosine kinase receptor, is essential for vascular integrity by regulating cellular adhesion between pericytes and endothelial cells. The aim of this study was to identify sites of expression of Tie‐2 other than the vasculature. Tie‐2 expression was first detected in human colon by Western blotting and reverse‐transcription‐polymerase chain reaction (RT‐PCR) in tissue extracts. The presence of the Tie‐2 mRNA and protein was detected by immunohistochemistry and in situ hybridization in cells of the colon myenteric and submucosal plexus, in both neuronal and Schwann cells. Tie‐2 protein was also found in the nervous system of the female urogenital tract. In the human sciatic nerve and schwannoma, RT‐PCR, Western blotting and immunohistochemistry analysis further confirmed the presence of Tie‐2 mRNA and protein in non‐autonomic peripheral nervous tissue. In conclusion, using several approaches and tissues we have demonstrated the presence of Tie‐2 in human peripheral and autonomic nervous tissue, suggesting a role for Tie‐2 in neural tissue. Thus, attempts to disrupt the tumour vessels by manipulation of the Tie‐2 system in tumours may result in side‐effects in peripheral nerves.
AMOG/β2 and glioma invasion: does loss of AMOG make tumour cells run amok?
Authors: Senner V. , Schmidtpeter S. , Braune S. , Püttmann S. , Thanos S. , Bartsch U. , Schachner M. , Paulus W.
Published: July 2003

The β2 subunit of Na,K‐ATPase, initially described as adhesion molecule on glia (AMOG), has been shown to mediate neurone‐astrocyte adhesion as well as neural cell migration in vitro. We have investigated the expression of AMOG/β2 in human gliomas and its effect on glioma cell adhesion and migration. Compared to normal astrocytes of human brain, AMOG/β2 expression levels of neoplastic astrocytes were down‐regulated in biopsy specimens and inversely related to the grade of malignancy. One rat and four human glioma cell lines showed complete loss of AMOG. To investigate the function of AMOG/β2, its expression was re‐established by transfecting an expression plasmid into AMOG/β2‐negative C6 rat glioma cells. In vitro assays revealed increased adhesion and decreased migration on matrigel of AMOG/β2‐positive cells as compared to their AMOG/β2‐negative counterparts. We conclude that increasing loss of AMOG/β2 during malignant progression parallels and may underlie the extensive invasion pattern of malignant gliomas.
Relationship of Nef‐positive and GFAP‐reactive astrocytes to drug use in early and late HIV infection
Authors: Anderson C. E. , Tomlinson G. S. , Pauly B. , Brannan F. W. , Chiswick A. , Brack‐Werner R. , Simmonds P. , Bell J. E.
Published: July 2003

Reactive astrocytosis is a well‐documented feature of HIV encephalitis (HIVE), but it is unclear whether restricted infection of astrocytes contributes to this phenomenon. In addition, the part played by reactive and/or infected astrocytes in AIDS‐related dementia is not fully understood. In this study of patients at different stages of the human immunodeficiency virus (HIV) infection, who had been treated at most with one antiretroviral drug, reactive astrocytes were identified by immunopositivity for glial fibrillary acidic protein (GFAP) and infected astrocytes by positivity for HIV Nef protein. Results were compared for drug‐using AIDS patients with (n = 9) and without (n = 7) HIVE, for presymptomatic HIV‐positive drug users (n = 12) and for control HIV‐negative subjects (n = 20), including a group who used drugs (n = 10). GFAP‐reactive astrocytes in both grey and white matter were significantly more numerous in HIVE subjects than in each of the other groups but did not correlate with viral load. Nef‐positive astrocytes were confined to HIVE cases and to white matter, but were numerous in only one subject who was treatment‐naive. Nef‐positive microglia were identified in all HIVE cases and in occasional AIDS and presymptomatic subjects who did not have HIVE. The results suggest that astrocytes may form an additional viral reservoir in late HIV infection and may contribute to HIVE. However, the number of GFAP‐positive astrocytes was neither increased in pre‐AIDS nor in drug abuse, in contrast with microglia which we have shown previously to be up‐regulated in both states.
The allograft inflammatory factor‐1 in Creutzfeldt–Jakob disease brains
Authors: Deininger M. H. , Weinschenk T. , Meyermann R. , Schluesener H. J.
Published: July 2003

