The purpose of this study was to investigate diethylnitrosamine (DEN)-induced liver damage in zebrafish. Zebrafish larvae were divided into five groups after seventy-two hours fertilization: group 1 (G1) as control, group 2 (G2) as probe control, groups 3, 4, and 5 (G3, G4, and G5) as DEN treated at doses of 25, 50, and 100 μg/mL, respectively. At twenty-two hours after DEN treatment, groups 2, 3, 4, and 5 were treated with ApoFlamma H 675 at a dose of 100 μM/zebrafish. They were examined by fluorescence stereomicroscope at twenty-four hours after DEN treatment. After fixation, the zebrafish were processed, embedded, sectioned and stained with hematoxylin and eosin (HE) and terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL) staining. Fluorescence intensity of the livers of G3, G4, and G5 was significantly increased compared with those of G1 (p<0.01). Furthermore, fluorescence intensity of the livers of G3 and G5 was significantly increased compared with those of G2 (p<0.05 and p<0.01). HE staining showed cell deaths in the livers of DEN-treated zebrafish and TUNEL staining confirmed cell death in the same location. Taken together, in vivo fluorescence bioimaging detected cell death in the liver of DEN-treated zebrafish. This outcome was confirmed with histopathological examination. The results of this study provide confidence for using zebrafish as a liver carcinogenesis model.

Acute myocardial infarction (AMI) is considered the major cause of mortality in the world. Tremendous animal studies are performed to develop novel therapeutics, and this study aimed to induce porcine myocardial infarction model by using polyethylene terephthalate (PET). Coronary guidewire was placed in left anterior descending artery (LAD). The balloon angioplasty catheter was inserted at the back of the PET. The balloon catheter was carefully pushed forward, until the balloon marker was located in mid-LAD. Coronary angiography was performed pre- and post-occlusion at 28 days by C-arm. Histologic analysis of heart tissue was performed 28 days after inducing AMI. Thirty three pigs were anesthetized and underwent percutaneous coronary catheterization. All pigs were successfully embolized in mid-LAD by PET. Fifteen pigs died due to ventricular fibrillation during post-anesthetic recovery time, and overall experiment mortality was 45.5%. In 2,3,5- triphenyl tetrazolium chloride staining, gross finding of the ischemic heart lesion showed firm and white area of infarction associated with the apex and left ventricular posterior wall. Infarct on H&E-stained sections demonstrated a region without myocytes and rich with cardiomyocyte with atypical nuclei. Successful induction of AMI by using PET may provide the pathophysiological information of ischemic heart disease and improvement of therapy development for AMI.

This study investigated the prokinetic effect of metoclopramide and mirtazapine on gastric transit time (GTT), small bowel transit time (SBTT) and gastrointestinal transit time (GITT) during capsule endoscopy in four healthy beagle dogs. Four beagle dogs participated in the experiment as four groups at intervals of more than three days as the following: Control group 1 (capsule alone), Control group 2 (capsule alone), Metoclopramide administered group (metoclopramide + capsule) and Mirtazapine administered group (mirtazapine + capsule). The results of this study demonstrated there was no significant difference in GTT ([min] control group 1: 105 ± 90, control group 2: 172.5 ± 102 vs metoclopramide administered group: 247.5 ± 93, p = 0.07, 0.10) and SBTT ([min] control group 1: 120 ± 88, control group 2: 75 ± 39 vs metoclopramide administered group: 37.5 ± 15, p = 0.20, 0.18) for capsule only administered groups (control group 1 & 2) compared to metoclopramide administered group. In addition, there was no significant difference in GTT ([min] control group 1: 105 ± 90, control group 2: 172.5 ± 102 vs mirtazapine administered group: 127.5 ± 45, p = 0.56, 0.36) and SBTT ([min] control group 1: 120 ± 88, control group 2: 75 ± 39 vs mirtazapine administered group: 157.5 ± 38, p = 0.29, 0.07) between capsule only administered groups (control group 1 & 2) and mirtazapine administered group. In this study, the fact that metoclopramide might be ineffective and administration of mirtazapine might be inadequate in dogs were confirmed.

