Genetic variants of chemokine receptor CCR7 in patients with systemic lupus erythematosus, Sjogren's syndrome and systemic sclerosis

<p>Abstract</p> <p>Background</p> <p>The chemokine receptor CCR7 is a key organizer of the immune system. Gene targeting in mice revealed that Ccr7-deficient animals are severely impaired in the induction of central and peripheral tolerance. Due to these defects, Ccr7-deficient mice spontaneously develop multi-organ autoimmunity showing symptoms similar to those observed in humans suffering from connective tissue autoimmune diseases. However, it is unknown whether mutations of <it>CCR7 </it>are linked to autoimmunity in humans.</p> <p>Results</p> <p>DNA samples were collected from 160 patients suffering from connective tissue autoimmune disease (Sjogren's syndrome, n = 40; systemic lupus erythematosus, SLE, n = 20 and systemic sclerosis, n = 100) and 40 health subjects (n = 40). All participants in this study were of German descent. Samples were screened for single nucleotide polymorphisms (SNP) by sequencing the coding region of the <it>CCR7 </it>gene as well asthe exon flaking intron sites and parts of the regions encoding for the 5'- and 3'-UTR. <it>CCR7 </it>variants were rare. We identified six different sequence variants, which occurred in heterozygosis. The identified SNP were observed at position -60 C/T (observed 1x), +6,476 A/G (7x), +6,555 C/T (15x), +6,560 C/T (6x), +10,440 A/G (3x) and +11,475 C/A (1x). Four of these variants (+6,476 A/G, +6,555 C/T, +6,560 C/T and +10,440 A/G) display allelic frequencies between 1% and 5 % and were present in both patients and control groups. The variants +6,476 A/G, +6,555 C/T, +6,560 C/T are located in the intron 2, while the +10,440 A/G variant corresponds to a silent mutation in exon 3. The variants -60 C/T and +11,475 C/A which are located at the 5'-UTR and 3-UTR respectively, display allelic frequencies below 1%. No correlation between these variants and the autoimmune diseases investigated could be observed. However, reporter gene expression assay demonstrated that the mutation at the -60 C/T position in homozygosis leads to reduced luciferase activity.</p> <p>Conclusion</p> <p>These results suggest that variants of <it>CCR7 </it>gene occur at an extremely low frequency in the German population and that neither Sjogren's syndrome, systemic lupus erythematosus, nor systemic sclerosis are associated with these variants. Nevertheless, the decreased luciferase activity observed in cells transfected with the promoter region bearing the -60 C/T mutation suggests that this <it>CCR7 </it>variant could potentially lead to increased susceptibility to autoimmunity.</p

Ethical Issues Associated with the Use of Animal Experimentation in Behavioral Neuroscience Research

This chapter briefly explores whether there are distinct characteristics in the field of Behavioral Neuroscience that demand specific ethical reflection. We argue that although the ethical issues in animal-based Behavioral Neuroscience are not necessarily distinct from those in other research disciplines using animal experimentation, this field of endeavor makes a number of specific, ethically relevant, questions more explicit and, as a result, may expose to discussion a series of ethical issues that have relevance beyond this field of science. We suggest that innovative research, by its very definition, demands out-of-the-box thinking. At the same time, standardization of animal models and test procedures for the sake of comparability across experiments inhibits the potential and willingness to leave well-established tracks of thinking, and leaves us wondering how open minded research is and whether it is the researcher’s established perspective that drives the research rather than the research that drives the researcher’s perspective. The chapter finishes by introducing subsequent chapters of this book volume on Ethical Issues in Behavioral Neuroscience

Animal Welfare Considerations: Should Context Matter?

Much consideration of animal welfare issues, as well as the bulk of welfare legislation, is closely related to context, with clear distinction between defined responsibilities towards farm-, laboratory-, companion- and wild animals. It has however been recognised that such distinction leads to inconsistencies, in that it might lead to different welfare management practices for the same rat (for example) as a pet, laboratory animal or pest. We suggest that if legal provisions recognize animals as having moral status, then that implies certain responsibilities towards the animal that cannot be context-dependent. Although we argue the moral responsibility to consider an animal’s welfare is context-independent, we suggest that the requirement to take action (and the form of that action) may alter with context. From a biological perspective, we argue that there exists an obligation to take action to address potential welfare problems only when a challenge exceeds the animal’s adaptive capacity, such that it cannot adequately respond by changes of its behaviour. The obligation to intervene further depends on whether or not there are practical (and economically feasible) options available for mitigation. Beyond this, additional constraints may be posed by anthropocentric aims or objectives, where any suffering may be adjudged avoidable (in theory), but necessary (because of specific human interests). Finally, whatever may be determined necessary or unavoidable by the actual manager or agent himself/herself, such judgement is inevitably subjective and possibly subject to self-interest. Thus any decision must be further evaluated within the context of what is considered by a wider society to be (morally) acceptable or unacceptable

Animal Welfare Considerations: Should Context Matter?

Much consideration of animal welfare issues, as well as the bulk of welfare legislation, is closely related to context, with clear distinction between defined responsibilities towards farm-, laboratory-, companion- and wild animals. It has however been recognised that such distinction leads to inconsistencies, in that it might lead to different welfare management practices for the same rat (for example) as a pet, laboratory animal or pest. We suggest that if legal provisions recognize animals as having moral status, then that implies certain responsibilities towards the animal that cannot be context-dependent. Although we argue the moral responsibility to consider an animal’s welfare is context-independent, we suggest that the requirement to take action (and the form of that action) may alter with context. From a biological perspective, we argue that there exists an obligation to take action to address potential welfare problems only when a challenge exceeds the animal’s adaptive capacity, such that it cannot adequately respond by changes of its behaviour. The obligation to intervene further depends on whether or not there are practical (and economically feasible) options available for mitigation. Beyond this, additional constraints may be posed by anthropocentric aims or objectives, where any suffering may be adjudged avoidable (in theory), but necessary (because of specific human interests). Finally, whatever may be determined necessary or unavoidable by the actual manager or agent himself/herself, such judgement is inevitably subjective and possibly subject to self-interest. Thus any decision must be further evaluated within the context of what is considered by a wider society to be (morally) acceptable or unacceptable

Sex Differences in Physiological Acclimatization after Transfer in Wistar Rats

Simple Summary: This study in laboratory rodents shows a sex specific effect of breeder to research facility transfer on several physiological parameters, such as heart rate and blood pressure. We recommend at least 8 days of acclimatization time after transfer in male rats and at least two weeks in female rats, before using these animals in research. Abstract: Most laboratory animals used in research are vendor-bred and transferred to research facilities. Transfer procedures might have considerable and unintended effects on research results. In the present study we compared physiological and behavioral parameters before and after external and internal transfer, as well as between transferred and non-transferred Wistar rats. The impact of both external and internal transfer on body weight, plasma corticosterone levels, heart rate, blood pressure, and locomotor activity was studied in both male and female Wistar rats, taking into account the sex differences in stress responsivity. External transfer was found to decrease body weight, increase plasma corticosterone, increase activity, increase heart rate in female rats, but decrease heart rate in male rats. Parameters showed differences between the sexes and light phases. This study shows that acclimatization after transfer is sex-specific and researchers should take the sex into consideration when determining the acclimatization period. It is recommended to allow for acclimatization of at least 8 days in males and two weeks in females after external transfer and timely (2 days before starting experiments) transfer the animals internally to the testing room

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal's behavioral profile can be investigated in one single test. The apparatus consists of a box, representing the 'protected' area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the 'unprotected' area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related-and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals' adaptive capacities. Due to testing order effects in a behavioral test battery, naive animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress