Experimental Animal Research Research Articles

Effects of Acupuncture on Heart Rate Variability in Beagles; Preliminary Results

Evidence-based animal experimental research concerning the effects of acupuncture on autonomic function was performed by two research teams from China and Austria. This study describes measurements in beagles. Heart rate variability (HRV) recordings were performed under stable conditions in Beijing, China, and the data analysis and interpretation were completed in Graz, Austria. The electrocardiograms were recorded during bilateral body acupuncture (PC6, Neiguan). Power of the low frequency (LF), high frequency (HF), and the ratio (LF/HF) changed significantly during acupuncture stimulation in beagles after injection of atropine and β-blocker. However, there was no significant change in HF power after needling the Neiguan acupoint when a cervical vagotomy has been performed. Our findings show that acupuncture can mediate the HRV even after pharmaceutical blocking of autonomic function. Acupuncture effects on HRV should rely not only on autonomic nervous system but on complete central nervous system.

Role of complement and NK cells in antibody mediated rejection

Despite extensive research on T cells and potent immunosuppressive regimens that target cellular mediated rejection, few regimens have been proved to be effective on antibody-mediated rejection (AMR), particularly in the chronic setting. C4d deposition in the graft has been proved to be a useful marker for AMR; however, there is an imperfect association between C4d and AMR. While complement has been considered as the main player in acute AMR, the effector mechanisms in chronic AMR are still debated. Recent studies support the role of NK cells and direct effects of antibody on endothelium cells in a mechanism suggesting the presence of a complement-independent pathway. Here, we review the history, currently available systems and progress in experimental animal research. Although there are consistent findings from human and animal research, transposing the experimental results from rodent to human has been hampered by the differences in endothelial functions between species. We briefly describe the findings from patients and compare them with results from animals, to propose a combined perspective.

Isoflurane induces endothelial apoptosis of the post-hypoxic blood-brain barrier in a transdifferentiated human umbilical vein endothelial cell model.

Isoflurane is a popular volatile anesthetic agent used in humans as well as in experimental animal research. In previous animal studies of the blood-brain barrier (BBB), observations towards an increased permeability after exposure to isoflurane are reported. In this study we investigated the effect of a 2-hour isoflurane exposure on apoptosis of the cerebral endothelium following 24 hours of hypoxia in an in vitro BBB model using astrocyte-conditioned human umbilical vein endothelial cells (AC-HUVECs). Apoptosis of AC-HUVECs was investigated using light microscopy of the native culture for morphological changes, Western blot (WB) analysis of Bax and Bcl-2, and a TUNEL assay. Treatment of AC-HUVECs with isoflurane resulted in severe cellular morphological changes and a significant dose-dependent increase in DNA fragmentation, which was observed during the TUNEL assay analysis. WB analysis confirmed increases in pro-apoptotic Bax levels at 4 hours and 24 hours and decreases in anti-apoptotic Bcl-2 in a dose-dependent manner compared with the control group. These negative effects of isoflurane on the BBB after a hypoxic challenge need to be taken into account not only in experimental stroke research, but possibly also in clinical practice.

In vitro cultivation of Anaplasma marginale and A. phagocytophilum in tick cell lines: a review

Continuous cell lines have been established from several ixodid and argasid tick species, representing an excellent tool suitable for the isolation of pathogens and their subsequent propagation, which in turn allows the production of antigenic material for diagnostic tests, antibody and vaccine production, and also for studies on host-vector-pathogen relationships. This paper reviews the use of tick cells for culture initiation and maintenance of two obligate intracellular bacterial pathogens, Anaplasma marginale and Anaplasma phagocytophilum. These in vitro cultivation systems have been used in a wide range of studies, covering morphological ultrastructural analysis, genetics, proteomics and biological differences between strains, including genome transcriptional and protein expression approaches, enabling comparisons between host and vector cells. Thus, such systems open a new window for a better understanding of interactions between pathogens and tick cells. Last but not least, such systems contribute to the reduction in usage of animals for experimental research, as antigenic material can be produced in reasonably large quantities without the use of in vivo species-specific systems.