The occurrence of allergic disease has increased harmfully in the last few decades. Atopic dermatitis (AD) is an allergic inflammation disorder characterized by itchy, red, swollen, cracked skin. Although the pathogenesis of AD is not fully understood, it is assumed that deregulation of T-helper 1 (Th1) and T-helper 2 (Th2) immune responses, a predominance of allergen-specific IgE, and interrupted epidermal barrier function are keys to the pathogenic mechanism. Activated T helper 2 (Th2) immune function is hallmark of various allergic diseases. Oxidative stress implicated in cutaneous damage in various inflammatory skin diseases. We investigated the effect of fermented soybean (SCGB1) on the improvement of AD. Soybean fermentation was carried out using B. amyloliquefaciense SCGB1 (KCCM11964P), which is known to produce of natural antibiotics. And then, we experiment of SCGB1 and soybean powder (NC) in DNCB-induced AD model. Mice were respectively oral administration of variety dose for 14 days. As a results, it was confirmed that serum Immunoglobulin E (IgE) expression was dose-dependently decreased in SCGB1 and NC compared to negative control, and it was reduced the skin pruritus inducing factor that Interleukin-31 (IL-31) mRNA level. In addition, the inflammatory cells were infiltration in skin for histological analysis. As a result, it reduced that epidermal hyperplasia, cancellation and aveolarization compared to negative control. These results suggest that SCGB1 may be effective for prevention and treatment of AD.

Nicotine is a component of tobacco products and is one of the most commonly abused substances that leads to addiction. Therefore, the present study was performed to investigate the behavioral pattern and toxicity by nicotine exposure in planarians. Basically, planarians were exposed to different concentration of nicotine for 5 min. To investigate detoxification effect, planarians were exposed to nicotine for 5 min, and treated glycyrrhizin for 5 min, then motility and seizure-like behavior were observed for 5 min. As a result, the motility of nicotine-exposed planarians decreased approximately more than 50% compared to freshwater control. However, the motility of glycyrrhizin-exposed planarians recovered than nicotineexposed planarians. In the assessment of seizure-like behavioral pattern, planarians exposed to nicotine showed head-bop or c-like type rather than screw-like or snake-like patterns. However, planarians exposed to glycyrrhizin showed no seizure-like behavior. To examine the oxidative stress response, planarians were cultured in fresh water containing 1 mM nicotine for 1 day. Planarians were homogenized and extracted to assay the contents of reactive oxygen species (ROS), lipid hydroperoxides (LH), glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD). The result showed that a significantly higher level of ROS, LH indicated in planarians exposed to nicotine, on the other hand, glycyrrhizin-exposed planarians were significantly decreased ROS, LH levels. In conclusion, the motility decreased when planarians were exposed to nicotine, in a dose-dependent manner, whereas seizurelike behavior increased. Nicotine induced behavioral disturbances and cell toxicity in planarians were recovered by glycyrrhizin, suggesting a candidate substance for nicotine addiction treatment.

The incidence of stomach cancer has been found to be gradually decreasing; however, it remains one of the most frequently occurring malignant cancers in Korea. According to statistics of 2017, stomach cancer is the top cancer in men and the fourth most important cancer in women, necessitating methods for its early detection and treatment. Considerable research in the field of bioinformatics has been conducted in cancer studies, and bioinformatics approaches might help develop methods and models for its early prediction. We aimed to develop a classification method based on deep learning and demonstrate its application to gene expression data obtained from patients with stomach cancer. Data of 60,483 genes from 334 patients with stomach cancer in The Cancer Genome Atlas were evaluated by principal component analysis, heatmaps, and the convolutional neural network (CNN) algorithm. We combined the RNA-seq gene expression data with clinical data, searched candidate genes, and analyzed them using the CNN deep learning algorithm. We performed learning using the sample type and vital status of patients with stomach cancer and verified the results. We obtained an accuracy of 95.96% for sample type and 50.51% for vital status. Despite overfitting owing to the limited number of patients, relatively accurate results for sample type were obtained. This approach can be used to predict the prognosis of stomach cancer, which has many types and underlying causes.

