Detection of pathogenic microorganisms takes several days by conventional methods. It is necessary to assess microorganisms in a timely manner to reduce the risk of spreading infection. For this purpose, bacteriophages are chosen for use as a biosensing tool due to their host specificity, wide abundance, and safety. However, their lytic cycle limits their efficacy as biosensors. Phage proteins involved in binding to bacteria could be a robust alternative in resolving this drawback. Here, a fragment of tail protein J (residues 784 to 1,132) of phage lambda fused with 6X His-tag (6HN-J) at its N-terminus was cloned, overexpressed, purified, and characterized for its binding with microorganisms. The purified protein demonstrated a size of about 38 kDa in sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) and bound with anti-His monoclonal antibodies. It bound specifically to Escherichia coli K-12, and not Salmonella typhimurium, Bacillus subtilis, or Pseudomonas aeruginosa in dot blotting. Binding of the protein to E. coli K-12 inhibited about 50% of the in vivo adsorption of the phage lambda to host cells at a concentration of $1{\mu}g/ml$ 6HN-J protein and almost 100% at $25{\mu}g/ml$ 6HN-J. The results suggest that a fusion viral protein could be utilized as a biosensing element (e.g., protein chips) for detecting microorganisms in real time.
Small Tail Han Sheep has significant characteristics of high prolificacy and non-seasonal ovulatory activity and is an excellent local sheep breed in P. R. China. Recently a novel member of the transforming growth factor $\beta$ (TGF$\beta$) superfamily termed bone morphogenetic protein 15 (BMP15) was shown to be specifically expressed in oocytes and to be essential for female fertility. Therefore, BMP15 is a candidate gene for reproductive performance of Small Tail Han Sheep. The whole genomic nucleotide sequence of BMP15 gene in Small Tail Han Sheep was searched for polymorphisms by PCR-SSCP and direct sequencing, and only one polymorphism was found. The polymorphism was a result of a 3 base pair deletion, which eliminated a single Leu codon (CTT). The allelic frequencies for A (without deletion) and B (with a codon deletion) are 0.73 and 0.27 respectively. The effects of BMP15 genotype on litter size were evaluated using the least squares model. This indicated that there was a significant association between litter size of Small Tail Han Sheep and a deletion in BMP15 gene (p=0.02<0.05). Small Tail Han Sheep ewes with AA and AB genotype produce on average 0.5 and 0.3 more lambs per litter than those ewes with BB genotype.
Cho, Hyung Taek;Kim, Jun Ho;Lee, Jin Hyup;Kim, Young Jun
Journal of Ginseng Research
/
v.41
no.3
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pp.347-352
/
2017
Background: Panax ginseng (PG) has a long history of use in Asian medicine because of its multiple pharmacological activities. It has been considered that PG in a type of white ginseng may induce undesirable thermogenic effects, but not in a type of red ginseng. However, there is a lack of evidence about the correlation between ginsenoside and thermogenesis. Methods: We investigated the effects of PG with different ginsenoside compositions on body temperature, blood pressure, and thermogenesis-related factors in rats. Results: With increasing steaming time (0 h, 3 h, 6 h, and 9 h), the production of protopanaxadiol ginsenosides increased, whereas protopanaxatriol ginsenosides decreased in white ginseng. In both short- and long-term studies, administration of four ginseng extracts prepared at different steaming times did not induce significant changes in body temperature (skin, tail, and rectum) and blood pressure of rats compared to saline control. In addition, there were no significant differences in the molecular markers related to thermogenesis (p > 0.05), mRNA expressions of peroxisome proliferator-activated receptor-gamma coactivator-$1{\alpha}$ and uncoupling protein 1 in brown adipose tissue, as well as the serum levels of interleukin-6, inducible nitric oxide synthase, and nitrite among the treatment groups. Conclusion: These observations indicate that the potential undesirable effects of PG on body temperature could not be explained by the difference in ginsenoside composition.