Effects of immunosuppression by cyclosporine A on allogenic uterine transplant in the rat

Research on uterine transplantation (UTx) is conducted in preparation for its introduction in the human as a treatment for absolute uterine factor infertility. A major area of research in experimental animals is to ascertain that immunosuppressants that will be used at UTx do not negatively affect the potential of the uterus to implant an embryo and to carry a pregnancy to term. This study investigates the effects on a uterine transplant in the rat of the calcineurin inhibitor, cyclosporine A (CsA), on uterine morphology and expression patterns of some mediators involved in implantation/inflammation. Donor rats were of Brown Norway strain and recipients were of Lewis strain. The uterus was transplanted to an orthotopic site by vascular anastomosis. The recipients were given CsA (10mg/kg) sc once daily or no CsA until they were sacrificed at postoperative day 7. Syngenic transplanted Lewis rats were used as controls. Uteri were analyzed regarding histology, immunohistochemistry against T-cells and mRNA levels of the implantation/inflammation-related markers leukaemia inhibitory factor (LIF), galectin-1, CD200, interleukin (IL)-1α, and IL-15. There was pronounced inflammation with abundance of CD8-lymphocytes in uterine grafts of non-CsA-treated animals and only mild inflammation in treated animals. The uterine mRNA levels of IL-1α were decreased after CsA in comparison to uteri of non-treated transplanted animals. The mRNA levels of galectin-1 were decreased in the rejected uteri and were higher in the CsA-treated. The levels of mRNA of IL-15 were lower in the syngenic transplanted group compared to the CsA-treated transplanted. There was no difference between the groups concerning mRNA levels of CD200, or LIF, with wide variation of the levels of the two latter mediators in all groups. Cyclosporine A suppresses rejection of an allogenic rat uterine transplant, with normalization of mRNA levels of the proinflammatory cytokine IL-1α and the glycan-binding protein galectin-1.

Technical guidelines for porcine liver

Experimental animal research has been pivotal in the development of clinical liver transplantation (LTx). Results obtained in these experiments have been applied in clinic and clinical challenges have been scrutinized in animal laboratories. Porcine model is an optimal model in the field of experimental LTx research. Here, we present the various techniques of experimental LTx in the porcine model in detail. Different methods and modifications have been described. The following major steps have been discussed in detail: donor liver preparation, recipient operation including recipient hepatectomy, and reconstruction phase, including the reconstruction of suprahepatic inferior vena cava (SHIVC), portal vein (PV), infrahepatic inferior vena cava (IHIVC), hepatic artery (HA) and bile duct (BD). IHIVC and SHIVC are anastomosed end to end directly or with the use of prosthesis anastomosed side to side. The PV anastomosis is performed end to end between donor and recipient PV, Cuff method or Stump method. Arterialization has been accomplished via carrel patch or donor HA end to end with recipient HA. There are three major methods for reconstruction of BD: end to end or end to side choledochocholedochostomy or choledojejunostomy with Roux-en-Y jejunal loop. Each method has advantages and disadvantages regarding the objectives of the study; the most physiological techniques may be preferred for long-term survival studies, while the faster techniques may be selected for experimentations aiming the direct postoperative phase.

Drugs during Pregnancy and Lactation: New Solutions to Serious Challenges

Drugs during Pregnancy and Lactation: New Solutions to Serious Challenges

A hyperbaric oxygen chamber for animal experimental purposes

Facilities for hyperbaric oxygen therapy that are suitable for animal experimental research are scarce. In this paper, the authors introduce a hyperbaric oxygen chamber that was developed specifically for animal experimental purposes. The hyperbaric oxygen chamber was designed to meet a number of criteria regarding safety and ease of use. The hyperbaric oxygen chamber conforms to 97/23/EC (Pressure Equipment Directive), Conformity Assessment Module G Product Group 1. It provides easy access, and can be run in manual mode, semi-automatic mode and full-automatic mode. Sensors for pressure level, oxygen level, temperature, humidity and carbon dioxide level allow full control. This state-of-the-art hyperbaric oxygen chamber for animal experimental purposes permits the investigation of the biological mechanisms through which hyperbaric oxygen therapy acts at a fundamental level.

