Expanding our scientific horizons: utilization of unique model organisms in biological research

During the past century, research studies using animal models have contributed to numerous scientific discoveries and have been vital for the understanding of numerous biological processes, including disease. Over the past decades, the scientific community has defined a small number of model organisms that includes a few mammals, fish (mainly zebrafish), birds (mainly chicken), frogs, flies, and nematodes. Rodents are by far the most commonly employed laboratory animals in biomedical research. Mice share many biological similarities to humans and can be genetically manipulated to express mutations linked to human diseases. Mice and rats reproduce relatively quickly and have a short life span, which allows scientists to study progressive disorders, including aging. A large range of inbred mice strains enables accurate and reproducible experiments by decreasing the variability often associated with animal models and biological systems in general. Finally, mice are cost‐effective, small, and relatively easy to handle, transport, and house. All of these advantages combine to make mice the major species for recapitulating and studying human diseases. However, focusing exclusively on one or very few animal models may lead researchers to lose sight of other species with vastly different biology that might inform and affect our understanding of disease pathogenesis.     

Cell response to the exposure to chitosan-TPP//alginate nanogels

Hydrophilic nanocarriers formed by electrostatic interaction of chitosan with oppositely charged macromolecules have a high potential as vectors in biomedical and pharmaceutical applications. However, comprehensive information about the fate of such nanomaterials in biological environment is lacking. We used chitosan from both animal and fungal sources to form well-characterized chitosan-pentasodium triphosphate (TPP)//alginate nanogels suitable for comparative studies. Upon exposure of human colon cancer cells (HT29 and CaCo2), breast cancer cells (MDA-MB-231 and MCF-7), glioblastoma cells (LN229), lung cancer cells (A549), and brain-derived endothelial cells (HCEC) to chitosan-(TPP)//alginate nanogels, cell type-, nanogel dosage-, and exposure time-dependent responses are observed. Comparing chitosan-TPP//alginate nanogels prepared from either animal or fungal source in terms of nanogel formation, cell uptake, reactive oxygen species production, and metabolic cell activity, no significant differences become obvious. The results identify fungal chitosan as an alternative to animal chitosan in particular if biomedical/pharmaceutical applications are intended.     

Human hepatic cell cultures: in vitro and in vivo drug metabolism

Drug metabolism is the major determinant of drug clearance, and the factor most frequently responsible for inter-individual differences in drug pharmacokinetics. The expression of drug metabolising enzymes shows significant interspecies differences, and variability among human individuals (polymorphic or inducible enzymes) makes the accurate prediction of the metabolism of a new compound in humans difficult. Several key issues need to be addressed at the early stages of drug development to improve drug candidate selection: a) how fast the compound will be metabolised; b) what metabolites will be formed (metabolic profile); c) which enzymes are involved and to what extent; and d) whether drug metabolism will be affected directly (drug-drug interactions) or indirectly (enzyme induction) by the administered compound. Drug metabolism studies are routinely performed in laboratory animals, but they are not sufficiently accurate to predict the metabolic profiles of drugs in humans. Many of these issues can now be addressed by the use of relevant human in vitro models, which speed up the selection of new candidate drugs. Human hepatocytes are the closest in vitro model to the human liver, and they are the only model which can produce a metabolic profile of a drug which is very similar to that found in vivo. However, the use of human hepatocytes is restricted, because limited access to suitable tissue samples prevents their use in high throughput screening systems. The pharmaceutical industry has made great efforts to develop fast and reliable in vitro models to overcome these drawbacks. Comparative studies on liver microsomes and cells from animal species, including humans, are very useful for demonstrating species differences in the metabolic profile of given drug candidates, and are of great value in the judicious and justifiable selection of animal species for later pharmacokinetic and toxicological studies. Cytochrome P450 (CYP)-engineered cells (or microsomes from CYP-engineered cells, for example, Supersomes) have made the identification of the CYPs involved in the metabolism of a drug candidate more straightforward and much easier. However, the screening of compounds acting as potential CYP inducers can only be conducted in cellular systems fully capable of transcribing and translating CYP genes.     