Chen, Lin;Li, Ruimei;Chen, Feiyan;Zhang, Hantao;Zhu, Zhu;Xu, Shuyi;Cheng, Yao;Zhao, Yunan
Journal of Ginseng Research
/
v.46
no.5
/
pp.666-674
/
2022
Background: Ginsenosides and their metabolites have antidepressant-like effects, but the underlying mechanisms remain unclear. We previously identified 14-3-3 ζ as one of the target proteins of 20 (S)-protopanaxadiol (PPD), a fully deglycosylated ginsenoside metabolite. Methods: Corticosterone (CORT) was administered repeatedly to induce the depression model, and PPD was given concurrently. The tail suspension test (TST) and the forced swimming test (FST) were used for behavioral evaluation. All mice were sacrificed. Golgi-cox staining, GSK 3β activity assay, and Western blot analysis were performed. In vitro, the kinetic binding analysis with the Biolayer Interferometry (BLI) was used to determine the molecular interactions. Results: TST and FST both revealed that PPD reversed CORT-induced behavioral deficits. PPD also ameliorated the CORT-induced expression alterations of hippocampal Ser9 phosphorylated glycogen synthase kinase 3β (p-Ser9 GSK 3β), Ser133 phosphorylated cAMP response element-binding protein (p-Ser133 CREB), and brain-derived neurotrophic factor (BDNF). Moreover, PPD attenuated the CORT-induced increase in GSK 3β activity and decrease in dendritic spine density in the hippocampus. In vitro, 14-3-3 ζ protein specifically bound to p-Ser9 GSK 3β polypeptide. PPD promoted the binding and subsequently decreased GSK 3β activity. Conclusion: These findings demonstrated the antidepressant-like effects of PPD on the CORT-induced mouse depression model and indicated a possible target-based mechanism. The combination of PPD with the 14-3-3 ζ protein may promote the binding of 14-3-3 ζ to p-GSK 3β (Ser9) and enhance the inhibition of Ser9 phosphorylation on GSK 3β kinase activity, thereby activating the plasticity-related CREBeBDNF signaling pathway.
Kim, Yunna;Lee, Hwa-Young;Choi, Yu-Jin;Cho, Seung-Hun
Journal of Ginseng Research
/
v.44
no.4
/
pp.603-610
/
2020
Background: Depression is a common neuropsychiatric disease that shows astrocyte pathology. Ginsenoside Rf (G-Rf) is a saponin found in Panax ginseng which has been used to treat neuropsychiatric diseases. We aimed to investigate antidepressant properties of G-Rf when introduced into the L-alphaaminoadipic acid (L-AAA)-infused mice model which is representative of a major depressive disorder that features diminished astrocytes in the brain. Methods: L-AAA was infused into the prefrontal cortex (PFC) of mice to induce decrease of astrocytes. Mice were orally administered G-Rf (20 mg/kg) as well as vehicle only or imipramine (20 mg/kg) as controls. Depression-like behavior of mice was evaluated using forced swimming test (FST) and tail suspension test (TST). We observed recovery of astroglial impairment and increased proliferative cells in the PFC and its accompanied change in the hippocampus by Western blot and immunohistochemistry to assess the effect of G-Rf. Results: After injection of L-AAA into the PFC, mice showed increased immobility time in FST and TST and loss of astrocytes without significant neuronal change in the PFC. G-Rf-treated mice displayed significantly more decreased immobility time in FST and TST than did vehicle-treated mice, and their immobility time almost recovered to those of the sham mice and imipramine-treated mice. G-Rf upregulated glial fibrillary acidic protein (GFAP) expression and Ki-67 expression in the PFC reduced by L-AAA and also alleviated astroglial change in the hippocampus. Conclusion: G-Rf markedly reversed depression-like behavioral changes and exhibited protective effect against the astrocyte ablation in the PFC induced by L-AAA. These protective properties suggest that G-Rf might be a therapeutic agent for major depressive disorders.