The allograft inflammatory factor‐1 (AIF‐1) is a 17‐kDa IFN‐γ inducible Ca2+‐binding EF‐hand protein that is encoded within the HLA class III genomic region and is involved in immune dysfunction and smooth muscle cell activation. We used immunohistochemistry double labelling experiments to analyse the spatial distribution and cell‐type‐specific localization of AIF‐1 in the brains of patients who died as a result of sporadic Creutzfeldt–Jakob disease (CJD) and neuropathologically unaltered controls. Significantly more AIF‐1 immunoreactive macrophages/microglial cells and, interestingly, neurones were observed in CJD patients compared to controls. Western blotting confirmed more prominent AIF‐1 immunoreactive bands of approximately 50 kDa in four CJD patients compared to three controls. Chaotropic SDS‐PAGE of the recombinant AIF‐1 resulted in almost complete reduction of the 50 kDa band and mass spectrometry revealed only AIF‐1‐specific tryptic protein fragments suggesting that trimerized AIF‐1 is the predominant form in vivo. Finally, we analysed mechanisms of neuronal AIF‐1 induction. Following H2O2 challenge, a model of general cell stress, we observed the gradual induction of AIF‐1 and, more interestingly, release to the supernatant of SKNSH neurones. Parallel reverse transcriptase polymerase chain reaction and sequencing was used to confirm AIF‐1 mRNA expression.
Glycohistochemical characterization of vascular muscle cell destruction in CADASIL subjects by lectins, neoglycoconjugates and galectin‐specific antibodies
Authors: Brulin‐Fardoux P. , Godfrain C. , Maurage C.‐A. , De Reuck J. , Hauw J.‐J. , Kaltner H. , Bovin N. V. , Gabius H.‐J. , Ruchoux M.‐M. , Kiss R. , Camby I.
Published: July 2003

CADASIL (Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy) is a type of small‐artery stroke and vascular dementia‐inducing pathology of the brain. In order to explain the molecular mechanisms behind the alterations to the blood vessels in CADASIL subjects, we scrutinized the expression of glycan and glycan‐binding sites in the wall of vessels taken from five such subjects (vs.five control subjects matched for age and sex). Specimens were taken from the brain, heart, kidney, liver and lung. Although the main vessel lesions were observed in the tissues depending on the blood–brain barrier, alterations to systemic vessels were also observed despite the absence of any symptoms. The histochemical expression of a panel of 10 biotinylated neoglycoconjugates [Gal‐β(1‐4)‐d‐Glc, Galβ(1‐3)GalNAc, α‐d‐GalNAc, β‐d‐GalNAc, GalNAcα(1‐3)‐d‐GalNAcα, GalNAcα(1‐3)‐d‐GalNAcβ, β‐d‐Glc, α‐d‐Man,l‐Fucose andd‐Glcα(1‐4)‐d‐Glc], eight plant lectins (PNA, MAA, SNA, DBA, WGA, ConA, GNA and UEA‐1) and two antigalectin antibodies was monitored by means of semiquantitative and quantitative computer‐assisted microscopy. The data show the altered histochemical binding of plant lectins, such as UEA‐1 and ConA, in the vessel walls of CADASIL subjects. The present work, based upon staining by a panel of neoglycoconjugates, provides a biochemical characterization of the alteration of vessel walls in the brain compared to other organs including the heart, kidney, lung and liver in CADASIL as opposed to control subjects. These glycohistochemical results suggest a functional relevance of protein–carbohydrate interactions in this disease.
The pathology and pathogenesis of retinal vasculitis
Authors: Hughes E. H. , Dick A. D.
Published: July 2003

