Mice and minipigs with compromised expression of the Alzheimer's disease gene SORL1 show cerebral metabolic disturbances on hyperpolarized [1-13C]pyruvate and sodium MRI.

The sortilin-related receptor 1 (SORL1) gene, encoding the cellular endosomal sorting-related receptor with A-type repeats (SORLA), is now established as a causal gene for Alzheimer's disease. As the latest addition to the list of causal genes, the pathophysiological effects and biomarker potential of SORL1 variants remain relatively undiscovered. Metabolic dysfunction is, however, well described in patients with Alzheimer's disease and is used as an imaging biomarker in clinical diagnosis settings. To understand the metabolic consequences of loss-of-function SORL1 mutations, we applied two metabolic MRI technologies, sodium (23Na) MRI and MRI with hyperpolarized [1-13C]pyruvate, in minipigs and mice with compromised expression of SORL1. At the age analysed here, both animal models display no conventional imaging evidence of neurodegeneration but show biochemical signs of elevated amyloid production, thus representing the early preclinical disease. With hyperpolarized MRI, the exchange from [1-13C]pyruvate to [1-13C]lactate and 13C-bicarbonate was decreased by 32 and 23%, respectively, in the cerebrum of SORL1-haploinsufficient minipigs. A robust 11% decrease in the sodium content was observed with 23Na-MRI in the same minipigs. Comparably, the brain sodium concentration gradually decreased from control to SORL1 haploinsufficient (-11%) to SORL1 knockout mice (-23%), suggesting a gene dose dependence in the metabolic dysfunction. The present study highlights that metabolic MRI technologies are sensitive to the functional, metabolic consequences of Alzheimer's disease and Alzheimer's disease-linked genotypes. Further, the study suggests a potential avenue of research into the mechanisms of metabolic alterations by SORL1 mutations and their potential role in neurodegeneration.

Nuclear Medicine Preclinical Research: The Role of Cell Cultures.

Cell lines are essential in biomedical research due to their adaptability and precise simulation of physiological and pathophysiological conditions. Cell culture techniques have greatly advanced our understanding of biology in various fields and are widely regarded as a reliable and durable tool. Their diverse applications make them indispensable in scientific research. Radiation-emitting compounds are commonly used in cell culture research to investigate biological processes. Radiolabeled compounds are utilized to study cell function, metabolism, molecular markers, receptor density, drug binding and kinetics, as well as to analyze the direct interaction of radiotracers with target organ cells. This allows for the examination of normal physiology and disease states. The In Vitro system simplifies the study and filters out nonspecific signals from the In Vivo environment, leading to more specific results. Moreover, cell cultures offer ethical advantages when evaluating new tracers and drugs in preclinical studies. While cell experiments cannot entirely replace animal experiments, they reduce the need for live animals in experimentation.

Preclinical Imaging Studies: Protocols, Preparation, Anesthesia, and Animal Care.

Today preclinical PET imaging connects laboratory research with clinical applications. Here PET clearly bridges the gap, as nearly identical imaging protocols can be applied to both animal and humans. However, some hurdles exist and researchers must be careful, partly because the animals are usually anesthetized during the scans, while human volunteers are awake. This review is based on our own experiences of some of the most important pitfalls and how to overcome them. This includes how studies should be designed, how to select the right anesthesia and monitoring. The choice of anesthesia is quite crucial, as it may have a greater influence on the results than the effect of the tested procedures. Monitoring is necessary, as the animals cannot fully maintain homeostasis during anesthesia, and reliable results are dependent on a stable physiology. Additionally, it is important to note that rodents, in particular, are prone to rapidly becoming hypothermic. Thus, the selection of an appropriate anesthetic and monitoring protocol is crucial for both obtaining accurate results and ensuring animal welfare. Prior to imaging, catheters for tracer administration and, if necessary, blood sampling should be implanted. The administration of tracers should be done in a manner that minimizes interference with the scans, and the same applies to any serial blood sampling. The limited blood volume and organ size of rodents should also be taken into consideration when planning experiments. Finally, if the animal needs to be awakened after the scan, proper care must be taken to ensure their welfare.

Effect of atropine and propofol on the minimum anaesthetic concentration of isoflurane in the freshwater turtle Trachemys scripta (yellow-bellied slider).

