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Adipogenesis: usefulness of in vitro and in vivo experimental models.

The use of experimental models is the foundation of experimental biology, so it is important to know how much the models can tell us about actual animals. Inconsistent or contradictory results from in vitro models are often associated with the perception that a particular model or results are somehow wrong and therefore cannot tell us anything important about how an animal works. In fact, in vitro conditions do not create new biology. Differences between in vitro and in vivo behavior can only result from the actual cellular repertoire, which provides a powerful tool to uncover new information. Adipose tissue research provides a useful context for examining this issue because the regulation of adipose growth and metabolism has important economic implications for livestock production. Examples are discussed in which either excess skepticism or narrow interpretation of results slowed progress toward our current understanding of adipose biology. Similarly, contemporary examples using genomics are used to suggest that large inconsistencies are still apparent with in vitro methods. Careful consideration of these inconsistencies may provide new insights.

Adipocytes↗

[Efficacy of extramucosal myotomy in an experimental model of intramural colonic obstruction. An experimental study in rats].

An experimental study over the efficacy of myotomy in the treatment of intramural colon obstruction is presented. Forty Wistar rats were used. Four groups were made with ten animals each: Group I: control; Group II: myenteric denervation of four cm segment of transverse colon; Group III: myenteric denervation of 4 cm segment of transverse colon and myotomy; Group IV: myotomy of segment of transverse colon. Myenteric denervation was produced by serosal application of 0.1% benzalkonium chloride during 30 minutes. Animals were evaluated by clinical parameters (survival, weight change, food intake, nutritional state), radiological (barium enema) and Histomorphometry. The myotomy is an effective technique for the treatment of the experimental model of colon obstruction used.

Animals↗

Electrical bone-growth stimulation in an experimental model of delayed union.

An experimental model has been devised for the consistent production of delayed bone healing of the tibia in adult dogs. A double-blind trial, with bias eliminated, was used to evaluate the use of a commercially available direct-current bone-growth stimulator with this model. The stimulator produced a statistically significant acceleration of bone healing at four weeks in the experimental model. Osteogenesis was normal, and no dysplastic, inflammatory, or neoplastic changes were found. This research has shown that electrical stimulation of bone is safe and augments bone formation. The bone-growth stimulator unit remains on trial, but in future it may alter the management of many difficult orthopaedic problems.

Animals↗

Experimental models of acute pancreatitis.

Various models of experimental acute pancreatitis are described. An ideal model would be reproducible, have a similar natural history to that of the human disease and a similar response to treatment. Furthermore, it should be relatively cheap and simple. Few, if any, of the available experimental models completely satisfy these criteria. The appropriate choice of model is discussed, as are possible future developments in this field.

Acute Disease↗

The pathobiology of moderate diffuse traumatic brain injury as identified using a new experimental model of injury in rats.

Experimental models of traumatic brain injury have been developed to replicate selected aspects of human head injury, such as contusion, concussion, and/or diffuse axonal injury. Although diffuse axonal injury is a major feature of clinical head injury, relatively few experimental models of diffuse traumatic brain injury (TBI) have been developed, particularly in smaller animals such as rodents. Here, we describe the pathophysiological consequences of moderate diffuse TBI in rats generated by a newly developed, highly controlled, and reproducible model. This model of TBI caused brain edema beginning 20 min after injury and peaking at 24 h post-trauma, as shown by wet weight/dry weight ratios and diffusion-weighted magnetic resonance imaging. Increased permeability of the blood-brain barrier was present up to 4 h post-injury as evaluated using Evans blue dye. Phosphorus magnetic resonance spectroscopy showed significant declines in brain-free magnesium concentration and reduced cytosolic phosphorylation potential at 4 h post-injury. Diffuse axonal damage was demonstrated using manganese-enhanced magnetic resonance imaging, and intracerebral injection of a fluorescent vital dye (Fluoro-Ruby) at 24-h and 7-day post-injury. Morphological evidence of apoptosis and caspase-3 activation were also found in the cerebral hemisphere and brainstem at 24 h after trauma. These results show that this model is capable of reproducing major biochemical and neurological changes of diffuse clinical TBI.