Dioscorea japonica Thunb its effects has been used in f31k remedies f9r various purposes including treatment of diabetes, on hypoglycemic actiity and energy metabolism were investigated. The plant was extracted with methanol(MeOH) and fractionated into four layers hexane, chloroform(CHCI$_3$), butanol(BuOH), and $H_2O$. Diabetes mellitus was induced in male Sprague-Dawley rats by the injection of streptozotocin(STZ) into tail vein at a dose of 45mg/kg body weight(BW). Sprague-Dawley male rats weighing 160-200g were divided into five groups a diabetic control and four experimental groups such as hexane group, CHCl$_3$ group, BuOH group, and $H_2O$ group. The rats of all groups were fed on a AIN-76 diet and the four experimental groups were orally administered each fraction(500mg/kg BW) for 12 days. The diabetic control group was orally administered 5% carboxymethyl cellulose. The body weights were monitored and the concentrations of blood glucose were determined. The levels of glycogen and protein in liver were also measured. The plasma levels of cholesterol, triglyceride (TG), and fee fatty acid(FFA) were also analysed. The body weight gain was higher in the $H_2O$ group than in the control group. Heart weight was significantly reduced by administrations of Dioscorea japonica Thunb. The extents of blood glucose decrement in BuOH and $H_2O$ group were greater than that found in the control group. The muscle protein levels showed significantly higher amounts in all experimental groups. Glycogen levels were higher in the BuOH group than in the control group. The levels of TG were decreased in all experimental groups and the levels of plasma FFA were lower in the BuOH group. The plasma cholesterol levels were not influenced by these four fractions in diabetic rats. These results suggest that the orally administered H2O fraction of Dioscorea japonica Thunb exhibited hypoglycemic effects in STZ induced diabetic rats. (Korean J Nutrition 31(7) : 1093-1099, 1998)
Background: Pseudotyped virus systems that incorporate viral proteins have been widely employed for the rapid determination of the effectiveness and neutralizing activity of drug and vaccine candidates in biosafety level 2 facilities. We report an efficient method for producing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus with dual luciferase and fluorescent protein reporters. Moreover, using the established method, we also aimed to investigate whether Korean Red Ginseng (KRG), a valuable Korean herbal medicine, can attenuate infectivity of the pseudotyped virus. Methods: A pseudovirus of SARS-CoV-2 (SARS-2pv) was constructed and efficiently produced using lentivirus vector systems available in the public domain by the introduction of critical mutations in the cytoplasmic tail of the spike protein. KRG extract was dose-dependently treated to Calu-3 cells during SARS2-pv treatment to evaluate the protective activity against SARS-CoV-2. Results: The use of Calu-3 cells or the expression of angiotensin-converting enzyme 2 (ACE2) in HEK293T cells enabled SARS-2pv infection of host cells. Coexpression of transmembrane protease serine subtype 2 (TMPRSS2), which is the activator of spike protein, with ACE2 dramatically elevated luciferase activity, confirming the importance of the TMPRSS2-mediated pathway during SARS-CoV-2 entry. Our pseudovirus assay also revealed that KRG elicited resistance to SARS-CoV-2 infection in lung cells, suggesting its beneficial health effect. Conclusion: The method demonstrated the production of SARS-2pv for the analysis of vaccine or drug candidates. When KRG was assessed by the method, it protected host cells from coronavirus infection. Further studies will be followed for demonstrating this potential benefit.
This investigation was conducted to evaluate the shrimp flesh (SF) and shrimp by-product (SB) of the most abundant shrimp species (Metapenaeus affinis, Penaeus semisulcatus and Penaeus monodon) caught in Central Vietnam, with the emphasis on yield, gross and amino acid (AA) composition and effect of heat treatment. The results showed that the mean edible SF and SB (head and shells with tail) yields of the three shrimp species averaged 56.7 and 43.3%, respectively, of the total wet body weight, with the M. affinis generating the highest by-product yield (45.7%) and P. semisulcatus (40.6%) the lowest. Significant differences in dry matter (DM), crude protein (CP) and ash content were found between SF and SB. The DM content of SF (21.5%) was lower than of SB (24.9%) and the ash content (on a DM basis) of the SB in all shrimp species was more than three times that of the SF (p<0.05), whereas the CP content was almost twice as high in the SF as compared with the SB (p<0.05). The SB of the three species contained (on a DM basis) between 44.0 and 49.8% CP (p<0.05) and between 13.5 and 18.1% chitin (p<0.05). The Ca content of SB differed also between species (p<0.05). On average, the sum of AA in SB corresponded to 89.3% of the CP and essential AA accounted for about 50% of the total AA. The most abundant AA were arginine, aspartic and glutamic acids, which accounted for 33% of the total AA. Minor, but significant differences in some AA concentrations of SB between species were observed (p<0.05). With the exception of the DM and ether extract content, all other chemical constituents of entire shrimp, SF and SB were not significantly affected by heat treatment (p>0.05).
