Scratching is a main behavioral response accompanied by acute and chronic itch conditions, and has been quantified as an objective correlate to assess itch in studies using laboratory animals. Scratching has been counted mostly by human annotators, which is a time-consuming and laborious process. It has been attempted to develop automated scoring methods using various strategies, but they often require specialized equipment, costly software, or implantation of device which may disturb animal behaviors. To complement limitations of those methods, we have adapted machine learning-based strategy to develop a novel automated and real-time method detecting mouse scratching from experimental movies captured using monochrome cameras such as a webcam. Scratching is identified by characteristic changes in pixels, body position, and body size by frame as well as the size of body. To build a training model, a novel two-step J48 decision tree-inducing algorithm along with a C4.5 post-pruning algorithm was applied to three 30-min video recordings in which a mouse exhibits scratching following an intradermal injection of a pruritogen, and the resultant frames were then used for the next round of training. The trained method exhibited, on average, a sensitivity and specificity of 95.19% and 92.96%, respectively, in a performance test with five new recordings. This result suggests that it can be used as a non-invasive, automated and objective tool to measure mouse scratching from video recordings captured in general experimental settings, permitting rapid and accurate analysis of scratching for preclinical studies and high throughput drug screening.
Animals ; Animals, Laboratory ; Behavior, Animal ; Body Size ; Complement System Proteins ; Decision Trees ; Drug Evaluation, Preclinical ; Humans ; Injections, Intradermal ; Machine Learning ; Methods ; Mice ; Motion Pictures as Topic ; Pruritus ; Research Design ; Sensitivity and Specificity ; Video Recording

Preclinical neuroimaging allows for the assessment of brain anatomy, connectivity, and function in laboratory animals, such as mice and this imaging field has been a rapidly growing aimed at bridging the translation gap between animal and human research. The progress in the animal research could be accelerated by high-resolution in vivo optical imaging technologies. Optical coherence tomography-based angiography (OCTA) estimates the scattering from moving red blood cells, providing the visualization of functional micro-vessel networks within tissue beds in vivo without a need for exogenous contrast agents. Recent advancement of OCTA methods have expanded its application to neuroimaging of small animal models of brain disorders. In this paper, we overview the recent development of OCTA techniques for blood flow imaging and its preclinical applications in neuroimaging. In specific, a summary of preclinical OCTA studies for traumatic brain injury, cerebral stroke, and aging brain on mice is reviewed.
Aging ; Angiography ; Animal Experimentation ; Animals ; Animals, Laboratory ; Brain ; Brain Diseases ; Brain Injuries ; Contrast Media ; Erythrocytes ; Humans ; Mice ; Models, Animal ; Neuroimaging ; Optical Imaging ; Stroke ; Tomography, Optical Coherence

No abstract available.
Animals, Laboratory

Efficient behavioral assays are crucial for understanding the neural mechanisms of cognitive functions. Here, we designed a high-throughput automatic training system for spatial cognition (HASS) for free-moving mice. Mice were trained to return to the home arm and remain there during a delay period. Software was designed to enable automatic training in all its phases, including habituation, shaping, and learning. Using this system, we trained mice to successfully perform a spatially delayed nonmatch to sample task, which tested spatial cognition, working memory, and decision making. Performance depended on the delay duration, which is a hallmark of working memory tasks. The HASS enabled a human operator to train more than six mice simultaneously with minimal intervention, therefore greatly enhancing experimental efficiency and minimizing stress to the mice. Combined with the optogenetic method and neurophysiological techniques, the HASS will be useful in deciphering the neural circuitry underlying spatial cognition.
Animals ; Automation, Laboratory ; instrumentation ; Behavior, Animal ; Equipment Design ; Habituation, Psychophysiologic ; Male ; Memory, Short-Term ; Mice, Inbred C57BL ; Spatial Memory

