Inappropriate modeling of chronic and complex disorders: How to reconsider the approach in the context of predictive, preventive and personalized medicine, and translational medicine
Preclinical investigations such as animal modeling make the basis of clinical investigations and subsequently patient care. Predictive, preventive, and personalized medicine (PPPM) not only highlights a patient-tailored approach by choosing the right medication, the right dose at the right time point but it as well essentially requires early identification, by the means of complex and state-of-the-art technologies of unmanifested pathological processes in an individual, in order to deliver targeted prevention early enough to reverse manifestation of a pathology. Such an approach can be achieved by taking into account clinical, pathological, environmental, and psychosocial characteristics of the patients or an individual who has a suboptimal health condition. Inappropriate modeling of chronic and complex disorders, in this context, may diminish the predictive potential and slow down the development of PPPM and consequently modern healthcare. Therefore, it is the common goal of PPPM and translational medicine to find the solution for the problem we present in our review. Both, translational medicine and PPPM in parallel, essentially need accurate surrogates for misleading animal models. This study was therefore undertaken to provide shreds of evidence against the validity of animal models. Limitations of current animal models and drug development strategies based on animal modeling have been systematically discussed. Finally, a variety of potential surrogates have been suggested to change the unfavorable situation in medical research and consequently in healthcare.
KeywordsPredictive preventive personalized medicine Future healthcare Animal modeling Disease modeling Clinical trial failure Translational medicine Chronic diseases Cardiovascular disorders Cancer Toxicology Drug discovery Drug development
American College of Surgeons
Acquired immunodeficiency syndrome
American Joint Committee on Cancer
Amyotrophic lateral sclerosis
Amyloid precursor protein
Advanced trauma life support
Azidothymidine (now renamed zidovudine, but still best known by the abbreviation AZT)
Coronary artery bypass grafting
Computer-aided drug design
Coronary heart disease
Chronic unpredictable mild stress model
The European Association for Predictive, Preventive and Personalised Medicine
U.S. Food and Drug Administration
High-density lipoprotein cholesterol
Human epidermal growth factor receptor 2
Human immunodeficiency virus
Idiopathic pulmonary fibrosis
Lethal dose, 50%, median lethal dose
Neurological, neuropsychiatric, and neurodegenerative diseases
Predictive, preventive, and personalized medicine
Quantitative structure activity relationships
Randomized controlled trial
Severe acute respiratory syndrome
Superoxide dismutase 1
Effector memory T cell
Theralizumab (also known as TGN1412, CD28-SuperMAB, and TAB08), a humanized monoclonal antibody that not only binds to, but is a strong agonist for, the CD28 receptor of the immune system’s T cells
Tumor, nodes, and metastases
Transactive response DNA-binding protein 43
X-linked severe combined immunodeficiency
Authors would like to thank Dr. Hilda Samimi and Dr. Mahmood Naderi for their scientific advices.
Idea: Soroush Seifirad (SS) and Vahid Haghpanah (VH)
Literature review: SS
Drafting article: SS except for suggestions which were written by both SS and VH
Final review and approval: SS and VH
Compliance with ethical standards
Consent for publication
Not applicable. This is a theoretical appraisal; neither patients nor animals were involved in this research.
The authors declare that they have no competing interests.
- 1.Golubnitschaja O, Baban B, Boniolo G, Wang W, Bubnov R, Kapalla M, et al. Medicine in the early twenty-first century: paradigm and anticipation—EPMA position paper 2016. EPMA J. 2016;7:23. https://doi.org/10.1186/s13167-016-0072-4.
- 3.Seifirad S. An emerging need for developing new models for myocardial infarction as a chronic complex disease: lessons learnt from animal vs. human studies on cardioprotective effects of erythropoietin in reperfused myocardium. Front Physiol. 2014;5:44. https://doi.org/10.3389/fphys.2014.00044.Google Scholar
- 5.Hackam DG. Translating animal research into clinical benefit. BMJ. 2007;334(7586):163–4. https://doi.org/10.1136/bmj.39104.362951.80.
- 7.Mak IW, Evaniew N, Ghert M. Lost in translation: animal models and clinical trials in cancer treatment. Am J Transl Res. 2014;6(2):114–8.Google Scholar
- 8.Arrowsmith J. Trial watch: phase III and submission failures: 2007-2010. Nat Rev Drug Discov 2011;10(2):87. doi: https://doi.org/10.1038/nrd3375.