Animals↗

Experimental model of psychic stress.

An experimental model for the induction of an informational psychic stress in the rat, comparable with the psychic stress in man, is described. The method consists in the production of the emission of significant acoustic signals by an "emitting" group of animals, subjected in an aleatory manner to electroshocks. The acoustic nociceptive signals are heard by a "receiving" group of animals. In this "receiving" group are studied the behaviour and the neuroendocrine changes. This experimental model may be useful for studies of psychopharmacology, for the research of psychotropic drugs and for the determination of their action upon the harmful effect of informational aggressions and psychic stress.

Animal Communication↗

[Use of experimental models in osteoarticular infections].

Animal experimental models are necessary to study a certain number of factors affecting the course and treatment of bone and joint infections: virulence of the bacteria, presence of foreign bodies or blood-stained effusion, administration of corticosteroids, mode of action of antibiotics. Whereas the penetration of antibiotics within the joint is excellent, it is variable during bony infections and the association of two bactericidal antibiotics and prolonged treatment are necessary to sterilise the infected bone. Although the information supplied by experimental models may be fundamental, it cannot be extrapolated to man without certain precautions.

Adrenal Cortex Hormones↗

[Inhibition of the infectivity of Plasmodium gametocytes by the serum of the parasite host. Perfecting an experimental model].

In the course of experimental malarial infections the infectivity of the gametocytes falls abruptly and at an early stage of the infection. This phenomenon is independent of the production of circulating antibodies. With Plasmodium yoelii nigeriensis infecting the white mouse, the maximum infectivity of gametocytes for Anopheles stephensi occurs on day 2, and on day 5 no more oocysts develop in the mosquito. The behaviour of the plasmodial strain in the mouse and particularly the "crisis" phenomenon were studied in detail in standardized experimental conditions. The decrease of the infectivity for the Anopheles begins just before the peak of parasitaemia, and a concomitant increase in the number of circulating gametocytes is observed. These events are very shortly followed by the crisis. The more severe is the crisis and virulent the infection, the earlier and more abrupt is the loss of infectivity. Previous studies having shown that this inhibition of the gametocytes infectivity was linked to a serum factor, our research aimed at setting up an experimental model allowing the identification of this factor. The intravenous injection of 5th day serum to mice harbouring infective gametocytes did not determine any loss of the gametocytes infectivity in the receiving mice. In order to study in vitro the effect of the 5th day serum, this one was added to blood from mice with highly infective gametocytes, and mosquitoes were fed on this through a membrane. As a result, a significant decrease of the infectivity of gametocytes was observed. This inhibition is immediate and does not appear to be "dose-dependent". Inversely, serum from mice still infective to the mosquito did not reduce the infectivity of the gametocytes. The experimental model set up thus comprises: a) mouse blood containing P. y. nigeriensis infective gametocytes to which is added the serum to be tested; b) Anopheles stephensi fed through a membrane; this allows quantifying the infectivity of the gametocytes in the presence of normal serum (control) or inhibitory serum. This model proved to be reliable; it should promote the study of any factor likely to modify the infectivity of Plasmodium gametocytes to the mosquito.

Animals↗

Experimental models of hepatic fibrosis: a review.