Retinal vasculitis is a rare, but potentially blinding intraocular inflammatory condition with diverse aetiology. Although commonly idiopathic, it has a strong association with systemic inflammatory diseases known to involve other areas of the central nervous system, most notably Behcet's disease, sarcoidosis, systemic lupus erythematosis and multiple sclerosis. This article describes the clinicopathologic features of retinal vasculitis and its visually damaging sequelae, reviewing available human histopathologic studies and work with experimental models to discuss the pathogenesis and immunopathology. Evidence indicates that noninfective retinal vasculitis is an autoimmune condition that may be induced by antecedent infection with microbes cross–reacting with putative autoantigens, influenced by genetic susceptibility of both HLA associations and cytokine polymorphisms. The growing understanding of the cellular mechanisms involved in the effector immune response is already providing a rationale for more specific therapeutic approaches.
Antigen Presentation by Murine Dendritic Cells is Nuclear Factor‐kappa B Dependent Both In Vitro and In Vivo
Authors: Yoshimura S. , Bondeson J. , Brennan F. M. , Foxwell B. M. J. , Feldmann M.
Published: July 2003

Antigen presentation is a key rate‐limiting step in the immune response. Dendritic cells (DCs) have been reported to be the most potent antigen‐presenting cells for naïve T cells, but little is known about the biochemical pathways that regulate this function. We here demonstrate that mature murine DC can be infected with adenovirus at high efficiency (>95%) and that an adenovirus transferring the endogenous inhibitor IκBα blocks nuclear factor‐kappa B (NF‐κB) function in murine DC. This result indicates that antigen presentation in the mixed leucocyte reaction is NF‐κB dependent, confirming data with human DC in vitro. However, the importance of this finding depends on verifying that this is true also in vivo. Using delayed type hypersensitivity with allogeneic cells, we show that NF‐κB inhibition had a marked immunosuppressive effect in vivo. These results thus establish NF‐κB as an effective target for blocking DC antigen presentation and hence inhibiting T‐cell‐dependent immune responses. This finding has potential implications for the development of therapeutic agents for use in various pathological conditions of the immune system, including allergy and autoimmunity, and also in transplantation.
Inclusion Body Myositis: Clonal Expansions of Muscle‐Infiltrating T Cells Persist Over Time
Authors: Müntzing K. , Lindberg C. , Moslemi A.‐R. , Oldfors A.
Published: July 2003

Inclusion body myositis (IBM) is a chronic inflammatory myopathy. The muscle histology is characterized by infiltration of T cells, which invade and apparently destroy muscle fibres. This study was performed to investigate whether predominant clones of muscle‐infiltrating T cells are identical in different muscles and whether they persist over time in IBM. By reverse transcriptase‐polymerase chain reaction, 25 T‐cell receptor (TCR) variable β (Vβ) chain families and the complementarity‐determining region 3 (CDR3) of the TCR were analysed in two different muscle biopsies of four patients with IBM. In two of the patients, the muscle biopsies were obtained from different muscles at one time point, whereas in two patients, the second biopsy was obtained 9 years after the first biopsy. T cells expressing predominant Vβ families were analysed for clonality by fragment length analysis of the CDR3. Predominant Vβ families were analysed by DNA sequencing to identify identical clones. Immunohistochemical staining of Vβ families was performed to study the distribution of T cells expressing identified predominant Vβ families. The muscle‐infiltrating lymphocytes showed restricted expression of TCR Vβ families. DNA sequencing proved that clonally expanded T cells were identical in different muscles and persisted 9 years after the first biopsy. Immunohistochemical analysis with Vβ family‐specific antibodies demonstrated the endomysial localization of these T cells in inflammatory cell infiltrates. Our results show that in IBM there is clonal restriction of TCR expression in muscle‐infiltrating lymphocytes. Identical T‐cell clones predominate in different muscles, and these clones persist for many years. These results indicate an important, continuous, antigen‐driven inflammatory reaction in IBM.
Age‐Related Increase in Resistance to Acute Trypanosoma cruzi Infection in Rats is Associated with an Appropriate Antibody Response
Authors: Pascutti M. F. , Bottasso O. A. , Hourquescos M. C. , Wietzerbin J. , Revelli S.
Published: July 2003

