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Effect of gentamicin and dexamethasone on the natural history of the rat Escherichia coli brain abscess model with histopathological correlation.

In a rodent model of Escherichia coli brain abscess, the natural history of the infection was studied and the influence of a glucocorticoid (dexamethasone) and an antibiotic (gentamicin) on the development of brain abscess and the survival of abscess-bearing animals was evaluated. The administration of steroids using three different dosage schedules suppressed the macrophage and glial response, decreased collagen formation, increased the number of pathologically evident bacteria, and decreased host survival. The administration of antibiotics by parenteral routes decreased the number of viable bacteria in the abscess. The simultaneous administration of systemic gentamicin and dexamethasone resulted in increased host survival to a level that was intermediate between that of animals treated with dexamethasone alone and that of those treated with gentamicin alone. Thus, some of the adverse effect of corticosteroids on host survival could be mitigated by the simultaneous administration of antibiotics. Finally, it was observed that the abscess in this model tends to expand along white matter tracks. This path of least resistance may be responsible for the observation that brain abscesses tend to rupture into the ventricle rather than into the subarachnoid space via the cortex.

Animals↗

Evaluation of the oncogenic potential of man-made vitreous fibres: the inhalation model.

A rodent inhalation model has been developed for the evaluation of the eoncogenic potential of man-made vitreous fibres. It is successful in delivering a quantified dose of well-characterized fibres to the lungs of rodents, and with it sufficiently high fibre aerosol concentrations were lofted to enable a maximum tolerated dose to be achieved. Fischer 344 male rats were exposed to a well-defined rat-respirable aerosol at concentrations for MMVF of 30, 16 or 3 mg m-3, 6 h per day, 5 days per week for 104 weeks with final sacrifice at 20% survival. A control group was exposed to filtered air. The high dose was chosen based upon a 28-day maximum tolerated dose study with refractory ceramic fibres (RCF). The fibre aerosol generation system lofted fibres without breaking, grinding or contaminating the bulk material. Exposure was by flow-past nose-only systems which provided fresh fibre in a laminar stream to each animal individually. The study was performed according to the Good Laboratory Practice regulations. Fibre count, fibre diameter and length distribution, aerosol mass and chemical composition were determined throughout the study. Interim sacrifices were performed at 3 or 6 month intervals for 24 months. At each sacrifice, full necropsy was performed, the accessory lobe removed for subsequent digestion to determine the fibre lung burden and the remaining lobes inflated with fixative for histopathological evaluation. The lungs were evaluated by a pathologist and graded for the degree of macrophage infiltration, bronchiolization, fibrosis and pleural thickening, and were also scored according to the Wagner scale. Lesions were evaluated according to the number of adenomas, carcinomas and mesotheliomas. The accessory lobe was digested by low-temperature plasma ashing and the number, size distribution and chemical composition of the fibres determined. This model provides a sensitive and reproducible method for evaluating existing and new fibres. A variety of different of ceramic, glass, rockwool and slagwood fibres have been evaluated with this model.

Aerosols↗

Central nervous system mechanisms in alcohol relapse.

BACKGROUND: Alcohol relapse is a major problem in the treatment of alcohol abuse and alcoholism. Understanding the long-term neuronal alterations that promote relapse of alcohol drinking is critical for the development of pharmacotherapies to treat alcoholism and alcohol abuse. The major objectives of this workshop were to present recent findings, by using rodent models, on behavioral and neurobiological factors that may underlie alcohol relapse and the results of clinical and pharmacological treatment strategies for preventing relapse. METHODS: Prolonged ethanol drinking and repeated periods of alcohol deprivation were studied in nonselected rats and in rats selectively bred for high alcohol preference (P rat). The expression of a robust alcohol deprivation effect (ADE) was used as a model for alcohol relapse in rodents. Operant techniques were used to examine responding for ethanol after deprivation in both rats and C57BL/6J mice. Environmental cues and stress were used to assess their effects on reinstatement of alcohol responding. Microdialysis and [14C]-2-deoxyglucose techniques were used to examine neuronal alterations associated with alcohol relapse. RESULTS: Prolonged free-choice ethanol drinking followed by deprivation produced an ADE in both stock and P rats. These rats demonstrated loss of control on reinstatement after chronic drinking and after prolonged deprivation. Acamprosate and naltrexone effectively reduced the ADE in stock rats. Stress reinstated operant responding for alcohol, and rats trained to associate a cue with ethanol presentation increased responding on the ethanol-associated lever in the absence of ethanol. After repeated deprivations, P rats shifted their preference toward drinking higher concentrations of ethanol, which increased the magnitude and duration of the ADE. Stock rats also shifted their preference toward solutions with higher ethanol concentrations and demonstrated a loss of control after prolonged ethanol drinking and deprivation. Long-lasting alterations in neuronal activity, serotonin-3 receptor function, and dopamine neurotransmission within the mesolimbic system were evident after chronic drinking that followed a prolonged deprivation period. CONCLUSIONS: The ADE is a useful model for studying alcohol relapse in both rats and mice. The potential validity of this model is supported by the findings that acamprosate and naltrexone are effective in preventing the ADE in rodents, and both compounds have gained recognition for their therapeutic effects in clinical populations. Genetics, stress, and environmental cues are all important factors that influence alcohol relapse. Long-term alterations in neuronal activity within the mesolimbic system, which possibly involve dopamine and serotonin, may underlie alcohol relapse.

