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Optimal conditions, experimentation and drug testing Lab Anim. (IF 5.9) Pub Date : 2024-07-15 Hippokratis Kiaris
Optimal conditions represent the golden standard in experimental biology studies. Yet, the definition of ‘optimal’ is rather vague and optimal conditions are non-existent in nature, which operates at suboptimal conditions. This apparent paradox has implications in both curiosity-driven research that seeks answers to fundamental biological questions and in preclinical research that aims to identify
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A new approach for Drosophila cardiac analysis Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Alexandra Le Bras
Drosophila melanogaster has emerged as a useful model to study cardiovascular diseases, including both developmental abnormalities and adult functional impairments. Despite being much simpler, the Drosophila heart displays several developmental and functional similarities to the vertebrate heart. The fruit fly heart, located ~100 μm below the dorsal surface, has also the advantage of being easily accessed
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zAvatars pass the test Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Alexandra Le Bras
Surgery is the primary form of treatment for colorectal cancer, resulting in cure for approximately 50% of patients. However, recurrence after surgery is a major problem and patients can greatly benefit from post-surgical systemic therapies. However, finding the optimal therapy for a patient can be challenging, leading to unnecessary side effects and loss of valuable time. In a new study, Rita Fior’s
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Pet and lab zebrafish microbes Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Jorge Ferreira
Zebrafish are not only a common research model but also a popular pet fish. Laboratory-bred zebrafish have well-characterized associated microbes. Pet shop zebrafish are frequently used in the laboratory setting. However, given that their associated microbes are still uncharacterized, the ramifications of introducing them to a laboratory environment are unknown. A study in PLOS Biology identifies a
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Using chemogenetics to reduce rat obesity Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Jorge Ferreira
Chemogenetics is a gene therapy approach in development where designer receptors activated exclusively by designer drugs (DREADD) are introduced via genetic manipulation to modulate specific pathways and, in the case of the nervous system, specific neurons or brain areas. Despite their potential interest in human therapeutics, the lack of proper preclinical validation impedes their advancement. A study
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Brain rodent surgery preserves memory Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Jorge Ferreira
Episodic memory refers to the unique set of personal experiences lived. The hippocampus is an essential hub for episodic memory processing. Studies focusing on the role of the hippocampus in learning and the contribution of adult hippocampal neurogenesis to cognition have mostly involved manipulating the brain bilaterally. However, current imaging methods, such as in vivo calcium imaging in freely
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Tracking peroxisomes Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Alexandra Le Bras
Peroxisomes are membrane-bound organelles that are critical for lipid metabolism and redox homeostasis. The importance of these organelles for human health and development is underlined by the existence of several inherited diseases, so called peroxisomal disorders. Existing approaches for imaging peroxisomes in animal models and patient samples are not suitable to study peroxisome dysfunction and
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Gland regeneration Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Alexandra Le Bras
Cell-based therapy for generating salivary glands is a promising future therapy for patients suffering from dry mouth, Sjögren’s syndrome or the side effects of radiotherapy for head and neck cancers. In Cell Reports, researchers used a conditional blastocyst complementation (CBC) technique to generate fully functional salivary glands from pluripotent stem cells in mice. Given that CBC is based on
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JNK1 role in zebrafish anxiety Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Jorge Ferreira
Anxiety and depression are prevalent mental disorders involving abnormal neural function and dysregulated fear response circuits. Drugs used for the treatment of mental disorders are not always effective and produce side effects. Developing a high-throughput assay to test behavior alterations in animal models after drug exposure would improve the detection of therapeutic components and the understanding
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3R centers tap into the human mindset to bolster replacement, reduction and refinement uptake Lab Anim. (IF 5.9) Pub Date : 2024-07-02 Charlotte Harrison
3R centers around the world are confronting the challenges of 3R implementation in animal research by sharing outcomes, resources and training with scientists.