- 10.Domingo RT, Fries CC, Sawyer PN, Wesolowski SA. Peripheral arterial reconstruction: transplantation of autologous veins. Trans Am Soc Artif Intern Organs. 1963;9:305–11.Google Scholar
- 13.Dowdle WR, Birmingham ME. The biologic principles of poliovirus eradication. J Infect Dis. 1997;175(Suppl 1):S286–92.Google Scholar
- 14.Hackam DG, Hackam AS. Translation of genetic discoveries into clinical therapies. Ann Intern Med. 2008;148(3):246–7.Google Scholar
- 15.Marwick C. FDA halts gene therapy trials after leukaemia case in France. BMJ. 2003 Jan 25;326(7382):181.Google Scholar
- 16.Marshall E. FDA halts all gene therapy trials at Penn. Science. 2000;28;287(5453):565,567.Google Scholar
- 17.Juni P, Altman DG, Egger M. Systematic reviews in health care: assessing the quality of controlled clinical trials. BMJ. 2001;323(7303):42–6.Google Scholar
- 18.van der Worp HB, Howells DW, Sena ES, Porritt MJ, Rewell S, O'Collins V, et al. Can animal models of disease reliably inform human studies? PLoS Med. 2010;7(3):e1000245. https://doi.org/10.1371/journal.pmed.1000245.
- 24.Morgan P, Van Der Graaf PH, Arrowsmith J, Feltner DE, Drummond KS, Wegner CD, et al. Can the flow of medicines be improved? Fundamental pharmacokinetic and pharmacological principles toward improving phase II survival. Drug Discov Today. 2012;17(9–10):419–24. https://doi.org/10.1016/j.drudis.2011.12.020.Google Scholar
- 26.Ennever FK, Lave LB. Implications of the lack of accuracy of the lifetime rodent bioassay for predicting human carcinogenicity. Regul Toxicol Pharmacol. 2003;38(1):52–7.Google Scholar
- 27.Omenn GS, Stuebbe S, Lave LB. Predictions of rodent carcinogenicity testing results: interpretation in light of the Lave-Omenn value-of-information model. Mol Carcinog. 1995;14(1):37–45.Google Scholar
- 30.Dwan K, Altman DG, Arnaiz JA, Bloom J, Chan AW, Cronin E, et al. Systematic review of the empirical evidence of study publication bias and outcome reporting bias. PLoS One. 2008;3(8):e3081. https://doi.org/10.1371/journal.pone.0003081.
- 31.Hakem R, Mak TW. Animal models of tumor-suppressor genes. Annu Rev Genet. 2001;35:209–41. https://doi.org/10.1146/annurev.genet.35.102401.090432.Google Scholar
- 32.Polivka J Jr, Kralickova M, Polivka J, Kaiser C, Kuhn W, Golubnitschaja O. Mystery of the brain metastatic disease in breast cancer patients: improved patient stratification, disease prediction and targeted prevention on the horizon? EPMA J. 2017;8(2):119–27. https://doi.org/10.1007/s13167-017-0087-5.Google Scholar
- 35.Giuliano AE, Connolly JL, Edge SB, Mittendorf EA, Rugo HS, Solin LJ, et al. Breast cancer—major changes in the American Joint Committee on Cancer eighth edition cancer staging manual. CA Cancer J Clin. 2017;67(4):290–303. https://doi.org/10.3322/caac.21393.
- 37.Golubnitschaja O, Debald M, Yeghiazaryan K, Kuhn W, Pesta M, Costigliola V, et al. Breast cancer epidemic in the early twenty-first century: evaluation of risk factors, cumulative questionnaires and recommendations for preventive measures. Tumour Biol. 2016;37(10):12941–57. https://doi.org/10.1007/s13277-016-5168-x.
- 39.Ellison GD. Animal models of psychopathology. The low-norepinephrine and low-serotonin rat. Am Psychol. 1977;32(12):1036–45.Google Scholar
- 40.Paterson NE, Markou A. Animal models and treatments for addiction and depression co-morbidity. Neurotox Res 2007;11(1):1–32, 1.Google Scholar
- 41.Czeh B, Fuchs E, Wiborg O, Simon M. Animal models of major depression and their clinical implications. Prog Neuropsychopharmacol Biol Psychiatry. 2016;64:293–310. https://doi.org/10.1016/j.pnpbp.2015.04.004.