Acamprosate↗

Seed-cache exchange promotes coexistence and coupled consumer oscillations: a model of desert rodents as resource processors.

Most models of resource competition assume that coexistence of consumers depends on tradeoffs in their abilities to exploit shared resources along dimensions of environmental heterogeneity generated by factors external to the consumers. However, consumers may create heterogeneity themselves by modifying resources that they do not immediately consume; such "resource processing" is predicted to allow coexistence if consumers vary in use of resources in primary vs. modified form. To explore whether external food storage (caching) represents a form of resource processing that contributes to observed patterns of species coexistence, we developed a biologically explicit simulation model of competition for a well-studied system, seed-eating desert heteromyid rodents. Here we present the model, compare competitive outcomes with and without inter-specific exchange of cached food, and describe population dynamics of coexisting competitors. The model predicts stable coexistence only when there is exchange of cached seeds via scavenging of caches left undefended by mortality or by pilferage of defended caches. Net interactions between coexisting consumers ranged from competition (10% of cases) to host-parasite (77%), commensalism (12%), and mutualism (1%). Population dynamics of coexisting consumers often showed strong periodicity and coupled synchronous or slightly lagged cycles, a possibility not previously anticipated for desert rodents occupying constant environments. Our model confirms that caching does represent a form of resource processing likely to play a significant role in the dynamics and diversity of communities of desert rodents and other caching animals.

Animals↗

Anti-inflammatory effects of a novel, potent inhibitor of poly (ADP-ribose) polymerase.

OBJECTIVE AND DESIGN: Oxygen- and nitrogen-derived free radicals and oxidants play an important role in the pathogenesis of various forms of inflammation. Recent work emphasizes the importance of oxidant-induced DNA strand breakage and activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) in the pathogenesis of various inflammatory diseases. We have recently demonstrated the efficacy of PJ34, a novel, potent phenanthridinone derivative PARP inhibitor, in rodent models of diabetic vascular dysfunction and stroke. Here we tested the efficacy of PARP inhibition in various models of local inflammation in rodents. MATERIALS AND METHODS: PJ34 (at doses of 0.03-30 mg/kg) was tested in rats and mice subjected to standard models of inflammation, with relevant parameters of inflammation measured using standard methods. RESULTS: PJ34 treatment (s.c, i.p. and i.v.) dose-dependently suppressed neutrophil infiltration and nitric oxide (but not KC and IL-1beta) production in peritonitis. In a model of systemic endotoxemia, PJ34 pretreatment significantly reduced plasma levels of TNF-alpha, IL-1beta and nitrite/nitrate (breakdown products of nitric oxide) production. PJ34 treatment (oral gavage) induced a significant suppression of the inflammatory response in dextran sulfate colitis, multiple low dose streptozotocin diabetes and cyclophosphamide-accelerated autoimmune diabetes in the non-obese diabetic mice, and reduced the degree of mononuclear cell infiltration into the iris in an endotoxin-induced uveitis model. Delaying the start of PJ34 administration in the colitis model conferred significant protective effects, while in the arthritis model the post-treatment paradigm lacked protective effects. CONCLUSIONS: PJ34 provides significant, dose-dependent, anti-inflammatory effects in a variety of local inflammation models. Some of its actions are maintained in the post-treatment regimen and/or after discontinuation of treatment. We conclude that PARP inhibition offers a powerful means for reducing the severity of various forms of local inflammatory responses.