The use of non-therapeutic antibiotics as animal feed additives has raised public health concerns due to the increasing resistance of pathogens to antibiotics. It is therefore required to develop safe and effective alternative feed additives to replace non-therapeutic antibiotics. The aim of this study was to assess the effects of the multiherbal compound, KIOM-C, on growth performance and immune response of growing-to-finishing pigs under farm conditions. The experimental trials were performed in a Korean commercial swine growing-to-finishing complex, and a total of 70-day-old 160 pigs were selected. Eighty pigs were treated with KIOM-C at the level of 2 kg/tonne until slaughter age (KT group), while another 80 pigs were not treated with KIOM-C (NT group). All animals were vaccinated against foot-and-mouth disease (FMD) at 60 and 110 days of age. During the trial period, average daily weight gain (ADWG), average daily feed intake (ADFI), feed conversion ratio (FCR), survival rates, and average slaughter ages were measured. The serum concentrations of tumor necrosis factor-a (TNF-α), interferon-γ (IFN-γ), and IgA were also evaluated. In order to evaluate specific humoral immune responses, the foot-and-mouth disease virus (FMDV) serotype O-specific antibody was measured. The ADWG, ADFI, and FCR of the KT group were significantly greater than those of the NT group (p<0.05). Serum concentrations of IgA in the KT group was statistically higher than the NT group. The antibody levels of the KT group against FMDV serotype O was higher than the NT group, and 86.67% of the KT group tested positive for anti-FMDV antibodies. Overall, these findings suggest that KIOM-C improves growth performance and immune response of pigs under growing-to-finishing farm conditions, and implies that the herbal compound may be used as a suitable alternative feed additive.

Inflammation is an important protective response mechanism that occurs against microbial invasion or injury. However, excessive inflammation may lead to cause of morbidity and mortality in diseases. The activated macrophages plays a vital role in inflammatory response by stimulation of lipopolysaccharide (LPS) and tumor necrosis factor-α (TNF-α). This activation further damages the host by inducing certain pro-inflammatory mediators such as nitric oxide (NO), interleukin-1β (IL-1β), interleukin- 6 (IL-6), TNF-α, inducible nitrous oxide (iNOS) and cyclooxygenase (COX-2). Flavonoids are bioactive compounds with potential effects as anti-cancer, anti-inflammation, anti-viral and anti-bacterial activities. Polymethoxyflavones (PMFs) are unique to citrus plants which are of specific interest owing to their biological effects that includes lipoprotein metabolism and anti-inflammatory activity. Sinensetin is one of the PMFs having five methoxy groups on the basic benzo-γ-pyrone skeleton with a carbonyl group at the C4 position. Sinensetin have been known for exerting various pharmacological activities including anti-angiogenesis, anti-diabetic and anti-inflammatory activities. However, there are no studies focused on the anti-inflammatory effects of sinensetin on skeletal muscle cells. In the present study, we investigated the antiinflammatory effect of flavonoids isolated from Sinensetin on the production of pro-inflammatory mediators mediated by nuclear factor-kappa B (NF-κB) by inhibition of signal transduction in LPS - induced L6 skeletal muscle cells.

A one-year-old, intact female, Maltese dog was presented with a history of anorexia and regurgitation. Thoracic radiographs and ultrasonography scans suggested an abnormal mass in the cranial mediastinal region, and computed tomography confirmed the origin of this mass. Ultrasound-guided fine needle aspiration cytology showed the presence of intermediate to large lymphoid cells showing mitotic figures. B-cell lymphoma was confirmed by the result of a polymerase chain reaction assay for antigen receptor rearrangement, therefore the patient was diagnosed with primary mediastinal large B-cell lymphoma (PMBL). The patient underwent L-CHOP (L-asparaginase, cyclophosphamide, doxorubicin, vincristine, prednisolone)-based chemotherapy, and showed complete tumoral remission from the beginning of chemotherapy. Seventytwo weeks after the completion of chemotherapy, the patient is still alive without any evidence of metastasis or relapse. A standardized treatment protocol has yet to be established for primary mediastinal lymphoma in dogs. This case report describes the complete remission of PMBL by an L-CHOP-based chemotherapy protocol in a young Maltese. Clinicians should consider that L-CHOP based chemotherapy may be useful against PMBL in dogs.