Inoculation at weaning with Trypanosoma cruzi in inbred ‘l’ rats resulted in a self‐resolving acute infection characterized by marked parasitaemias, whereas challenge to adult rats revealed a mild disease with extremely low parasitaemias. To explore the mechanisms underlying such age‐associated differences in disease outcome, we analysed the in vitro replication of T. cruzi, nitric oxide and tumour necrosis factor‐α (TNF‐α) production in peritoneal macrophages (PMs), the serum concentrations of the specific immunoglobulins (Igs) IgM and IgG, antibodies exhibiting lytic activity against bloodstream forms of T. cruzi and circulating levels of nitrate, TNF‐α and interferon‐γ (IFN‐γ). Macrophages from young rats were as effective as their adult counterparts for restraining intracellular parasite replication. When stimulated with IFN‐γ, culture supernatants from young PMs contained higher amounts of nitrite and TNF‐α. Serum samples from 4 and 7 days post infection revealed easily detectable amounts of nitrate, with values being further augmented by day 7 post infection and significantly higher in the young group. TNF‐α levels were only detected in the young group by day 7 post infection. Both groups had increased amounts of IFN‐γ in their sera, although in adult rats, this trend was followed by a significant drop at day 7, with young rats showing values still higher by the same time point evaluation. In contrast, young rats presented significantly lower levels of IgM and IgG antibodies during the first week of infection. Increased resistance in adult rats seems to be the result of a more appropriate antibody production.
Increased Production of Interferon‐γ by Leishmania Homologue of the Mammalian Receptor for Activated C Kinase‐Reactive CD4+ T cells among Human Blood Mononuclear Cells: An Early Marker of Exposure to Leishmania?
Authors: Bourreau E. , Pascalis H. , Prévot G. , Kariminia A. , Jolly N. , Milon G. , Buffet P. , Michel R. , Meynard J.‐B. , Boutin J.‐P. , Aschimoff D. , Launois P.
Published: July 2003

A prospective study was undertaken to define early predictive immunological marker(s) of exposure to Leishmania in naïve subjects who have never been exposed to any Leishmania and who were also free of any cutaneous leishmaniasis lesions. These naïve subjects could have been exposed to Leishmania in a rain forest where Leishmania guyanensis and their natural vectors and mammalian host are cocirculating. The production of interferon (IFN)‐γ in response to the Leishmania homologue of the mammalian receptor for activated c kinase (LACK), a candidate for vaccine against leishmaniasis was analysed. At the end of their stay in the rain forest, LACK‐specific CD8+ T cells were detected in subjects whose peripheral blood mononuclear cell (PBMC) produced IFN‐γ in response to soluble Leishmania antigens (SLA) and in those whose PBMC remained unresponsive to SLA. However, LACK‐specific CD4+ T cells were detected only in PBMCs from individuals who became IFN‐γ responders to SLA. In subjects whose PBMC became positive to SLA, LACK‐reactive CD4+ T cells producing high level of IFN‐γ were detectable before the SLA‐reactive IFN‐γ producing CD4+ T cells, suggesting that the former readout assay could be used as an early predictive immunological marker of exposure to Leishmania in subjects who remained disease free.
Immobilized MICA Could Expand Human Vδ1 γδ T Cells In Vitro that Displayed Major Histocompatibility Complex Class I Chain‐Related A‐Dependent Cytotoxicity to Human Epithelial Carcinomas
Authors: Qi J. , Zhang J. , Zhang S. , Cui L. , He W.
Published: July 2003

