• Journal of Microbiology and Biotechnology
• /
• v.29 no.8
• /
• pp.1310-1315
• /
• 2019

Hyaluronidases enhance therapeutic drug transport by breaking down the hyaluronan barrier to lymphatic and capillary vessels, facilitating their tissue absorption. Commercially available hyaluronidases are bovine in origin; however, they pose risks such as bovine spongiform encephalopathy. The present study aimed to develop a novel, highly active hyaluronidase and assess its function. Therefore, in order to find the most efficient active hyaluronidase, we produced several shortened hyaluronidases with partial removal of the N- or C-terminal regions. Moreover, we created an enzyme that connected six histidines onto the end of the hyaluronidase C-terminus. This simplified subsequent purification using $Ni^{2+}$ affinity chromatography, making it feasible to industrialize this highly active recombinant hyaluronidase which exhibited catalytic activity equal to that of the commercial enzyme. Therefore, this simple and effective isolation method could increase the availability of recombinant hyaluronidase for research and clinical purposes.

• Biomolecules & Therapeutics
• /
• v.29 no.5
• /
• pp.545-550
• /
• 2021

Chemotherapy-induced alopecia and hair loss can be stressful in patients with cancer. The hair grows back, but sometimes the hair tends to stay thin. Therefore, understanding mechanisms regulating hair regeneration may improve the management of chemotherapy-induced alopecia. Previous studies have revealed that chemotherapeutic agents induce a hair follicle vascular injury. As hair growth is associated with micro-vessel regeneration, we postulated that the stimulation of angiogenesis might enhance hair regeneration. In particular, mice treated with 5-fluorouracil (5-FU) showed delayed anagen initiation and reduced capillary density when compared with untreated controls, suggesting that the retardation of anagen initiation by 5-FU treatment may be attributed to the loss of perifollicular micro-vessels. We investigated whether the ETS transcription factor ETV2 (aka ER71), critical for vascular development and regeneration, can promote angiogenesis and hair regrowth in a 5-FU-induced alopecia mouse model. Tie2-Cre; Etv2 conditional knockout (CKO) mice, which lack Etv2 in endothelial cells, presented similar hair regrowth rates as the control mice after depilation. Following 5-FU treatment, Tie2-Cre; Etv2 CKO mice revealed a significant reduction in capillary density, anagen induction, and hair restoration when compared with controls. Mice receiving lentiviral Etv2 injection after 5-FU treatment showed significantly improved anagen induction and hair regrowth. Two-photon laser scanning microscopy revealed that enforced Etv2 expression restored normal vessel morphology after 5-FU mediated vessel injury. Our data suggest that vessel regeneration strategies may improve hair regrowth after chemotherapeutic treatment.

• Journal of Ginseng Research
• /
• v.46 no.3
• /
• pp.337-347
• /
• 2022

Coronavirus disease 2019 (COVID-19) is currently a pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 are directly associated with hyper-activation of innate immune response that excessively produce pro-inflammatory cytokines and induce cytokine storm, leading to multi-organ-failure and significant morbidity/mortality. Currently, several antiviral drugs such as Paxlovid (nirmatrelvir and ritonavir) and molnupiravir are authorized to treat mild to moderate COVID-19, however, there are still no drugs that can specifically fight against challenges of SARS-CoV-2 variants. Panax ginseng, a medicinal plant widely used for treating various conditions, might be appropriate for this need due to its anti-inflammatory/cytokine/viral activities, fewer side effects, and cost efficiency. To review Panax ginseng and its pharmacologically active-ingredients as potential phytopharmaceuticals for treating cytokine storm of COVID-19, articles that reporting its positive effects on the cytokine production were searched from academic databases. Experimental/clinical evidences for the effectiveness of Panax ginseng and its active-ingredients in preventing or mitigating cytokine storm, especially for the cascade of cytokine storm, suggest that they might be beneficial as an adjunct treatment for cytokine storm of COVID-19. This review may provide a new approach to discover specific medications using Panax ginseng to control cytokine storm of COVID-19.

