Mercury and its compounds are globally managed chemicals as risks to the human body and ecosystem. It mainly enters the human body through ingestion of seafood, especially, methylmercury causes serious diseases such as central nervous system (CNS) disorder and renal dysfunction. In this study, total mercury (Hg) and methyl mercury (MeHg) were determined in seafood (16 species, n = 97) commonly consumed in Daejeon, using a gold amalgamation method. The average concentration (Hg/MeHg mean ± S.D. (minimum–maximun) mg/kg) of total Hg and MeHg in the samples was as follows; Fish 0.038 ± 0.058 (0.004 – 0.272) / 0.028 ± 0.047 (N.D. – 0.236), Crustacea 0.023 ± 0.021 (0.003 – 0.078) / 0.016 ± 0.018 (N.D. – 0.055), Mollusks 0.015 ± 0.015 (0.002 – 0.056) / 0.008 ± 0.013 (N.D. – 0.040). The concentration of MeHg in seafood were significantly correlated with total Hg concentration (p<0.001). The species with the highest average concentration of Hg was the Korean rockfish, but there was no sample that exceeded the maximum residual limit. The total %provisional tolerable weekly intake (%PTWI) value of MeHg for all of the samples was 3.76%, compared with the JECFA’s reference value, which indicates that there is almost no health risk from heavy MeHg intake through the consumption of seafood distributed in Daejeon.

The rapid development of computer vision and deep learning has enabled these technologies to be applied to the automated classification and counting of microscope images, thereby relieving of some burden from pathologists in terms of performing tedious microscopic examination for analysis of a large number of slides for pathological lesions. Recently, the use of these digital methods has expanded into the field of medical image analysis. In this study, the Inception-v3 deep learning model was used for classification of chondrocytes from knee joints of rats. Knee joints were extracted, fixed in neutral buffered formalin, decalcified, processed and embedded in paraffin, and hematoxylin and eosin (H&E) stained. The H&E stained slides were converted into whole slide imaging (WSI), and the images were cropped to 79 × 79 pixels. The images were divided into training (60.42%) and test (39.58%) sets (46,349 and 30,360 images, respectively). Then, images containing chondrocytes were classified by Inception-v3 and accuracy was calculated. We visualized the images containing chondrocytes in WSIs by adding colored dots to patches. When images of chondrocytes in knee joints were evaluated, the accuracy was within the range of 91.20 ± 8.43%. Therefore, it is considered that the Inception-v3 deep learning model was able to distinguish chondrocytes from non-chondrocytes in knee joints of rats with a relatively high accuracy. The above results taken together confirmed that this deep learning model could classify the chondrocytes and this promising approach will provide pathologists a fast and accurate analysis of diverse tissue structures.

A 5-year-old female fennec fox weighing 1 kg presented to the Veterinary Medical Center, Chungbuk National University with swelling of the mammary glands and hematuria. There were no significant complete blood count (CBC) or serum chemistry findings. The uterus obtained by ovariohysterectomy was filled with blood-clot-like material. The uterine fluid contained erythrocytes, a few leukocytes, and bilirubin crystals without bacteria. The progesterone concentration in the blood serum the day before ovariohysterectomy was 7.3 ng/ mL, which is higher than anestrus and lower than a true pregnant condition in a fennec fox. Despite the low blood progesterone concentration and absence of a fetus in the uterus, the histopathological zonary placental findings indicated that the fennec fox had been pregnant. This is the first case report of hematometra associated with fetal death in a captive fennec fox.

Testicular germ cell tumors (GCT) arise from embryonic or extraembryonic differentiated totipotential germ cells. The tumor cells can differentiate into germ cells as well as, other lineages including yolk sac tumors, choriocarcinomas, and teratomas. Mixed GCT is composed of more than one GCT component including one or more nonseminomatous elements in a tumor, accounting for one-third of GCT. Herein we report the case of a collision tumor with two distinct and separated GCTs in the testis, adjacent to each other. A 48-year-old, previously healthy man showed the hard swelling and heterogeneous enhancing mass in the right scrotum, and right orchiectomy was performed. Grossly, the lesion was two distinct and well-circumscribed masses in the testis. Microscopically, a larger tumor was immature teratoma (prepubertal type) and another smaller tumor was seminoma. These two tumors can be from a common precursor, germ cell neoplasia in situ (GCNIS); however, they are two distinct pathological entities. Given that teratomas can evolve from seminomas by additional genetic alterations, seminomas are also a precursor for postpubertal-type teratomas. Two distinct GCNISs may occur at different times. Because GCNIS is patchy distributed, a close gross examination must be performed in GCNIS or GCT to miss other GCTs with poor prognosis and to prevent under-treatment.