- 42.Yan HC, Cao X, Das M, Zhu XH, Gao TM. Behavioral animal models of depression. Neurosci Bull. 2010 Aug;26(4):327–37.Google Scholar
- 43.Fuchs E, Fliugge G. Experimental animal models for the simulation of depression and anxiety. Dialogues Clin Neurosci. 2006;8(3):323–33.Google Scholar
- 45.Jesberger JA, Richardson JS. Animal models of depression: parallels and correlates to severe depression in humans. Biol Psychiatry. 1985;20(7):764–84.Google Scholar
- 48.McKinney WT. Animal models of depression: an overview. Psychiatric developments. 1984;2(2):77–96.Google Scholar
- 49.Neumann ID, Wegener G, Homberg JR, Cohen H, Slattery DA, Zohar J, et al. Animal models of depression and anxiety: what do they tell us about human condition? Prog Neuropsychopharmacol Biol Psychiatry. 2011;35(6):1357–75. https://doi.org/10.1016/j.pnpbp.2010.11.028.
- 52.Ma L, Xu Y, Wang G, Li R. What do we know about sex differences in depression: a review of animal models and potential mechanisms. Prog Neuropsychopharmacol Biol Psychiatry. 2018;89:48–56. https://doi.org/10.1016/j.pnpbp.2018.08.026.
- 53.Palanza P. Animal models of anxiety and depression: how are females different? Neurosci Biobehav Rev. 2001;25(3):219–33.Google Scholar
- 54.Renaud A. Animal models of depression. Soins Psychiatr. 1988(88):10–3.Google Scholar
- 55.Scott S, Kranz JE, Cole J, Lincecum JM, Thompson K, Kelly N, et al. Design, power, and interpretation of studies in the standard murine model of ALS. Amyotroph Lateral Scler. 2008;9(1):4–15. https://doi.org/10.1080/17482960701856300.
- 59.Roberts NA, Martin JA, Kinchington D, Broadhurst AV, Craig JC, Duncan IB, et al. Rational design of peptide-based HIV proteinase inhibitors. Science. 1990;248(4953):358–61.Google Scholar
- 60.Sweet A, Erickson RP, Huntington C, Dawson D. A potential animal model for studying CF heterozygote advantage: genetic variation in theophylline-inducible colonic chloride currents among inbred strains of mice. Biochem Med Metab Biol. 1992;47(1):97–102.Google Scholar
- 61.Barinaga M. Knockout mice offer first animal model for CF. Science. 1992;257(5073):1046–7.Google Scholar
- 62.Knight A, Bailey J, Balcombe J. Animal carcinogenicity studies: 3. Alternatives to the bioassay. Altern Lab Anim. 2006;34(1):39–48.Google Scholar
- 63.Knight A, Bailey J, Balcombe J. Animal carcinogenicity studies: 1. Poor human predictivity. Altern Lab Anim. 2006;34(1):19–27.Google Scholar
- 66.Jerie P. New catastrophe in pharmacological treatment—the crisis of clinical studies? Acute organ failure after administration of TGN1412. Casopis lekaru ceskych. 2006;145(6):426.Google Scholar
- 68.Enterline PE. Early animal research on asbestos cancer. Am J Ind Med. 1993;24(6):783–5 author reply 7-91.Google Scholar
- 69.Enterline PE, Hartley J, Henderson V. Asbestos and cancer: a cohort followed up to death. Br J Ind Med. 1987;44(6):396–401.Google Scholar
- 70.Stewart A. Alternative sources of risk estimates for cancer effects of radiation. Mt Sinai J Med. 1995;62(5):380–5.Google Scholar
- 71.Gardner MJ, Snee MP, Hall AJ, Powell CA, Downes S, Terrell JD. Results of case-control study of leukaemia and lymphoma among young people near Sellafield nuclear plant in West Cumbria. BMJ. 1990;300(6722):423–9.Google Scholar
- 72.Ainley CC, Senapati A, Brown IM, Iles CA, Slavin BM, Mitchell WD, et al. Is alcohol hepatotoxic in the baboon? J Hepatol. 1988;7(1):85–92.Google Scholar
- 73.Zbinden G, Flury-Roversi M. Significance of the LD50-test for the toxicological evaluation of chemical substances. Arch Toxicol. 1981;47(2):77–99.Google Scholar
- 74.Ekwall B, Barile FA, Castano A, Clemedson C, Clothier RH, Dierickx P, et al. MEIC evaluation of acute systemic toxicity: part VI. The prediction of human toxicity by rodent LD50 values and results from 61 in vitro methods. Altern Lab Anim. 1998;26(Suppl 2):617–58.Google Scholar
