Traditional medicine and herbal remedies are gaining popularity worldwide, comprising a significant portion of healthcare research, advancements, and market demand. Growing scientific evidence supports their substantial efficacy as pharmaceutical ingredients and dietary supplements in preventive healthcare. When developing pharmaceuticals, it is crucial to ensure that ingredients are free from side effects and toxicity in order to prioritize safety. Geckos, known as shou gong, are a diverse group of lizards that are widely utilized for treating various diseases in Korean Medicine. This study was conducted to assess the potential acute toxicity of a water extract Gekko gecko by a single oral dose in Sprague-Dawley rats. Twenty rats of each sex were randomly assigned to four groups (5 rats each). Test articles were administrated once by oral gavage to rats at dose levels of 0, 500, 1,000, or 2,000 mg/kg body weight. Mortality, changes of body weight, and clinical signs of gross observation were monitored for 14 days after dosing. At the end of a 14-day observation period, all animals were sacrificed and complete macroscopic and hematological examinations were performed. There was no dead animal or test article-related effect on clinical signs, body weight, or gross finding. Other specific changes were not found between control and treated groups in hematology. Results showed no adverse effect at a dose of 500, 1,000, or 2,000 mg/kg in rats. The minimal lethal dose was considered to be over 2,000 mg/kg body weight in rats.

Drug-induced liver injury (DILI) is considered to be a significant cause of drug wastage. To mitigate clinical DILI risks, assessing drugs using human liver models is crucial since animal studies may fall short due to species-specific liver pathway variations. Cell-based preclinical hepatotoxicity testing is often pertinent. In the present study, cells from a human liver cancer line (HepG2 and HepaRG) were cultured in both formats of 2D and 3D spheroids to explore their responses to drugs. Liver-specific marker expressions across cell lines and culture formats were also examined to assess disparities in DILI marker expressions. After treating each cell with the drugs, cytotoxicity and liver injury markers aspartate aminotransferase and alanine aminotransferase were increased. In addition, liver specific markers albumin and urea decreased in a drug concentration-dependent manner. These findings were consistent with drug sensitivity. Additionally, mRNA expression levels of cytochrome P450 enzymes (CYPs) involved in hepatocellular drug metabolism were compared following treatment with enzyme inducers. CYP1A2 and CYP2C9 were not epxressed in HepG2 cells. HepaRG cells exhibited significantly increased expression of CYP1A2, 2C9, and 3A4 post-treatment. Notably, enzyme expression was notably higher in 3D cultures than in 2D cultures. Collectively, these findings suggest that HepaRG cells and 3D cultures hold promise for evaluating DILI during early-stage drug development.

Caprine cryptosporidiosis mainly occurs in young goats, with morbidity rates of 80%–100% and mortality over 50% in goat kids. However, limited research has been conducted on the impact of Cryptosporidium parvum, a diarrhea-causing pathogen, on the intestinal microbiota of goat kids. In this study, 16S rRNA-based metataxonomic analysis was performed to compare the microbial diversity and abundance of the gut microbiota between C. parvum-infected and uninfected goat kids. In total, 12 goat fecal samples were collected, including seven naturally C. parvum-infected and five uninfected goats from Chungcheongbuk-do, Korea. After amplification of the V3–V4 hypervariable region of the bacterial 16S rRNA, high-throughput sequencing was performed. The results showed differences in the microbial composition between C. parvum-infected and uninfected groups based on beta diversity. Firmicutes and Bacteroidetes were the most dominant phyla in both groups. However, no significant difference was observed in the Bacteroidetes/Firmicutes ratio between the two groups. Compared with the uninfected group, the C. parvum-infected group showed significantly higher abundances of Tyzzerella nexillis, Lactobacillus johnsonii, Butyricicoccus pullicaecorum, Enterococcus raffinosus, Enterococcus faecalis, and Negativicoccus massiliensis, and significantly reduced abundances of Aerococcus vaginalis, Faecalicoccus pleomorphus, Oribacterium parvum, and Coprococcus comes. These findings indicate that C. parvum infection, which is associated with diarrhea in neonatal goats, induces alterations in the caprine gut microbiota.