Leiomyosarcoma is malignant mesenchymal tumor of smooth muscle and commonly encountered in the gastrointestinal tract of dogs. However, primary canine leiomyosarcoma in oral cavity is rare due to lack of smooth muscle in the oral tissue. A 13-year-old, neutered male Poodle presented a hard and immobile mass on the left maxilla. Imprinting cytology from the mass as well as fine-needle aspirated cytology from the left scapular lymph node revealed predominant spindle cells met malignancy criteria of the tumors, including coarse chromatin, high N/C ratio, nuclear molding, macro/multi-nucleoli with cigar-shaped nucleus. Radiography of the skull showed lysis of the nasopalatine bone, and mineral radiopacity in the mass. Computed tomography showed soft tissue attenuating mass from the left incisor teeth to the left retrobulbar space with loss of nasopalatine bone and medial wall of orbit. The histopathological examination showed irregularly arranged malignant spindle-shaped cells with oval or elongated nuclei. The nucleolus is distinct and moderate cellular polymorphism is observed. Mitotic figures are occasionally observed. The tumor cells are positive to vimentin, desmin, α-smooth muscle actin when immunohistochemistry was performed, and in Masson’s trichrome stain, tumor cells are stained as red. Overall, histopathologic exam and immunohistochemistry confirmed canine oral leiomyosarcoma. Because of the poor prognosis, the owner did not consent further treatment.

Colorectal cancer causes the most cancer-associated death worldwide, having a high cancer incidence. Pectin is a complex polysaccharide present in various fruits, emerging as an anti-carcinogenic candidate. Although pectin has a suppressive capacity for colon carcinogenesis, the effect of reactive oxygen species (ROS) generation and colonic aberrant foci formation in the colon carcinogenesis mouse model remains unclear. Therefore, this study investigates the regulatory effect of pectin supplementation on colon carcinogenesis induced by azoxymethane (AOM) and dextran sodium sulfate (DSS) in mice. In an animal experiment, thirty male institute for cancer research (ICR) mice were divided into two experimental groups; AOM/DSS (control group) and AOM/DSS + pectin (5% in drinking water). Furthermore, the number of aberrant crypt foci (ACF) and aberrant crypt (AC) on colonic mucosa were counted, and thiobarbituric acid-reactive substances (TBARS) assay was performed to estimate lipid peroxidation in feces. Pectin treatment significantly decreased the number of ACF and AC per colon compared with the control. Additionally, fecal TBARS level in the pectin group was significantly lower than those in the control group. Conclusively, these findings indicate that pectin-inhibited hyperplastic alteration and oxidative stress suppress colitis-associated colon carcinogenesis.

We conducted diagnostic investigations to analyze the causes of abortions (46 cases, 65.7%), deaths (22 cases, 31.4%) and muscular lesions (2 cases, 2.9%) occurred after foot and mouth disease (FMD) vaccination in livestock farms in Korea. Bacterial culture, enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) were performed to detect the causative agents of abortion in bovine and caprine. The diagnostic results showed that 36 (51.4%) cases, referring as “Identified”, were occurred by influence of underlying disease including bovine viral diarrhea (12 cases, 17.1%), neosporosis (7 cases, 10.0%), septicemic colibacillosis (5 cases, 7.1%), Q fever (4 cases, 5.7%) and other abnormal conditions (8 cases, 11.4%) not by vaccination. Other 2 (3.0%) cases were suspected to be vaccine-associated adverse reaction on the basis of pathological findings (shock lung, oil-component-induced granuloma) and clinical symptoms (dyspnea with pulmonary edema). The other 32 (45.7%) cases were determined “Unknown” because any pathogens and pathological changes were not identified. However, many of the “Unknown” cases were presumptive to be the vaccine-related adverse reaction based on epidemiological investigation, especially, the cases which showed the clinical signs within 2 days after the vaccination. It is important to conduct pathological, microbilogical and epidemiological investigation to diagnose whether the cases are from vaccine-associated adverse reaction or not.