• Journal of Animal Reproduction and Biotechnology
• /
• v.38 no.4
• /
• pp.199-208
• /
• 2023

Background: Canine induced pluripotent stem cells (iPSCs) are an attractive source for veterinary regenerative medicine, disease modeling, and drug development. Here we used vitamin C (Vc) to improve the reprogramming efficiency of canine iPSCs, and its functions in the reprogramming process were elucidated. Methods: Retroviral transduction of Oct4, Sox2, Klf4, c-Myc (OSKM), and GFP was employed to induce reprogramming in canine fetal fibroblasts. Following transduction, the culture medium was subsequently replaced with ESC medium containing Vc to determine the effect on reprogramming activity. Results: The number of AP-positive iPSC colonies dramatically increased in culture conditions supplemented with Vc. Vc enhanced the efficacy of retrovirus transduction, which appears to be correlated with enhanced cell proliferation capacity. To confirm the characteristics of the Vc-treated iPSCs, the cells were cultured to passage 5, and pluripotency markers including Oct4, Sox2, Nanog, and Tra-1-60 were observed by immunocytochemistry. The expression of endogenous pluripotent genes (Oct4, Nanog, Rex1, and telomerase) were also verified by PCR. The complete silencing of exogenously transduced human OSKM factors was observed exclusively in canine iPSCs treated with Vc. Canine iPSCs treated with Vc are capable of forming embryoid bodies in vitro and have spontaneously differentiated into three germ layers. Conclusions: Our findings emphasize a straightforward method for enhancing the efficiency of canine iPSC generation and provide insight into the Vc effect on the reprogramming process.

• Progress in Medical Physics
• /
• v.19 no.2
• /
• pp.95-101
• /
• 2008

In this study, we observed the alteration of choline signal intensity in hippocampus region of the depressive rat model induced by forced swimming test (FST). The purpose of this study was to evaluate the antidepressant efficacy in the depressive animal model using MR spectroscopy. Fourteen experimentally naive male Sprague-Dawley rats weighting $160{\sim}180\;g$ were used as subjects. Drug injection group was exposed to the FST except for control group. The drugs were administered subcutaneously (SC) in a volume equivalent to 2ml/kg. And three injections were administered 23, 5, and 1h before beginning the given test. 1H MR spectra were obtained with use of a point resolved spectroscopy (PRESS) localization sequence performed according to the following parameters: repetition time, 2500 ms; echo time, 144 ms; 512 average; 2048 complex data points; voxel dimensions, $1.5{\times}2.5{\times}2.5\;mm^3$ ; acquisition time, 25min. There were no differences in NAA/Cr and Cho/Cr ratio between the right and the left hippocampus both normal control rats and antidepressant-injected rats. Also, no differences were observed in NAA/Cr and Cho/Cr ratio between the normal control rats and the antidepressant-injected rats both the right and the left hippocampus. In this study, we found the recovery of choline signals in the depressive animal model similar to normal control groups as injecting desipramine-HCl which was antidepressant causing anti-immobility effects. Thus, we demonstrated that MR spectroscopy was able to aid in evaluating the antidepressant effect of desipramine-HCl.

• Journal of Food Hygiene and Safety
• /
• v.29 no.3
• /
• pp.202-206
• /
• 2014

In this study, the bacteriological survey was examined on ice creams at manufacturing factories in Korea during the summer season of 2011. The nineteen selected among 166 samples by preliminary test were collected from 11 different manufacturing factories in four major manufacturers in May 2011. Samples from ice milk, ice creams, sherbets, and non milk fat ice creams were tested for the total aerobic bacteria, coliform bacteria, and five food borne pathogens, respectively. The results showed that the coliforms including E. coli O157:H7, Salmonella spp., Staphylococcus aureus, Clostridium perfringens, and Listeria monocytogenes were not detected on all the ice creams. The total aerobic bacteria of the packed samples examined ranged between $2.5{\times}10^3$ and $5.5{\times}10^5cfu/g$. One ice cream, two sherbets, and four ice milk samples exceeded the acceptable limits of total aerobic bacteria according to the Korean standards for ice cream ($1.0{\times}10^5cfu/g$) and others ($5.0{\times}10^4cfu/g$). The levels of these microorganisms from ice creams were higher in three original equipment manufacturers than seven self-manufacturers. Three of ten ice creams (30.0%), three of six ice milks (50.0%), and one of two sherbets (50%) exceeded the acceptable limits of total aerobic bacteria, respectively. The personnel hygiene procedures with chocolate and vanilla chip addition from the manufacturing process were the main sources of the microbial contamination of stick-bar type ice creams when being produced in a factory. Improvement of the hazard analysis critical control points (HACCP) system should be introduced into the ice cream factory to improve the microbial quality of the ice cream products in Korea.