This study aimed to analyze a high-performance liquid chromatography (HPLC) separation using a pentafluorophenyl column of parent drug hydroxychloroquine (HCQ) and its active metabolite, desethylhydroxchloroquine (DHCQ) applying to determine bioequivalence of two different formulations administered to patients. A rapid, simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for bioanalysis of HCQ and its metabolite DHCQ in human whole blood using deuterium derivative hydroxychloroquine-D4 as an internal standard (IS). A triple-quadrupole mass spectrometer was operated using electrospray ionization in multiple reaction monitoring (MRM) mode. Sample preparation involves a two-step precipitation of protein techniques. The removed protein blood samples were chromatographed on a pentafluorophenyl (PFP) column (50 mm × 4.6 mm, 2.6 μm) with a mobile phase (ammonium formate solution containing dilute formic acid) in an isocratic mode at a flow rate of 0.45 mL/min. The standard curves were found to be linear in the range of 2 – 500 ng/mL for HCQ; 2 – 2,000 ng/mL for DHCQ in spite of lacking a highly sensitive MS spectrometry system. Results of intra- and inter-day precision and accuracy were within acceptable limits. A run time of 2.2 min for HCQ and 2.03 min for DHCQ in blood sample facilitated the analysis of more than 300 human whole blood samples per day. Taken together, we concluded that the assay developed herein represents a highly qualified technology for the quantification of HCQ in human whole blood for a parallel design bioequivalence study in a healthy male.

Dopaminergic neurons are one of the major neuronal components in the brain. Mesencephalon dopamine (DA) neurogenesis takes place in the ventricular zone of the floor plate, when DA progenitors divide to generate postmitotic cells. These cells migrate through the intermediate zone while they differentiate and become DA neurons on reaching the mantle zone. However, neurogenesis and neuronal migration on dopaminergic neurons remain largely unexplored in the mesencephalon development. This study presents neurogenesis and neuronal migration patterns of dopaminergic neurons during mesencephalic development of the mouse. Neurons from embryonic day (E) 10–14 were labelled by a single injection of 5-bromodeoxyuridine and immunohistochemistry was performed. The neurogenesis occurred mainly at the E10 and E11, which was uniformly distributed in the mesencephalic region, but neurons after E13 were observed only in the dorsal mesencephalon. At the postnatal day 0 (P0), E10 generated neurons were spread out uniformly in the whole mesencephalon whereas E11-originated neurons were clearly depleted in the red nucleus region. DA neurons mainly originated in the ventromedial mesencephalon at the early embryonic stage especially E10 to E11. DA neurons after E12 were only observed in the ventral mesencephalon. At E17, E10 labelled neurons were only observed in the substantia nigra (SN) region. Our study demonstrated that major neurogenesis occurred at E10 and E11. However, neuronal migration continued until neonatal period during mesencephalic development.

Korean Native Pig (KNP) has a uniform black coat color, excellent meat quality, white colored fat, solid fat structure and good marbling. However, its growth performance is low, while the western origin Yorkshire pig has high growth performance. To take advantage of the unique performance of the two pig breeds, we raised crossbreeds (KNP × Yorkshire to make use of the heterotic effect. We then analyzed the liver transcriptome as it plays an important role in fat metabolism. We sampled at two stages: 10 weeks and at 26 weeks. The stages were chosen to correspond to the change in feeding system. A total of 16 pigs (8 from each stage) were sampled and RNA sequencing was performed. The reads were mapped to the reference genome and differential expression analysis was performed with edgeR package. A total of 324 genes were found to be significantly differentially expressed (|log2FC| > 1 & q < 0.01), out of which 180 genes were up-regulated and 144 genes were down-regulated. Principal Component Analysis (PCA) showed that the samples clustered according to stages. Functional annotation of significant DEGs (differentially expressed genes) showed that GO terms such as DNA replication, cell division, protein phosphorylation, regulation of signal transduction by p53 class mediator, ribosome, focal adhesion, DNA helicase activity, protein kinase activity etc. were enriched. KEGG pathway analysis showed that the DEGs functioned in cell cycle, Ras signaling pathway, p53 signaling pathway, MAPK signaling pathway etc. Twenty-nine transcripts were also part of the DEGs, these were predominantly Cys2His2-like fold group (C2H2) family of zinc fingers. A protein-protein interaction (PPI) network analysis showed that there were three highly interconnected clusters, suggesting an enrichment of genes with similar biological function. This study presents the first report of liver tissue specific gene regulation in a cross-bred Korean pig.