Animal ethics in Cardiovascular Research

More than 100 million animals are used for research purposes around the world annually, most of them in Europe and the USA. Rats, mice, and other rodents make up 85–90% of all research animals. 1–1.5% correspond to dogs and cats and less than 1% is represented by non-human primates. Despite a significant decrease in the numbers of animals used in biomedical research and testing in the last two decades, in part due to more restrictive policies and development of alternative methods ( in vitro tests, availability of human cells and tissues, computational models, and others), the magnitude of these numbers, together with an increasing social sensitivity towards animal pain and suffering, makes the debate about the use of animals in experimental research an important and indisputable ethical issue among scientists, scientific journals, regulatory committees, and local, national, and international agencies. The use of animals in research has historically represented an irresolvable moral dilemma between the value of the expected benefits and the suffering, pain, confinement, and death to which it subjects sensitive living beings. Scientists using animals have to work on the assumption that the resulting increase in knowledge or improvement in human health outweighs all these associated costs, which in some cases has been arguable. The Cartesian view that prevailed up to the second half of the last century tried to circumvent this ethical conflict by depriving the animals of any sensitive and intellectual capacity. This view of animal status was extended throughout Europe for centuries due to the important impact the French philosopher Descartes (1596–1650) had on our concept about animal consciousness. Non-human living beings were considered to lack cognitive abilities and could not feel pain or distress but only automatic reactions in response to external stimuli. This doctrine had a pernicious effect on animal welfare and gave rise …

Specific biomarkers of receptors, pathways of inhibition and targeted therapies: clinical applications

A deeper understanding of the role of specific genes, proteins, pathways and networks in health and disease, coupled with the development of technologies to assay these molecules and pathways in patients, promises to revolutionise the practice of clinical medicine. In particular, the discovery and development of novel drugs targeted to disease-specific alterations could benefit significantly from non-invasive imaging techniques assessing the dynamics of specific disease-related parameters. Here we review the application of imaging biomarkers in the management of patients with brain tumours, especially malignant glioma. This first part of the review focuses on imaging biomarkers of general biochemical and physiological processes related to tumour growth such as energy, protein, DNA and membrane metabolism, vascular function, hypoxia and cell death. These imaging biomarkers are an integral part of current clinical practice in the management of primary brain tumours. The second article of the review discusses the use of imaging biomarkers of specific disease-related molecular genetic alterations such as apoptosis, angiogenesis, cell membrane receptors and signalling pathways. Current applications of these biomarkers are mostly confined to experimental small animal research to develop and validate these novel imaging strategies with future extrapolation in the clinical setting as the primary objective.

“Personality” in laboratory mice used for biomedical research: A way of understanding variability?

The mouse, including countless lines of transgenic and knockout mice, has become the most prominent model organism in biomedical research. Behavioral characterization is often conducted in batteries of short tests on locomotion, anxiety, learning and memory, etc. In such tests, any individual differences within groups are usually considered to be disturbing variance. In order to reduce variance in experimental animal research enormous efforts of standardization have been made. While a substantial reduction of variability has been reached compared to the earlier years of experimental animal studies a considerable amount of inter-individual differences still seems to escape standardization. This effect is demonstrated and evaluated by re-analyzing data from two experiments conducted in our laboratory with inbred mice. Interestingly, behavioral patterns of individual animals seem to be correlated across context and time. In evolutionary biology, "animal personalities" have been discussed recently to comprise such stable patterns. We argue here, that nonrandom behavioral correlations across contexts and time might underlie the variability commonly found in biomedical mouse studies.

Cardioprotection of Insulin-Like Growth Factor-1 During Reperfusion Therapy

In the setting of acute myocardial infarction, reperfusion of ischemic myocardium carried out early after coronary occlusion can salvage reversibly injured, viable myocardium. Although still controversial, however, reperfusion itself may cause a population of reversibly injured cells to die, a phenomenon termed lethal reperfusion injury .1 A variety of pharmacological agents have been studied to limit ischemia/reperfusion injury as an adjunct to current reperfusion, and some beneficial effects have been demonstrated in experimental animal research, but their clinical applications have not been achieved.2–4 Article see p 327 Among the various agents claimed to have cardioprotective effects is insulin-like growth factor-1 (IGF-1), which is a hormone produced primarily by the liver. Its molecular structure is similar to that of insulin. IGF-1 plays an important role in cellular survival and growth by binding to its specific receptor (IGF1R), which is present on many cell types, including cardiac cells. Activation of IGF1R, a receptor tyrosine kinase, has been demonstrated to attenuate both apoptotic and necrotic cell death induced by ischemia/reperfusion injury through stimulating both the intracellular phosphoinositide 3-kinase/protein kinase B signaling pathway and extracellular signal-regulated kinase 1/2 signaling cascades.5 Buerke at al6 administered IGF-I 1 hour before ischemia in a murine model of 20 minutes of myocardial ischemia followed by 24 hours of reperfusion. IGF-I preserved ischemic/reperfused myocardium through inhibition of polymorphonuclear leukocyte-induced cardiac necrosis and inhibition of reperfusion-induced apoptosis of cardiac myocytes. Davani et al7 subjected isolated murine hearts to 20 minutes of global ischemia followed by 2 hours of reperfusion with either modified Kreb's solution alone or …