- 75.Ekwall B, Clemedson C, Crafoord B, Ekwall B, Hallander S, Walum E, et al. MEIC evaluation of acute systemic toxicity: part V. Rodent and human toxicity data for the 50 reference chemicals. Altern Lab Anim. 1998;26(Suppl 2):571–616.Google Scholar
- 76.Kelly JT, Abuzzahab FS Sr. The antiparkinson properties of amantadine in drug-induced parkinsonism. J Clin Pharmacol New Drugs. 1971;11(3):211–4.Google Scholar
- 77.Danielczyk W. Twenty-five years of amantadine therapy in Parkinson’s disease. J Neural Transm Suppl. 1995;46:399–405.Google Scholar
- 78.Hubsher G, Haider M, Okun MS. Amantadine: the journey from fighting flu to treating Parkinson disease. Neurology. 2012 Apr 3;78(14):1096–9.Google Scholar
- 79.Ban TA. Fifty years chlorpromazine: a historical perspective. Neuropsychiatr Dis Treat. 2007;3(4):495–500.Google Scholar
- 80.Anisimov VN. Age and dose-dependent carcinogenic effects of N-nitrosomethylurea administered intraperitoneally in a single dose to young and adult female mice. J Cancer Res Clin Oncol. 1993;119(11):657–64.Google Scholar
- 81.Anisimov VN. Carcinogenesis and aging. III. The role of age in initiation and promotion of carcinogenesis. Exp Pathol. 1982;22(3):131–47.Google Scholar
- 82.Shanks N, Greek R, Greek J. Are animal models predictive for humans? Philos Ethics Humanit Med. 2009;4(2):2. https://doi.org/10.1186/1747-5341-4-2.
- 84.Lidegaard O. Smoking and use of oral contraceptives: impact on thrombotic diseases. Am J Obstet Gynecol. 1999;180(6 Pt 2):S357–63.Google Scholar
- 85.Formenty P, Boesch C, Wyers M, Steiner C, Donati F, Dind F, et al. Ebola virus outbreak among wild chimpanzees living in a rain forest of Cote d’Ivoire. J Infect Dis. 1999;179(Suppl 1):S120–6. https://doi.org/10.1086/514296.
- 86.Pennisi E. Monkey virus DNA found in rare human cancers. Science. 1997;275(5301):748–9.Google Scholar
- 87.Reinhardt V, Roberts A. The African polio vaccine-acquired immune deficiency syndrome connection. Med Hypotheses. 1997;48(5):367–74.Google Scholar
- 88.Lucas S. The river: a journey back to the source of HIV and AIDS. BMJ. 2000;320(7247):1481A.Google Scholar
- 89.Folks TM. Chimpanzees as original source for HIV. JAMA. 2000;283(3):310.Google Scholar
- 90.Horowitz LG. Murder and cover-up could explain the Florida dental AIDS mystery. Br Dent J. 1994 Dec 10–24;177(11–12):423–7.Google Scholar
- 91.Hayflick L. The choice of the cell substrate for human virus vaccine production. Lab Pract. 1970;19(1):58–62.Google Scholar
- 92.Hayflick L. Human virus vaccines: why monkey cells? Science. 1972;176(4036):813–4.Google Scholar
- 94.Kitazawa M, Medeiros R, Laferla FM. Transgenic mouse models of Alzheimer disease: developing a better model as a tool for therapeutic interventions. Curr Pharm Des. 2012;18(8):1131–47.Google Scholar
- 95.LaFerla FM, Green KN. Animal models of Alzheimer disease. Cold Spring Harb Perspect Med. 2012;2(11). https://doi.org/10.1101/cshperspect.a006320.
- 97.Roher AE, Kuo YM, Kokjohn KM, Emmerling MR, Gracon S. Amyloid and lipids in the pathology of Alzheimer disease. Amyloid. 1999;6(2):136–45.Google Scholar
- 98.Kuo YM, Beach TG, Sue LI, Scott S, Layne KJ, Kokjohn TA, et al. The evolution of a beta peptide burden in the APP23 transgenic mice: implications for A beta deposition in Alzheimer disease. Mol Med. 2001;7(9):609–18.Google Scholar
- 99.Wen PH, Hof PR, Chen X, Gluck K, Austin G, Younkin SG, et al. The presenilin-1 familial Alzheimer disease mutant P117L impairs neurogenesis in the hippocampus of adult mice. Exp Neurol. 2004;188(2):224–37. https://doi.org/10.1016/j.expneurol.2004.04.002.