Spontaneous tumours of pet dog as models for human cancers: searching for adequate guidelines.

Despite the ongoing search for "replacement" alternatives, animal models continue to play a crucial role in bio-medical research. However, both in vivo and in vitro models usually employed, such as rodents and/or cell lines, display intrinsic limits related to the specific characteristics of the biological systems used, whose management is very complex, and whose pathology, usually induced under artificial laboratory conditions, is frequently dissimilar to the studied human spontaneous disease. It has been suggested that carrying out clinical trials based on pre-clinical data obtained after a screening on animal models developing the neoplastic disease in a more similar way to human beings, as represented by spontaneous canine tumours, could accelerate the entry of new effective drugs into the clinical practice. In this paper the authors discuss the scientific foundation as well as the ethic and legal concerns related to the use of this "pet model" in comparative oncology, suggesting some "key words" as the necessary starting point for a correct approach to the matter.     

Development and evaluation of a new methodology for the fast generation of patient-specific Finite Element models of the buttock for sitting-acquired deep tissue injury prevention

The occurrence and management of Pressure Ulcers remain a major issue for patients with reduced mobility and neurosensory loss despite significant improvement in the prevention methods. These injuries are caused by biological cascades leading from a given mechanical loading state in tissues to irreversible tissue damage. Estimating the internal mechanical conditions within loaded soft tissues has the potential of improving the management and prevention of PU. Several Finite Element models of the buttock have therefore been proposed based on either MRI or CT-Scan data. However, because of the limited availability of MRI or CT-Scan systems and of the long segmentation time, all studies in the literature include the data of only one individual. Yet the inter-individual variability can’t be overlooked when dealing with patient specific estimation of internal tissue loading. As an alternative, this contribution focuses on the combined use of low-dose biplanar X-ray images, B-mode ultrasound images and optical scanner acquisitions in a non-weight-bearing sitting posture for the fast generation of patient-specific FE models of the buttock. Model calibration was performed based on Ischial Tuberosity sagging. Model evaluation was performed by comparing the simulated contact pressure with experimental observations on a population of 6 healthy subjects. Analysis of the models confirmed the high inter-individual variability of soft tissue response (maximum Green Lagrange shear strains of 213 ± 101% in the muscle). This methodology opens the way for investigating inter-individual factors influencing the soft tissue response during sitting and for providing tools to assess PU risk.     

Natural Products: An Alternative to Conventional Therapy for Dermatophytosis?

The increased incidence of fungal infections, associated with the widespread use of antifungal drugs, has resulted in the development of resistance, making it necessary to discover new therapeutic alternatives. Among fungal infections, dermatophytoses constitute a serious public health problem, affecting 20–25 % of the world population. Medicinal plants represent an endless source of bioactive molecules, and their volatile and non-volatile extracts are clearly recognized for being the historical basis of therapeutic health care. Because of this, the research on natural products with antifungal activity against dermatophytes has considerably increased in recent years. However, despite the recognized anti-dermatophytic potential of natural products, often advantageous face to commercial drugs, there is still a long way to go until their use in therapeutics. This review attempts to summarize the current status of anti-dermatophytic natural products, focusing on their mechanism of action, the developed pharmaceutical formulations and their effectiveness in human and animal models of infection.     

Defining biological communication

Communication is ubiquitous in biology, and agreement on terms essential for scientific progress. Yet there is no agreed definition of biological communication. Definitions couched in terms of adaptation are often used, but there is significant variability in exactly which criteria are invoked. An alternative is to define communication in terms of information transfer. This article reviews the merits of these approaches, and argues that the former is to be preferred, so long as we demand that both the signal and the response be adaptive, rather than just one or the other, as is common. Specific concerns with the definition are addressed, and it is then explained why an account of communication predicated on information transfer is necessarily derivative upon such an approach. Other alternatives and some variants of the adaptationist definition are also briefly discussed.     