Adenosine Triphosphate↗

[A new model for the study of filarial physiopathology: Molinema dessetae in its definitive natural host, Proechimys oris. 1].

The filarial rodent model Molinema dessetae/Proechimys oris, recently adapted in the laboratory allows studies on the host/parasite relationships. Development of filarial worm was described elsewhere, and the model standardized. Biological, parasitological data and antifilarial drug response were determined. Natural models can seldom be used in laboratory; in non natural models many informations on the host-parasite relationships are lost and pathological studies can only be fragmentary and punctual. With inbred Proechimys oris we have the possibility to investigate with a natural filarial model. Three rodent groups have been studied for over a period of one year. The first group comprises eleven parasite free animals; the second one, eight rodents infected once by L3, and the last with eight rodents regularly re-infected during the period of experiment. Several biological parameters were considered: weight, total red and white blood cell counts, haemoglobinemia, mean cell volume, differential leucocyte count, blood urea nitrogen, total serum proteins and electrophoretic pattern, endogenous creatinine clearance, circulating immune complexes. On the whole, more than 19,700 biological data were obtained and several multivariate statistical analysis were necessary for a valid interpretation. The Principal Component Analysis (ACP) shows in each infected rodent distinct trajectories related either with the age of the host or with the filarial infection itself. Three graphic methods were used: individual graphics of each rodent, graphics of each rodent group and, individual and group statistical center of gravity. At the end of the experiments, the rodents were autopsied and a detailed histological study of the whole body was made. Different lesions and stages of inflammatory processes found were analysed and classified as follows: granulomatous nodule with eosinophilic and neutrophilic polymorphs; foreign-body granulomatous nodule; sclerocicatricial nodule; mononucleate cell infiltrate. Secondary pathological alterations, such as irritative lesions and blocage of lymph and blood vessels, were found. A comprehensive study of all our qualitative histological results was performed by a second multivariate analysis: the Correspondence Analysis (AFC). This analysis confirmed the heterogeneity of each Proechimys infected and multi-infected groups. The cross checking between the ACP and AFC allows the separation of two physiopathological filarial groups; the first group (A) is characterized by moderate biological variations and the absence of hepatosplenic lesions.(ABSTRACT TRUNCATED AT 400 WORDS)

Aedes↗

A model of controlled acute hyperglycemia in rats: Effects of insulin and glucagon-like peptide-1 analog.

A rodent model of controlled acute hyperglycemia that is sensitive to glucose-lowering agents insulin and glucagon-like peptide-1 (GLP-1) analog has been developed. The studies show that anesthesia could be induced in fasted rats with ketamine (100 mg/kg) plus a low dose of xylazine (5 mg/kg) without inducing the acute hyperglycemia typically associated with these agents. Under these conditions, continuous infusion of glucose (10 and 20%) via the jugular vein for 30 to 150 min induced hyperglycemia in a time-dependent fashion. Administration of "loading" boluses of glucose (0.2-0.6 ml of a 20% solution) prior to continuous infusion of 10% glucose produced more immediate and sustained hyperglycemia. Plasma levels of a variety of glucoregulatory and stress hormones such as insulin, growth hormone, glucagon, and corticosterone were determined. Only glucagon levels changed significantly during induction and maintenance of hyperglycemia. The infusion of insulin (0.1 U/kg/h) or GLP-1 analog (10 microg/kg/h) effectively lowered blood glucose from its elevated levels. Insulin produced a significant increase in glucagon levels, and GLP-1 analog produced a significant increase in insulin levels without any change in other glucoregulatory and stress hormone levels. In conclusion, the present studies identified a novel approach for the induction of anesthesia and surgical manipulations without inducing hyperglycemia and further defined an approach for producing acute hyperglycemia in a controlled fashion in rodents. This model will be beneficial to study the influence of hyperglycemia in acute models of critical illness where hyperglycemia develops following the precipitating event. This model was responsive to insulin and GLP-1 analog, both of which were effective in ameliorating hyperglycemia.

Acute Disease↗

Failure of neuronal protection by inhibition of glial activation in a rat model of striatonigral degeneration.