Happy People Live Longer: Subjective Well-Being Contributes to Health and Longevity

Seven types of evidence are reviewed that indicate that high subjective well-being (such as life satisfaction, absence of negative emotions, optimism, and positive emotions) causes better health and longevity. For example, prospective longitudinal studies of normal populations provide evidence that various types of subjective well-being such as positive affect predict health and longevity, controlling for health and socioeconomic status at baseline. Combined with experimental human and animal research, as well as naturalistic studies of changes of subjective well-being and physiological processes over time, the case that subjective well-being influences health and longevity in healthy populations is compelling. However, the claim that subjective well-being lengthens the lives of those with certain diseases such as cancer remains controversial. Positive feelings predict longevity and health beyond negative feelings. However, intensely aroused or manic positive affect may be detrimental to health. Issues such as causality, effect size, types of subjective well-being, and statistical controls are discussed.

The neuropathological contribution of prenatal inflammation to schizophrenia

Evaluation of: Ellman LM, Deicken RF, Vinogradov S et al. Structural brain alterations in schizophrenia following fetal exposure to the inflammatory cytokine interleukin-8. Schizophr. Res. 121, 46–54 (2010).Maternal infection and/or inflammation during pregnancy has been repeatedly shown to elevate the risk of schizophrenia and related psychotic disorders in offspring. However, the relative contribution of such immune-related prenatal insults to specific neuropathological outcomes in schizophrenia remains essentially unknown. The study by Ellman et al. is the first to explore whether prenatal levels of the proinflammatory cytokine, IL-8, are associated with the volume of specific brain regions in patients diagnosed with schizophrenia spectrum disorder. The authors provide new data to support a significant association between higher prenatal IL-8 levels in the second/third trimester of pregnancy and greater ventricular cerebrospinal fluid volume in adult schizophrenia spectrum cases. In addition, the results provide evidence for a significant relationship between higher prenatal IL-8 levels and lower volumes in the left entorhinal cortex and right posterior cingulate. The findings provided by Ellman et al., together with the results obtained in other epidemiological studies and experimental animal research, should encourage basic researchers and clinicians alike to make efforts towards the investigation of maternal immunomodulatory interventions that may help alleviate abnormal brain development and long-term psychotic illness in offspring.

Developmental Vitamin D Deficiency and Risk of Schizophrenia: A 10-Year Update

There is an urgent need to generate and test candidate risk factors that may explain gradients in the incidence of schizophrenia. Based on clues from epidemiology, we proposed that developmental vitamin D deficiency may contribute to the risk of developing schizophrenia. This hypothesis may explain diverse epidemiological findings including season of birth, the latitude gradients in incidence and prevalence, the increased risk in dark-skinned migrants to certain countries, and the urban-rural gradient. Animal experiments demonstrate that transient prenatal hypovitaminosis D is associated with persisting changes in brain structure and function, including convergent evidence of altered dopaminergic function. A recent case-control study based on neonatal blood samples identified a significant association between neonatal vitamin D status and risk of schizophrenia. This article provides a concise summary of the epidemiological and animal experimental research that has explored this hypothesis.

The Developing Brain: A largely overlooked health endpoint in risk assessments for synthetic chemical substances

A large body of experimental animal research on the neurotoxic effects of certain environmental chemicals provides evidence of a cascade of neurobehavioural effects including learning deficits, hyperactivity, anxiety, depression, lack of motivation, increased aggressiveness, altered maternal care and bonding, and an over‐reaction to small stressors. For a number of years, scientific meetings and reports have recommended that assessing risk in chemical safety evaluations should include tests for developmental neurotoxicity (DNT) to uncover the potential of chemical substances to affect the developing brain. However, DNT testing has been required only in rare circumstances, and for some classes of neurotoxic pesticides. The role of developmental neurotoxicants as causative or contributory factors in children’s learning and behavioural disorders warrants more attention in both research and policy.

