Establishing veterinary graduation competencies and its impact on veterinary medical education in Korea.

Competencies are defined as an observable and assessable set of knowledge, skills, and attitudes. Graduation competencies, which are more comprehensive, refer to the required abilities of students to perform on-site work immediately after graduation. As graduation competencies set the goal of education, various countries and institutions have introduced them for new veterinary graduates. The Korean Association of Veterinary Medical Colleges has recently established such competencies to standardize veterinary education and enhance quality levels thereof. The purpose of this study is to describe the process of establishing graduation competencies as well as their implication for veterinary education in Korea. Graduation competencies for veterinary education in Korea comprise 5 domains (animal health care and disease management, one health expertise, communication and collaboration, research and learning, and veterinary professionalism). These are further divided into 11 core competencies, and 33 achievement standards, which were carefully chosen from previous case analyses and nation-wide surveys. Currently, graduation competencies are used as a standard for setting clear educational purposes for both instructors and students. Establishing these competencies further initiated the development of detailed learning outcomes, and of a list of basic veterinary clinical performances and skills, which is useful for assessing knowledge and skills. The establishment of graduation competencies is expected to contribute to the continuous development of Korean veterinary education in many ways. These include curriculum standardization and licensing examination reform, which will eventually improve the competencies of new veterinary graduates.

Adrenergic Control of Primo Tissue Size in Rats.

Background: Hyperplastic morphological changes associated with erythropoiesis have been reported in the primo vascular system (PVS) tissue on the surface of abdominal organs in rats with heart failure (HF) or hemolytic anemia (HA). Objectives: Since adrenergic activity is commonly activated in both HF and HA, we investigated whether adrenergic signaling mediates the abovementioned morphological changes. Methods: We compared the effects of adrenolytic treatments (exercise training and 6-hydroxydopamine) on the gross morphology of the PVS tissues isolated from organ surfaces in HF or HA rats. HF and HA were induced by ligating the left coronary artery and injecting phenylhydrazine, respectively. We further compared the effects of norepinephrine and norepinephrine plus α- or ß-adrenoceptor blockers. Results: The number of samples per rat, PN size, and proportion of red-colored samples in the PVS tissue increased in the HF and HA rats. These changes were reversed by adrenolytic treatments. Interestingly, 6-hydroxydopamine also reversed phenylhydrazineinduced hemolytic changes in erythrocytes. Subcutaneous administration of norepinephrine (3 mg/kg/d) increased the sampling frequency per rat and the PN size, but these effects were blunted at a higher dose (10 mg/kg/d). Norepinephrine administration had little effect on the proportion of red-colored tissues. Norepinephrine-induced morphological changes were completely blocked by a ß-adrenoceptor antagonist (propranolol) but increased slightly by an α-adrenoceptor antagonist (phentolamine). Conclusion: Adrenergic signaling controls hyperplastic changes in the organ surface PVS in rats. These findings may explain the morphological dynamics of the PVS tissues proposed by Bong Han Kim and further clarify the pathophysiological roles of the PVS.

Microglial activation induced by LPS mediates excitation of neurons in the hypothalamic paraventricular nucleus projecting to the rostral ventrolateral medulla.

Microglia are known to be activated in the hypothalamic paraventricular nucleus (PVN) of rats with cardiovascular diseases. However, the exact role of microglial activation in the plasticity of presympathetic PVN neurons associated with the modulation of sympathetic outflow remains poorly investigated. In this study, we analyzed the direct link between microglial activation and spontaneous firing rate along with the underlying synaptic mechanisms in PVN neurons projecting to the rostral ventrolateral medulla (RVLM). Systemic injection of LPS induced microglial activation in the PVN, increased the frequency of spontaneous firing activity of PVN-RVLM neurons, reduced GABAergic inputs into these neurons, and increased plasma NE levels and heart rate. Systemic minocycline injection blocked all the observed LPS-induced effects. Our results indicate that LPS increases the firing rate and decreases GABAergic transmission in PVN-RVLM neurons associated with sympathetic outflow and the alteration is largely attributed to the activation of microglia. Our findings provide some insights into the role of microglial activation in regulating the activity of PVN-RVLM neurons associated with modulation of sympathetic outflow in cardiovascular diseases. [BMB Reports 2021; 54(12): 620-625].

Basic reproduction number of African swine fever in wild boars (Sus scrofa) and its spatiotemporal heterogeneity in South Korea.