• Journal of Food Hygiene and Safety
• /
• v.28 no.3
• /
• pp.217-221
• /
• 2013

This study developed predictive models for the kinetic behavior of Staphylococcus aureus on processed cheeses. Mozzarella slice cheese and cheddar slice cheese were inoculated with 0.1 ml of a S. aureus strain mixture (ATCC13565, ATCC14458, ATCC23235, ATCC27664, and NCCP10826). The inoculated samples were then stored at $4^{\circ}C$ (1440 h), $15^{\circ}C$ (288 h), $25^{\circ}C$ (72 h), and $30^{\circ}C$ (48 h), and the growth of all bacteria and of S. aureus were enumerated on tryptic soy agar and mannitol salt agar, respectively. The Baranyi model was fitted to the growth data of S. aureus to calculate growth rate (${\mu}_{max}$; ${\log}CFU{\cdot}g^{-1}{\cdot}h^{-1}$), lag phase duration (LPD; h), lower asymptote (log CFU/g), and upper asymptote (log CFU/g). The growth parameters were further analyzed using the square root model as a function of temperature. The model performance was validated with observed data, and the root mean square error (RMSE) was calculated. At $4^{\circ}C$, S. aureus cell growth was not observed on either processed cheese, but S. aureus growth on the mozzarella and cheddar cheeses was observed at $15^{\circ}C$, $25^{\circ}C$, and $30^{\circ}C$. The ${\mu}_{max}$ values increased, but LPD values decreased as storage temperature increased. In addition, the developed models showed acceptable performance (RMSE = 0.3500-0.5344). This result indicates that the developed kinetic model should be useful in describing the growth pattern of S. aureus in processed cheeses.

• Asian-Australasian Journal of Animal Sciences
• /
• v.19 no.11
• /
• pp.1566-1573
• /
• 2006

The study was conducted on 30 true acyclic Sahiwal cows (15 cows, ${\geq}90$ days postpartum; 15 postpubertal heifers, ${\geq}30$ months of age) and a similar 20 untreated controls (10 cows, 10 heifers). An 'Eazi' breed Controlled Internal Drug Release (CIDR) device (containing 1.38 g progesterone) was inserted intravaginally for 7 days (days 0 to 7) followed by 500 IU eCG i.m. at CIDR removal in all the treated animals. Heifers also received 5 mg oestradiol valerate i.m at CIDR insertion. The reproductive performance of these animals was recorded in terms of oestrus induction response, conception and pregnancy rates. Plasma progesterone ($P_4$) and oestradiol-$17{\beta}$ ($E_2$) profiles of 4 representative animals from each treatment group before, during and after CIDR treatment were also monitored. An oestrus induction response of 100% was observed in treated cows and heifers. The majority of cows (53.3%) and heifers (60%) were induced to oestrus within 24-36 and 36-48 h, respectively after CIDR withdrawal; with mean intervals of $44{\pm}3.18$ and $48{\pm}2.35h$, respectively. The conception rate at induced oestrus was higher in cows (40%) than heifers (20%). The final pregnancy rates after 2 subsequent oestruses were 80 and 60% in cows and heifers, respectively (overall 70% for all treated animals). In comparison, only 10% of control animals (2 cows only, 2/20) showed oestrus and become pregnant (10%) during theentire study period. The pretreatment (day 0) mean plasma P4 levels were statistically (p>0.05) similar in cows and heifers ($0.40{\pm}0.04$ and $0.49{\pm}0.11ng/ml$, respectively). The peak $P_4$ levels were observed on day 1 in cows ($13.94{\pm}1.41ng/ml$) and day 2 in heifers ($19.15{\pm}3.30ng/ml$) with a progressive decline up to the day of CIDR withdrawal ($3.35{\pm}0.92$ and $8.79{\pm}1.71ng/ml$, respectively). Mean $P_4$ levels on day 9 and 10 in cows and heifers did not differ significantly from their respective day 0 values and the lowest values were recorded on day 10 both in cows and heifers ($0.13{\pm}0.03$ and $0.14{\pm}0.02ng/ml$, respectively). Wide variations in individual pretreatment $E_2$ levels were observed both in the cows (range = 4-26, mean = $13.00{\pm}4.65pg/ml$) and heifers (range = 10-14, mean = $11.50{\pm}0.96pg/ml$). Thereafter also, $E_2$ levels in cows showed variation and reached a peak level ($53.50{\pm}2.99pg/ml$) on day 8. In heifers, peak mean $E_2$ level ($111.25{\pm}39.81pg/ml$) was recorded on day 1, followed by a non-significant decline on day 2, a significant fall on day 6 and a non-significant increase on day 9 and 10. However, mean $E_2$ levels on days 7 (p<0.05), 8 and 9 (p<0.01) were significantly higher in cows compared to heifers. The post-CIDR withdrawal mean highest $P_4$ and lowest $E_2$ levels coincided with the period when the majority of animals were induced to oestrus. CIDR and eCG treatment resulted in effective induction of oestrus with satisfactory pregnancy rates in true acyclic Sahiwal cows and heifers.