Patient-controlled sedation systems, in which the patient has control of sedative administration, have been shown to be effective and safe for patients undergoing minor procedures. Safety depends on the adequacy of the step-wise increase in dose of the sedative, the lockout time and patient’s ability to adjust the effective activation of the handset. While there is no patient-controlled sedation (PCS) system available domestically, we have developed a new purpose-built PCS pump, and determined its degree of precision using a microweight measuring scale. The PCS system was designed to be applied to most intravenous sedatives and work through a newly developed operating software algorithm that titrates the doses and lockout times. Our system titrates the doses through software modification, and for experimental purposes, the number of steps for handset activation by the patient was set to four, and the lockout time was set to 30 s. The degree of precision of the PCS system was tested using an electronic weighing scale. The comparison of the amount infused via the PCS system with weight on the scale showed nearly same patterns at the four infusion stages, and the difference was minimal (3% on average at each infusion step). We designed and manufactured a pilot button-type PCS pump capable of use by the patient for optimal sedation, and confirmed the precise working mechanism of the pump, thereby reducing the margin of error for infusion accuracy to 3% or less on average

Declined salivary gland function is commonly observed in patients with diabetes. Advanced glycation end products (AGEs) are believed to contribute to the pathogenesis of diabetes-induces hypofunction of the salivary glands. Polydatin (resveratrol-3-O-β-mono-D-glucoside) is a polyphenol that can be easily accessed from peanut, grape, and red wines. Although polydatin is known to have anti-glycation, anti-oxidation, and anti-inflammation effects, its effect in the salivary gland is not known. In the present study, we evaluated the AGEs burden in the salivary gland in streptozotocin (STZ)-induced diabetic rats and the protective effect of polydatin on diabetes-related salivary hypofunction. Polydatin (50 and 100 mg/kg/day) was orally administered in the STZ-induced diabetic rats for 4 weeks. The results showed that the salivary flow rate of the STZ-induced diabetic rats was reduced compared with that in the normoglycemic control rats. The circulating AGEs in serum and secreted AGEs in saliva increased in the STZ-induced diabetic rats. The reactive oxygen specises (ROS) were highly generated in the salivary gland in these diabetic rats. The salivary gland from the diabetic rats showed increased acinar cell apoptosis compared to normoglycemic control mice. However, polydatin suppressed all of these diabetes-related salivary changes. Overall, polydatin could provide a beneficial option for diabetes-related salivary hypofunction.

The natural relics excavated from the archeological site reveal the life, hunting, fishing, and gathering of mankind at the time, and the animal bones excavated from the remains become valuable data for the study of geography and bone diseases at the time. This study investigated the morphological structure, size, and osteometry of Sus scrofa bones excavated from the well area of the Gasan-Ri archaeological site in Jinju, presumed to be a relic of the Three Kingdoms period. The total weight of identified animal bones was 975.30 g and the number of bone fragments was 447. Among these 447 animal bones identified, the bone weight of Sus scrofa was 468.00 g (47.99%) and the number of bone fragments was 204 (45.64%). These 204 bone fragments of Sus scrofa consisted of 35 skulls (17.16%), 94 vertebrate axial skeletons (46.08%), 28 forelimb bones (13.73%), and 47 hindlimb bones (23.04%). The bones of Sus scrofa have been identified as those of two animals. The estimated age of these bones of Sus scrofa was 5-7 weeks. There was a fracture presumably caused by an artificial blow to the parietal bone located near the bregma in the skull.

A 19-year-old male Siberian tiger was presented with inappetence and paralysis of hind limbs. In a computed tomography (CT) scan, intervertebral disk disease at L3–L4 was detected. Cardiac arrest occurred during the surgery. At autopsy, myxomatous mitral valve degeneration (MMVD) and eccentric hypertrophy of the left heart were noted. The diagnosis was congestive heart failure caused by MMVD. Microscopically, myocardial and pulmonary fibrosis were observed in addition to the disintegration of the fibrosa layer and accumulation of glycosaminoglycans and proteoglycans in the spongiosa layer of the mitral valve. This is the first case of congestive heart failure with MMVD in a Siberian tiger.