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Model matchmaking via the Solve-RD Rare Disease Models & Mechanisms Network (RDMM-Europe) Lab Anim. (IF 5.9) Pub Date : 2024-06-24 Kornelia Ellwanger, Julie A. Brill, Elke de Boer, Stephanie Efthymiou, Ype Elgersma, Marynelle Icmat, François Lecoquierre, Amanda G. Lobato, Manuela Morleo, Michela Ori, Ashleigh E. Schaffer, Antonio Vitobello, Sara Wells, Binnaz Yalcin, R. Grace Zhai, Marc Sturm, Birte Zurek, Holm Graessner, Eva Bermejo-Sánchez, Teresinha Evangelista, Nicoline Hoogerbrugge, Vincenzo Nigro, Rebecca Schüle, Alain Verloes
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The love maze allows female marmosets to choose their partner Lab Anim. (IF 5.9) Pub Date : 2024-06-17 Jorge Ferreira
Marmosets (Callithrix jacchus) have been increasingly used in biomedical research, but commercial vendors have been struggling to provide enough animals for large projects. Many laboratories have therefore resorted to breeding animals to create their own colonies. However, breeding can be rather difficult and time-consuming, with female marmosets failing to reproduce efficiently. The current system
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Social transmission of inflammation Lab Anim. (IF 5.9) Pub Date : 2024-06-17 Jorge Ferreira
In many species, members of a group respond to their conspecifics’ physiological and emotional states. In some situations, affective mirroring is present, in which the affective state seems to be socially transmitted from one conspecific to another. However, it is unclear if the same phenomenon occurs in inflammatory states. In a recent preprint (not peer-reviewed), Quintana et al. explored the potential
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Crossing species boundaries in regenerative neuroscience with rat–mouse brain chimeras Lab Anim. (IF 5.9) Pub Date : 2024-06-17 Stefano Pluchino, Ivan Lombardi
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Naturally sterile Mus spretus hybrids are suitable for the generation of pseudopregnant embryo transfer recipients Lab Anim. (IF 5.9) Pub Date : 2024-06-17 Chris Preece, Daniel Biggs, Edward Grencis, Maj Simonsen Jackson, Sue Allen, Martin Fray, Antony Adamson, Benjamin Davies
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Screening new opioids with metabolomics Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Jorge Ferreira
The psychoactive drug market is ever-evolving and difficult to track as new designer drugs are rapidly released into the market each year. These drugs are characterized by various structures and unknown metabolic profiles that make them difficult to detect in traditional tests. Novel synthetic opioids are one of the most dangerous drugs and have been involved in around 80% of opioid abuse-related deaths
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Diet, dysbiosis and disease Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Alexandra Le Bras
Dysbiosis – defined by an imbalance in the gut microbiome – is an important feature of many chronic diseases and involves the outgrowth of pathobionts such as Klebsiella pneumoniae in the gut. Given that diet can alter microbiota composition, identifying the dietary components that contribute to dysbiosis is critical. A study in JCI showing that dietary carbohydrates are critical for K. pneumoniae
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Nrf2 protects from neuroinflammation Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Jorge Ferreira
Systemic inflammation might be connected to the cognitive impairment reported in patients with sepsis or after major surgery. Dexmedetomidine (DEX), a drug that acts as an agonist of α2-adrenergic receptors, has been hypothesized to be neuroprotective, with reports showing that DEX reduces neuroinflammation in lipopolysaccharide (LPS) mouse models. Previous work has shown that DEX enhances the expression
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Microbiome acidity protects flies from pathogens Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Jorge Ferreira
Throughout life, we are exposed to a huge variety of microorganisms, be they exogenous or endogenous, such as our gut microbiome. The gut microbiome is involved in many physiological functions, but the influence of our internal microbiome on external microbes is unknown. A study in Cell Reports using gnotobiotic Drosophila melanogaster, a model with a simple gut microbiome consisting of one or two
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Role of Evi5 in Drosophila iron metabolism Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Alexandra Le Bras
Iron is biologically essential and participates in a wide variety of metabolic processes including oxygen transport, oxidative metabolism and cellular proliferation. However, iron is also potentially toxic and abnormal iron levels have been associated with several disorders, including iron deficiency anemia, porphyrias and neurodegenerative disorders such as multiple sclerosis (MS). Iron level must
