Retrospective review of anesthetic and analgesic regimens used in animal research proposals

Pain has a profound effect on an animal’s wellbeing. In Germany, researchers using animals have been legally required to reduce any possible pain, suffering, distress or lasting harm to an absolute minimum since 1972. To evaluate how these provisions have been implemented in practice, an assessment of refinements to experimental techniques was conducted by retrospectively reviewing 684 surgical interventions described in 506 animal research applications that were sent to the German competent authorities for approval in 2010. This paper focuses on the efficacy of proposed anesthesia and peri- and postoperative analgesia. Postoperative analgesia was not proposed for 30% of surgeries. Following 10% of procedures, animals were to be given pain relieving medication if the investigators decided this was necessary; however, structured assessments to detect pain were absent. Consequences of unalleviated pain and omission of pain assessment techniques are discussed, and some recommendations to improve anesthesia and analgesia are given. The findings of this review highlight the need for improvement, both to fulfil legal requirements and to improve animal welfare. To monitor compliance with animal welfare regulations and ensure good veterinary and scientific practices, education and training need to be intensified. Adherence to the items listed in the PREPARE and ARRIVE guidelines and the Gold Standard Publication Checklist (GSPC) should become legally binding

Lack of evidence for an association of Epstein–Barr virus infection with breast carcinoma

BACKGROUND: Epstein–Barr virus (EBV) is a ubiquitous human γ-herpes virus infecting more than 90% of the population worldwide. EBV is associated with certain malignancies (e.g. Burkitt lymphoma, Hodgkin lymphoma and nasopharyngeal carcinoma). Recent studies have raised the possibility that EBV may also be involved in the pathogenesis of breast carcinoma, the most common carcinoma of females. If substantiated, this finding would have major implications regarding prevention and therapy of the disease. The studies published so far have employed diverse methods, however, and the results have been controversial. METHODS: Using the EBV DNA PCR, EBV DNA in situ hybridisation and in situ hybridisation for the detection of the EBV-encoded RNAs, and using immunohistochemistry for the demonstration of the EBV-encoded nuclear antigen 1, we have studied a series of 59 invasive breast carcinomas for evidence of EBV infection. RESULTS: EBV-encoded RNA-specific in situ hybridisation and EBV-encoded nuclear antigen 1 immunohistochemistry were negative in all cases. Using the PCR, EBV DNA was detected in four out of 59 cases. These cases were further studied by EBV DNA in situ hybridisation, showing an absence of viral DNA from the tumour cells. CONCLUSION: These results indicate that breast carcinoma is not an EBV-associated tumour

Isolation-dependent enhancement of 14C-2-deoxyglucose uptake in the forebrain of zebra finch males

Bischof H-J, Herrmann K. Isolation-dependent enhancement of 14C-2-deoxyglucose uptake in the forebrain of zebra finch males. Behavioral and Neural Biology. 1988;49(3):386-397.In a previous study (H. J. Bischof and K. Herrmann (1986), Behavioral Brain Research, 21, 215-221) we demonstrated that four forebrain areas of the zebra finch male are activated in situations which arouse the animal, for example when the birds are chased around the cage or when they are exposed to a female. These areas, the hyperstriatum accessorium-dorsale (HAD), a part of the medial neo-hyperstriatum (MNH), the lateral neo-hyperstriatum (LNH), and a portion of the caudal archi-neostriatum (ANC), show enhanced 2-[14C]deoxyglucose (2-DG) uptake according to the experimental situation. On the basis of these experiments, we examined whether the activation of the areas is correlated with motor activity and is influenced by different isolation times prior to a 2-DG experiment, where courtship of the male birds is elicited by exposing them to a female zebra finch. For this purpose, we isolated male zebra finches for 1 day, 1 week, or 8 weeks, respectively, before we injected the 2-DG and exposed the birds to a female. During the experiment, besides other activities, the number of song motifs performed by the bird and the frequency of changing perches was recorded. Our experiments demonstrate that there is a weak negative correlation between motor activity and 2-DG uptake, and a positive correlation between isolation time and 2-DG uptake. We suggest that long isolation blocks courtship behavior by some unknown mechanisms, and that the "internal drive" of the animal, which possibly corresponds with the activity of the four forebrain areas, is enhanced by isolation and by the fact that the birds do not perform the consummatory behavior. Our results also demonstrate that the 2-DG method can show up small differences in the internal state of an animal, which cannot easily be detected by behavioral measurements

Symplectic Partitioned Runge‐Kutta Methods for High‐Order Approximation in Linear‐Quadratic Optimal Control of Hamiltonian Systems