Transient ischemic attack (TIA) indicates high risk for major stroke and is considered a medical emergency. Diffusion-weighted imaging (DWI) enables detection of acute ischemic lesions. The clinical significance of DWI positive lesions in TIA is obscure and its prevalence, clinical features are not established. Therefore, we performed a clinical, etiological and prognostic analysis through a cross-sectional analysis of 235 TIA patients, grouped according to presence of DWI lesion. Clinical features, underlying risk factors for stroke, outcome and rate of recurrence were analyzed. 3 months follow-up of modified Rankin Scales (mRS) were done with telephone survey. DWI positive lesions were present in 14.0% of patients. Etiological factors significantly associated with DWI lesions in TIA patients were male sex (p = 0.038), stroke history (p = 0.012) and atrial fibrillation (p < 0.001). Presence of at least one medium or high risk of cardioembolism from TOAST classification were not associated with lesions when excluding association to atrial fibrillation (p = 0.108). Clinical features showed no significant difference. Whether the patients had lesion-positive DWI was not related to an increase in mRS score during the hospital stay or at the 3-month follow-up after discharge. Future studies should include multi-center samples with large numbers, considering each unique medical environment. Routine acquisition of follow-up DWI for proper evaluation of the tissue-based definition of TIA should also be considered.

Aspergillus luchuensis 34-1 was inoculated into wheat pellets with different conditions of raw materials to produce nuruk. The degree of substrate reactivity improvement of steam treated raw materials compared with that of non-heat treated was analyzed. The water content of the pellet was adjusted to 25% and 35%, and steam treatment for 10 minutes improved the substrate reactivity at 2.1-fold and 3.1-fold, and sterilization was also possible. The characteristics of improvement pellet nuruk were investigated according to the degree of crushing and water content of raw materials according to the temperatures and humidities (23°C, 30°C and RH 60%, RH 80%). The pH of the pellet nuruk was higher depending on the temperature, humidity and moisture content of the koji were lower, and the pH of the flour-pellet nuruk was lower than that of 2 mm milling wheat-pellet nuruk according to milling degree. It can be seen that the milling degree affects the growth of mold. The acidity and amino acid were generally higher as fermentation time increased. Also, the higher the incubation temperature, humidity and moisture content, the higher the value. Glucoamylase activity was significantly the highest in moisture content 35% D2b nuruk, cultured at 30 °C and 80% RH for 38 hours. This is higher than the previous reports on glucoamylase of rice-koji or commercial nuruk using fungi isolated from traditional nuruk. From these study, it is expected that making of improvement pellet nuruk would save the fermentation time considerably compared with traditional nuruks.