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More activity helps mice with migraines Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Jorge Ferreira
Migraines are one of the most common debilitating neurological disorders affecting people’s lives. Evidence has emerged indicating that the environment can influence the onset and treatment of various neurological disorders and influence pain. However, the effect of the environment on migraine is still unknown. In a new study, the team analyzed behavioral responses and alterations in protein expression
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A microminipig model of DMD Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Alexandra Le Bras
Duchenne muscular dystrophy (DMD) is a X-linked genetic disorder characterized by progressive muscle degeneration caused by DMD gene mutations leading to the absence of dystrophin in the muscle membrane. The clinical course of human DMD is severe, with gradual inability to walk and premature death due to respiratory and cardiac failure. Although rodent and dog models have been useful to study DMD pathogenesis
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Role of REM sleep in fear extinction Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Alexandra Le Bras
Rapid eye movement (REM) sleep is known to facilitate fear extinction, and REM sleep deprivation after a traumatic event increases the risk of developing post-traumatic stress disorder (PTSD). However, the mechanisms by which REM sleep protects against fearful memories are largely unknown. In a new study published in Current Biology, Hong and colleagues identified the infralimbic cortex (IL) – a brain
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Humane endpoints, defined Lab Anim. (IF 5.9) Pub Date : 2024-06-04 Lauren Danridge, Bill Greer, Samantha Sullivan, Obed Rutebuka, Louis DiVincenti, Axel Wolff
Animals used in biomedical research may experience pain or distress as part of the experimental protocol. By implementing humane endpoints, pain or distress can be prevented or alleviated whilst still meeting scientific aims and objectives. We invited experts from Arizona State University (Samantha Sullivan) and Loma Linda University (Obed Rutebuka), along with USDA and OLAW representatives (Louis
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Temperature-dependent differences in mouse gut motility are mediated by stress Lab Anim. (IF 5.9) Pub Date : 2024-05-28 Alvin Han, Courtney Hudson-Paz, Beatriz G. Robinson, Laren Becker, Amanda Jacobson, Julia A. Kaltschmidt, Jennifer L. Garrison, Ami S. Bhatt, Denise M. Monack
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ELVAs: The new ‘super-organelles’ of the oocyte Lab Anim. (IF 5.9) Pub Date : 2024-05-23 Helena Fulka
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The pig as an optimal animal model for cardiovascular research Lab Anim. (IF 5.9) Pub Date : 2024-05-21 Hao Jia, Yuan Chang, Jiangping Song
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Sex differences in lean-NAFLD mice Lab Anim. (IF 5.9) Pub Date : 2024-05-03 Alexandra Le Bras
Although regularly associated with obesity, non-alcoholic fatty liver disease (NAFLD) is increasingly identified in patients with normal body mass index. Compared with obese individuals with NAFLD, lean individuals with NAFLD have a lower prevalence of diabetes, hypertension, hypertriglyceridemia and metabolic syndrome, but higher fibrosis scores and cardiovascular morbidity. New studies with relevant
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Effects of microbiome transplants on fly microbiome Lab Anim. (IF 5.9) Pub Date : 2024-05-03 Jorge Ferreira
The gut microbiome greatly influences the host organism. However, it is still unknown how microbial diversity affects systemic functions and how the microbiome content and functions change across the host lifespan. A study in Scientific Reports studying Drosophila shows that transplanting microbiomes from flies of different ages affects the host in an unexpected and complex manner. When the researchers
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Disrupted circadian rhythms affect cardiovascular health Lab Anim. (IF 5.9) Pub Date : 2024-05-03 Jorge Ferreira
End-stage heart disease is associated with severe tissue loss and fat wasting, with obesity being a risk factor. However, obese patients have a higher likelihood of surviving heart failure compared to normal-weight individuals. While the circadian rhythm is closely linked to lipid metabolism, the relationship between the circadian rhythm, fat loss and heart failure remains unknown. A study in Scientific
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Effects of Kombucha Tea on C.elegans Lab Anim. (IF 5.9) Pub Date : 2024-05-03 Alexandra Le Bras