No experimental model reproduces exactly human liver fibrosis by etiology. Nonetheless, each of the models reviewed in this article has served to enhance our understanding of pathogenetic mechanisms of liver fibrosis. There have been important common findings derived from several different models. The best example is the role of Ito cells in liver fibrogenesis. Involvement of Ito cells was consistently seen in the experimental models regardless of whether the fibrogenic stimulus was nutritional, hepatotoxic, or immunologic. Cellular and molecular mechanisms of Ito cell activation have begun to be explored in different models. Another example is the role of TGF beta in liver fibrogenesis. In both murine schistosomiasis model and Tsukamoto-French rat model, TGF beta was shown to be closely associated with fibrogenesis. With both in vitro and in vivo experimental approaches using cellular and molecular techniques, the experimental model of liver fibrosis will continue to provide data on the pathogenetic mechanisms of liver fibrogenesis. Future genetic and molecular approaches may allow development animal models with liver fibrosis that is inducible and genetically similarity to that of man.

Animals↗

Fertility in cryptorchidism: further development of an experimental model.

Further development of an experimental model for evaluating fertility in cryptorchidism led to studies of unilateral cryptorchidism, endocrinological cryptorchidism, and the effects of treatment. The results demonstrate that rats with unilateral mechanical cryptorchidism have a significant diminution in the ability to impregnate females (impregnation rate, 45%) when compared with sham-operated controls (84%) and rats undergoing unilateral orchiectomy (88%). In addition, we demonstrated that lower doses of estradiol caused cryptorchidism and resulted in infertility of approximately the same degree as higher doses (impregnation rates, 18% and 0%, respectively), but avoided obvious side effects. Treatment of estradiol-induced cryptorchidism with human chorionic gonadotropin resulted in testicular descent, but did not significantly improve the ability to bear offspring (10% with hCG vs. 0% without). Surgical orchiopexy after surgically induce mechanical cryptorchidism resulted in improved fertility (30% vs. 0%); however, the improvement was still significantly less than the control rate. In summary, this experimental model demonstrates the effects of various aspects of cryptorchidism and its treatment on fertility and can easily be adapted to evaluate important clinical problems.

Animals↗

Experimental models of pulmonary infection.

Experimental models of pulmonary infection are being discussed, focused on various aspects of good experimental design, such as choice of animal species and infecting strain, and route of infection/inoculation techniques (intranasal inoculation, aerosol inoculation, and direct instillation into the lower respiratory tract). In addition, parameters to monitor pulmonary infection are being reviewed such as general clinical signs, pulmonary-associated signs, complication of the pulmonary infection, mortality rate, and parameters after dissection of animals. Examples of pulmonary infection models caused by bacteria, fungi, viruses or parasites in experimental animals with intact or impaired host defense mechanisms are shortly summarized including key-references.

Animals↗

[Therapeutic effect of aqueous-extract from a traditional Chinese medical herb Drynaria fortunei on rat experimental model of alveolar bone resorption].

OBJECTIVE: To establish a rat model of experimental alveolar bone resorption and understand the therapeutic effect of a traditional Chinese medical herb Drynaria fortunei (DFS) on the rats suffering from alveolar bone resorption. METHOD: A SD rat model with experimental alveolar bone resorption was established by using injection with E. coli Lipopolysaccharide (E-LPS) in local tissue of the animals. A DFS preparation (DFS aqueous-extract) was extracted with distilled water. The modeled rats were administrated by perfusion with DFS aqueous-extract at a dosage of 15 g crude drug/kg once a day for 10, 20 and 30 d, respectively. The effects of DFS aqueous extract on experimental alveolar bone resorption were estimated by detections of serum alkali phosphate (ALP) activity, Ca2+ and osteocalcin (OC) levels, and TRAP stained osteoclast count, bone mineral density (BMD) measurement and HE stained histopathological examination of the tooth-periodontal samples. RESULT: In comparison with the controls, BMD values of the alveolar bones at experimental tooth sites of the tested rats with DFS aqueous-extract for 10 d administration were obviously increased (P < 0.05). In the tooth-periodontal samples from the rats for 10 d administration, disappearance of the osteoclast, decrease of Howship's lacuna numbers and formation of new non-calcified bone-like matrix attached by osteoblasts in alveolar bone at the root furcation from most of the samples occurred. Similar examining results in the tooth-periodontal samples from the rats for 20 and 30 d administrations were obtained, respectively. However, no statistically significant differences of the serum ALP activity, Ca2+ and OC levels among the tested rats for 10-30 d treatment and the controls (P > 0.05) were found. CONCLUSION: The DFS aqueous extract has exact therapeutic effect on rat experimental alveolar bone resorption through suppressing bone resorption and promoting bone regeneration. Serum ALP activity, Ca2+ and osteocalcin (OC) levels can not be used as the effective index for examining alveolar bone resorption and regeneration in animal models.