OBJECTIVE: To determine if the administration of atropine would reduce the measured minimum anaesthetic concentration of isoflurane (MACisoflurane) in freshwater turtles - the yellow-bellied slider (Trachemys scripta scripta). STUDY DESIGN: Paired, blinded, randomized, prospective studies of 1) the effect of atropine in isoflurane anaesthetized freshwater turtles (T. scripta scripta) and 2) the effect of atropine in yellow-bellied sliders in which anaesthesia was induced with propofol and maintained with isoflurane. ANIMALS: T. scripta scripta (n = 8), female, adult. METHODS: Atropine (2 mg kg-1) or an isovolumetric control injection of saline was administered intraperitoneally 15 minutes prior to induction of anaesthesia with isoflurane. Individual MACisoflurane was then determined by end-tidal gas analysis in a bracketing design by an experimenter blinded to the administered drug, with a 2 week washout period. The experiment was repeated, with atropine (2 mg kg-1) or saline administered intravascularly in combination with propofol for anaesthetic induction. Linear mixed modelling was used to determine the effects of atropine and propofol on the individual MACisoflurane. Data are presented as mean ± standard deviation. RESULTS: Premedication with atropine significantly reduced MACisoflurane (p = 0.0039). In isoflurane-induced T. scripta scripta, MACisoflurane decreased from 4.2 ± 0.4% to 3.3 ± 0.8% when atropine had been administered. Propofol as an induction agent had a MAC-sparing effect (p < 0.001) such that MACisoflurane following propofol and a control injection of saline was 2.3 ± 1.0%, which decreased further to 1.5 ± 0.8% when propofol was combined with atropine. CONCLUSIONS AND CLINICAL RELEVANCE: Atropine, presumably by inhibiting parasympathetically mediated pulmonary artery constriction, decreases right-to-left cardiac shunting and the MACisoflurane in yellow-bellied sliders, and thereby may facilitate control of inhalant anaesthesia. Propofol can be used for induction of anaesthesia and reduces the required concentration of inhaled anaesthesia assessed 1.5 hours following induction.

The CRISPR/Cas9 Minipig-A Transgenic Minipig to Produce Specific Mutations in Designated Tissues.

The generation of large transgenic animals is impeded by complex cloning, long maturation and gastrulation times. An introduction of multiple gene alterations increases the complexity. We have cloned a transgenic Cas9 minipig to introduce multiple mutations by CRISPR in somatic cells. Transgenic Cas9 pigs were generated by somatic cell nuclear transfer and were backcrossed to Göttingen Minipigs for two generations. Cas9 expression was controlled by FlpO-mediated recombination and was visualized by translation from red to yellow fluorescent protein. In vitro analyses in primary fibroblasts, keratinocytes and lung epithelial cells confirmed the genetic alterations executed by the viral delivery of single guide RNAs (sgRNA) to the target cells. Moreover, multiple gene alterations could be introduced simultaneously in a cell by viral delivery of sgRNAs. Cells with loss of TP53, PTEN and gain-of-function mutation in KRASG12D showed increased proliferation, confirming a transformation of the primary cells. An in vivo activation of Cas9 expression could be induced by viral delivery to the skin. Overall, we have generated a minipig with conditional expression of Cas9, where multiple gene alterations can be introduced to somatic cells by viral delivery of sgRNA. The development of a transgenic Cas9 minipig facilitates the creation of complex pre-clinical models for cancer research.

Set sustainable goals for the Arctic gateway coordinated international governance is required to resist yet another tipping point.

Global warming is reducing the Arctic sea-ice and causing energetic stress to marine key predatory species such as polar bears and narwhals contributing to the ongoing pollution already threatening the biodiversity and indigenous people of the vulnerable region. Now, the opening of the Arctic gateway and in particular the increase in shipping activities causes further stress to marine mammals in the region. These shipping activities are foreseen to happen in the Northwest and Northeast Passage, Northern Sea Route and Transpolar Sea Route in the Arctic Ocean, which could be yet another step towards a crucial tipping point destabilizing global climate, including weathering systems and sea-level rise. This calls for international governance through the establishment of Arctic International National Parks and more Marine Protected Areas through the Arctic Council and UN's Law of the Sea to ensure sustainable use of the Arctic Ocean and adjacent waters.

A recent global review of hazardous chlorpyrifos pesticide in fruit and vegetables: Prevalence, remediation and actions needed.