- 101.Dunn L, Prosser HC, Tan JT, Vanags LZ, Ng MK, Bursill CA. Murine model of wound healing. J Vis Exp. 2013;(75):e50265. https://doi.org/10.3791/50265.
- 102.Balls M. Replacement of animal procedures: alternatives in research, education and testing. Lab Anim. 1994;28(3):193–211.Google Scholar
- 104.Arora T, Mehta AK, Joshi V, Mehta KD, Rathor N, Mediratta PK, et al. Substitute of animals in drug research: an approach towards fulfillment of 4R’s. Indian J Pharm Sci. 2011 Jan-Feb;73:1): 1–6.Google Scholar
- 105.Gordon S, Daneshian M, Bouwstra J, Caloni F, Constant S, Davies DE, et al. Non-animal models of epithelial barriers (skin, intestine and lung) in research, industrial applications and regulatory toxicology. Altex. 2015;32(4):327–78. https://doi.org/10.14573/altex.1510051.
- 107.Doke SK, Dhawale SC. Alternatives to animal testing: a review. Saudi Pharm. J. 2015;23(3):223–9.Google Scholar
- 108.Lilienblum W, Dekant W, Foth H, Gebel T, Hengstler J, Kahl R, et al. Alternative methods to safety studies in experimental animals: role in the risk assessment of chemicals under the new European Chemicals Legislation (REACH). Arch Toxicol. 2008;82(4):211–36.Google Scholar
- 109.Gruber FP, Hartung T. Alternatives to animal experimentation in basic research. Altex. 2004;21:3–31.Google Scholar
- 111.Freires IA, Sardi JCO, de Castro RD, Rosalen PL. Alternative animal and non-animal models for drug discovery and development: bonus or burden? Pharm Res. 2017;34(4):681–6.Google Scholar
- 112.Mosig AS. Organ-on-chip models: new opportunities for biomedical research. Future Sci OA. 2016 Jun;3(2):FSO130. Published online 2016 Jul 6. https://doi.org/10.4155/fsoa-2016-0038.
- 113.Beeson PB. The growth of knowledge about a disease: hepatitis. Am J Med. 1979;67(3):366–70.Google Scholar
- 114.Kannel WB, Castelli WP, McNamara PM, McKee PA, Feinleib M. Role of blood pressure in the development of congestive heart failure. N Engl J Med. 1972;287(16):781–7. https://doi.org/10.1056/NEJM197210192871601.
- 115.Sytkowski PA, Kannel WB, D'Agostino RB. Changes in risk factors and the decline in mortality from cardiovascular disease. The Framingham Heart Study. N Engl J Med. 1990;322(23):1635–41. https://doi.org/10.1056/NEJM199006073222304.
- 117.Mendelson MM, Marioni RE, Joehanes R, Liu C, Hedman AK, Aslibekyan S, et al. Association of body mass index with DNA methylation and gene expression in blood cells and relations to cardiometabolic disease: a Mendelian randomization approach. PLoS Med. 2017;14(1):e1002215. https://doi.org/10.1371/journal.pmed.1002215.
- 118.Holmes MV, Lange LA, Palmer T, Lanktree MB, North KE, Almoguera B, et al. Causal effects of body mass index on cardiometabolic traits and events: a Mendelian randomization analysis. Am J Hum Genet. 2014;94(2):198–208. https://doi.org/10.1016/j.ajhg.2013.12.014.
- 119.Shah S, Casas JP, Drenos F, Whittaker J, Deanfield J, Swerdlow DI, et al. Causal relevance of blood lipid fractions in the development of carotid atherosclerosis: Mendelian randomization analysis. Circ Cardiovasc Genet. 2013;6(1):63–72. https://doi.org/10.1161/CIRCGENETICS.112.963140.
- 121.Holmes MV, Asselbergs FW, Palmer TM, Drenos F, Lanktree MB, Nelson CP, et al. Mendelian randomization of blood lipids for coronary heart disease. Eur Heart J. 2015;36(9):539–50. https://doi.org/10.1093/eurheartj/eht571.