This study was conducted to collect the patents of microbiome-based treatment technology for pets. An electronic search for microbiome or probiotics in brain nervous system disease was studied using the WINTELIPS database. Patent Cooperation Treaty of Korea, Japan, the EU, the US, and China that were registered by October 31, 2022 were selected in this study. A total of 206 patents were included for final analysis. Since 2016, patent activity has shown an explosive increase in recent years. China is the leading market in this technology field, and Korea has the second-highest market share. To provide the groundwork for the next research and development, we examined the industrial trend of microbiome for brain nervous system diseases in this study using an analysis of patents that have been applied for and registered up to this point. Looking at the overall patent trends by year in the technology field related to treating of brain and nervous system diseases using the microbiome, there was a tendency to repeat increasing and decreasing trends. However, considering 2021 and 2022, which have undisclosed sections, it can be seen that patent activity has tended to increase explosively in recent years, starting in 2016. If related studies use the patent analysis data constructed in this way strategically, it is expected that it will lead to patent registration and the development of new products, ultimately contributing to the revitalization of the companion animal industry.

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease by John Cunningham virus (JC virus) infection in oligodendrocytes. The radiographic and clinical features, along with the identification JC in cerebrospinal fluid polymerase chain reaction, are sufficient for the diagnosis of PML in immunodeficiency. However, it is difficult to suspect PML without the patient history of immunodeficiency. A 32-year-old man presented with headache for a month without any medical history. Based on clinical and image features, the differential diagnoses included demyelinating lesion and neoplasms. Microscopically, biopsy specimen showed multifocal demyelinating and degenerative white matter, consistent with PML. Oligodendrocytes cells with increased nuclei and plum-colored inclusions were admixed with perivascular lymphocytic and histiocytic infiltration, and loss of myelin. Atypical astrocytes had large or multiple nuclei. After brain biopsy, human immunodeficiency virus infection was confirmed by serum chemiluminescent immunoassay. It is unlikely that PML would be considered without the information of immunosuppression. Therefore, it is very important to be aware of the histological features of PML.

In response to the expanding landscape of the biotechnology industry and the increasing demand for comprehensive drug development as well as the conduct of preclinical and clinical trials, there is a growing need for employment of diverse animal models, including both small and large animals. The focus of this study was on refining ex vivo culture techniques for bioluminescence imaging following administration of intradermal injections in large animals. To examine the feasibility of our approach, varying concentrations of the rFluc protein were administered to rats and live imaging was employed to validate the corresponding levels of expression. Subsequently, following administration of rFluc to mini-pigs, ex vivo analyses were performed on sample tissues to assess the levels of protein expression across different concentrations. In particular, optimal culturing conditions that facilitated the sustained expression of the protein in samples post-euthanasia were identified. Moreover, by employing small animal imaging devices, we were able to capture clear images of the sample plates, which provided evidence of the successful application of our experimental techniques. The findings from this research represent a significant effort toward refining bioluminescence imaging methods tailored for use with large animal models—an imperative facet of contemporary drug development and biomedical research.

Canine hyperadrenocorticism, a prevalent endocrine disorder characterized by excessive cortisol production. Notably, hypercoagulability leading to pulmonary thromboembolism (PTE) poses a substantial concern. PTE may be underestimated because of the rapid dissolution of canine thrombi postmortem. However, traditional coagulation assays face challenges in early detection of hypercoagulability. Therefore, this study explored the use of thromboelastography (TEG) as a diagnostic tool for identifying hypercoagulability in dogs with hyperadrenocorticism. A total of 31 dogs visited the Gyeongsang Animal Medical Center between 2018 and 2022, comprising 21 dogs with hyperadrenocorticism and 10 controls who underwent clinical and coagulation analyses. Hyperadrenocorticism was diagnosed using a low-dose dexamethasone stimulation test or adrenocorticotropin hormone stimulation test, and conventional laboratory parameters and coagulation parameters, such as the prothrombin time, activated partial thromboplastic time, fibrinogen, and TEG results, were compared between the groups. Clinical data revealed significantly elevated monocyte, platelet, alanine aminotransferase, alkaline phosphatase, triglyceride, and cholesterol concentrations in dogs with hyperadrenocorticism, which were attributed to excess cortisol secretion (p<0.05). TEG analysis demonstrated significantly decreased K values and increased α and MA values in hyperadrenocorticism dogs (p<0.05), indicating a shortened clotting time and enhanced clot strength, suggestive of hypercoagulability. TEG effectively highlights hypercoagulability in dogs with hyperadrenocorticism and provides valuable insights in predicting blood clot formation. Although predicting clot formation in dogs remains complex owing to multifactorial influences, this study underscores the potential utility of TEG in enhancing such predictions for dogs with hyperadrenocorticism.