Naturally occurring left ventricular hyperplasia is a rare but lethal disease. There are very few reports of this cardiac disease in captive nonhuman primates. In a colony of Macaca mulatta (Rhesus monkey) at California National Primate Research Center, a large number of rhesus macaques were diagnosed by autopsy with naturally occurring left ventricular hypertrophy without obvious underlying diseases over a 22-year period. The confirmatory diagnosis of ventricular hypertrophy was based on findings of notable left ventricular concentric hypertrophy with reduced left ventricular lumen, which is very similar to human ventricular hypertrophy cases. This report discusses an 11-year-old Macaca fascicularis monkey (Cynomolgus monkey, crab-eating macaque), weighing 2.95 kg, that was presented for enrollment in a pharmacokinetic (PK) study. During the PK experiment, the monkey died following a sudden decrease in percutaneous oxygen saturation and heart rate. Gross and histological examinations of the heart were performed. On gross pathology, the left ventricular wall was thickened, and the chamber lumen was reduced. In histopathological examination using hematoxylin- eosin and Masson-trichrome stains, fibrosis and myocyte disarray were not observed, but an increased cell density, compared to the normal heart, was confirmed. The autopsy results confirmed left ventricular hyperplasia as the major cause of death.

Alzheimer’s disease (AD) is an irreversible and progressive neurodegenerative disease accompanied by aging, followed by memory impairment and cognitive decline. Although numerous attempts have been made to develop treatments for AD, most clinical trials have failed to delay or stop the progression of AD. Electroacupuncture (EA) is a complementary alternative medicine technique widely used to treat pain, inflammation, and neurodegenerative diseases. Additionally, blood-brain barrier (BBB) disruption is a known pathophysiology of neurodegenerative diseases, including AD. Moreover, amyloid beta deposition increases BBB permeability and produces inflammatory cytokines induced by glial activation. However, our previous study revealed that EA treatment at the Taegye acupoints (KI3) improves memory impairment through anti-neuroinflammation and increases glucose metabolism in 5XFAD mice. Therefore, we evaluated whether EA treatment at KI3 regulates BBB dysfunction in the prefrontal cortex of 5XFAD mice. For this study, 6.5-month-old 5XFAD mice were treated with EA stimulation at KI3 three times a week for two weeks. Western blotting, immunohistochemistry, and flow cytometry were used to evaluate the effects of EA treatment on BBB dysfunction. We found that EA stimulation attenuates BBB integrity by protecting BBB tight junction proteins (CD31, aquaporin 4, occludin, and claudin 5) in the prefrontal cortex of 5XFAD mice. In addition, EA treatment regulated inflammatory cytokines (IL-1α, IL-1β, IL-17, IL-23, IFN-ɣ, monocyte chemoattractant protein 1 (MCP-1), granulocyte-macrophage colony stimulating factors [GM-CSF], and IL-10) in the peripheral circulation of 5XFAD mice. Therefore, our data suggest that EA treatment could be a therapeutic agent for enhancing BBB dysfunction in AD.

Colon cancer has been considered a leading cause of cancer-associated death. Folic acid is a vitamin necessary for cellular physiological functions and cell viability. However, the association between folic acid intake and colon cancer has been examined in several prospective cohort studies are controversial. This study investigated the effects of folate intake on colon carcinogenesis and oxidative stress in an azoxymethane (AOM)/dextran sodium sulfate (DSS) institute for cancer research (ICR) mouse model. Thirty male ICR mice (5 weeks old) were divided into the control group and the experimental group supplied 0.03% folic acid via drinking water (50 mL/week/mouse) for 6 weeks. To induce colonic pre-neoplastic lesions, the animals were subcutaneously injected three times weekly with AOM (10 mg/kg body weight), followed by 2% DSS in drinking water for a week. Folic acid supplementation significantly suppressed the total number of aberrant crypt foci and aberrant crypts. Histological image data showed that folic acid supplementation attenuated neoplastic change. In addition, we measured the thiobarbituric acid reactive substances concentration of dry feces samples to identify the effect of folic acid on reactive oxygen accumulation. The folic acid supplementation group had reduced reactive oxygen species levels in dry feces compared to the control group. In conclusion, these findings indicate that folic acid suppresses colon carcinogenesis and oxidative stress in an AOM/DSS mouse model.