Kombucha – a type of tea that has been fermented – is purported to confer health benefits, including lowering blood pressure and protecting against metabolic disease. However, the mechanisms underlying the beneficial effects of the Kombucha Tea-associated microbes (KTM) on host metabolism remain unknown. Using the nematode Caenorhabditis elegans as an animal model system to study the host physiological
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Wild mice for research Lab Anim. (IF 5.9) Pub Date : 2024-05-03 Alexandra Le Bras
Standardized mouse inbred strains have been central to biomedical research for decades, ensuring experimental reproducibility across laboratories and experiments. However, the lack of genetic diversity within and between inbred mouse strain is an important limitation. A study in Plos Genetics reports the development of a new set of 11 inbred strains derived from wild-caught Mus musculus domesticus
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Naked mole-rats hearts resist hypoxic stress Lab Anim. (IF 5.9) Pub Date : 2024-05-03 Jorge Ferreira
The naked mole-rat (NMR), known for its extreme longevity, has emerged as a promising new model for studying aging. Understanding the mechanisms behind NMR’s resistance to hypoxia and cardiovascular injury is important for cardiovascular research. A study in Nature Communications identifies the unique NMR’s genetic and metabolic adaptations to their underground low-oxygen environment. Comparing cardiac
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A single-cell atlas of sex differences and androgen effects Lab Anim. (IF 5.9) Pub Date : 2024-05-03 Alexandra Le Bras
Hormones, especially androgens and estrogens, are most likely contributing substantially to sex-biased phenotypes. To explore sex differences in mice and their modulation by androgens at the cellular and molecular levels, researchers performed single-cell transcriptome profiling of 17 tissues in Mus musculus (C57BL/6 mice), using four different groups of mice: male mice that underwent castration, male
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HIV infection affects cocaine dependence Lab Anim. (IF 5.9) Pub Date : 2024-05-03 Jorge Ferreira
HIV infection and substance abuse are often co-occurring, and cocaine is one of the most commonly used drugs. However, the relationship between cocaine use and HIV infection needs to be better characterized. A study in Communications Biology utilizes humanized mouse models engrafted with human hematopoietic stem and progenitor cells to study the interaction between HIV infection and cocaine-related
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Paper towel shredding as a novel, affordable, noninvasive method for detecting arousals in hibernating rodents Lab Anim. (IF 5.9) Pub Date : 2024-04-18 Amalie J. Hutchinson, Brynne M. Duffy, Lauren H. Rego, James F. Staples
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Streamlining mouse genome editing by integrating AAV repair template delivery and CRISPR-Cas electroporation Lab Anim. (IF 5.9) Pub Date : 2024-04-10 Natalia Moncaut
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Water restriction still has a place Lab Anim. (IF 5.9) Pub Date : 2024-04-09 Pamela Reinagel
The citric acid (CA) method was recently proposed as an alternative to water restriction, which is commonly used to motivate rodents to perform behavioral tasks. As the person who first developed the CA method1, I read with great interest your Protocol Review article “The weights of refinement and flexibility”2. I am gratified to see that this approach is already so widely adopted and discussed, and
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Humanized mouse models of drug metabolism Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Alexandra Le Bras
Accumulating evidence suggests that species differences between mice and humans can compromise drug efficacy studies. In humans, the pharmacological activity of many small molecules is altered by activity of the cytochrome P450 (CYP) system. Humans have eight genes within the CYP1A, CYP2C, CYP2D and CYP3A subfamilies, while mice have 34 genes within the CYP system. These species differences lead to
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AAV gene therapy to treat Friedreich’s ataxia cardiomyopathy Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Jorge Ferreira
Friedreich’s ataxia (FRDA), a common inherited ataxia, affects the nervous system and the heart, causing cardiomyopathy. The disease is associated with a reduced expression of the mitochondrial protein frataxin (FXN), and adeno-associated viral vector (AAV)-mediated gene transfer can be a potential therapeutic approach to induce frataxin expression. While this approach has been effective in cardiac
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The risk of oversight Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Lauren Danridge, Bill Greer, Michelle Aparicio, Sarah Viranda, Elysse Ann Orchard, Louis DiVincenti, Axel Wolff