Human major histocompatibility complex class I chain‐related A (MICA) is a human leucocyte antigen‐related polymorphic molecule, which is expressed on many kinds of epithelial tumours and can be recognized by the Vδ1 subset of γδ T cells. In the present study, monoclonal antibodies (MoAbs) were produced in mice immunized with recombinant MICA (rMICA)*008. It was found that MICA was expressed on ovarian and colonic tumour tissues and could be detected by these anti‐MICA MoAbs. The immobilized rMICA could induce the proliferation of human ovarian epithelial carcinoma‐ or colonic carcinoma‐derived γδ T cells of the Vδ1 phenotype in vitro. These Vδ1 T cells displayed a strong, broad‐range cytolytic activity towards tumour cell lines positive for MICA. The efficiency of this cytolytic activity depended greatly on the level of MICA expressed on the cell surface and could be inhibited by anti‐MICA MoAbs. Therefore, MICA may play an important role in immune responses against epithelial tumours and function as a stimulating factor for the growth of Vδ1 γδ T cells, whereas MICA‐reactive Vδ1 γδ T cells might serve as a new candidate for adoptive cellular therapy of tumours.
Mucosal Vaccination Against Encapsulated Respiratory Bacteria – New Potentials for Conjugate Vaccines?
Authors: Jakobsen H. , Jonsdottir I.
Published: July 2003

Polysaccharide (PS)‐encapsulated bacteria such as Haemophilus influenzae type b (Hib), Streptococcus pneumoniae (pneumococcus), Neisseria meningitides (meningococcus) and group B streptococcus (GBS), cause a major proportion of disease in early childhood. Native PS vaccines are immunogenic and provide protection against disease in healthy adults but do not induce immunological memory. PSs are T‐cell‐independent antigens and do not elicit antibodies in infants and young children, but by conjugating PS to proteins they become T‐cell dependent and immunogenic at an early age. Despite excellent efficacy of PS–protein conjugate vaccines against invasive disease, protection against mucosal infections such as pneumococcal otitis media has been less efficacious. Circulating PS‐specific antibodies may protect against infections at mucosal sites, but mucosal immunoglobulin A antibodies may also contribute significantly to protection against mucosal infections. Mucosal immunization of experimental animals with conjugate vaccines against Hib, pneumococcus, meningococcus and GBS induces systemic and mucosal immune responses, which provide protection against carriage, otitis media and invasive disease in a variety of challenge models, providing new means for protection against encapsulated bacteria. In addition, mucosal immunization of neonatal mice with a pneumococcal conjugate and the nontoxic adjuvant LT‐K63 has been superior to parenteral immunization in eliciting protective antibodies and PS‐specific memory, and thus circumventing the limitations of antibody responses to PS that are responsible for enhanced susceptibility of neonates and infants to infections caused by encapsulated bacteria. Through T‐cell dependent enhanced immunogenicity of PS–protein conjugate vaccines, mucosal immunization could be an attractive approach for early life immunization against encapsulated bacteria.
Positional Cloning of Ncf1– a Piece in the Puzzle of Arthritis Genetics
Authors: Olofsson P. , Holmdahl R.
Published: July 2003

Positional cloning of susceptibility genes in complex diseases like rheumatoid arthritis in humans is hampered by aspects like genetic heterogeneity and environmental variations, while genetic studies in animal models contain several advantages. With animal models, the environment can be controlled, the genetic complexity of the disease is minimized and the disease onset can be predicted, which simplify diagnosis and characterization. We use pristane‐induced arthritis in rats to investigate the inheritance of arthritis. Until now, we have identified 15 loci that significantly predispose rats to the development of arthritis. One of these arthritis loci has been isolated and confirmed to be caused by a polymorphism in the Ncf1 gene. In this review, we outline the methods used to identify Ncf1 as one single susceptibility gene in a complex puzzle of inherited factors that render susceptibility to a complex autoimmune disorder like arthritis.
Regulation of Adhesion and Motility in B Lymphocytes
Authors: Severinson E. , Westerberg L.
Published: July 2003