• Korean Journal of Veterinary Service
• /
• v.29 no.3
• /
• pp.347-364
• /
• 2006

This study is concerned with assessment of diethylnitrosamine (DEN 0.01 %) induced liver cell carcinogenesis by measurement of changes preceding the development of neoplasms. Therefore, it was undertaken to investigate changes of liver-specific enzyme activities in Sprague-Dawley (SD) rats by ad libitum feeding of DEN. And also. the changes of hepatic morphology in SD rats were detected by haematoxylineosin stain and immunohistochemistry (PCNA). 5- Fluorouracil (5- FU) is one of the most widely used anticancer agents for digestive cancers including hepatocellular carcinoma, and is known to affect the cell cycle and induce apoptosis of cancer cells. In the present study, SD rats were given drinking water containing 0.01% diethylnitrosamine (DEN) for 8 weeks. Minor behavioral change, brittleness of hair and decreased amount of water and diet intake were observed in rats 4 weeks after DEN administration. The body and liver weights were significantly (p < 0.05) decreased in rats 11 weeks after DEN administration. The liver weight ratio to body weight was rather stable and not significantly decreased in the all treatment groups. The liver specific enzyme activities (AST, ALT, ${\gamma}$-GTP) were significantly increased in all treatment groups compared to control group (p < 0.05). Variable size of liver tumor and hepatomegaly were observed in rats treated with DEN after 10 weeks. Numerous vacuoles were seen on the midzonal and or peripheral areas of hepatic lobules. The large and polymorphological hepatocytes with eosinophilic cytoplasm or densely basophilic mitotic nucleoli were seen. Several proliferative small round cells were seen on vacuolated and necrotic areas in peripheral hepatic lobules or portal areas. PCNA-positive cells were seen on the vacuolated portal areas and peripheral areas of hepatic lobules in the areas of small round cells. We examined functional and morphological changes of livers by 5 - FU treatments on DEN -treated rat. The DEN -treated rats compared to 5 - FU -treated rats after DEN treatment for 8 weeks. The serum total protein and triglyceride were significantly (p < 0.05) decreased, and the liver enzyme activities of AST and ALT were significantly(p < 0.05) increased. After 8 weeks, in the non-5-FU -treated group, the size of liver tumor were varied and hepatomegaly were observed, hepatocellular vacuolization, necrosis and steatosis were observed on the midzonal and peripheral areas of hepatic lobules. The large and polymorphological hepatocytes were seen, the interlobular connective tissues were proliferated. PCNA positive cells were seen in the portal areas and peripheral areas of hepatic lobules in the non-5-FU-treated group. In hepatocytes, condensation of nuclear chromatin and vacuolization were observed, shape of the nuclei were irregular, the degraded nuclei and organelles were observed. The livers of rats in the 5 - FU treatment group were seen grossly brilliant, red-brown color, and the vacuolated and degenerated regions, hyperplastic nodules were not nearly observed. In the electron microscope, the cytoplasm of the hepatocytes contained a large number of mitochondria, rough endoplasmic reticulum, developed organelles surrounding nuclei. The above findings suggest that 5 - FU will be effective as anti -liver tumor drug.

• Parasites, Hosts and Diseases
• /
• v.31 no.1
• /
• pp.57-66
• /
• 1993

The potential of fumagillin dicyclohexylamine salt to treat and prevent intestinal giant-cystic disease in Israel carp, Cyprinus carpio nudus, was monitored in field experimental studies. In experiment 1 (therapeutic), most fish were already naturally infected with more advanced stage of Relohqnellu: kitnuet. Fumagillin was administered to ash (mean body weiht of 830 g for a Penod of one month at a dose of 10.62 mg in the first group and 5.3 mg in the second group per fi sh per day. In experiment 2 (fprophylactlcl), most flesh also were already naturally infected with an early developmental stage of the protozoa and fish (average body weight of 484 g) were administered fumagillln for 45 days at a dose of 3.95 mg per fish per day. In both experiments, the cumulative mortalities of fish and the extrusion rates of the polar filaments of the spores were significantly decreased in a dose-independent fashion. In experiment 2 no dead fish were observed. No adverse side effects of the drug were observed among fish from any dosage group. In experiment 2, an oval or dot-like concave lesion of most cysts developed at the 7th day and the vegetative form was never observed at the 17th day postmedication and the cysts were grossly reduced in size as compared with the control group, beginning at the 24th day until the end of the study. In contrast, it was scarcely effective to the cysts in experiment 1. Taking the seasonal development of the protozoa into consideration, the above results revealed that oral administration of fumagillin at 3.95 mg/500 g body weight/day for a month Is the optimal dose for the treatment and prevention of thelohanellosls caused by T kitnuei among C. carpio nudus.