Symplectic partitioned Runge-Kutta (SPRK) methods are known to be a good choice in forward simulations of Hamiltonian systems due to their structure-preserving properties. Recent works study the application of SPRK methods to nonlinear and linear-quadratic optimal control problems howing various advantages of these methods compared to standard non-symplectic integration schemes. Now, our focus is on extending the comparison to SPRK and RK methods of higher orders. For linear-quadratic optimal control problems, we consider the discrete-time Riccati feedback as well as the feedforward control implementation. For applications in which computational power or computation time is limited, low sampling rates are of particular interest. Hence we study this case for the n-fold harmonic oscillator

The sensitive period for the morphological effects of monocular deprivation in two nuclei of the tectofugal pathway of zebra finches

Herrmann K, Bischof H-J. The sensitive period for the morphological effects of monocular deprivation in two nuclei of the tectofugal pathway of zebra finches. Brain Research. 1988;451(1-2):43-53.Previous experiments with 2-deoxyglucose (2-DG) suggested the existence of a critical previous termperiodnext term for the effects of monocular deprivation in the nucleus rotundus of zebra finches. The present study concerns the time course of this sensitive period for the morphological effects of monocular deprivation in two areas of the tectofugal visual pathway of zebra finches, the nucleus rotundus of the thalamus and the telencephalic ectostriatum. Cell size and volume changes were measured in birds subjected to 40 days of unilateral eye closure starting at ages spaced regularly throughout the first 70 days of life. The results show that monocular deprivation markedly affects cell size in both areas if the treatment starts at one or 10 days posthatch. The differences between deprived and non-deprived neurons decline monotonically with increasing visual experience prior to deprivation. However, deprivation onset at day 40 again causes as severe effects as early monocular closure. Deprivation as from day 50 or later no longer leads to abnormalities. The measurements of the volume of the nucleus rotundus parallel the cell size measurements, with the exception that the second increase in sensitivity occurs with deprivation onset at day 50 instead of day 40. These data indicate that the time course of the sensitive period for the effects of monocular deprivation may be double-peaked: the sensitivity for external stimuli declines from hatch until day 30, but has another peak at 40–50 days of life. The definite end of the sensitive period, as determined with this method, can therefore be assumed to be at around day 50–60

Animal Experimentation: Working Towards a Paradigm Change

Animal experimentation has been one of the most controversial areas of animal use, mainly due to the intentional harms inflicted upon animals for the sake of hoped-for benefits in humans. Despite this rationale for continued animal experimentation, shortcomings of this practice have become increasingly more apparent and well-documented. However, these limitations are not yet widely known or appreciated, and there is a danger that they may simply be ignored. The 51 experts who have contributed to Animal Experimentation: Working Towards a Paradigm Change critically review current animal use in science, present new and innovative non-animal approaches to address urgent scientific questions, and offer a roadmap towards an animal-free world of science.https://www.wellbeingintlstudiesrepository.org/ebooks/1018/thumbnail.jp

A brief patient-reported outcome instrument for primary care: German translation and validation of the Measure Yourself Medical Outcome Profile (MYMOP)

Background: Measure Yourself Medical Outcome Profile (MYMOP) is a patient-generated outcome instrument capable of measuring effects from a wide range of health care interventions. This paper reports the translation of this instrument into German (MYMOP-D) and the assessment of validity and sensitivity to change for the MYMOP-D. The instrument was piloted in a German primary care context. Methods: The translation process was conducted according to international guidelines. Recruited patients of both general practitioners and non-medical Complementary and Alternative Medicine (CAM) practitioners (“Heilpraktiker”) in the German state of Baden-Wuerttemberg completed a questionnaire comprised of the MYMOP-D and the EQ-5D. Responses were analysed to assess construct validity. For assessing the instrument’s sensitivity to change, patients received the MYMOP-D again after four weeks at which point they were also asked for their subjective views on change of symptoms. Correlation between MYMOP-D and EQ-5D and sensitivity to change as gradient in score change and as standardized response mean (SRM) were calculated. Results: 476 patients from general practices and 91 patients of CAM practitioners were included. Construct validity of the MYMOP-D was given with a correlation of r = .47 with the EQ-5D. Sensitivity to change for subjective change of symptoms could only be analysed for improvement or no change of symptoms, as only 12 patients reported deterioration of symptoms. Results showed the expected smooth gradient with 2.2, 1.3, and 0.5 points of change for large, little improvement and no change, respectively. SRM for MYMOP-D Profile Score was 0.88. Conclusions: The MYMOP-D shows excellent construct validity. It is able to detect changes when symptoms in patients improve or remain unchanged. Deterioration of symptoms could not be evaluated due to too few data. With its brevity and simplicity, it might be an important tool for enhancing patient-centred care in the German health care context