- 122.Ahnen DJ. Are animal models of colon cancer relevant to human disease. Dig Dis Sci. 1985;30(12 Suppl):103S–6S.Google Scholar
- 123.Pories SE, Ramchurren N, Summerhayes I, Steele G. Animal models for colon carcinogenesis. Arch Surg. 1993;128(6):647–53.Google Scholar
- 125.Gilbart MK, Hutchison CR, Cusimano MD, Regehr G. A computer-based trauma simulator for teaching trauma management skills. Am J Surg. 2000;179(3):223–8.Google Scholar
- 128.Burt T, John CS, Ruckle JL, Vuong LT. Phase-0/microdosing studies using PET, AMS, and LC-MS/MS: a range of study methodologies and conduct considerations. Accelerating development of novel pharmaceuticals through safe testing in humans—a practical guide. Expert Opin Drug Deliv. 2017;14(5):657–72. https://doi.org/10.1080/17425247.2016.1227786.Google Scholar
- 130.Xu K-P, Li X-F, F-SX Y. Corneal organ culture model for assessing epithelial responses to surfactants. Toxicol Sci. 2000;58(2):306–14.Google Scholar
- 131.Shay JW, Wright WE. The use of telomerized cells for tissue engineering. Nat Biotechnol. 2000;18(1):22–3.Google Scholar
- 132.Hill AJ, Teraoka H, Heideman W, Peterson RE. Zebrafish as a model vertebrate for investigating chemical toxicity. Toxicol Sci. 2005;86(1):6–19.Google Scholar
- 133.Peterson RT, Nass R, Boyd WA, Freedman JH, Dong K, Narahashi T. Use of non-mammalian alternative models for neurotoxicological study. Neurotoxicology. 2008;29(3):546–55.Google Scholar
- 134.Lagadic L, Caquet T. Invertebrates in testing of environmental chemicals: are they alternatives? Environ Health Perspect. 1998;106(Suppl 2):593–611.Google Scholar
- 135.Wilson-Sanders SE. Invertebrate models for biomedical research, testing, and education. ILAR J. 2011;52(2):126–52.Google Scholar
- 136.Gilbert LI. Drosophila is an inclusive model for human diseases, growth and development. Mol Cell Endocrinol. 2008;293(1–2):25–31.Google Scholar
- 137.Barr MM. Super models. Physiol Genomics. 2003;13(1):15–24.Google Scholar
- 138.Madeo F, Engelhardt S, Herker E, Lehmann N, Maldener C, Proksch A, et al. Apoptosis in yeast: a new model system with applications in cell biology and medicine. Curr Genet. 2002;41(4):208–16.Google Scholar
- 139.Karathia H, Vilaprinyo E, Sorribas A, Alves R. Saccharomyces cerevisiae as a model organism: a comparative study. PLoS One. 2011;6(2):e16015.Google Scholar
- 140.Hedges SB. The origin and evolution of model organisms. Nat Rev Genet. 2002;3(11):838–49.Google Scholar
- 141.Höfer T, Gerner I, Gundert-Remy U, Liebsch M, Schulte A, Spielmann H, et al. Animal testing and alternative approaches for the human health risk assessment under the proposed new European chemicals regulation. Arch Toxicol. 2004;78(10):549–64.Google Scholar
- 142.Creech JL Jr, Johnson MN. Angiosarcoma of liver in the manufacture of polyvinyl chloride. J Occup Med. 1974;16(3):150–1.Google Scholar
- 143.Davies MR, Hruska KA. Pathophysiological mechanisms of vascular calcification in end-stage renal disease. Kidney Int. 2001;60(2):472–9.Google Scholar
- 144.Hendriksen CF. Replacement, reduction and refinement alternatives to animal use in vaccine potency measurement. Expert Rev Vaccines. 2009;8(3):313–22.Google Scholar
- 145.Dezfulian M, Bartlett JG. Selective isolation and rapid identification of Clostridium botulinum types A and B by toxin detection. J Clin Microbiol. 1985;21(2):231–3.Google Scholar
- 146.Flaten GE, Dhanikula AB, Luthman K, Brandl M. Drug permeability across a phospholipid vesicle based barrier: a novel approach for studying passive diffusion. Eur J Pharm Sci. 2006;27(1):80–90.Google Scholar