C-reactive protein (CRP) is an acute-phase protein produced by hepatocytes. The plasma concentration of CRP peaks around 48 hours following stimulation. A rapid decrease in CRP levels is observed when the stimuli end. Considering these characteristics of CRP, it is used to evaluate acute inflammatory responses in clinics. In addition, as it reflects the degree of surgical trauma, measuring the CRP concentration is an effective method of evaluating tissue damage depending upon the surgical method. As CRP is also an acute-phase protein in dogs, the purpose of the present study was to compare the factors related to acute inflammatory responses between dogs undergoing elective general surgery (GS) and orthopedic surgery (OS). A total of 44 healthy dogs were included in this study. The sex ratio of dogs that underwent OS was similar. Both groups showed increased CRP levels and white blood cell (WBC) counts on post-operative day (POD) 1-2. The CRP level in the OS group on POD 1-2 was significantly higher than that of the GS group and it was increased about 16-fold compared to the pre-operative value. No significant difference in WBC counts was observed between the two groups. Although a slight increase in the WBC count was observed on POD 1-2 in both groups, it was near the reference range during follow-up. Thus, measuring CRP levels can be more effective in evaluating acute inflammatory responses than WBC counts. Although a high level of post-operative serum CRP concentrations was observed in the OS group, it rapidly decreased to a level similar to the pre-operative value if there were no post-operative complications.

An 8-year-old, spayed female Maltese dog was presented with a one-month history of erythema, swelling and alopecia of periocular region with pruritus. The skin lesions were first detected at the age of three years, but this was the first time that symptoms had appeared in the eyes. Physical examination revealed markedly swollen and erythematous eyelids and conjunctiva including Meibomian glands. In addition, periocular alopecia and tears were identified. Slit lamp microscopy revealed erythema of conjunctiva and swelling of Meibomian glands. The Schirmer’s tear test was normal. Impression smear cytology of eyes revealed sterile neutrophils and corneal epithelial cells. There were no virus or bacterial infections in the eyes. On skin examination, generalized erythema was detected but there were no other skin lesions. A case of allergic blepharoconjunctivitis associated with canine atopic dermatitis was diagnosed based on history taking and skin examination. Treatment included cetirizine, cyclosporine, prednisolone and Forus eye drops®. The owner was instructed to wear an Elizabethan collar around his dog's neck at all times and restrict walking. And the diet was changed to hypoallergenic dog food. Medications and environmental restrictions significantly reduced erythema, edema and swelling of meibomian glands. Pruritus was also decreased. Six weeks later, the edema lesions of eyes disappeared.

Aluminum (Al) is one of the most widely applied metals in various industries. Anthropogenic activities of industrial waste result in increased accumulation of Al in natural water resources and aqueous organisms, leading to increased heavy metal pollution in the environment. This will ultimately associate with health risks to all living beings including humans. The present study addresses the possible toxic effects of Al on the motility and regeneration of planarians, using Dugesia japonica as the experimental animal model. Planarians were exposed to varying concentrations of Al (50–1,200 mg/L) for 1 hr, and subsequently evaluated for their motility, seizure-like behaviors, regeneration and alterations of the cell-organelles in their body. Results of the study exhibit that increasing Al concentrations lead to decreased motility and increased seizure-like behaviors, especially the c-type and head bob behaviors in planarians (p<0.05). The regeneration ability in the tail segments was reduced by increased Al concentrations, and the eyespot and optic nerves were more clearly observed in the control groups than planarians exposed to Al. Organelle analysis revealed morphological abnormalities in the mitochondria, golgi apparatus, endoplasmic reticulum, and cilia in the head region of planarians, as compared to the control groups. Taken together, our results indicate that exposure to Al alters the behavior of planarians and delays the regeneration of amputated body parts. Therefore, we suggest that Al exposure adversely can affect the lives of aquatic organisms, and induce toxicity such as cell abnormalities in animals.