Salivary gland dysfunction is a common complication of diabetes. Decreased saliva production and changes in saliva composition may cause oral diseases. Reactive oxygen species (ROS) generation in the salivary glands results in the loss of acinar cells and decreased saliva secretion. Glucagon-like peptide 1 (GLP-1) is the incretin hormone that regulates blood glucose level and can suppress ROS production and inflammation through its antioxidant effects. Dipeptidyl peptidase-4 (DPP-4) is an enzyme that breaks down GLP-1. In this study, we evaluated the pathological role of DPP-4 and GLP-1 on salivary gland dysfunction in type 2 diabetic db/db mice. We observed reduced salivary secretion and histopathological alteration of salivary glands in the db/db mice. The increased DPP-4 and decreased GLP-1 levels in the salivary glands were also detected in the db/db mice. Furthermore, the db/db mice had increased apoptosis and oxidative injury in salivary glands. There was an accumulation of advanced glycation end products and mucus in the salivary glands of the db/db mice. In conclusion, these results showed the possible involvement of DPP-4 and GLP-1, leading to increased ROS-induced apoptosis in diabetes-related salivary gland dysfunction. DPP-4 and GLP-1 may be a pharmacological target for patients with diabetes-related salivary gland dysfunction.

Animal bones excavated from historic sites provide valuable data for identifying the lifestyles of people and the distribution of animals at that time. In this study, we investigated the morphological structure, size, and measurements of Cervidae bones excavated from a well at the Gasan-ri archaeological site in Jinju, which are believed to be relics from the Three Kingdoms period. The total number of excavated animal bones was 447, of which 102 (22.82%) were classified as Cervidae bones. The weight of Cervidae bones was 453.79 g, accounting for 46.53% of the total weight of the identified bones (975.30 g). The Cervidae bones were identified as those of two animals with an estimated age of 5–6 months. The Cervidae bones are divided into skull bones, vertebrae, ribs, sternum, hip bones, forelimb bones, and hindlimb bones. The 102 Cervidae bones consisted of 19 skull bones (18.63%), 14 vertebral axial skeletons (13.72%), 28 ribs and sternum (28.43%), 16 forelimb bones (15.69%), and 19 hindlimb bones (18.63%). The remaining six were difficult to distinguish. A fracture of the parietal bone located near the bregma of a skull was observed and was presumed to have been caused by an artificial blow. This study can be used as basic data to estimate the types of animals and human culture at the time through Cervidae bones believed to be relics from the Three Kingdoms period.

Iron is an essential nutrient for mammalian cells. Most iron absorption occurs in the duodenal epithelial cells and is regulated by hepcidin, which is produced and secreted in the liver. High hepcidin levels can cause iron deficiency anemia due to iron absorption failure. Inside the cell, iron conjugates with a porphyrin ring and is placed with an iron coordinated to heme. One of the heme-binding proteins, known as progesterone receptor membrane component 1 (Pgrmc1), is a non-canonical progesterone receptor associated with diverse molecular gene regulation. Previous studies showed that Pgrmc1 is related to iron homeostasis via hepcidin; however, these mechanisms remain to be elucidated. In the present study, to investigate the role of Pgrmc1 in mammalian iron metabolism, we introduced Pgrmc1 knockout (KO) mice and performed molecular biological analyses using qPCR and western blotting. Pgrmc1 deficiency decreased Hamp mRNA expression and hepcidin protein levels. However, Pgrmc1 deficiency failed to decrease Hamp transcript expression and hepcidin protein levels in siPGRMC1-transfected HepG2 cells and primary Pgrmc1 KO hepatocytes, respectively. PGRMC1 knockdown cells revealed low HAMP mRNA levels upon cyclic AMP (cAMP) activation, suggesting that PGRMC1 promotes HAMP mRNA transcription via cAMP activation. It has been implicated that hepatic Pgrmc1 cannot control hepcidin directly; however, the internal environment caused by the lack of Pgrmc1 may potentially cause low hepcidin levels.

