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Improved experimental model for measuring skin degerming activity on the human hand.

A gloved-hand method is presented for evaluating the interaction of antimicrobial agents with the normal resident bacterial flora of human skin. One of the key features of the experimental model is a simplified technique for sampling the skin, which involves the addition of eluting fluid to the gloved hand. As with other skin sampling techniques, the number of bacteria recovered from the hands showed considerable variation from subject to subject. However, no significant differences were observed between the numbers of bacteria recovered from the right and left hands of individual subjects. The mean number of bacteria recovered from the hand before and after washing with nonmedicated soap was consistent and reproducible over a period of at least 5 consecutive days. The number of recoverable bacteria from the hand was greatly reduced by a single treatment with a surgical scrub preparation containing hexachlorophene. The extent of skin degerming achieved was little affected by the use of a surgical brush, and was maximal at approximately 30 min after contact with the hexachlorophene-containing formulation. It was determined that the level of transient bacteria on the hands could be controlled by a simple wash with nonmedicated soap, resulting in a stabilized base-line level from which treatment interactions with the resident microflora could be measured more precisely. The basic elements of the method presented fulfill the requirements of a satisfactory experimental model for the in vivo evaluation of skin-degerming agents on the hand. The selection of appropriate experimental designs allows treatment comparisons to be made with a high degree of statistical confidence.

Anti-Infective Agents, Local↗

[Experimental models for studying the effects induced by staphylococcal toxins A and B on human keratinocytes in culture].

The toxic effects of the two serotypes of staphylococcal exotoxin: exfoliatin A (ETA) and B (ETB) on two experimental models: organotypic cultures of the skin and cellular cultures of the epidermis reconstructed "in vitro" have been studied. The results show that, in both cases, purified ETB (Fig. 4a, 6-6a, b, c) reproduces the characteristics of Lyell's Staphylococcal Syndrome that is the intraepidermal cleavage either between the granulosa and the spinous layers or at the granulosa layer of the epidermis. As the images of the LM and EM demonstrate, purified ETA behaves differently in the two experimental models; in fact, it produces the same effect as purified toxin B on the organotypic cultures (Fig. 3a, b), whereas it causes no alterations in the epithelial cultures reproduced "in vitro" (Fig. 5a, b).

Bacterial Toxins↗

[Contribution of experimental models to the treatment of respiratory tract infections].

The need for more effective and potentially less toxic antimicrobial agents than aminoglycosides for treatment of gram negative bacilli pneumonia specially Pseudomonas aeruginosa, explains the interest for experimental models of pneumonia. These models allow to study the efficacy and safety of antibiotics alone or in combination. Aminoglycosides and new quinolones are more effective than beta-lactams for life threatening infection with high inoculum, the only exception to this finding has been for N-formimidoyl thienamycin, whereas for less severe infections aminoglycosides and beta-lactams are equivalent in efficacy. In contrast to high inocula pneumonia, combining beta-lactams with aminoglycosides give additive of synergic benefits for treating low inocula. Experimental models allow to compare continuous versus intermittent administration of antibiotics, different regimens of the same antibiotics (single daily dose of aminoglycosides versus conventional administration) and new antiinfectious agents as anti-pseudomonas hyperimmune globulins.

4-Quinolones↗

An experimental model for the study of canine flexor tendon adhesions.

An experimental model for the study of canine flexor tendon adhesions was designed using a standardized crush-abrasion injury, meticulous sheath closure, and three-week limb immobilization. Ten animals in the experimental protocol were evaluated for visible adhesion formation. With use of a flexor tendon adhesion rating scale, consistent adhesion formation was documented with an average score of 10.4 +/- 2.1 (range, 0 to 12). Five additional animals had biomechanical testing. Applying an increasing load to the proximally divided profundus tendon (0 to 10 Newtons), it was found that the angle of distal interphalangeal joint motion and the displacement of the toe were significantly decreased, and the work generated significantly increased in the experimental versus control digits (p less than 0.05). The consistent production of visible adhesions thus correlates with biomechanical impairments in toe motion and work. The study of agents aimed at diminishing flexor tendon adhesions will thus be facilitated by this reliable model. Systemic or intrasheath administration of agents will be possible.

Animals↗

Splenic dendritic cells induced by oral antigen administration are important for the transfer of oral tolerance in an experimental model of asthma.