A clear organizational reporting structure within the Animal Care and Use Program and a collegial working relationship between the Institutional Official, Attending Veterinarian and Institutional Animal Care and Use Committee are essential to maintain the highest standards of animal wellbeing and research quality. We invited experts from The Feinstein Institutes for Medical Research (Michelle Aparicio)
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Environmental enrichment improves mice motor performance Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Jorge Ferreira
Environmental enrichment (EE) is a well-established concept that shows benefits for both brain development and cognitive function. However, rodents are commonly raised in poorly enriched conditions, which can affect neuroscience data. A study in Scientific Reports shows that providing EE to mice improves conditioned responses and motor performance. EE mice showed slower acquisition of eyeblink-conditioned
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Effect of chronic stress on metastasis Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Jorge Ferreira
Chronic stress, a common issue in daily life, is particularly pronounced in patients with cancer. Chronic stress affects the whole body and perturbs systemic homeostasis, promoting cancer growth in mice. Whether and why chronic stress increases metastasis rates is still unknown. A study in Cancer Cell shows that chronic stress promotes metastasis in a metastatic lung cancer mouse model by increasing
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Cancer resistance in bats Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Alexandra Le Bras
Laboratory mice and rats are traditionally used as model organisms to study the molecular mechanisms of carcinogenesis. However, these cancer-prone models have limitations in understanding the mechanisms of cancer resistance. Certain long-lived mammalian species such as naked mole rats seem to have evolved unique anti-cancer mechanisms. Understanding how these animals are protected from cancer could
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Tracking marmoset 3D movement and social behavior Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Jorge Ferreira
A broad range of research areas use behavioral analyses. Despite behavior being extensively used, it is quite uncommon for researchers to analyze long-term continuous behavior, especially in social groups, as it is challenging to accurately track individuals and their behaviors. This is even more pressing in social animals such as marmosets, which show many social close contact behaviors while moving
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Toward phage therapy for tuberculosis Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Alexandra Le Bras
Tuberculosis (TB) is a highly infectious disease caused by Mycobacterium tuberculosis (Mtb), which primarily affects the lungs to cause pulmonary TB, but can also affect other tissues to cause extra-pulmonary TB. Mtb is an intracellular pathogen that can evade the immune system and proliferate within host cells, making antibiotic treatment more challenging and potentially resulting in antimicrobial
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Rhesus macaques for human norovirus studies Lab Anim. (IF 5.9) Pub Date : 2024-04-03 Alexandra Le Bras
Norovirus is a common cause of acute gastrointestinal illness worldwide. Norovirus vaccine development is a priority for both public-health and economic reasons, but the search for a vaccine has been hampered by a lack of suitable and reliable animal models. In Nature Microbiology, Rimkute et al. show that rhesus macaques are susceptible to oral infection with human noroviruses from two different genogroups
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Animal models of postpartum hemorrhage Lab Anim. (IF 5.9) Pub Date : 2024-03-25 Sarah E. Hargett, Elaine F. Leslie, Hector O. Chapa, Akhilesh K. Gaharwar
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A primate pregnancy metabolome atlas Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Jorge Ferreira
Pregnancy comes with many bodily changes during fetus development. However, the extent of the metabolic changes is still largely unknown. A study in Cell now presents an atlas of the metabolomes of 23 different maternal tissues of pregnant cynomolgus monkeys, which were analyzed at various pregnancy stages. The study revealed signs of a dynamic metabolic evolution across different tissues and pregnancy
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A new window to image liver spheroids Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Alexandra Le Bras
Recent advancements in 3D in vitro systems such as liver organoids have opened new avenues for studying liver diseases, but the lack of an in vivo environment can limit the physiological relevance of these models. Previous studies have reported the transplantation of liver spheroids into recipient mice to better mimic physiological conditions, but the transplantation sites (under the kidney capsule