A 14-year-old castrated male Shih Tzu presented with acute hemorrhagic vomiting. The initial medical records indicated a probable diagnosis of acute gastritis due to inappropriate food intake. Although gastrointestinal (GI) endoscopy was the preferred diagnostic approach, the client declined anesthesia because of the dog’s underlying heart condition. Therefore, we opted for anesthesia-free capsule endoscopy. The procedure identified severe gastritis with no detectable abnormalities in the other GI regions. Following diagnosis, dietary modifications and omeprazole treatment were initiated, resulting in the resolution of clinical symptoms. Follow- up capsule endoscopy 3 weeks later verified a significant improvement in gastritis. This case highlights the potential of capsule endoscopy as a valuable diagnostic tool in patients presenting with acute vomiting.

A one-year-old, female, Maltese dog was presented with head tilting, horizontal nystagmus, and tetraparesis. Blindness was first identified, and magnetic resonance imaging (MRI) scanning revealed diffuse lesion which was hyperintense on T2-weighted image over the cerebellum and brainstem. The immunosuppressive therapy had been administered, but the patient had no improvement. Re-performed MRI revealed the progression of the pre-existed inflammatory lesions. Treatment with prednisolone, leflunomide, cyclosporine, and cytosine arabinoside was initiated. However, neurological signs had been progressive, and the patient was euthanized. The histopathological examination revealed the disseminated granulomatous meningoencephalomyelitis (GME). This GME case suggests the importance of initiation of treatment at the appropriate time.

Mucosa-associated lymphoid tissue (MALT) lymphoma (ML) is a type of non-Hodgkin’s lymphoma involving MALT, commonly the stomach or salivary glands, although virtually any mucosal site can be affected. ML originates from B cells in the marginal zone of MALT, and is also called extranodal marginal zone B cell lymphoma. It is a slow-growing cancer that usually responds well to treatment. A 59-year-old female presented with a 1-day history of quadriparesis and dysarthria. Up arrival at the hospital, motor power in the right upper and lower extremities was grade 3/5 according to the Medical Research Council scale, while that in the left leg was 4/5. The patient had been diagnosed with gastric ML 1 year prior, and had received antibiotics during the previous 2 weeks. The emergency magnetic resonance imaging of the brain performed at the time of presentation showed multifocal embolic infarction in the cerebral hemisphere bilaterally, which did not have a cardiac origin. Magnetic resonance angiography revealed no stenotic or occlusive lesions. Secondary prophylaxis with daily administration of 300 mg aspirin was prescribed. The patient was discharged with residual right hemiparesis 2 weeks after the onset of symptoms. Herein, we present a rare case of multifocal cerebral infarction in a gastric ML patient.

A 12-year-old intact female Schnauzer was referred for the evaluation of poorly controlled diabetes mellitus: despite insulin therapy, blood glucose concentration was consistently high, indicating a decreased insulin sensitivity. Laboratory analyses revealed persistent hyperglycemia, glucosuria, and ketonuria. Diagnostic approaches were performed to identify concurrent disorders that can cause insulin resistance. The dog was found to have concurrent hyperadrenocorticism, hyperlipidemia, pancreatitis, and vaginal cytology indicating diestrus in the estrus cycle. Trilostane administration for hyperadrenocorticism improved the insulin response; however, the dog remained hyperglycemic. Eventually, the dog showed complete remission without insulin administration 1 week after the ovariohysterectomy. The dog remained in remission for approximately 4 months, but eventually relapsed and the condition was permanent. Diestrus in intact females and hyperadrenocorticism are known to be the two main causes of insulin resistance in dogs. After the management of these conditions, the dog achieved diabetes remission, which rarely achieves in dogs. In cases of insulin resistance, such as hormonal imbalances or inflammatory conditions, remission can be achieved by addressing the underlying cause. Hence, it is important to assess the presence of comorbidities associated with insulin resistance in dogs with poorly controlled diabetes mellitus and to treat each condition as soon as possible.