Inbred mice are an essential animal strain for research as they can improve the reproducibility and reliability of study results. The establishment of new inbred lines is continuing, and new inbred lines are being used in many research fields. C57BL/6 is a mouse laboratory animal that has been developed and used as an inbred strain since early stage of mouse strain development, and, in the 1950s, C57BL/6 was separated into substrains by the Jackson Laboratory (C57BL/6J) and the National Institutes of Health (C57BL/6N). C57BL/6 mice have been used in immunology and antitumor activity studies since the early strain development stage. After the mouse genome was fully described, C57BL/6 mice use in many areas of research has expanded. In particular, immunological characteristics such as those related to cell-mediated immunity and NK cell activity are relatively higher in C57BL/6 mice than in other mice. The C57BL/6NKorl is a stock of C57BL/6N established as part of a localization of experimental animal strategy of the Korean Food and Drug Administration. Based on analysis of single nucleotide polymorphisms (SNPs), C57BL/6NKorl is considered a genetically distinct inbred stock from other C57BL/6N. Various research efforts have been made to describe the characteristics and increase knowledge of the characteristics of C57BL/6Nkorl. The results obtained through these efforts are expected to increase the utilization of C57BL/6Nkorl as a domestic laboratory animal resource and to enhance the reliability of mouse based studies.
Allergy and Immunology ; Animals ; Animals, Laboratory ; Genome ; Immunity, Cellular ; Killer Cells, Natural ; Mice* ; National Institutes of Health (U.S.) ; Polymorphism, Single Nucleotide ; United States Food and Drug Administration

Mouse is a commonly used animal in life science studies and is classified as outbred if genetically diverse and inbred if genetically homogeneous. Outbred mouse stocks, are used in toxicology, oncology, infection and pharmacology research. The National Institute of Food and Drug Safety Evaluation (NIFDS; former the Korea National Institute of Health) have bred ICR mice for more than 50 years. We investigated to provide users with information and promote accountability to the Korl:ICR. To secure the indigenous data, biological characteristics of Korl:ICR were identified by comparing with other ICR stocks. This domestic ICR stock was denominated as ‘Korl:ICR’. Phylogenetic analysis using SNPs indicated that the population stratification of the Korl:ICR was allocated different area with other ICR. In addition, we measured litter size, body weight, body length, various organ weight, hematology and clinical blood chemistry of the Korl:ICR compared to other ICR. Otherwise, there are no significant differences among the biological phenotypes of Korl:ICR and other ICR. These results suggest that as a genetically indigenous source colony, the Korl:ICR is seperated (or independent) stock with other ICR. Also, we confirmed that there is no difference among the Korl:ICR and other ICR on biological phenotypes. Therefore, the Korl:ICR source colony might be a new stock in distinction from other ICR, it is a good milestone in securing ownership of the national laboratory animal resource. The NIFDS expects that the Korl:ICR mice will be useful animal resource for our domestic researchers.
Animals ; Animals, Laboratory ; Biological Science Disciplines ; Body Weight ; Chemistry ; Hematology ; Korea ; Litter Size ; Mice ; Mice, Inbred ICR ; Organ Size ; Ownership ; Pharmacology ; Phenotype ; Polymorphism, Single Nucleotide ; Population Characteristics ; Rodentia ; Social Responsibility ; Toxicology

Several studies have reported a good correlation between levels of serum hepatitis B virus surface antigen (HBsAg) and covalently closed circular DNA (cccDNA) before and after antiviral therapy. As a result, the quantification of HBsAg levels has attracted much attention in recent years as an important approach to evaluate viral activity. In this study, mAbs against HBsAg were generated and 9 mAbs (H17, H30, H31, H67, H73, H97, H101, H118, and H128) were investigated for optimization of HBsAg quantitation ELISA. Determination of the best combinations of mAbs for sandwich ELISA identified H17 and H31 mAbs as the ideal capture and horseradish peroxidase (HRP) conjugate mAbs, respectively. A standard curve for the current assay system exhibited linearity up to 40 ng/ml of HBsAg while a detection limit of approximately 1 ng/ml of HBsAg was also estimated, which was comparable to that of the other commercial ELISA kits. The ELISA system established in this study is particularly differentiated from other commercial kits in using mAbs for both capture and HRP conjugate, which provides a solution to inconsistency of quality and ethical issues in polyclonal antibodies production using laboratory animals.
Animals, Laboratory ; Antibodies ; Antigens, Surface ; DNA, Circular ; Enzyme-Linked Immunosorbent Assay* ; Ethics ; Hepatitis B Surface Antigens ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Horseradish Peroxidase ; Limit of Detection