BACKGROUND: African swine fever (ASF) is a hemorrhagic fever occurring in wild boars (Sus scrofa) and domestic pigs. The epidemic situation of ASF in South Korean wild boars has increased the risk of ASF in domestic pig farms. Although basic reproduction number (R0) can be applied for control policies, it is challenging to estimate the R0 for ASF in wild boars due to surveillance bias, lack of wild boar population data, and the effect of ASF-positive wild boar carcass on disease dynamics. OBJECTIVES: This study was undertaken to estimate the R0 of ASF in wild boars in South Korea, and subsequently analyze the spatiotemporal heterogeneity. METHODS: We detected the local transmission clusters using the spatiotemporal clustering algorithm, which was modified to incorporate the effect of ASF-positive wild boar carcass. With the assumption of exponential growth, R0 was estimated for each cluster. The temporal change of the estimates and its association with the habitat suitability of wild boar were analyzed. RESULTS: Totally, 22 local transmission clusters were detected, showing seasonal patterns occurring in winter and spring. Mean value of R0 of each cluster was 1.54. The estimates showed a temporal increasing trend and positive association with habitat suitability of wild boar. CONCLUSIONS: The disease dynamics among wild boars seems to have worsened over time. Thus, in areas with a high elevation and suitable for wild boars, practical methods need to be contrived to ratify the control policies for wild boars.

Electrophysiological Properties of Ion Channels in Ascaris suum Tissue Incorporated into Planar Lipid Bilayers.

Ion channels are important targets of anthelmintic agents. In this study, we identified 3 types of ion channels in Ascaris suum tissue incorporated into planar lipid bilayers using an electrophysiological technique. The most frequent channel was a large-conductance cation channel (209 pS), which accounted for 64.5% of channels incorporated (n=60). Its open-state probability (Po) was ~0.3 in the voltage range of -60~+60 mV. A substate was observed at 55% of the main-state. The permeability ratio of Cl- to K+ (PCl/PK) was ~0.5 and PNa/PK was 0.81 in both states. Another type of cation channel was recorded in 7.5% of channels incorporated (n=7) and discriminated from the large-conductance cation channel by its smaller conductance (55.3 pS). Its Po was low at all voltages tested (~0.1). The third type was an anion channel recorded in 27.9% of channels incorporated (n=26). Its conductance was 39.0 pS and PCl/PK was 8.6±0.8. Po was ~1.0 at all tested potentials. In summary, we identified 2 types of cation and 1 type of anion channels in Ascaris suum. Gating of these channels did not much vary with voltage and their ionic selectivity is rather low. Their molecular nature, functions, and potentials as anthelmintic drug targets remain to be studied further.

Dexamethasone reduces infectious bursal disease mortality in chickens.

Very virulent infectious bursal disease virus (vvIBDV) causes high mortality in chickens but measures to reduce the mortality have not been explored. Chickens (8-9 weeks) were treated with 3 agents before and during vvIBDV inoculation. Dexamethasone treatment reduced the mortality of infected chickens (40.7% vs. 3.7%; p < 0.001), but treatment with aspirin or vitamin E plus selenium did not affect the mortality. The bursa of Fabricius appeared to have shrunk in both dead and surviving chickens (p < 0.01). The results indicate that dexamethasone can reduce mortality in vvIBDV-infected chickens and may provide therapeutic clues for saving individual birds infected by the virus.

The role of the voltage-gated potassium channel, Kv2.1 in prostate cancer cell migration.

Voltage-gated potassium (Kv) channels are involved in many important cellular functions and play pivotal roles in cancer progression. The expression level of Kv2.1 was observed to be higher in the highly metastatic prostate cancer cells (PC-3), specifically in their membrane, than in immortalized prostate cells (WPMY-1 cells) and comparatively less metastatic prostate cancer cells (LNCaP and DU145 cells). However, Kv2.1 expression was significantly decreased when the cells were treated with antioxidants, such as N-acetylcysteine or ascorbic acid, implying that the highly expressed Kv2.1 could detect reactive oxygen species (ROS) in malignant prostate cancer cells. In addition, the blockade of Kv2.1 with stromatoxin-1 or siRNA targeting Kv2.1 significantly inhibited the migration of malignant prostate cancer cells. Our results suggested that Kv2.1 plays an important role as a ROS sensor and that it is a promising therapeutic molecular target in metastasis of prostate cancer. [BMB Reports 2021; 54(2): 130-135].

Primo Bundles Identified by Microcomputed Tomography in Primo Vascular Tissue on the Surface of Rat Abdominal Organs.