Previous studies in rodent models of neurodegenerative disorders have demonstrated that minocycline exerts neuroprotective effects unrelated to its antimicrobial action. The purpose of the present study was to analyze whether minocycline exhibits neuroprotective activity in a rat model of striatonigral degeneration (SND), the core pathology underlying levodopa-unresponsive parkinsonism associated with multiple system atrophy (MSA). We observed no significant effect of minocycline on locomotor impairment in double-lesioned SND rats. Minocycline significantly suppressed astroglial and microglial activation (P < 0.01); however, 3'5'-monophosphate-regulated phosphoprotein (DARPP 32) immunohistochemistry revealed no significant differences in striatal lesion volume of minocycline-treated versus untreated control SND rats. Furthermore, there was no protection of nigral dopaminergic neurons in the double-lesion model. We conclude that despite its astrocytic and microglial suppression, minocycline failed to attenuate lesion-induced neuronal damage in the SND rat model.

Animals↗

Neurochemical and electrophysiological events underlying stress-induced depression in an animal model.

This paper has described an animal (rodent) model of depression in which depression-like characteristics are produced by exposure of animals to stressful events that they cannot control. This model, called "stress-induced depression," appears to be mediated by stress-induced changes in brain norepinephrine (NE), and evidence now indicates that a large depletion of NE in the Locus Coeruleus region of the brain stem (LC) is critical for producing the behavioral disturbance seen in this model. To explain the functional significance of NE depletion in the LC, it has been suggested that this change results in decreased stimulation, (i.e., functional blockade) of alpha-2 receptors that normally inhibit firing of LC neurons. Consequently, LC neurons should be disinhibited in stress-induced depression. The final part of this paper describes findings from electrophysiological measurement of LC activity. Prior to measuring changes occurring in stress-induced depression, studies examined the basic role played by alpha-2 receptors in LC activity. These receptors were found to regulate the responsivity of LC neurons to excitatory stimuli, so that pharmacological blockade of these receptors increased the firing of LC neurons to excitatory input. When LC activity was then measured in stressed animals showing behavioral depression, responsivity of LC neurons to excitatory input was elevated in comparison to animals not exposed to stress. Also, administration of an alpha-2 blocking drug could not increase responsivity of LC neurons in stressed animals, which further indicated that alpha-2 receptors in the LC region were functionally blocked in these animals. Finally, the amount that LC neurons increased their firing to an excitatory stimulus (i.e., an index of alpha-2 receptor blockade) was highly correlated with the amount of behavioral depression seen in an activity test conducted just prior to electrophysiological measurement. These electrophysiological findings indicate that LC neurons are disinhibited in stress-induced depression, and that this disinhibition is particularly characterized by increased responsivity of LC neurons to excitatory input, which is indicative of alpha-2 receptor blockade. These findings further support the view that the LC is involved in stress-induced depression, and are consistent with a proposed mechanism that attributes behavioral disturbance in the model to disinhibition of LC neurons arising from subnormal activation of inhibitory alpha-2 receptors.

Animals↗

Pharmacological evaluation of the selective spinal nerve ligation model of neuropathic pain in the rat.

Rodent models of neuropathic pain are used to investigate the underlying mechanisms of pain associated with damage to peripheral nerves and to evaluate the efficacy of novel compounds. However, few models have been adequately characterized and the validity of many models remains unclear. The present experiment examined the activity of known anti-allodynic compounds in the L5 spinal nerve ligation (SNL) model of peripheral mononeuropathy in the rat, a modified version of the L5/L6 SNL model [S.H. Kim, J.M. Chung, An experimental model for peripheral neuropathy produced by segmental spinal nerve ligation in the rat, Pain 50 (1992) 355-363]. Tactile sensitivity was measured 7-21 days post-surgery using von Frey monofilaments before and following treatment with gabapentin (30, 60 and 120 mg/kg), morphine (1, 3 and 6 mg/kg), amitriptyline (1.5, 3 and 10 mg/kg), fluoxetine (3, 10 and 30 mg/kg), WIN55,212-2 (0.5, 1 and 2.5 mg/kg), indomethacin (1 and 5 mg/kg) or U-50,488H (3 and 6 mg/kg). Compared to sham-operated control animals, L5 SNL animals displayed significant tactile allodynia in the ipsilateral hindpaw that was completely reversed by treatment with gabapentin, morphine, and WIN55,212-2, partially reversed by amitriptyline and fluoxetine, and unaffected by U-50,488H or indomethacin. The robust effects of the non-selective cannabinoid receptor agonist WIN55,212-2 and morphine support reports in the literature that systemic cannabinoid receptor agonists and opioids are active in neuropathic pain. These results suggest that the L5 SNL model can be utilized to determine the anti-allodynic activity of novel compounds.