Pollution with pesticides is a widespread global problem and biomonitoring of the environment and human populations is necessary to assess potential harmful biological effects. One of the pesticides that are showing up in vegetables and fruit is chlorpyrifos (CPS). CPS is a nerve-poisoning organophosphorus insecticide, which is in up to 1/3 of all conventionally produced citrus fruits. Our review shows that CPS is a hazardous material that poses risks to human health and also pollutes the environment. There is numerous risk assessment of CPS reported, however, the assessment is easily affected by factors such as climate change, exposure period and CPS concentration. Therefore, rigorous update of the hazardous level of CPS is needed to determine the threshold level safe for humans and animals. There is a need for remediation using for example photoreactive nanoparticle methods and microbial degeneration possessing high degradation efficiency (73-97%). In addition, stringent biomonitoring of food, environment and human exposure should occur to avoid exposure to chemicals via citrus fruits and vegetables. This is necessary to assess health risks and socioeconomic impacts which also require collaboration between private and public sectors to facilitate the growth, sale and manufacturing of biopesticides.

Environmental management of two of the world's most endangered marine and terrestrial predators: Vaquita and cheetah.

Two of the world most endangered marine and terrestrial species are at the brink of extinction. The vaquita (Phocoena sinus) is the smallest existing cetacean and the population has declined to barely 22 individuals now remaining in Mexico's Gulf of California. With the ongoing decline, it is likely to go extinct within few years. The primary threat to this species has been mortality as a result of by-catch from gillnet fishing as well as environmental toxic chemicals and disturbance. This has called for the need to establish a National Park within the Gulf of California to expand essential habitat and provide the critical ecosystem protection for vaquita to thrive and multiply, given that proper conservation enforcement and management of the park are accomplished. In the terrestrial environment, the cheetah (Acinonyx jubatus) is reduced to a low number worldwide with the Iran subpopulation currently listed as Critically Endangered and the Indian subpopulation already extinct. There is a need for conservation efforts due to habitat loss, but also an indication of the conspicuous threat of illegal trade and trafficking from Africa and Arab countries in the Middle East. Funds have also been set up to provide refuges for the cheetah by working directly with farmers and landowners, which is a critical movement in adaptive management. These are the potential options for the preservation and possibly the expansion of the overall vaquita and cheetah populations.

First predatory journals, now conferences: The need to establish lists of fake conferences.

Science of the Total Environment recently discussed how open access and predatory journals affect the flow of scientific knowledge in an unfortunate way. Now, South Korea's Ministry of Education is intervening to establish a system that will help its researchers avoid the growing global number of fake conferences of low academic and scientific merit. Here, we discuss solutions to this problem with respect to what is needed. Particularly, a list similar to that of Beall's for predatory conferences, without restricting researchers' academic freedom.

Response to comments on "Factors affecting global flow of scientific knowledge in environmental sciences" by Pourret et al.

We have read the response to our Discussion about Open Access and predatory journals entitle Factors affecting global flow of scientific knowledge in environmental sciences. The purpose of a Discussion in STOTEN is to inspire for debating and we therefore thank Pourret et al. (2020) for their response to our Discussion about Open Access, Plan S and Open Access Predatory Journals - three topics that concern us as researchers (Sonne et al., 2020).

Factors affecting global flow of scientific knowledge in environmental sciences.

Here we present our view on the current Open Access debate, predatory journals and the on-going publication and promotion strategy of some countries and research institutions. We urge the world's researchers, journals and grant holders in collaboration to carefully consider how best to ensure continuous high-quality scientific publications in the future in a way so that limited funding results in important data and information being unpublished.

The ongoing cut-down of the Amazon rainforest threatens the climate and requires global tree planting projects: A short review.

The Amazon rainforest has sustained human existence for more than 10,000 years. Part of this has been the way that the forest controls regional climate including precipitation important for the ecosystem as well as agroforestry and farming. In addition, the Amazon also affects the global weather systems, so cutting down the rainforest significantly increases the effects of climate change, threatening the world's biodiversity and causing local desertification and soil erosion. The current fire activities and deforestation in the Amazon rainforest therefore have consequences for global sustainability. In the light of this, the current decisions made in Brazil regarding an increase in Amazon deforestation require policy changes if the global ecosystems and biodiversity are not to be set to collapse. There is only one way to move forward and that is to increase efforts in sustainable development of the region including limitation in deforestation and to continuously measure and monitor the development. The G7 countries have offered Brazil financial support for at least 20 million euros for fighting the forest fires but the president denies receiving such financial support and says that it is more relevant to raise new forests in Europe. In fact, this is exactly what is happening in Denmark and China in order to reduce climate change. Such activities should be global and include South America, Europe, Africa and Asia where deforestation is important issue. Forest restoration reduces climate change, desertification, and preserves both the regional tropical and global environment if the wood is not burned at a later stage but instead used in e.g. roads as filling material. Changes are therefore needed through improved international understanding and agreements to better avoid the global climate changes, from cutting down the precious rainforest before it is too late as rainforest cannot be re-planted.