Peripheral tolerance can be induced after the feeding of Ag, which is referred to as oral tolerance. We demonstrated in this study that the oral administration of OVA induced tolerance in an experimental model of asthma in mice, and investigated which cells function as the regulatory cells in the transfer of this oral tolerance. In OVA-fed mice, the percentage of eosinophils in bronchoalveolar lavage fluid, serum IgE levels, airway hyperresponsiveness, and mRNA levels of IL-13 and eotaxin were significantly lower than found in nonfed mice. Histological examination of lung tissue showed a suppression of the accumulation of inflammatory cells in the peribronchial area of OVA-fed mice. Feeding after the first immunization or between the first and the second immunization suppressed these findings, whereas feeding just before the airway Ag challenge did not. The suppression of disease in OVA-fed mice was successfully transferred by injection of whole spleen cells of OVA-fed mice. When CD11c+ dendritic cells (DCs) were removed from splenocytes, this transfer of suppression was completely abolished. The injection of splenic DCs purified from OVA-fed mice alone transferred the suppression, whereas the injection of splenic DCs from naive mice that were cocultured with OVA in vitro did not. These data suggest that not only CD4+ T cells, but also CD11c+ DCs induced by Ag feeding are important for the active transfer of oral tolerance in this murine experimental model of asthma.

Administration, Oral↗

[Biopathology of craniocerebral injuries: experimental models].

INTRODUCTION: In traumatic brain injury the secondary damage is responsible for the majority of the clinical and pathological sequels. This circumstance together with the difficulties of acquiring well preserved brain material for specialized neuropathological study impair the understanding of processes implied in their pathophysiology. DEVELOPMENT AND CONCLUSIONS: The development of experimental models has played an outstanding role for the better knowledge of these processes. For the development of standardized experimental models the biophysical and anatomical characteristics of brain and skull of the various utilized species have to be considered, but the physiological, neurochemical and molecular peculiarities must also be taken into account. Various in vivo models have been developed, inducing direct impact to the skull or to the brain, or mimicking lesions subsequent to craniocerebral trauma, such as subarachnoid hemorrhage, hematoma or necrosis. To analyze the molecular mechanisms of cellular response in vitro models have been developed. A great amount of cells in the contusional area die immediately following traumatic brain injury, but a still higher number of cells will undergo in the following hours. This second wave of neuronal death is induced by phenomena such as secondary axotomy and brain edema. In the pathogenesis of both the damage of cytoplasmic membrane is involved.

Animals↗

Effects of heparin in experimental models of acute pancreatitis and post-ERCP pancreatitis.

BACKGROUND: Acute pancreatitis (AP) is a complication of diagnostic or therapeutic endoscopic retrograde cholangiopancreatography (ERCP). In a recent clinical trial, a decreased rate of post-ERCP pancreatitis was shown after prophylactic heparin treatment. The aim of this study was to evaluate the effects of prophylactic heparin application in various experimental models of AP and pancreatic duct obstruction and to assess the underlying mechanisms. METHODS: In various experimental models, pancreatic injury of graded severity was induced in Wistar rats: (1) mild pancreatitis by IV cerulein infusion over 6 hours; (2) severe pancreatitis by infusion of glycodeoxycholic acid into the pancreatic duct plus IV cerulein application over 6 hours. The clinical ERCP situation was imitated in groups (3) obstruction of the pancreatic duct and (4) infusion of contrast medium into the pancreatic duct plus obstruction. In every group the animals received either no heparin (n=six per group) or continuous IV heparin (n=six per group) starting before pancreatic injury. Histologic changes, amylase, and lipase in plasma were evaluated 12 hours after induction of pancreatic injury. Additional animals were treated to investigate pancreatic microcirculation by intravital microscopy (n=six per group). RESULTS: In groups 1, 3, and 4 (mild AP/duct obstruction/duct obstruction plus contrast medium), IV heparin-treated animals showed reduced edema, inflammation, and peak amylase values compared with the corresponding non-heparin-treated animals (P<.05). Moreover, mean erythrocyte velocity was significantly higher and leukocyte-endothelium interaction was reduced in these groups after prophylactic administration of heparin. In contrast, group 2 (severe AP) did not show any difference between control animals and animals that received heparin as assessed by histology and intravital microscopy. CONCLUSIONS: Prophylactic systemic application of heparin provides a protective effect in mild AP and in experimental post-ERCP pancreatitis. The mechanism of the protective effects of heparin seems to be the reduction of leukocyte-endothelium interaction and the normalization of pancreatic microcirculation.