Analgesics↗

Microinjections of muscimol into lateral superior colliculus disrupt orienting and oral movements in the formalin model of pain.

An important reaction in rodent models of persistent pain is for the animal to turn and bite/lick the source of discomfort (autotomy). Comparatively little is known about the supraspinal pathways which mediate this reaction. Since autotomy requires co-ordinated control of the head and mouth, it is possible that basal ganglia output via the superior colliculus may be involved; previously this projection has been implicated in the control of orienting and oral behaviour. The purpose of the present study was therefore, to test whether the striato-nigro-tectal projection plays a significant role in oral responses elicited by subcutaneous injections of formalin. Behavioural output from this system is normally associated with the release of collicular projection neurons from tonic inhibitory input from substantia nigra pars reticulata. Therefore, in the present study normal disinhibitory signals from the basal ganglia were blocked by injecting the GABA agonist muscimol into different regions of the rat superior colliculus. c-Fos immunohistochemistry was used routinely to provide regional estimates of the suppressive effects of muscimol on neuronal activity. Biting and licking directed to the site of a subcutaneous injection of formalin (50 microliters of 4%) into the hind-paw were suppressed in a dose-related manner by bilateral microinjections of muscimol into the lateral superior colliculus (10-50 ng; 0.5 microliter/side); injections into the medial superior colliculus had little effect. Bilateral injections of muscimol 20 ng into lateral colliculus caused formalin-treated animals to re-direct their attention and activity from lower to upper regions of space. Muscimol injected unilaterally into lateral superior colliculus elicited ipsilateral turning irrespective of which hind-paw was injected with formalin. Oral behaviour was blocked when the muscimol and formalin injections were contralaterally opposed; this was also true for formalin injections into the front foot. Interestingly, when formalin was injected into the perioral region, injections of muscimol into the lateral superior colliculus had no effect on the ability of animals to make appropriate contralaterally directed head and body movements to facilitate localization of the injected area with either front- or hind-paw. These findings suggest that basal ganglia output via the lateral superior colliculus is critical for responses to noxious stimuli which entail the mouth moving to and acting on the foot, but not when the foot is the active agent applied to the mouth. The data also suggest that pain produces a spatially non-specific facilitation of units throughout collicular maps, which can be converted into a spatially inappropriate signal by locally suppressing parts of the map with the muscimol.

Animals↗

Animal models of intestinal and joint inflammation.

Recent rodent models have been exploited to explore mechanisms of intestinal and joint inflammation. HLA-B27 transgenic rats develop colitis, gastritis, and arthritis when raised in a conventional environment, but have no evidence of inflammation under germfree (sterile) conditions. Metronidazole treatment attenuates gastrointestinal inflammation, suggesting that anaerobic bacteria are important. Experimental bacterial over-growth of predominantly anaerobic bacteria reactivates arthritis in Lewis rats which have been previously injected intra-articularly with bacterial cell wall polymers. Reactivation arthritis is mediated by interleukin-1, tumour necrosis factor-alpha, and can be blocked by metronidazole. Intramural injection of the bacterial cell wall polymer, peptidoglycan-polysaccharide, leads to biphasic, chronic granulomatous enterocolitis and peripheral arthritis in Lewis rats, but only transient intestinal inflammation and no arthritis in Buffalo or MHC-matched Fischer rats. Chronic granulomatous inflammation is mediated by T lymphocytes and interleukin-1 and is dependent on persistent antigenic stimulation by poorly biodegradable bacterial polymers. Results in these models firmly incriminate resident normal enteric flora (especially anaerobes), bacterial products, and host genetic susceptibility in the pathogenesis of spondyloarthropathies. We suggest that increased uptake of luminal bacterial components across the inflamed mucosa leads to systemic distribution of these arthropathic products. The genetically susceptible host develops reactive arthritis due to defective downregulation of inflammation in response to immunologically active bacterial components.

Animals↗

Beneficial effects of mercaptoethylguanidine, an inhibitor of the inducible isoform of nitric oxide synthase and a scavenger of peroxynitrite, in a porcine model of delayed hemorrhagic shock.