Collagen peptides have garnered significant attention as functional foods across multiple fields due to their capacity to regulate physiological and hormonal processes, offering numerous advantages. However, despite their broad range of applications, comprehensive research on the potential toxicity of these substances remains lacking. Therefore, this study sought to assess the acute oral toxicity of a collagen peptide derived from skate (Raja kenojei) skin (CPSS) in both rats and dogs. In the rat model, CPSS was orally administered at doses of 300 and 2,000 mg/kg to Sprague-Dawley rats. An escalating single-dose oral toxicity assessment at doses of 500, 1,000, and 2,000 mg/kg was carried out in beagle dogs with 3-day intervals between doses. Throughout the 14-day post-administration assessment period, clinical signs, mortality rates, changes in body weight, and necropsy observations were closely monitored. After oral administration, no signs of toxicity associated with CPSS were observed in either rats or dogs. Therefore, the oral LD50 (approximate lethal dose for 50% mortality) for CPSS in rats was determined to exceed 5,000 mg/kg, and the maximum tolerated dose for dogs was estimated to be above 2,000 mg/kg. Consequently, this study offers safety data on the use of CPSS in functional foods and medicinal applications.

1,2-Dichlorobenzene (1,2-DCB) is in various industries as solvents in herbicides, pesticides, and wax, and as degreasers or in the production of dyes. Studies on the hazards and risks of 1,2-DCB showed that this substance can cause skin corrosion, skin irritability, respiratory irritability, and certain target organ toxicity. Industrial workers are exposed to 1,2-DCB by inhalation or skin exposure and there is a lack of information on human hazards even though they can be exposed to organic compounds such as benzene or other DCB complexes rather than a single substance. In this study, we investigated the specific organ toxicity of 1,2- DCB and sex differences using whole-body inhalation in laboratory mice. Male and female mice were exposed to 0–120 ppm of the test substance for 13 weeks. After euthanization, the organs were collected, histopathological assessments and immunohistochemistry (IHC) were performed, and lipid peroxidation was analyzed. Macro and microscopic lesions were observed in the livers of male mice exposed to the test substance, and microscopic alterations were observed in the nasal cavities of male and female mice. IHC analysis of the liver confirmed a greater increase in cytochrome P450 induction in males than in female mice, and malondialdehyde and 4-hydroxynonenal were increased in both sexes by 1,2-DCB inhalation. Based on the relevant literature and experimental results, 1,2-DCB is believed to cause specific organ toxicity in the livers of male mice and the nasal cavities of both sexes of mice, which is supposed to be related to sex differences in cytochrome P450 induction and changes in lipid and oxidative products associated with the early metabolites of the test substance.

Babesiosis is a tick-borne disease caused by intraerythrocytic protozoa. Despite the increasing acknowledgement that babesiosis represents a threat to animal and human health, to date there have been few studies focusing on the disease in the Republic of Korea (ROK). In the present study, we report a Babesia capreoli infection in an Ixodes nipponensis tick obtained from a Korean water deer (Hydropotes inermis argyropus). The tick was identified with polymerase chain reaction analysis as I. nipponensis (Japanese hard tick). A phylogenetic analysis based on the 18S rRNA gene sequences revealed that the isolate found in I. nipponensis belonged to the B. capreoli lineage and was distinct from the Asian, European, and North American lineages of Babesia divergens. Although our isolate belonged to the B. capreoli lineage it did not form a cluster with others isolates in the same lineage; this may be due to differences in the tick species that transmit B. capreoli or in the host species. We were unable to identify the reservoir host for our case of B. capreoli transmission, though regional ticks may be the primary vector. This study confirms the presence of B. capreoli in the ROK, and its presence suggests that further study is warranted to determine its prevalence and pathogenicity in wild and domesticated animals.