Acute Disease↗

Chymase inhibitor suppresses adhesion formation in a hamster experimental model.

To clarify the role of chymase produced by mast cells in adhesion formation, we investigated the preventive effect of a specific chymase inhibitor, Suc-Val-Pro-Phe(p) (OPh)2, on adhesion formation in a hamster experimental model. Hamsters underwent resection of the right uterine body and then 10 microM Suc-Val-Pro-Phe(p) (OPh)2 or placebo was injected into the abdomen. Two weeks after the operation, the scores for adhesion formation in the chymase inhibitor-treated group were significantly lower than that in the placebo-treated group (placebo-treated group, 3.60+/-0.22; chymase inhibitor-treated group, 2.10+/-0.22; P<0.01). This specific chymase inhibitor, Suc-Val-Pro-Phe(p) (OPh)2, significantly suppressed the scores for adhesion formation in a hamster experimental model. Thus, chymase may play an important role in the adhesion formation.

Animals↗

[Animal experimental models for acute pancreatitis].

In acute pancreatitis, multiple organ failure in the early phase and infectious complications in the late phase are contributors to mortality. To analyze the mechanism of aggravation of acute pancreatitis is to investigate the mechanism of organ dysfunction and infection. As strategy to elucidate the mechanism, various animal experimental models are utilized. Caerulein-induced pancreatitis and bile salt-induced pancreatitis (duct injection model) are frequently employed for mild edematous pancreatitis and severe necrotizing pancreatitis, respectively. It is important to select an appropriate experimental model that corresponds to the purpose of study.

Acute Disease↗

Establishment of an experimental model for continuous intraportal infusion of chemo/immunotherapeutic agents in the rat.

The results of adjuvant continuous intraportal chemotherapy in preventing hepatic metastasis, following curative colorectal surgery, are still equivocal. To improve the efficacy of this treatment we tried to establish an experimental model for continuous intraportal infusion in Wistar male rats. Using a drug infusion balloon catheter, which can be fixed on the back of the rat, the continuous intraportal administration of chemo/immunotherapeutic agents was carried out in 47 rats for 5 days. Of them, 23 received chemotherapeutic (Mitomycin C/5-fluorouracil) and 24 received chemoimmunotherapeutic (Mitomycin C/5-fluorouracil/Lentinan) agents. The overall success rate of complete 5-day infusion of the agents was 70.2%. The concentration of 5-fluorouracil in the peripheral blood ranged from 5ng/ml to 17ng/ml (mean 10.4ng/ml) and in the portal blood from 192ng/ml to 610ng/ml (mean 312.0ng/ml) following the perfect continuous infusion of the agents. It can thus be suggested that our established experimental model can be used for the study of continuous intraportal infusion in unrestrained rats.

Animals↗

[A technical experimental model for esophageal anastomosis in rabbits].

This article has as its main objective to divulge the development and optimization of surgical-technical training procedures for pediatric surgeons required to correct esophageal atresias. Included are the esophageal anastomosis techniques used in New Zealand rabbits, proposed as an experimental model, to later be reproduced and therefore decrease morbiletality as a result of insufficient surgical in this field. The approach, diameter and consistency of the esophagus of the experimental model, shows very similar characteristics as those seen in the newborn with esophageal atresia.

Anastomosis, Surgical↗

Importance of the autonomic nervous system in an experimental model of commotio cordis.

BACKGROUND: Young athletes may die suddenly when they are struck in the chest (commotio cordis). Proposed mechanisms of sudden death in commotio cordis include hypervagatonia and activation of the sympathetic nervous system. In an experimental model of commotio cordis, the importance of the sympathetic and parasympathetic nervous system in the initiation of ventricular fibrillation was evaluated. MATERIAL/METHODS: Juvenile swine weighing between 8 and 12 kg were anesthetized with ketamine and isoflurane and placed prone in a sling. Twenty animals were randomized to pretreatment with placebo or sympathetic and parasympathetic blockade. Chest blows were guided by echocardiography to the center of the left ventricle and animals received 1-3 strikes with a regulation baseball propelled at 30 mph and timed to impact 10-30 ms prior to the peak of the T-wave. RESULTS: With 17 impacts in 10 autonomically blocked animals, 6 episodes of ventricular fibrillation were seen; with 15 impacts in 10 control animals, ventricular fibrillation occurred 6 times. There was also no significant difference between the groups in occurrence of nonsustained polymorphic ventricular tachycardia or ST elevation. Transient complete heart block was less commonly seen in animals treated with autonomic blockade, although this did not achieve statistical significance. CONCLUSIONS: In this experimental model of commotio cordis, autonomic blockade did not affect the frequency of sudden cardiac death, polymorphic ventricular tachycardia or ST segment elevation. Thus, vagotonic and sympathetic surges likely do not contribute to the syndrome of sudden death due to chest blows in young people and athletes.