During the differentiation process of B lymphocytes, they go through changes in adhesion and motility. In order to investigate the molecular mechanism of such changes, in vitro culture systems are necessary. When B cells are activated by various stimuli, they form different types of homotypic aggregates. In addition, they might also spread and express microvilli and/or become polarized, the latter being a sign of motility. In this review, we summarize our own research in this area. We give evidence for involvement of different adhesion and signalling molecules, and by the end, we speculate on the in vivo significance of our findings.
A Human Leucocyte Antigen‐DR1 Transgene Confers Susceptibility to Experimental Allergic Encephalomyelitis Elicited by an Epitope of Myelin Basic Protein
Authors: Sireci G. , Dieli F. , Caccamo N. , Barera A. , Carta P. , Di Sano C. , Meraviglia S. , Bonanno C. T. , Salerno A.
Published: July 2003

Much evidence now indicates that human leucocyte antigen (HLA) class I and class II transgenic (Tg) mice can be of value in analysing HLA‐restricted presentation of T‐cell epitopes relevant to experimental models of autoimmune diseases. One area where this has been applied is the characterization of myelin epitopes presented by HLA class II molecules in experimental model of multiple sclerosis (experimental allergic encephalomyelitis (EAE)). As a first step towards humanized disease models in HLA Tg mice, we have analysed immune response of lymph node cells of HLA‐DR1 Tg mice immunized with the human myelin basic protein (MBP) peptides 13–33, 87–106 and 139–154 bound by HLA‐DR1. We report here that HLA‐DR1 Tg mice display a hierarchy of response in vivo and in vitro to MBP epitopes depending on the binding affinity to DRB*0101 molecule. In fact, the 13–33 epitope induced a strong T helper 1 (Th1) response accompanied by high T‐cell precursor frequency and caused mild EAE, while the two other epitopes gave poor (139–154) or no disease (87–106), and these data correlate with in vitro Th1 response. These data could prove a useful tool in understanding the role played by different MBP epitopes in EAE.
Transforming Growth Factor‐β as a Regulator of Site‐Specific T‐Cell Inflammatory Response
Authors: Lúðvíksson B. R. , Gunnlaugsdóttir B.
Published: July 2003

A common immunopathological hallmark of many autoimmune inflammatory diseases is a T‐cell invasion and accumulation at the inflamed tissue. Although the exact molecular and microenvironmental mechanisms governing such cellular invasion and tissue retention are not known, some key immunological principles must be at work. Transforming growth factor‐β (TGF‐β) is known to modulate some of these processes including homing, cellular adhesion, chemotaxis and finally T‐cell activation, differentiation and apoptosis. The chronicity of such T‐cell‐driven inflammation probably involves an innate immunological response leading to a T‐1 (Th/Tc), T‐2 or T‐3 (Th/Tr) T‐cell adaptive immune response. Several studies suggest that the key to T‐cell final destination resides on its and the antigen‐presenting cell's phenotype as well as the coreceptor expression pattern and their signalling intensity. Recent observations suggest other equally important regulatory elements of T‐cell inflammatory response that are sensitive to TGF‐β modulation. These include: (i) the stage of T‐cell activation/differentiation; (ii) the chemotactic/adhesion molecule expression pattern; and (iii) the conditioning at the immunological synapse determining their sensitivity to known regulators such as TGF‐β. In this article, we focus on how the phenotype of the responding T cell and the T‐cell receptor (TCR)‐signalling intensity could drive the given inflammatory response. In particular, we discuss how TGF‐β can influence the process of T‐cell migration and activation during such site‐specific inflammation.