Fowl adenovirus serotype-4 (FAdV-4) infection, also known as hydropericardium-hepatitis syndrome (HHS), is one of the most prevalent diseases in the Korean poultry industry. Therefore, an effective vaccination against FAdV-4 should be developed for prevention of HHS infection. However, a standard animal model with significant pathologic characteristics has not been established for evaluation of HHS vaccine efficacy. In this study, we comparatively evaluated FAdV-4 pathogenicity from specific- pathogen-free (SPF) chickens using FAdV-4 strain (ADL091024) isolated in laying chickens in Korea according to chicken age, passage number and virus titers. In brief, FAdV-4 passaged four times in yolk sac of SPF eggs with the highest viral titers could induce the most severe clinical signs in 1-day-old chickens, similar to natural HHS outbreaks. Furthermore, the efficacy of a newly developed attenuated live FAdV-4 vaccine candidate was successfully determined using the established 1-day-old SPF chickens. There was no significant pathogenicity based on the criteria such as mortality, body weight, gross lesions, histological lesions and virus detection rates in the vaccination and non-challenge control groups. However, the challenge group without vaccination showed significant pathogenicity including hepatic necrosis, histological lesion scores in the liver and heart, virus replication and decreased body weight gain. In conclusion, the chicken challenge model established in this report was proposed for consistent and reliable evaluation of FAdV- 4 vaccine efficacy.

The young shoots of Aralia elata, Chaenomeles sinensis fruit and Glycyrrhizae radix are edible and traditionally used as anti-inflammatory and antioxidant agents. The present study was performed to investigate the protective effect of an ethanol extract mixture of these three medicinal plants (ACG) against amyloid β protein (Aβ) (25– 35)-induced memory impairment in an ICR mouse model. Memory impairment was induced by intracerebroventricular microinjection of 15 nmol Aβ (25–35) and assessed using the passive avoidance test and the Morris water maze test. The step-through latency in the passive avoidance test was decreased and the latency to reach the hidden platform in the Morris water maze test was increased in mice treated with Aβ (25–35), indicating memory impairment. This memory impairment induced by Aβ (25–35) was significantly prevented by chronic treatment with ACG (10, 25, and 50 mg/kg, p.o., 8 days). In memory impaired mice brain, cholinesterase activity and concentration of thiobarbituric acid reactive substance, a lipid peroxidation marker, were increased and glutathione level was decreased. These biochemical changes in Aβ (25–35)-treated mice were reversed by chronic administration of ACG. The present results suggest that antioxidant and anti-cholinesterase activities of ACG might be responsible for the inhibition of Aβ (25– 35)-induced memory impairment and that ACG preparation may have a therapeutic role in preventing the progression of Alzheimer’s disease.

Lead (Pb) is a major heavy metal that augments environmental pollution and is a health risk to living organisms. This study was performed to investigate the effect of lead on characteristics of planarian, Dugesia japonica. Briefly, planarians were cultivated in water containing different concentrations (0–400 mg/L) of Pb, and incubated for varying durations (1, 3, and 5 hr). After termination of the incubation time, motility and seizure-like behaviors (c-like, head-bop, snake-like, and screw-like) of the planarians were counted in fresh water. Results showed that increasing Pb concentration and time resulted in decreased motility of the planarians (p<0.05). Increasing concentrations of Pb also resulted in significant increase in the seizure-like behavioral patterns, in particular the c-like and head-bop behaviors. In order to examine eye regeneration, the head region was cut, and rest of the body was incubated in water in the absence or presence of Pb, until emergence of the eye extrusion. Formation of eye spots was initiated in amputated fragments of the control group (without Pb) on day 4 after incubation, whereas appearance of the eye spots was delayed in planarian exposed to 200 mg/L Pb. Moreover, immunohistochemistry revealed that formation of the optic nerve was delayed in planarians exposed to Pb. Thus, our studies determined that planarians exposed to high concentrations of Pb resulted in decreasing motility and induction of seizure-like behaviors, as well as delayed eye regeneration. Results of the current study therefore validate that exposure to lead has a negative effect on the lifespan of aquatic organisms and can cause disturbance of the nervous system in animals, thereby implying the possibility of threatening health.