BACKGROUND: The primo vascular system (PVS) is a novel network composed of primo nodes (PNs) and primo vessels (PVs). Currently, its anatomy is not fully understood. OBJECTIVES: The aim of this study was to elucidate the three-dimensional PN-PV structure. METHODS: Organ-surface PVS tissue was isolated from healthy and anemic rats. The tissues were analyzed by X-ray microcomputed tomography (CT), hematoxylin and eosin staining, and scanning electron microscopy. RESULTS: From CT images, we identified one or more bundles in a PV. In the PN, the bundles were enlarged and existed in isolation and/or in anastomosis. The transverse CT images revealed four areas of distinct intensities: zero, low, intermediate, and high. The first two were considered to be the sinuses and the subvessels of the PVS and were identified in the hematoxylin and eosin-stained PN sections. The enlargement of the PN from anemic rats was associated with an increase in the intermediate-intensity area. The high-intensity area demarcated the bundle and was overlapped with the mesothelial cells. In scanning electron microscopy, the PV bundles branched out, tapering down to a single bundle at some distance from the PN. Each bundle was composed of several subvessels (∼5 µm). Clustered round microcells (1-25 µm), scattered flat oval cells (∼15 µm), and amorphous extracellular matrix were observed on the surface of the PVS tissue. CONCLUSIONS: The results newly showed that the primo bundle is a structural unit of both PVs and PNs. A bundle was demarcated by high CT intensity and mesothelial cells and consisted of multiple subvessels. The PN bundles contained also sinuses.

Tentonin 3/TMEM150C senses blood pressure changes in the aortic arch.

The baroreceptor reflex is a powerful neural feedback that regulates arterial pressure (AP). Mechanosensitive channels transduce pulsatile AP to electrical signals in baroreceptors. Here we show that tentonin 3 (TTN3/TMEM150C), a cation channel activated by mechanical strokes, is essential for detecting AP changes in the aortic arch. TTN3 was expressed in nerve terminals in the aortic arch and nodose ganglion (NG) neurons. Genetic ablation of Ttn3 induced ambient hypertension, tachycardia, AP fluctuations, and impaired baroreflex sensitivity. Chemogenetic silencing or activation of Ttn3+ neurons in the NG resulted in an increase in AP and heart rate, or vice versa. More important, overexpression of Ttn3 in the NG of Ttn3-/- mice reversed the cardiovascular changes observed in Ttn3-/- mice. We conclude that TTN3 is a molecular component contributing to the sensing of dynamic AP changes in baroreceptors.

Transplantation of hMSCs Genome Edited with LEF1 Improves Cardio-Protective Effects in Myocardial Infarction.

Stem cell-based therapy is one of the most attractive approaches to ischemic heart diseases, such as myocardial infarction (MI). We evaluated the cardio-protective effects of the human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) stably expressing lymphoid enhancer-binding factor 1 (LEF1; LEF1/hUCB-MSCs) in a rat model of MI. LEF1 overexpression in hUCB-MSCs promoted cell-proliferation and anti-apoptotic effects in hypoxic conditions. For the application of its therapeutic effects in vivo, the LEF1 gene was introduced into an adeno-associated virus integration site 1 (AAVS1) locus, known as a safe harbor site on chromosome 19 by CRISPR/Cas9-mediated gene integration in hUCB-MSCs. Transplantation of LEF1/hUCB-MSCs onto the infarction region in the rat model significantly improved overall survival. The cardio-protective effect of LEF1/hUCB-MSCs was proven by echocardiogram parameters, including greatly improved left-ventricle ejection fraction (EF) and fractional shortening (FS). Moreover, histology and immunohistochemistry successfully presented reduced MI region and fibrosis by LEF1/hUCB-MSCs. We found that these overall positive effects of LEF1/hUCB-MSCs are attributed by increased proliferation and survival of stem cells in oxidative stress conditions and by the secretion of various growth factors by LEF1. In conclusion, this study suggests that the stem cell-based therapy, conjugated with genome editing of transcription factor LEF1, which promotes cell survival, could be an effective therapeutic strategy for cardiovascular disease.

Acute Anemia Induces Erythropoiesis in Rat Organ Surface Primo-Vascular Tissue.