Sucrose intake lowers µ-opioid and dopamine D2/3 receptor availability in porcine brain.

Excessive sucrose consumption elicits addiction-like craving that may underpin the obesity epidemic. Opioids and dopamine mediate the rewarding effects of drugs of abuse, and of natural rewards from stimuli such as palatable food. We investigated the effects of sucrose using PET imaging with [11C]carfentanil (µ-opioid receptor agonist) and [11C]raclopride (dopamine D2/3 receptor antagonist) in seven female anesthetized Göttingen minipigs. We then gave minipigs access to sucrose solution for one hour on 12 consecutive days and performed imaging again 24 hours after the final sucrose access. In a smaller sample of five minipigs, we performed an additional [11C]carfentanil PET session after the first sucrose exposure. We calculated voxel-wise binding potentials (BPND) using the cerebellum as a region of non-displaceable binding, analyzed differences with statistical non-parametric mapping, and performed a regional analysis. After 12 days of sucrose access, BPND of both tracers had declined significantly in striatum, nucleus accumbens, thalamus, amygdala, cingulate cortex and prefrontal cortex, consistent with down-regulation of receptor densities. After a single exposure to sucrose, we found decreased binding of [11C]carfentanil in nucleus accumbens and cingulate cortex, consistent with opioid release. The lower availability of opioid and dopamine receptors may explain the addictive potential associated with intake of sucrose.

Kinetic Modelling of [68Ga]Ga-DOTA-Siglec-9 in Porcine Osteomyelitis and Soft Tissue Infections.

BACKGROUND: [68Ga]Ga-DOTA-Siglec-9 is a positron emission tomography (PET) radioligand for vascular adhesion protein 1 (VAP-1), a protein involved in leukocyte trafficking. The tracer facilitates the imaging of inflammation and infection. Here, we studied the pharmacokinetic modelling of [68Ga]Ga-DOTA-Siglec-9 in osteomyelitis and soft tissue infections in pigs. METHODS: Eight pigs with osteomyelitis and soft tissue infections in the right hind limb were dynamically PET scanned for 60 min along with arterial blood sampling. The fraction of radioactivity in the blood accounted for by the parent tracer was evaluated with radio-high-performance liquid chromatography. One- and two-tissue compartment models were used for pharmacokinetic evaluation. Post-mortem soft tissue samples from one pig were analysed with anti-VAP-1 immunofluorescence. In each analysis, the animal's non-infected left hind limb was used as a control. RESULTS: Tracer uptake was elevated in soft tissue infections but remained low in osteomyelitis. The kinetics of [68Ga]Ga-DOTA-Siglec-9 followed a reversible 2-tissue compartment model. The tracer metabolized quickly; however, taking this into account, produced more ambiguous results. Infected soft tissue samples showed endothelial cell surface expression of the Siglec-9 receptor VAP-1. CONCLUSION: The kinetics of [68Ga]Ga-DOTA-Siglec-9 uptake in porcine soft tissue infections are best described by the 2-tissue compartment model.

Aviation, melting sea-ice and polar bears.

On 11 May 2019, the Mauna Loa, Hawaii, Earth System Research Laboratory reported the highest CO2 concentration in human meteorological history. Continuing CO2 rise will devastate ecosystems, and ice dependent species like polar bears ultimately will disappear. Commercial aviation is presently a relatively small CO2 contributor, but this CO2 intensive mode of transportation is projected to increase greatly. Scientists and conservationists are often among the most frequent of flyers, despite their recognition that emissions must be reduced. Here we illustrate the carbon footprint of air travel in terms of its impact on the sea ice habitat necessary for polar bear persistence, and suggest our colleagues reduce their air travel where-ever possible. Each metric ton of CO2 emitted melts ~3 m2 of arctic summer sea ice, and current air travel melts over 5000 m2 each year. Each scientist making the short flight from Copenhagen to Oslo to join an IUCN polar bear meeting will melt ~1 m2 of Arctic summer sea-ice. Annually hundreds of scientists and conservationists make frequent flights of much greater distances for AMAP, CAFF, IUCN, and other conservation related meetings. Much of this travel could be avoided with better planning and employing internet linkages for remote participation. When air travel, such as for necessary fieldwork, cannot be easily substituted by Web linkage, we all should search for routes and carriers allowing the lowest CO2 emissions. We encourage all of our colleagues to join 'No Fly Climate Sci' to show their commitment to CO2 reduction and learn more about doing so. As scientists, if we are serious about preserving polar bears and their Arctic sea ice habitat, we need to walk the talk and show an example for the rest of society by significantly reducing our air travel.