Patients experiencing out-of-hospital cardiac arrest (OOHCA) during the weekend are less likely to survive to hospital admission than those experiencing OOHCA on weekdays. We aimed to determine whether the survival rates of in-hospital cardiac arrest (IHCA) were lower on weekends than on weekdays. We comprehensively reviewed the records of patients who experienced IHCA at CBNUH between 2018 and 2020. A total of 861 IHCAs occurred during the study period; these data included recurrent IHCA cases, as some patients experienced more than one IHCA. Of these, 739 IHCA cases were included in the survival analysis, and the survival rate was 65.2%, which is higher than the rate reported in a previous study. There were no differences in the survival rate between weekdays and weekends. Additionally, the time of day at which IHCA occurred and pre-IHCA intubation status did not affect survival. Patients in wards were less likely to survive than those in intensive care units (60.0% vs. 66.0%). Although pre-IHCA intubation did not show any added value in preventing sudden cardiac arrests, meticulous patient care and monitoring in terms of intra- or extrapulmonary oxygen therapy is needed, as is the promotion of cardio-pulmonary-cerebral resuscitation (CPCR) equipment availability and quick rescuer responses for patients with IHCA. Our findings may be of value in improving CPCR guidelines and monitoring patients at risk of IHCA.

Toll-like receptor 4 (TLR4) is known to contribute to the modulation of insulin resistance and systemic inflammation seen in obesity and the metabolic syndrome. The present study was performed to investigate the fertility competence of TLR4 knock out male mice (TLR4 mice) on a high-fat diet (HFD), compared to a normal-chow diet (NCD). The controls included wildtype (WT) mice fed on a HFD or NCD. Six-week-old male mice were fed with either a NCD or HFD for 20 weeks. Body and organ weights, serum levels of glucose, triglycerides and hepatoxicity, sperm quality and spermatogenesis were observed after the sacrifice. Also, randomly selected male mice were mated with virgin female mice after feeding of 19 weeks. The weight of the body and organs increased in WT and TLR4 mice on a HFD compared to those of mice on a NCD. The weights of the reproductive organs did not vary among the treatment groups. The motility and concentration of the epididymal spermatozoa decreased in both WT and TLR4 mice fed a HFD. The pregnancy rate and litter size declined in the HFD-fed WT mice compared to the HFD-fed TLR4 mice. In conclusion, the HFD alters energy and steroid metabolism in mice, which may lead to male reproductive disorders. However, fertility competence was somewhat restored in HFD-fed TLR4 male mice, suggesting that the TLR4 is involved in testis dysfunction due to metabolic imbalance.

Macrophages secrete various cytokines and inflammatory mediators, resulting in playing critical roles in inflammation and immunity. In this study, we investigated anti-inflammatory and immune enhancing properties of PB203, which is a water-soluble extract powder from the fruit of Actinidia polygama, in macrophages. A. polygama is a medicinal plant traditionally known to treat abdominal pain, stroke and rheumatoid arthritis. However, the molecular mechanism for the immune modulation of PB203 is still unclear. Therefore, we assessed the effects of PB203 on the lipopolysaccharide (LPS)-induced inflammation and immune activation, and elucidated its action mechanism in mouse macrophage, RAW264.7 cells. PB203 significantly suppressed not only the levels of nitric oxide (NO), prostaglandin E2, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), but also the mRNA expression of inducible NO synthase, cyclooxygenase-2, TNF-α and IL-1β in LPS-stimulated RAW264.7 cells. We also found that these anti-inflammatory activities of PB203 were mediated through the inhibition of toll-like receptor 4 and nuclear factor kappa B (NF-κB) induced by LPS. On the other hand, in normal macrophages, PB203 dose-dependently elevated the gene expression of immunomodulators including granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor, monocyte chemoattractant protein-1 and TNF-α in a statistically significant manner. The expression of IL-10, IL-1β, IL-6, and interferon-γ were also remarkably upregulated by the treatment of 500 μg/mL PB203. In addition, PB203-mediated production of NO and TNF-α was attenuated by NF-κB inhibition in RAW264.7 cells. Interestingly, PB203 promoted the production of nuclear factor erythroid-2-related factor 2, resulting in the increased level of heme oxygenase-1, which is a representative antioxidant enzyme, in both LPS-stimulated and normal RAW264.7 cells. Taken all together, these results suggest that PB203 may have great potential as the candidate of anti-inflammatory agent for improving inflammatory diseases or immune enhancing agent for preventing infectious diseases. Keywords: Actinidia polygama extract (PB203); macrophages; immunomodulator; nuclear factor kappa B (NF-κB); heme oxygenase-1 (HO-1)