The objectives of this study were to find out the plant to enhance immune activity among 42 kinds of foods frequently consumed by the Korean elderly consisting of 5 food groups and 5 wild plants. Each sample was assessed the immunoactive effect by measuring NFkB/ AP1 gene expression, nitric oxide and cytokine production in RAW-Blue™ cell. Soybean sprouts of 47 plants showed the highest NF-κB/AP1 gene expression at the level of 1.13 ± 0.03 (O.D. 650 nm) and Soritae, sweet potato, banana, apple, garlic, crown daisy, cabbage and Ailanthus altissima also had high activity of NF-kB/AP1 gene in RAW-Blue™ cell stimulated by LPS. NO production of Ailanthus altissima was significantly higher than that of other plants and 16 plants of glutinous sorghum, black rice, Seoritae, Heuktae, sweet potato, banana, apple, garlic, mungbean sprouts, spinach, crown daisy, young pumpkin, cabbage, soybean sprouts, Actinidia arguta and Aster scaber were the next best activity. The above results selected 17 out of 47 plant samples. Moreover, soybean sprouts was significantly shown to increase TNF-α (1,509.55 ± 1.38 pg/mL) and IL-1β (54.56 ± 1.08 pg/mL) cytokines in comparison with RAW-Blue cell stimulated by LPS. According to the results of in vitro evaluation, the ethanol extract of soybean sprout increased the production of immune-enhancing cytokines by proliferation of macrophages. In addition, NF-kB transcription factor activity and NO production ability were excellent, and it was selected as a material having excellent immunological activity.

Angiotensin receptor blockers, such as telmisartan, are considered effective in the treatment of hypertension and proteinuria due to chronic kidney disease (CKD) in cats. It selectively blocks the AT1 receptor and does not affect the AT2 receptor, thus effectively blocking the activity of the renin angiotensin aldosterone system. This study aims to compare over time the changes in various indicators, including systemic hypertension and proteinuria, before and after the administration of telmisartan in cats with CKD. Decrease in blood pressure (BP) (p<0.001) and urine protein to creatinine (UP/C) ratio (p<0.001) were found to be statistically significant over time after the administration of telmisartan. BP and the UP/C ratio were 160 ± 22.2 and 0.50 ± 0.647 before telmisartan administration (Day 0), 150 ± 21.0 and 0.27 ± 0.487 on the 30th day (Day 30), 150 ± 17.0 and 0.25 ± 0.376 on the 60th day (Day 60), and 140 ± 17.8 and 0.15 ± 0.233 on the 90th day (Day 90) after administration, respectively. BP and UP/C were statistically significantly lower in cats with CKD over time at each time point from Day 0 to Day 90 at 30 day intervals. Especially after 90 days of telmisartan administration, the improvement of BP and UP/C were estimated to be about 20 mmHg and 0.35, respectively. In conclusion, the oral administration of telmisartan to cats with CKD is effective in improving BP and proteinuria, which has a positive effect on long-term survival in cats with CKD.

Echocardiography is one of the most useful diagnostic techniques for differentiating heart disease as well as mitral valve lesion. Forty client-owned small breed dogs (weight, 2.3−13.2 kg) aged between 8−17 years with myxomatous mitral valve degeneration (MMVD) were included in the present study. The diagnosis of MMVD in dogs was made based on the clinical signs, chest radiography data, and echocardiographic findings. Echocardiographic examinations were conducted in accordance with recommended standards for dogs. M-mode, Doppler, and 2D echocardiography were performed in left and right lateral recumbency. 2D echocardiography was used to measure LA and Ao diameter from 2D short axis at the level of the aortic valve. In the comparison of conventional echocardiography indices in dogs with different stages of heart failure with MMVD, significant differences were observed in E/A ratio (p=0.005), EDV (p<0.001), EDVI (p<0.001), E-peak velocity (p= 0.001), ESV (p=0.028), ESVI (p=0.004), LA (p<0.001), LA/Ao Ratio (p<0.001), LVIDd (p<0.001), LVIDd/Ao Ratio (p<0.001), LVIDs (p=0.036), LVIDs/Ao Ratio (p=0.002), and MR Velocity (p=0.026). In addition, distinct correlations were found in EDV (r=0.712), LA/Ao ration (r=0.830), LVIDd (r=0.724), and LVIDd/Ao ratio (r=0.759). This study found that known conventional echocardiographic indices, including EDV, LA/Ao ratio, LVIDd dimension, and LVIDd/Ao ratio correlated with the severity of MMVD in point of significant differences and distinct correlations.