The primo-vascular system (PVS) is a newly identified vascular tissue composed of primo-nodes (PNs) and primo-vessels (PVs). Previously, we reported erythropoietic activity in the organ-surface PVS (osPVS) tissue of rats with heart failure. In this study, we further investigated whether acute anemia could induce erythropoiesis in the PVS of rats, based on the hypothesis that erythropoiesis in osPVS tissue is due to anemia accompanying heart failure. Acute anemia was induced by an intraperitoneal injection of phenylhydrazine (PHZ). Circulating red blood cells (RBCs) and hematocrit decreased by 31.6%, whereas reticulocytes and white blood cells increased at day 3 and day 6 after PHZ treatment. All these parameters recovered to control levels at day 10. At days 3 and 6, we observed an increase in the size of the PNs (P < 0.05), the number of the osPVS tissue samples per rat (P < 0.01), and the proportion of osPVS tissue samples with red chromophore (P < 0.001), which was from the RBCs in the PVS tissue. The number of RBCs, estimated from the PN sections stained with hematoxylin and eosin, increased at day 6 in the rats with anemia (P < 0.01). All these anemia-induced changes in the osPVS tissue recovered to the control levels by day 10. Taken together, the results showed that the morphological and cytological changes in the osPVS tissue appear to be related to the erythropoietic activity induced by acute anemia in rats. This study confirmed the previous findings that the osPVS can exert erythropoietic activity in disease states accompanied by anemia, such as heart failure.

Mesothelial Cells Covering the Surface of Primo Vascular System Tissue.

The primo vascular system (PVS) is reported to have a periductium composed of cells with spherical or spindle-shaped nuclei and abundant cytoplasm. However, little is known about these periductium cells. In this study, we examined the morphological features of cells covering the PVS tissue isolated from the surface of abdominal organs of rats. By hematoxylin and eosin (H&E) staining, we observed a layer of dark nuclei on the basement membrane at the borders of the sections of primo node (PN), primo vessel (PV), and their subunits. The nuclei appeared thin and linear (10-14 µm), elliptical (8-10 × 3-4 µm), and round (5-7 µm). The borders of the PVS tissue sections were immunostained with a selective antibody for mesothelial cells (MCs). Areas of immunoreactivity overlapped with the flattened cells are shown by hematoxylin and eosin staining. By scanning electron microscopy, we further identified elliptical (11 × 21 µm) and rectangular squamous MCs (length, 10 µm). There were numerous stomata (∼200 nm) and microparticles (20-200 nm) on the surface of the PVS MCs. In conclusion, this study presents the novel finding that the PVS periductium is composed of squamous MCs. These cells tightly line the luminal surface of the PVS tissue, including PNs, PVs, and small branches of the PVs in the abdominal cavity. These results will help us to understand the physiological roles such as hyaluronan secretion and the fine structure of PVS tissue.

Physicochemical factors that affect electroporation of lung cancer and normal cell lines.

Electroporation is used for cancer therapy to efficiently destroy cancer tissues by transferring anticancer drugs into cancer cells or by irreversible tumor ablation without resealing pores. There is growing interest in the electroporation method for the treatment of lung cancer, which has the highest mortality rate among cancers. Improving the cancer cell selectivity has the potential to expand its use. However, the factors that influence the cell selectivity of electroporation are debatable. We aimed to identify the important factors that influence the efficiency of electroporation in lung cells. The electropermeabilization of lung cancer cells (H460, A549, and HCC1588) and normal lung cells (MRC5, WI26 and L132) was evaluated by the transfer of fluorescence dyes. We found that membrane permeabilization increased as cell size, membrane stiffness, resting transmembrane potential, and lipid cholesterol ratio increased. Among them, lipid composition was found to be the most relevant factor in the electroporation of lung cells. Our results provide insight into the differences between lung cancer cells and normal lung cells and provide a basis for enhancing the sensitivity of lung cancers cells to electroporation.

Exercise training normalizes elevated firing rate of hypothalamic presympathetic neurons in heart failure rats.

Exercise training (ExT) normalizes elevated sympathetic nerve activity in heart failure (HF), but the underlying mechanisms are not well understood. In this study, we examined the effects of 3 wk of ExT on the electrical activity of the hypothalamic presympathetic neurons in the brain slice of HF rats. HF rats were prepared by ligating the left descending coronary artery. The electrophysiological properties of paraventricular nucleus neurons projecting to the rostral ventrolateral medulla (PVN-RVLM) were examined using the slice patch-clamp technique. The neuronal firing rate was elevated in HF rats, and ExT induced a reduction in the firing rate ( P < 0.01). This ExT-induced decrease in the firing rate was associated with an increased frequency of spontaneous and miniature inhibitory postsynaptic current (IPSCs; P < 0.05). There was no significant change in excitatory postsynaptic current. Replacing Ca2+ with Mg2+ in the recording solution reduced the elevated IPSC frequency in HF rats with ExT ( P < 0.01) but not in those without ExT, indicating an increase in the probability of GABA release. In contrast, ExT did not restore the reduced GABAA receptor-mediated tonic inhibitory current in HF rats. A GABAA receptor blocker (bicuculline, 20 µM) increased the firing rate in HF rats with ExT ( P < 0.01) but not in those without ExT. Collectively, these results show that ExT normalized the elevated firing activity by increasing synaptic GABA release in PVN-RVLM neurons in HF rats. Our findings provide a brain mechanism underlying the beneficial effects of ExT in HF, which may shed light on the pathophysiology of other diseases accompanied by sympathetic hyperactivation.