Alkaline Phosphatase↗

Complement activation and inhibition in experimental models of arthritis.

Complement activation has been implicated as a pathological process in a number of inflammatory and autoimmune disorders including chronic rheumatoid arthritis (RA). Animal models of experimental arthritis have been widely used to investigate the pathogenesis of RA and also in the development of novel therapies. Many of these models are complement-dependent and both incidence and progression of disease can be influenced by complement inhibition. In certain situations, local inhibition is of greater therapeutic benefit than systemic decomplementation. An increasing awareness and availability of a wide range of naturally occurring complement regulatory proteins can now offer a more targeted approach to complement inhibition while the availability of novel engineering strategies has also improved the efficiency of this process. The success of complement inhibition in the experimental models described should offer a novel therapeutic approach to the treatment of human inflammatory arthritis.

Animals↗

Experimental models of hypersensitivity pneumonitis.

Experimental models of hypersensitivity pneumonitis are important tools for the study of the pathogenesis of this disease. In this paper we review the characteristics of the main animal models developed until now. The HP models in rats seem to be particularly appropriate for studying pigeon fancier's disease and the HP induced by chemicals, as well as for studying mediators of acute lesions induced by immunocomplexes. However, the HP models developed in rats are of less value in the evaluation of other aspects of the pathogenesis of this clinical entity in humans. The murine models of HP offer several advantages: the ease and simplicity of intranasal administration, the ability to produce acute and subacute pulmonary lesions similar to those found in humans, the possibility of reproducing lesions similar to those of nonaffected exposed subjects and the possibility of pharmacologically modulating the process. Their disadvantages lie in the different pulmonary lymphocyte response and the difficulty in reproducing a model of chronic fibrosis. The HP models in rabbits are extraordinarily useful for evaluating the immunological mechanisms through which subjects repeatedly exposed to the antigen do not develop clinical manifestations. However, the rabbit has several immunological differences when compared to humans, and the effect of some immunomodulators in this animal is different. The models of HP in guinea-pigs have as advantages the ease in handling the animals, the possibility of pharmacological manipulation, and the ability to induce an acute phase that is very similar to that observed in humans. The drawback, however, is the low lymphocyte response and the striking eosinophilic reaction that contrast with the bronchoalveolar data found in HP in humans. In conclusion, there is no ideal model to reproduce all the findings observed in humans, suggesting that the experimental animal and the method of developing HP should be selected on the basis of concrete research aims.

Alveolitis, Extrinsic Allergic↗

Experimental models in microsurgery.

The development of experimental microsurgery can be considered the natural evolution of a diffuse need to increase precision in many fields of surgery. Microsurgery accelerated the possibility of deepening many unclear aspects of pathophysiology, using miniaturized and reproducible experimental models. We report briefly on the fundamental principles of microsurgery and the most frequently performed and useful models of experimental microsurgery, especially to employ as training models for surgeons, but also as bases for developing new and always-welcome models.

Disease Models, Animal↗

An experimental model of Pseudomonas aeruginosa renal infection in rats.