OBJECTIVE: In rodent models, enhanced formation of nitric oxide and formation of peroxynitrite have been implicated in the pathogenesis of various forms of shock. Here we examined the effect of mercaptoethylguanidine (MEG), an inducible nitric oxide synthase inhibitor and peroxynitrite scavenger, in a severe hemorrhagic shock model. DESIGN: Randomized, placebo-controlled trial. SETTING: Animal laboratory. SUBJECTS: Twenty-one anesthetized immature Yorkshire pigs. INTERVENTIONS: Mechanical ventilation, sternotomy, continuous cardiac output (pulmonary artery flowmetry), and systemic and intracardial pressure measurements were taken. Pigs were bled to a cardiac index of 40 mL/kg/min for 2 hrs, which was followed by saline resuscitation (20 mL/kg). MEG was administered in the resuscitation fluid (15 mg/kg bolus plus 15 mg/kg/hr infusion). MEASUREMENTS AND MAIN RESULTS: Hemodynamic variables, systemic and mixed venous blood gas tensions and oxygenation, arterial lactate concentration, myeloperoxidase activity, malondialdehyde content, and histologic injury in the lung and intestine were measured. Reduction of cardiac output to 40 mL/kg/min led to the following changes during hypovolemia: decreases in mean arterial blood pressure (to 30-35 mm Hg), both atrial pressures, systemic oxygen consumption (by 35%), mixed venous saturation (by 65%), and lactic acidosis (5.5-6.0 mM). Fluid replacement failed to restore blood pressure and cardiac output during resuscitation and was followed by gradual hemodynamic decompensation. Hemorrhagic shock induced lipid peroxidation, neutrophil deposition, and severe histologic alterations in the lung and intestine. MEG significantly ameliorated the decrease in blood pressure and cardiac output during resuscitation, improved survival rate, reduced lipid peroxidation in the intestine, and ameliorated neutrophil accumulation in the lung and intestine. MEG prevented the reduction in oxygen consumption during resuscitation. CONCLUSIONS: When given during resuscitation, MEG exerted beneficial effects in a porcine model of severe hemorrhagic shock. We propose that the mode of MEG's action is related to improved cardiac contractility.

Analysis of Variance↗

A new porcine model of reperfusion injury after lung transplantation.

Rodent models have been described to investigate lung preservation and reperfusion injury but have significant disadvantages. In large animals single lung transplant studies are probably optimal but problems remain over the ability to rigorously separate the lungs for assessment while promoting medium to long-term animal survival for meaningful investigation. Our aim was to develop a novel and refined large animal model to assess reperfusion injury in the transplanted lung, overcoming the difficulties associated with existing models. Specifically, small animal models of lung transplantation usually have short perfusion times (often one hour) and include extracorporeal circuits while larger animal models often require the contralateral lung to be excluded after transplantation-an unphysiological situation under which to evaluate the graft. A porcine model of left lung allotransplantation was developed in which native and donor lungs are individually ventilated. Sampling catheters placed within the graft lung allowed specimen withdrawal without mixing of blood from the contralateral lung after reimplantation. The model permits a variety of clinical scenarios to be simulated with the native lung supporting the animal irrespective of function in the graft. This model has been used in over 60 transplant procedures with a postoperative survival time of 12 h being readily achieved. The mean operating time was 2.6 h. The mortality rate is 4% in our series. We have found the model to be reliable, reproducible and flexible. We propose this model as an adaptable investigation for evaluating lung reperfusion injury and preservation.

Animals↗

Antidiabetic actions of estrogen: insight from human and genetic mouse models.

There is increasing evidence both in humans and rodents linking the endogenous estrogen 17b-estradiol (E2) to the maintenance of glucose homeostasis. Postmenopausal women develop visceral obesity and insulin resistance and are at increased risk for type 2 diabetes mellitus, but hormone replacement therapy leads to a reduction in the incidence of diabetes. In various spontaneous rodent models of type 2 diabetes, female rodents are protected against hyperglycemia unless they are ovariectomized, and E2 perfusion reverses diabetes in male rodents. Finally, the study of transgenic mice and mice with genetic alteration of E2 secretion or E2 action has shed light on the antidiabetic properties of E2 at a tissue-specific level. Thus, E2 secretion and action in rodents seems to be implicated 1) in adipose tissue biology and the prevention of obesity, 2) in the stimulation of liver fatty acid metabolism and suppression of hepatic glucose production, and 3) in the protection of pancreatic b-cell function/survival and insulin secretion in conditions of oxidative stress.