Osteoporosis is a major worldwide public health problem that poses a great economic burden to society. Puerariae Radix, the dried root of Pueraria lobata (Wild.) Ohwi, has been widely used in Asia. This study investigated the effects of Puerariae Radix (PR) on bone loss in ovariectomized (OVX) mice. C3H/HeN mice (10 weeks old) were divided into sham and OVX groups. The OVX mice were treated with vehicle, 17β-estradiol (E2), PR (oral administration, 250 mg/ kg/day) or PR (intraperitoneal administration, 50 mg/ kg/every other day) for 6 weeks. Grip strength, uterus weight, serum alkaline phosphatase (ALP), estradiol concentration and osteoclast surface levels were measured. Tibiae were analyzed using microcomputed tomography. There were no significant differences in the degree of grip strength, body weight and uterine weight between OVX group and PR-treated group. As compared with the OVX group, the serum estradiol levels were significantly increased in the PR-treated group. PR (i.p.) significantly preserved trabecular bone volume, trabecular bone number, structure model index and bone mineral density of proximal tibiae metaphysic. The administration of PR lowered serum ALP and osteoclast surface levels in OVX mice, suggesting that PR can reduce the bone turnover rate in mice. The results indicate that the supply of PR can prevent OVX-induced bone loss in mice.

The purpose of this study was to investigate probiotic characteristics and immune enhancing effects of Bifidobacterium (B.) longum KBB1-26 and BIF-4, B. breve KBB5-22 isolated from human intestine for probiotic use in humans and animals. We measured acid, bile and heat tolerance, antimicrobial activity against pathogenic bacteria, Escherichia (E.) coli, Salmonella (S.) Enteritidis, Staphylococcus (S.) aureus, and Listeria (L.) monocytogenes. Immune enhancing effects of B. longum and B. breve were investigated by measuring nitric oxide (NO), nuclear factor (NF–κb), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-12 (IL-12) and tumor necrosis factor- α (TNF- α) in RAW 264.7 cells or RAW BLUE cells. B. longum KBB1-26 was survived at pH 2.0. B. longum KBB1-26 and BIF-4, B. breve KBB5-22 also showed tolerance to 0.3% of oxgall bile salt. B. longum KBB1-26 was able to survive at 70 ℃ and 80 ℃ for 20 min. KBB1-26 showed the antimicrobial inhibition zone to pathogenic bacteria such as E. coli (12 mm), S. Enteritidis (14 mm), S. aureus (14 mm) and L. monocytogenes (41 mm). The production of NO (4.5 ± 0.00μM/mL) and IL-1β (39.7 ± 0.55pg/mL) of KBB1-26 significantly higher than BIF-4 and KBB5-22, respectively. In addition, KBB1-26 and KBB5-22 induce the production of high level of TNF-α and IL-6 in macrophages. Collectively, B. longum KBB1-26 have acid, bile, heat tolerance, antimicrobial activity and immune enhancing effects. These results suggest that KBB1-26 can be used as probiotics for humans and animals.

Earthworms have been employed in traditional oriental medicine for the treatment of neurological disorders, as anticonvulsants, analgesics, and sedatives. In fact, earthworms are currently used as a medicinal agent in China, South Korea, Japan, Taiwan and North Korea. However, both the species and the genus of the earthworms registered in the pharmacopoeia of each country are different. Neural injury is induced by oxidative stress, inflammation, and apoptosis. The role of various synthetic chemicals of earthworms as antioxidant and anti-inflammatory agents have been studied and earthworm extract and its components have been shown to protect nerve cells and restore nerve function in various preclinical neuronal damage models. We employed earthworm extracts to provide prevention and treatment strategies for many neurodegenerative disorders including Parkinson's disease, mild cognitive impairment, cerebral infarction, and peripheral nerve damage. In this study, we investigated the effects of earthworm extracts and its components to explore their prophylactic and therapeutic effects in various neuropathic models. We used earthworm resources to provide prevention and treatment strategies for many neurodegenerative disorders including Parkinson's disease, mild cognitive impairment, cerebral infarction, and peripheral nerve damage. We summarized the protective effects of both earthworms and their extracts on neuropathies. The current study identified some earthworm components to be used in treatment and prevention strategies for nerve disorders and could be helpful for the development of new therapies for intractable diseases.