The model of experimental pyocyanic renal infection in rats based on direct inoculation of bacteria to the renal parenchyma is described. The authors describe in detail the procedure and evaluation of the results with respect to microbiology, histology and reproducibility. The method produced a focal supurrative inflammation with a significant bacteriologica finding in renal tissue for 8 weeks at the least. The inflammation has a tendency to form focal cicatrices not accompanied by diffuse shrinkage of the kidney. The method is well reproducible and its technique is relatively simple thus permitting to induce infection in a large series of animals within a short period of time. In the second part of the paper, the use of this method in testing the therapeutical efficacy of gentamicin, carbenicillin and their combination is reported. The results showed gentamicin to be more effective than carbenicillin and the combination of the two to have the highest therapeutical effect. Moreover, their combination made it possible to reduce the dosage of the antibiotics and thereby reduce their potential undesirable effect. Further, the results showed the described model of experimental renal infection to be well suited for testing the efficacy of chemotherapeutics and resolving some problems of the treatment of infectious diseases.

Bacteriuria↗

An experimental model of postnatal jaundice in the suckling rat. Suppression of induced hyperbilirubinemia by Sn-protoporphyrin.

A model of experimental postnatal hyperbilirubinemia in the rat has been developed utilizing the heme precursor delta-aminolevulinic acid (ALA) to produce jaundice during a selective time period after birth. This time period is defined as that between 7 d postnatally, when the initial postpartum alterations of serum bilirubin and heme metabolism in the neonate have subsided, and 21 d, when the hepatic conjugation mechanism for the bile pigment appears fully developed. Administration of ALA in this time period led to a rapid, consistent, and significant dose-dependent increase in serum bilirubin levels in the newborn animals. Heme administration produced a qualitatively similar but enhanced effect. Both compounds, in addition, induced a dose-dependent increase in hepatic heme oxygenase activity concomitant with the increase in serum bilirubin levels. Neither compound increased serum bilirubin levels significantly when administered at or after 21 d postnatally. Administration of the synthetic metalloporphyrin, Sn-protoporphyrin, to ALA-treated neonates resulted in a dose-dependent decrease in serum bilirubin levels and hepatic heme oxygenase activity. Mn- and Zn-protoporphyrin in comparable doses did not significantly inhibit ALA-induced hyperbilirubinemia. Sn-protoporphyrin also inhibited the hyperbilirubinemia produced by heme in the suckling animals. ALA administration to newborn rats during the specific postnatal period described provides a simple and convenient model of experimental jaundice in the developing neonate which permits an examination of the potential ability of synthetic metalloporphyrins or other compounds to suppress induced hyperbilirubinemia in the newborn animal. The ability to induce a consistent and significant degree of jaundice in the postnatal rat by the method described may also be useful for other types of studies concerned with the biological disposition and effects of endogenously formed bilirubin in the neonate. The results of this study confirm in another model system the potent ability of Sn-protoporphyrin to suppress jaundice in the neonate, and suggest that suppression of heme oxidation by synthetic heme analogues may represent a useful therapeutic approach to the problem of severe hyperbilirubinemia in human premature newborn.

Aging↗

Experimental models of heart failure.

Experimental models of heart failure can be used to address specific questions not easily answered in patients, but no single model can reproduce exactly any of the clinical syndromes of heart failure since these are dominated by fatigue and breathlessness. Heart failure may be induced experimentally by pressure loading, volume loading, myocardial infarction, or by the creation of other disease states within the myocardium. Pressure loading may be especially useful in the study of ventricular hypertrophy, cellular derangements and vascular changes. Volume loading may be useful when examining the pathogenesis of hormone and electrolyte disturbances. Models of myocardial infarction or destruction are likely to be the most suitable for assessing novel therapy provided that peripheral reflexes are maintained. Experimental cardiomyopathy can provide an important means of identifying pathological subcellular mechanisms. They may be of use in the evaluation of vasodilator drugs but caution should be exercised in the study of inotropic agents. Any one model may be useful if it permits study of a single factor or variable in isolation or at a time when information is not obtainable from patients. For greatest clinical relevance, studies should be made in conscious animals with intact reflexes.

Animals↗