Dissection and optimization of Adeno-associated virus (AAV) DNA family shuffling technology: The journey is the reward

Viral vectors based on Adeno-associated viruses (AAV) have a broad application spectrum including gene therapy and basic research. However, because naturally occurring AAV capsids are rarely sufficiently efficient and/or specific for a given application, techniques were developed to broaden the existing capsid repertoire. A prototype technology is DNA family shuffling where, in a first step, homologous cap genes encoding capsid subunits are fragmented and recombined, yielding a viral library which can then be subjected to selection in order to enrich promising variants. The aim of the present study was to dissect and improve four critical steps along this procedure. Firstly (1), two different methodologies for production of cap gene fragments were compared, resulting in the identification of DNase I based fragmentation as the most robust approach. Interestingly, cap DNA concatamer formation during nested PCR was observed, leading to amendment of the PCR purification protocol. Next (2), we studied the impact of chimerism on the essential assembly-activating protein (AAP) that is encoded in an alternative open reading frame within cap and is recombined as well during DNA family shuffling. Importantly, by performing a battery of complementary experiments, we were able to show that shuffling of AAP is not impairing its function, i.e. the support of particle assembly. Furthermore, no influence on titers was observed for wild-type and most chimeric vector productions, altogether relieving long-standing concerns about a potential rate-limiting role of AAP for AAV vector generation and evolution. Thirdly (3), we established a pioneering in vivo AAV library selection strategy in which, unlike most previously reported schemes, we selected novel capsids in specific cell types within an organ instead of the organ as a whole. Specifically, we were motivated by the facts that liver disease is wide-spread in humans and that hepatic stellate cells (HSC) are known to drive liver fibrosis, thus contributing to disease progression. Alas, tools to genetically manipulate HSCs are limited. Therefore, a library encompassing 10 capsid variants was selected in HSC by AAV injection into mice, HSC isolation and PCR rescue using purified total DNA. Following multiple selection rounds, in vivo bulk validation was performed based on next-generation sequencing. In total, 157 capsid variants were screened in parallel and again, the liver was segregated into the single cell types, i.e. hepatocytes, HSC, Kupffer cells and liver sinusoidal endothelial cells. Notably, this revealed that the selection was successful as hepatocyte-detargeted vectors were identified that showed a strong co-transduction of HSC and Kupffer cells. Intriguingly, we noted differences in vector specificity and efficiency on the DNA versus the RNA level. In order to even further restrict the new vectors to a given cell type, vector cassettes were generated bearing cell-type specific promoters and miRNA binding sites to suppress off-targeting in cells expressing these miRNAs. Testing of these constructs in vitro gave promising results especially for the miRNA-based detargeting strategy. Finally (4), we implemented improvements during the selection and analysis steps, including the use of PacBio/SMRT sequencing technology to monitor AAV sequence enrichments throughout the course of selection. Additionally, we managed to increase the stringency of the PCR rescue of cap genes, by incorporating sample-specific barcodes, i.e., short, unique nucleotide stretches, into the AAV library genomes. By using these barcodes as a primer during sample recovery, we could isolate single libraries out of a complex library mixture, as validated in vitro. In the future, this original strategy could be exploited to track individual libraries in vivo upon injection of a mixture of libraries, which should in turn help to accelerate the identification of top-performing variants for validation studies. In summary, different steps along the powerful methodology of DNA family shuffling were improved advancing future vector development and the lingering concern about AAP impairment upon shuffling was dispersed

Time to abolish the forced swim test in rats for depression research?

This publication was financed by Portuguese national funds within the CFCUL – Centro de Filosofia das Ciência da Universidade de Lisboa, research center strategic project, ref. UIDB/00678/2020, funded by FCT – Fundação para a Ciência e a Tecnologia. TAM is grateful for the partial support by CEAUL (funded by FCT – Fundação para a Ciência e a Tecnologia, Portugal, through the project UIDB/00006/2020).The forced swim test (FST) is a controversial rodent test that has been used for decades, mainly in depression studies. The severity of the procedure makes it ethically questionable and its validity has also been questioned. In this paper we contribute new data to this debate. We identified original research papers related to Major Depressive Disorder (MDD), using rats as models. We compared the citations received by studies that used the FST and by studies that did not, within subsequent human medical papers. The results show that the number of citations received by both groups was very low, but in the papers describing the FST data the median citation number was zero. Citation analysis indicates that the FST is not contributing significantly to the understanding or cure of MDD. We briefly review other approaches that overcome the ethical limitations of the FST, and which might also surpass its efficacy.Publisher PDFPeer reviewe

Does adult ADHD interact with COMT val 158 met genotype to influence working memory performance?

Peer reviewedPostprin