Animals↗

Temporal changes in intracranial pressure in a modified experimental model of closed head injury.

OBJECT: The authors describe an experimental model of closed head injury in rodents that was modified from one developed by Marmarou and colleagues. This modification allows dual control of the dynamic process of impact compared with impulse loading that occurs at the moment of primary brain injury. The principal element in this weight-drop model is an adjustable table that supports the rat at the moment of impact from weights positioned at different heights (accelerations). The aim was to obtain reproducible pathological intracranial pressure (ICPs) while maximally reducing the incidence of mortality and skull fractures. METHODS: Intracranial pressure was investigated in different experimental settings, including two different rat strains and various impact-acceleration conditions and posttrauma survival times. Identical impact-acceleration injuries produced a considerably higher mortality rate in Wistar rats than in Sprague-Dawley rats (50% and 0%, respectively). Gradually increasing severity of impact-acceleration conditions resulted in findings of a significant correlation between the degree of traumatic challenge and increased ICP at 4 hours (p < 0.001, R2=0.73). When the impact-acceleration ratio was changed to result in a more severe head injury, the ICP at 4, 24, and 72 hours was significantly elevated in comparison with that seen in sham-injured rats (4 hours: 19.7+/-2.8 mm Hg, p=0.004; 24 hours: 21.8+/-1.1 mm Hg, p=0.002; 72 hours: 11.9+/-2.5 mm Hg, p=0.009). Comparison of the rise in ICP between moderate and severe impact-acceleration injury at 4 and 24 hours revealed a significantly higher value after severe injury (4 hours: p=0.008; 24 hours: p=0.004). Continuous recordings showed that ICP mounted very rapidly to peak values, which declined gradually toward a pathological level dependent on the severity of the primary insult. Histological examination after severe trauma revealed evidence of irreversible neuronal necrosis, diffuse axonal injury, petechial bleeding, glial swelling, and perivascular edema. CONCLUSIONS: This modified closed head injury model mimics several clinical features of traumatic injury and produces reliable, predictable, and reproducible ICP elevations with concomitant morphological alterations.

Animals↗

Pathogenesis of bovine tuberculosis: the role of experimental models of infection.

In many countries, test-and-slaughter policies based on tuberculin skin testing have made a significant impact on the control of bovine tuberculosis (caused by infection with Mycobacterium bovis). However, in some countries these policies have not proved as effective and improved disease control strategies are required (including improved diagnostic tests and development of vaccines). The host pathogen interactions in bovine tuberculosis are very complex. While studies of the disease in naturally infected field cases of bovine tuberculosis have provided valuable information, detailed knowledge can also be gained through studies of disease models. A number of studies have developed M. bovis infection models employing a range of routes and challenge doses. An early objective was assessment of vaccine efficiency, and models of infection remain central to current work in this area. Development of the intra-nasal and intra-tracheal models have also advanced our understanding of the kinetics of the immune response. In many of these studies, understanding of pathogenesis has been improved by definition of the cells that respond to infection and those that are instrumental in modulation of host responses. Experimental models of infection have been adapted to study cattle to cattle transmission, modeling one of the fundamental routes of infection. This review provides a historical perspective on the types of experimental models used in over 100 years of research and outlines new opportunities to refine those methods for bovine and human tuberculosis and to contribute to improved diagnostics, advanced understanding of immunology and vaccine design.

Aerosols↗

[Orbital endoscopic surgery: experimental model for foreign bodies extraction].