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Age is not enough to predict delirium in mice Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Jorge Ferreira
Age is one of the proposed main triggers of neuropsychological disease. However, the effect of age is heterogeneous, and the mechanisms behind disease progression are not fully determined. A study in Communications Biology shows that neuroinflammation and white matter disruption have an important role in the progression of delirium in aged mice. When comparing 8- and 25-month-old mice, older mice were
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A minimal metadata set (MNMS) to repurpose nonclinical in vivo data for biomedical research Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Anastasios Moresis, Leonardo Restivo, Sophie Bromilow, Gunnar Flik, Giorgio Rosati, Fabrizio Scorrano, Michael Tsoory, Eoin C. O’Connor, Stefano Gaburro, Alexandra Bannach-Brown
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DYRK1A causes heart defects in Down syndrome Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Alexandra Le Bras
Down syndrome (DS) is a gene-dosage disorder caused by the presence of all or part of a third copy of chromosome 21 (Hsa21). DS is associated with different complex phenotypes, including congenital heart disease (CHD), but the causes of heart defects are unclear. A new study used a previously described mouse model of DS (Dp1Tyb mice) that recapitulates key features of DS and develops CHD to provide
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Cannabis activates hunger neurons in rodents Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Jorge Ferreira
A well-known effect of cannabis is the promotion of appetite. However, the neurobiological mechanism behind this stimulation is still unknown. A study in Scientific Reports demonstrates that neurons within the mediobasal hypothalamus, particularly in the arcuate nucleus, have a role in stimulating rat feeding behavior linked to cannabis exposure. When compared with animals exposed to air, rats exposed
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How mice make sounds Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Alexandra Le Bras
Mice mainly communicate in two ways: via human-audible squeaks when experiencing pain or fear; or through higher-frequency ultrasonic vocalizations (USVs) used during courtship and other social interactions. According to a new study, these two different vocalization types are controlled by distinct neural circuits. The researchers at Cornell University showed that TRAP2-mediated ablation of a specialized
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Inhalation-activated drug delivery in mice Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Jorge Ferreira
Chronic diseases are prevalent and pose a considerable challenge to the individual and the healthcare system. The most common route for continuous treatment is injection. However, this method poses a considerable burden on patients, creating an urgent need for improved drug delivery methods. A study in Nature Communications shows the effective treatment of mice with non-alcoholic fatty liver disease
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Mechanisms of gill regeneration in zebrafish Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Alexandra Le Bras
Vertebrate species have evolved efficient respiratory organs – such as gills and lungs – to mediate gas exchange and satisfy their O2 needs. While fish and amphibians can grow and regenerate gills throughout life, mammals only show partial, facultative regeneration of lung tissue following injury. A new study used adult zebrafish to characterize the signaling pathways involved in the early stages of
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The governance and use of genetically modified animals across countries Lab Anim. (IF 5.9) Pub Date : 2024-03-04 Lauren Danridge
In a 2023 Protocol Review, US respondents addressed a scenario1 wherein genetically modified animals (GMAs) were donated to an investigator and used for an unrelated study. In the US, the production and subsequent management of GMAs are governed by specific regulations (PHS Policy, NIH Guidelines), and GMAs that are not used for the intended research would not qualify for other research activities
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Statistical design of experiments: the forgotten component of Reduction Lab Anim. (IF 5.9) Pub Date : 2024-02-21 Penny Reynolds
A strategic and statistically based experimental design is a key component of Reduction, and the backbone of reproducible research. Design basics consist of formal structuring of input variables and clearly defined experimental units. Formal designs can evaluate two or more input factors simultaneously, identify and prioritise the most important inputs, and identify interactions where most discovery