Animals↗

PEGylated recombinant human soluble tumour necrosis factor receptor type I (r-Hu-sTNF-RI): novel high affinity TNF receptor designed for chronic inflammatory diseases.

The proinflammatory cytokine, tumour necrosis factor alpha (TNFalpha) has been shown to play a pivotal part in mediating acute and chronic inflammation. The activities of TNFalpha are modulated by the proteolytic shedding of the soluble extracellular domains of the two TNF receptors, p55 sTNF-RI and p75 sTNF-RII. Amgen Inc has cloned and expressed a recombinant form of a natural inhibitor of TNFalpha, referred to as recombinant human soluble TNF receptor type I (r-Hu-sTNF-RI, sTNF-RI). sTNF-RI is an E coli recombinant, monomeric form of the soluble TNF-type I receptor. A high molecular weight polyethylene glycol (PEG) molecule is attached at the N-terminus position to form the molecule intended for clinical evaluations (PEG sTNF-RI). Preclinical studies to date demonstrate that PEG sTNF-RI is efficacious in rodent models of chronic inflammatory disease including rheumatoid arthritis and Crohn's disease at doses as low as 0.3 mg/kg given every other day. This dose results in plasma concentrations of 0.3 to 0.5 microg/ml. Higher doses with correspondingly higher plasma concentrations yield higher efficacy. It has also demonstrated efficacy in E coli lipopolysaccharide, and Staphylococcus enterotoxin B mediated models of acute inflammation in rodents and primates. Pharmacokinetic studies in mice, rats, cynomolgus monkeys, baboons, and chimpanzees have been conducted with PEG sTNF-RI. Absorption from a subcutaneous dose was slow, with the time to reach maximal plasma concentrations of 24-48 hours in rats, and in monkeys, and 3-29 hours in chimpanzees. The initial volume of distribution of PEG sTNF-RI was essentially equivalent to that of plasma (40 ml/kg). This suggests the protein does not appear to extensively distribute from the systemic circulation with a volume of distribution at steady state (Vss) less than 200 ml/kg in all species studied. These results are consistent with previous experience with PEGylated proteins in which PEGylation decreases both the rate of absorption and the plasma clearance of human recombinant proteins in animals and humans. The use of a PEG molecule will probably provide a more advantageous dosing schedule (that is, less frequent dosing) for the patient compared with a non-PEG sTNF-RI.

Animals↗

The value of animal models for drug development in multiple sclerosis.

The rodent model for multiple sclerosis, experimental allergic (autoimmune) encephalomyelitis (EAE), has been used to dissect molecular mechanisms of the autoimmune inflammatory response, and hence to devise and test new therapies for multiple sclerosis. Clearly, artificial immunization against myelin may not necessarily reproduce all the pathogenetic mechanisms operating in the human disease, but most therapies tested in multiple sclerosis patients are nevertheless based on concepts derived from studies in EAE. Unfortunately, several treatments, though successful in pre-clinical EAE trials, were either less effective in patients, worsened disease or caused unexpected, severe adverse events, as we review here. These discrepancies must, at least in part, be due to genetic and environmental differences, but the precise underlying reasons are not yet clear. Our understanding of EAE pathogenesis is still incomplete and so, therefore, are any implications for drug development in these models. Here, we suggest some potential explanations based on new thinking about key pathogenic concepts and differences that may limit extrapolation from EAE to multiple sclerosis. To try to circumvent these rodent-human dissimilarities more systematically, we propose that pre-clinical trials should be started in humanized mouse models.

Animals↗

Persistence of rubrospinal projections following spinal cord injury in the rat.

Recent neurophysiological and magnetic resonance imaging studies of clinically "complete" human spinal cord injuries indicate that some patients have considerable subclinical function and substantial morphological integrity of descending spinal tracts. In contrast, extensive histopathological changes, including cell death, have been described in central nervous system nuclei projecting to the cord following experimental transection or hemisection of the spinal cord in animal models. We have used a rodent model of severe compression cord injury that more closely resembles the clinical pathophysiology to investigate the extent of the persistence of the rubrospinal projection. Counts of red nucleus neurons retrogradely labelled with Fluorogold demonstrate that in contrast to the results obtained with transection models, compression injuries of the spinal cord do not result in massive loss of rubrospinal projections, at least up to 8 weeks postinjury. The results also suggest that many of the axons persist distal to the lesion site and that they are functionally intact with respect to retrograde transport capabilities.

Animals↗