PURPOSE: Endoscopy is a useful technique that may be applied to study orbital pathology because it is safe. In this paper we describe the surgical technique using endoscopy for orbital foreign body extraction. METHODS: In an experimental model in the dog we inserted a spheric metallic foreign body into the intraconal portion of the orbit. After general anesthesia we performed a 2 mm. peritomy in the conjuntiva. Then we introduced the optic system connected to a TV monitor. Associated to the optic system we fixed a 14-G Abbocath to introduce a 1.7 mm foreign body forceps. RESULTS: After a complete endoscopic exam of the orbit we identified the metallic foreign body. With the 1.7 mm forceps we could take out the foreign body. The extractions were very simple and the technique was not traumatic for the orbit. CONCLUSIONS: Endoscopic surgery of the orbit is a useful and safe technique to extract orbital foreign bodies in this experimental model.

Animals↗

The rat brachial plexus and its terminal branches: an experimental model for the study of peripheral nerve regeneration.

Despite the introduction of microsurgical techniques into clinical practice, the results of surgical procedures involving the brachial plexus and peripheral nerves are still far from spectacular. We therefore studied the rat brachial plexus and its terminal branches in 203 rats. Detailed anatomic and morphologic analyses of the biceps brachii and musculocutaneous nerve, finger flexors, flexor carpi radialis, and the median nerve were performed. Various sources of conventional and vascularized nerve grafts were explored. After musculocutaneous nerve section or median nerve section, there were no articular contractures or automutilations, which constitutes an advantage for these experimental models over the sciatic nerve model. The brachial plexus and its terminal branches provide a good experimental model which can be used to assess the development and normal control of muscle function, examine the mechanisms underlying functional recovery, and test the effects of treatments to enhance recovery.

Animals↗

Mycophenolate mofetil reduces tissue damage and inflammation in an experimental model of colitis in rats.

BACKGROUND: Lymphocytes are widely believed to be responsible for persistent intestinal inflammation in inflammatory bowel diseases. Mycophenolate mofetil (MMF) is a potent immunosuppressant that inhibits lymphocyte proliferation and has been shown to be effective in preventing allograft rejection after organ transplantation. The purpose of this study was to assess the modulating effects of MMF on intestinal inflammation in an experimental model of colitis in rats. METHODS: Colitis was induced by rectal instillation of trinitrobenzenesulfonic acid (TNBS) in ethanol in male Sprague-Dawley rats. One group of rats (n = 10) was treated with MMF i.p. (25 mg/kg b.w.) daily for 1 week starting 24 h after induction of colitis. A second group of rats (n = 10) was treated with MMF at the same dose 2 days, I day and 1 h prior to induction of colitis. Control animals (n = 10) received vehicle only. After being killed, colonic tissue was macroscopically evaluated for necrosis and microscopically for ulcerations. Sections were stained and examined for the presence of granulocytes. RESULTS: Administration of MMF after induction of TNBS colitis reduced macroscopic injury by 62% compared to control animals (P = 0.01). Microscopic ulcerations were reduced by 64% compared to controls (P = 0.009). In addition, posttreatment significantly reduced the number of granulocytes. MMF pretreatment did not significantly prevent macroscopic or microscopic tissue damage, or change the number of granulocytes. CONCLUSION: Systemic administration of MMF significantly ameliorates tissue damage in a model of experimental colitis in rats suggesting that this compound may play an important role as an immunosuppressant in the therapy of inflammatory bowel diseases.

Animals↗

An experimental model for cerebral metastasis: preliminary light and ultrastructural studies.

An experimental model for hematogenously spread cerebral metastases by injection of a suspension of M3 fibrosarcoma cells into the carotid artery of C57 BL/6 mice was developed. Intracerebral metastatic tumor nodules were consistently produced by this method with subsequent death of the animals. Development of extracerebral metastatic disease was minimal. Light and electron microscopic studies were carried out at various time intervals postintracarotid injection of tumor cells to observe the morphologic events during the development of the brain metastases. Tumor cells were observed arrested in the cerebral capillaries from 15 minutes to 4 days post-injection. From 1 day to 4 days post-injection, individual tumor cells were also observed in the pericapillary spaces in the brains of the injected animals. From 5 days post-injection on, tumor cells were seen to be proliferating in peri-capillary spaces displacing the brain parenchyma and eventually formed tumor nodules with resulting death of the animals. Morphological changes were observed in the endothelial cells of the blood vessels which were surrounded by the growing metastatic tumors. This model, and modifications thereof, should prove to be valuable in the study of cerebral metastatic disease.

Animals↗