The Current Status of Animal Use and Alternatives in Korean Veterinary Medical Schools

Two new Korean laws regulating animal welfare and the humane use of animals in science came into effect in 2008 and 2009. Both these laws impose ethical committee review prior to the performance of animal experiments in research, testing and education. This study briefly summarises the new Korean laws, and investigates the current status regarding the numbers of animals used, the alternatives to animals which are used, the curricula relating to the humane use of animals, and ethical review practices in Korean veterinary education. Approximately 4,845 animals, representing 20 different species, were used in veterinary medical education in Korea in 2007. Korea has begun to introduce formal courses on animal welfare for the humane treatment of animals used in experiments, and an ethical protocol review system prior to animal use in education. Korea is moving toward better animal welfare, by incorporating practices consistent with international standards. The information presented represents the first such data gathered in Korea, which should prove useful for monitoring the implementation of replacement, reduction, and refinement measures in animal use for education purposes.

Randomized Intervention Studies in Human Polycystic Kidney and Liver Disease

Autosomal dominant polycystic disease (ADPKD) is the most common life-threatening genetic disease in the United States. ADPKD is more common than Huntington disease, hemophilia, cystic fibrosis, sickle cell disease, Down syndrome, and myotonic dystrophy combined .1 The increase in kidney cyst volume over time correlates with hypertension and progressive renal dysfunction, yet, to date, there is no established intervention to slow or prevent the renal cyst growth. Increased proliferation accompanies renal cyst growth, and recent research in nonorthologous experimental animals and patients with ADPKD suggests aberrant activation of the Ser/Thr kinase mammalian target of rapamycin (mTOR), which modulates cell growth and proliferation.2–5 Recently, rapamycin was shown to have dramatic effects on liver cyst volumes in humans6 and in an orthologous rodent model with conditional inactivation of the PKD1 gene.7 The results demonstrate that mTOR inhibition with rapamycin decreases cyst growth, fibrosis, and proliferation and improves renal function. Of interest, there was regression of cyst burden, which may have been due to increased apoptosis among the cystic epithelium. Perico et al. 8 in this issue of JASN performed a renal safety and efficacy study to examine further this potential pathogenetic ADPKD pathway in humans. This randomized crossover study examined the effect of sirolimus (rapamycin) over 6 months on progression of ADPKD compared with conventional therapy. The main efficacy variable was the effect on total kidney volume, but renal cyst and parenchymal volumes were also measured. The effect of sirolimus on GFR was also examined relative to kidney volume and structure …

Three Aspects of Darwin's Ethical Practice

Previous articleNext article No AccessGrasmere Summer Conference 2009Three Aspects of Darwin's Ethical PracticePeter W. GrahamPeter W. Graham Search for more articles by this author PDFPDF PLUS Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinkedInRedditEmail SectionsMoreDetailsFiguresReferencesCited by The Wordsworth Circle Volume 41, Number 1Winter 2010Grasmere Summer Conference 2009 Article DOIhttps://doi.org/10.1086/TWC24043682 Views: 10Total views on this site © 2010 by The University of Chicago. All rights reserved.PDF download Crossref reports no articles citing this article.

Picture Perception in Birds: Perspective from Primatologists.

In their target article, Weisman and Spetch (2010) question the validity of pictures to present real things to birds, mostly because pictures are primarily made for the human eye, and not for the eye of birds with different functional properties. Here, we argue that this issue of picture validity is similarly critical for primatologists, even when they study the higher nonhuman primates with a more similar visual system, and emphasize cognitive limitations in referential abilities that may be an important source of differences in picture processing modes between human and animals. In their target paper, Weisman and Spetch (2010) review the literature on picture perception in birds and adequately question the validity of pictorial stimuli (photographs, digitized pictures, and movies) to present real objects or scenes to these animals. In their review, they raise the important issue that with the current technology, pictures are primarily designed to be perceived by human eyes and might therefore, not be well adapted to birds possessing a different visual system. They document the risk for anthropocentric errors in experimental research using pictures with birds, and call for new empirical evidence showing a correspondence between the perception of pictures and the perception of objects in birds. Despite the widespread use of pictures in animal psychology, investigators have rarely questioned the realistic nature of pictures and their ecological validity. In that respect, Weisman and Spetch's paper (2010) is among the very few to explicitly do so (see also Cabe, 1976, Fagot 2000). They must be applauded for bringing back this important issue to the attention of the field. In general, we are supportive of their position that a greater care must be given to the use of pictures in experimental animal research. However, consideration of the primate literature suggests that