Parameter Redundancy in Multistate Capture‐Recapture Models

Multistate capture‐recapture models are a powerful tool to address a variety of biological questions concerning dispersal and/or individual variability in wild animal populations. However, biologically meaningful models are often over‐parameterized and consequently some parameters cannot be estimated separately. Identifying which quantities are separately estimable is crucial for proper model selection based upon likelihood tests or information criteria and for the interpretation of the estimates obtained. We show how to investigate parameter redundancy in multistate capture‐recapture models, based on formal methods initially proposed by Catchpole and his associates for exponential family distributions (Catchpole, Freeman and Morgan, 1996. Journal of the Royal Statistical Society Series B 58, 763–774). We apply their approach to three models of increasing complexity.     

Human tissue for in vitro research as an alternative to animal experiments: a charitable "honest broker" model to fulfil ethical and legal regulations and to protect research participants

Research with human tissue offers the possibility not only of improving preclinical pharmaceutical research and safety assessment, but also of the substitution of some animal experiments. Surgically removed human tissue is discarded after pathological evaluation. This tissue would be of enormous value for research, especially in the pharmaceutical branch, if it were readily available in an ethically and legally approved manner. But there are public concerns about the use of human tissue, especially for "commercial" purposes, such as in the pharmaceutical industry. The question is whether the ethical boundaries are sufficiently respected in the course of striving for industrial profit. To overcome this problem, a clear procedure for tissue donation, collection, supply and allocation must be established, which is guaranteed to be independent of special interests. The persisting problem seems to be the lack of an authority which asks for informed consent, coordinates tissue as well as blinded data collection, and supplies research facilities with tissue samples in a transparent manner. Therefore, a charitable, state-controlled foundation acting as an "honest broker" was initiated, to cover the ethical and legal aspects, as well as to protect the research participants in their use of human tissue as an alternative to animal experiments.     

Meta-analysis of variation suggests that embracing variability improves both replicability and generalizability in preclinical research

The replicability of research results has been a cause of increasing concern to the scientific community. The long-held belief that experimental standardization begets replicability has also been recently challenged, with the observation that the reduction of variability within studies can lead to idiosyncratic, lab-specific results that cannot be replicated. An alternative approach is to, instead, deliberately introduce heterogeneity, known as “heterogenization” of experimental design. Here, we explore a novel perspective in the heterogenization program in a meta-analysis of variability in observed phenotypic outcomes in both control and experimental animal models of ischemic stroke. First, by quantifying interindividual variability across control groups, we illustrate that the amount of heterogeneity in disease state (infarct volume) differs according to methodological approach, for example, in disease induction methods and disease models. We argue that such methods may improve replicability by creating diverse and representative distribution of baseline disease state in the reference group, against which treatment efficacy is assessed. Second, we illustrate how meta-analysis can be used to simultaneously assess efficacy and stability (i.e., mean effect and among-individual variability). We identify treatments that have efficacy and are generalizable to the population level (i.e., low interindividual variability), as well as those where there is high interindividual variability in response; for these, latter treatments translation to a clinical setting may require nuance. We argue that by embracing rather than seeking to minimize variability in phenotypic outcomes, we can motivate the shift toward heterogenization and improve both the replicability and generalizability of preclinical research.

A meta-analysis study of the variability in phenotypic outcomes in both control and experimental animal models of ischaemic stroke provides novel perspectives in which heterogeneity can be embraced to improve the reproducibility and translation of preclinical studies.     

SysFinder:A customized platform for search,comparison and assisted design of appropriate animal models based on systematic similarity

Animal models are increasingly gaining values by cross-comparisons of response or resistance to clinical agents used for patients.However,many disease mechanisms and drug effects generated from animal models are not transferable to human.To address these issues,we developed SysFinder(http://lifecenter.sgst.cn/SysFinder),a platform for scientists to find appropriate animal models for translational research.SysFinder offers a "topic-centered" approach for systematic comparisons of human genes,whose functions are involved in a specific scientific topic,to the corresponding homologous genes of animal models.Scientific topic can be a certain disease,drug,gene function or biological pathway.SysFinder calculates multi-level similarity indexes to evaluate the similarities between human and animal models in specified scientific topics.Meanwhile,SysFinder offers species-specific information to investigate the differences in molecular mechanisms between humans and animal models.Furthermore,SysFinder provides a userfriendly platform for determination of short guide RNAs(sgRNAs) and homology arms to design a new animal model.Case studies illustrate the ability of SysFinder in helping experimental scientists.SysFinder is a useful platform for experimental scientists to carry out their research in the human molecular mechanisms.     