The military medical system of the Korean People's Army (KPA) first appeared in August 1946 when a central military hospita was established at the headquarters. Inside the KPA, the military medical and veteran services were first established in February 1948. The military medical officers of the KPA were those who were initially engaged in North Korea's health care sector. Most of the early military medical officers were those who had been trained in the Japanese medical system before liberation and were surgeons. After the establishment of the government in September 1948, Lee Dongwha rapidly introduced the medical system of the Soviet army into the KPA. The KPA military medical system was a mix of Soviet, Japanese and Chinese military medical systems. The medical section of the KPA was similar to that of the Japanese army, and the medical section of the lower army was similar to that of the Soviet army. The stretcher platoon of the KPA were similar to those of the Japanese and Chinese armies. The KPA mainly used Japanese medical equipment at the beginning, and after the establishment of the North Korean regime in September 1948, they were gradually replaced with Soviet products. The military medical office of the KPA were equipped with treatment rooms, laboratories, hospitals, pharmacy, and inpatient rooms. The military medical office purchased medical journals and specimens for medical research and set up a separate research fund. In addition, the military medical office was equipped with a laboratory for medical experiments and raised laboratory animals. The KPA military medical system was specialized in the fields of infectious disease prevention and preventive medicine. At the time, infectious disease in North Korea was mainly caused by bacteria and viruses in unsanitary living environments. The KPA set up a special anti-infectious disease department in consideration of the soldiers living in the collective facilities. The second characteristic of the KPA military medical system is preventive medicine. Since early 1946, North Korea has been interested in preventive medicine and has established various medical facilities and personnel. In line with this history of preventive medicine, the preventive department was installed in the KPA military medical system.
Animals, Laboratory ; Asian Continental Ancestry Group ; Bacteria ; Communicable Diseases ; Democratic People's Republic of Korea ; Financial Management ; Health Care Sector ; Hospitals, Military ; Humans ; Inpatients ; Military Personnel* ; Pharmacy ; Preventive Medicine ; Surgeons ; Veterans

G-protein–coupled receptors (GPCRs) are one of the most attractive therapeutic target classes because of their critical roles in intracellular signaling and their clinical relevance to a variety of diseases, including cancer, infection and inflammation. However, high conformational variability, the small exposed area of extracellular epitopes and difficulty in the preparation of GPCR antigens have delayed both the isolation of therapeutic anti-GPCR antibodies as well as studies on the structure, function and biochemical mechanisms of GPCRs. To overcome the challenges in generating highly specific anti-GPCR antibodies with enhanced efficacy and safety, various forms of antigens have been successfully designed and employed for screening with newly emerged systems based on laboratory animal immunization and high-throughput-directed evolution.
Animals, Laboratory ; Antibodies* ; Epitopes ; Immunization ; Inflammation ; Mass Screening

Many researchers are using viruses to deliver genes of interest into the brains of laboratory animals. However, certain target brain cells are not easily infected by viruses. Moreover, the differential tropism of different viruses in monkey brain is not well established. We investigated the cellular tropism of lentivirus and adeno-associated virus (AAV) toward neuron and glia in the brain of cynomolgus monkeys (Macaca fascularis). Lentivirus and AAV were injected into putamen of the monkey brain. One month after injection, monkeys were sacrificed, and then the presence of viral infection by expression of reporter fluorescence proteins was examined. Tissues were sectioned and stained with NeuN and GFAP antibodies for identifying neuronal cells or astrocytes, respectively, and viral reporter GFP-expressing cells were counted. We found that while lentivirus infected mostly astrocytes, AAV infected neurons at a higher rate than astrocytes. Moreover, astrocytes showed reactiveness when cells were infected by virus, likely due to virus-mediated neuroinflammation. The Sholl analysis was done to compare the hypertrophy of infected and uninfected astrocytes by virus. The lentivirus infected astrocytes showed negligible hypertrophy whereas AAV infected astrocytes showed significant changes in morphology, compared to uninfected astrocytes. In the brain of cynomolgus monkey, lentivirus shows tropism for astrocytes over neurons without much reactivity in astrocytes, whereas AAV shows tropism for neurons over glial cells with a significant reactivity in astrocytes. We conclude that AAV is best-suited for gene delivery to neurons, whereas lentivirus is the best choice for gene delivery to astrocytes in the brain of cynomolgus monkeys.
Animals, Laboratory ; Antibodies ; Astrocytes ; Brain* ; Dependovirus* ; Fluorescence ; Haplorhini ; Hypertrophy ; Lentivirus* ; Macaca fascicularis* ; Neuroglia ; Neurons ; Putamen ; Tropism*