Comparison of electrophysiological properties of two types of pre-sympathetic neurons intermingled in the hypothalamic paraventricular nucleus.

The hypothalamic paraventricular nucleus (PVN) contains two types of neurons projecting to either the rostral ventrolateral medulla (PVNRVLM) or the intermediolateral horn (IML) of the spinal cord (PVNIML). These two neuron groups are intermingled in the same subdivisions of the PVN and differentially regulate sympathetic outflow. However, electrophysiological evidence supporting such functional differences is largely lacking. Herein, we compared the electrophysiological properties of these neurons by using patch-clamp and retrograde-tracing techniques. Most neurons (＞70%) in both groups spontaneously fired in the cell-attached mode. When compared to the PVNIML neurons, the PVNRVLM neurons had a lower firing rate and a more irregular firing pattern (p ＜ 0.05). The PVNRVLM neurons showed smaller resting membrane potential, slower rise and decay times, and greater duration of spontaneous action potentials (p ＜ 0.05). The PVNRVLM neurons received greater inhibitory synaptic inputs (frequency, p ＜ 0.05) with a shorter rise time (p ＜ 0.05). Taken together, the results indicate that the two pre-sympathetic neurons differ in their intrinsic and extrinsic electrophysiological properties, which may explain the lower firing activity of the PVNRVLM neurons. The greater inhibitory synaptic inputs to the PVNRVLM neurons also imply that these neurons have more integrative roles in regulation of sympathetic activity.

Kv3.1 and Kv3.4, Are Involved in Cancer Cell Migration and Invasion.

Voltage-gated potassium (Kv) channels, including Kv3.1 and Kv3.4, are known as oxygen sensors, and their function in hypoxia has been well investigated. However, the relationship between Kv channels and tumor hypoxia has yet to be investigated. This study demonstrates that Kv3.1 and Kv3.4 are tumor hypoxia-related Kv channels involved in cancer cell migration and invasion. Kv3.1 and Kv3.4 protein expression in A549 and MDA-MB-231 cells increased in a cell density-dependent manner, and the pattern was similar to the expression patterns of hypoxia-inducible factor-1α (HIF-1α) and reactive oxygen species (ROS) according to cell density, whereas Kv3.3 protein expression did not change in A549 cells with an increase in cell density. The Kv3.1 and Kv3.4 blocker blood depressing substance (BDS) did not affect cell proliferation; instead, BDS inhibited cell migration and invasion. We found that BDS inhibited intracellular pH regulation and extracellular signal-regulated kinase (ERK) activation in A549 cells cultured at a high density, potentially resulting in BDS-induced inhibition of cell migration and invasion. Our data suggest that Kv3.1 and Kv3.4 might be new therapeutic targets for cancer metastasis.

Alteration of MicroRNA Expression Profiles by Surface-Modified Gold Nanoparticles in Human Lung Adenocarcinoma Cells.

MicroRNAs that bind to mRNA are important post-transcriptional regulators that control gene expression by degradation or suppressing translation of target mRNAs. Several studies indicate that nanoparticles (NPs) induce alterations in microRNA expression relating to cell processes including cell development and progressive diseases. However, the alteration of microRNA expression by surface-modified gold nanoparticles (AuNPs) in A549 cells has not been reported. In order to investigate the patterns of microRNA expression, we analyzed data from microRNA arrays using cells treated with citrate- or chitosan-AuNPs. The results demonstrate that the expression of microRNA (hsa-miR-198) in cells treated with citrate-AuNPs significantly differed from non-treated cells, and the expression of 16 microRNAs in cells treated with chitosan-AuNPs significantly differed from non-treated cells. Furthermore, the predicted target genes of microRNAs were related to proliferation, apoptosis, migration, and cell differentiation, including the mitogen-activated protein kinase, ErbB, and Wnt signaling pathway. Thus, the alteration of microRNA expression profiles by citrate- and chitosan-AuNPs would mediate the regulation of the cell processes including cell survival, migration, and differentiation.