A two-centre evaluation of the human organotypic skin explant culture model for screening contact allergens

Animal models are considered to be the "gold standard" for determining the potential contact allergenicity of low molecular weight chemicals. However, governmental regulations and ethical considerations limit the use of animals for such purposes. There is therefore a need for in vitro alternative models. The human organotypic skin explant culture (HOSEC) model is reported to be a promising alternative method for the predictive testing of contact allergens. The accelerated migration of Langerhans cells from the epidermis upon exposure to contact allergens is used to identify chemicals that are potentially capable of inducing a delayed-type hypersensitivity. In the study described in this paper, the model was further refined, and used, in two independent laboratories, to screen 23 low molecular weight compounds of known classification for their allergenicity. Each laboratory was able to accurately detect the contact allergens, despite small variations in the protocols used. However, the classification of dermal irritants, which have often been falsely classified as allergens, varied between the two laboratories. Despite the current limitations of the HOSEC model, the accuracy of the predictions made (sensitiser or non-sensitiser) compare favourably with classifications obtained with commonly used animal models. The HOSEC model has the potential to be developed further as an in vitro alternative to animal models for screening for contact allergens.     

Gene electro-transfer of an improved erythropoietin plasmid in mice and non-human primates

BACKGROUND: Anemia due to impaired erythropoietin (EPO) production is associated with kidney failure. Recombinant proteins are commonly administered to alleviate the symptoms of this dysfunction, whereas gene therapy approaches envisaging the delivery of EPO genes have been tried in animal models in order to achieve stable and long-lasting EPO protein production. Naked DNA intramuscular injection is a safe approach for gene delivery; however, transduction levels show high inter-individual variability in rodents and very poor efficiency in non-human primates. Transduction can be improved in several animal models by application of electric pulses after DNA injection. METHODS: We have designed a modified EPO gene version by changing the EPO leader sequence and optimizing the gene codon usage. This modified gene was electro-injected into mice, rabbits and cynomolgus monkeys to test for protein production and biological effect. CONCLUSIONS: The modified EPO gene yields higher levels of circulating transgene product and a more significant biological effect than the wild-type gene in all the species tested, thus showing great potential in clinically developable gene therapy approaches for EPO delivery.     

The structural basis of inter-individual differences in human behaviour and cognition

Inter-individual variability in perception, thought and action is frequently treated as a source of 'noise' in scientific investigations of the neural mechanisms that underlie these processes, and discarded by averaging data from a group of participants. However, recent MRI studies in the human brain show that inter-individual variability in a wide range of basic and higher cognitive functions - including perception, motor control, memory, aspects of consciousness and the ability to introspect - can be predicted from the local structure of grey and white matter as assessed by voxel-based morphometry or diffusion tensor imaging. We propose that inter-individual differences can be used as a source of information to link human behaviour and cognition to brain anatomy.     

Gender differences in hemocyte immune parameters of bivalves: the Sydney rock oyster Saccostrea glomerata and the pearl oyster Pinctada fucata

Many authors have highlighted a high inter-individual variability in immune parameters of marine bivalves. A high number of studies have reported the impact of external factors on hemocytes immune parameters such as temperature, salinity, pollutants or pathogens. However, only a few of them considered the impact of intrinsic parameters such as sex. Therefore, the present study assessed the impact of gender on hemocytes functions on two marine bivalves. Our results led to the conclusion that the gender contributes to this inter-individual variability. When studying the impact of an environmental variable, a pathogen or a pollutant, the sex of each animal should be determined and taken into account in the analysis and interpretation of immune parameters.     

Possibilities of replacing and reducing animal tests in the pharmacopoeias: an evaluation of unused methods and proposals for change

The animal tests currently carried out according to the German and European pharmacopoeias are evaluated. The routine testing of biological medicines, excluding vaccines and pyrogen tests, is examined. The opportunities for replacing obligatory bioassays and safety tests are assessed, taking into account both pharmaceutical quality and animal welfare aspects. The aim is to encourage institutions, public authorities, and the pharmaceutical industry to investigate the importance and the necessity of the remaining tests. In addition, all parties should be encouraged to initiate integrated projects to develop, validate and establish alternative methods.     

Nonhuman primates and teratological research

Nonhuman primates were first recognized as models for the study of developmental toxicity (teratology) following the thalidomide tragedy. Since that time they have played important roles in both testing of drugs for human safety and as models for studying specific malformations commonly seen in children. Although in vitro and alternative test systems using lower animal forms or simplified test systems have been incorporated into developmental toxicity studies, whole animal testing will be required for the foreseeable future because of the complex relationship of the maternal/embryofetal/placental unit. The nonhuman primate will be particularly valuable where equivocal results are experienced in other commonly used laboratory species, when the drug/chemical is likely to be used during pregnancy, and for human-derived biotechnical products which often are not bioactive in nonprimate species.     

Progress toward generating a ferret model of cystic fibrosis by somatic cell nuclear transfer

Mammalian cloning by nuclear transfer from somatic cells has created new opportunities to generate animal models of genetic diseases in species other than mice. Although genetic mouse models play a critical role in basic and applied research for numerous diseases, often mouse models do not adequately reproduce the human disease phenotype. Cystic fibrosis (CF) is one such disease. Targeted ablation of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in mice does not adequately replicate spontaneous bacterial infections observed in the human CF lung. Hence, several laboratories are pursuing alternative animal models of CF in larger species such as the pig, sheep, rabbits, and ferrets. Our laboratory has focused on developing the ferret as a CF animal model. Over the past few years, we have investigated several experimental parameters required for gene targeting and nuclear transfer (NT) cloning in the ferret using somatic cells. In this review, we will discuss our progress and the hurdles to NT cloning and gene-targeting that accompany efforts to generate animal models of genetic diseases in species such as the ferret.     

The implications of microarray technology for animal use in scientific research

Microarray technology has the potential to affect the number of laboratory animals used, the severity of animal experiments, and the development of non-animal alternatives in several areas scientific research. Microarrays can contain hundreds or thousands of microscopic spots of DNA, immobilised on a solid support, and their use enables global patterns of gene expression to be determined in a single experiment. This technology is being used to improve our understanding of the operation of biological systems during health and disease, and their responses to chemical insults. Although it is impossible to predict with certainty any future trends regarding animal use, microarray technology might not initially reduce animal use, as is often claimed to be the case. The accelerated pace of research as a result of the use of microarrays could increase overall animal use in basic and applied biological research, by increasing the numbers of interesting genes identified for further analysis, and the number of potential targets for drug development. Each new lead will require further evaluation i n studies that could involve animals. In toxicity testing, microarray studies could lead to increases in animal studies, if further confirmatory and other studies are performed. However, before such technology can be used more extensively, several technical problems need to be overcome, and the relevance of the data to biological processes needs to be assessed. Were microarray technology to be used in the manner envisaged by its protagonists, there need to be efforts to increase the likelihood that its application will create new opportunities for reducing, refining and replacing animal use. This comment is a critical assessment of the possible implications of the application of microarray technology on animal experimentation in various research areas, and makes some recommendations for maximising the application of the Three Rs.     

Modeling sepsis in the laboratory: merging sound science with animal well-being

Despite impressive advances in biomedical research, few noteworthy breakthroughs have been made in the treatment of sepsis during the past several decades. This stalemate is primarily due to the intricate and heterogenic nature of the systemic immune responses characterized as the sepsis syndrome. In general, such complexity must be approached with in vivo models. Several animal models have been described, suggesting that none adequately address all of the pressing needs in sepsis research. The most clinically applicable models involve a localized infection, such as surgically induced polymicrobial sepsis, that gradually propagates a systemic immune response. Because relevant models must mimic a severe and chronic syndrome, animal well-being is often a concern in sepsis research. A balance between the needs of sepsis research and animal welfare can only be achieved through knowledge of the strengths and weaknesses of and alternatives to in vivo sepsis models.