Background: Recent studies have revealed that the properties Traditional Chinese Medicine is mostly associated with are substance and energy metabolism. Our study aimed to compare the effect of red ginseng (RG) (warm property) and ginseng leaves (GL; cold property) on the substance and energy metabolism of rats with hypothyroidism. Materials and methods: Rats were administered propylthiouracil intraperitoneally for 20 d to cause hypothyroidism. The reference group was orally administered Aconiti Lateralis Radix Praeparaia [FZ (Fuzi in Chinese)], while both the RG and GL groups were orally administrated crude drugs. The rectal, tail, toe, and axilla temperature of the rats were assayed every 3 d. Oxygen consumption, carbon dioxide production, heat production, and energy expenditure were measured via TSE phenoMaster/LabMaster animal monitoring system. Adenosine monophosphate-activated protein kinase, $Na^+-K^+$-ATPase, fumarase, pyruvic acid and cyclic adenosine monophosphate/cyclic guanosine monophosphate were determined. Results: The lower levels of triiodothyronine, tetraiodothyronine, and thyrotropin-releasing hormone and the higher level of thyroid stimulating hormone revealed the successful establishment of a hypothyroidism model. Oxygen consumption, carbon dioxide production, heat production, and energy expenditure in the FZ and RG groups were obviously increased. The activity of $Na^+-K^+$-ATPase and fumarase in the FZ and RG groups was significantly increased. The cyclic adenosine monophosphate/cyclic guanosine monophosphate level in the FZ and RG groups was increased, while the GL group showed the opposite. Conclusion: Our research provides a new way to explore the efficiency of Chinese medicine on the basis of the relationship between drug property and effects on substance and energy metabolism.
Choi, Jong Hee;Lee, Min Jung;Jang, Minhee;Kim, Hak-Jae;Lee, Sanghyun;Lee, Sang Won;Kim, Young Ock;Cho, Ik-Hyun
Journal of Ginseng Research
/
v.42
no.1
/
pp.107-115
/
2018
Background: Depression is one of the most commonly diagnosed neuropsychiatric diseases, but the underlying mechanism and medicine are not well-known. Although Panax ginseng has been reported to exert protective effects in various neurological studies, little information is available regarding its antidepressant effects. Methods: Here, we examined the antidepressant effect and underlying mechanism of P. ginseng extract (PGE) in a chronic restraint stress (CRS)-induced depression model in mice. Results: Oral administration of PGE for 14 d decreased immobility (depression-like behaviors) time in forced swim and tail suspended tests after CRS induction, which corresponded with attenuation of the levels of serum adrenocorticotropic hormone and corticosterone, as well as attenuated c-Fos expression in the amygdala. PGE enhanced messenger RNA expression level of brain-derived neurotrophic factor but ameliorated microglial activation and neuroinflammation (the level of messenger RNA and protein expression of cyclooxygenase-2 and inducible nitric oxide synthase) in the amygdala of mice after CRS induction. Interestingly, 14-d treatment with celecoxib, a selective cyclooxygenase-2 inhibitor, and $N_{\omega}$-nitro-L-arginine methyl ester hydrochloride, a selective inducible nitric oxide synthase inhibitor, attenuated depression-like behaviors after CRS induction. Additionally, PGE inhibited the upregulation of the nuclear factor erythroid 2 related factor 2 and heme oxygenase-1 pathways. Conclusion: Taken together, our findings suggest that PGE exerts antidepressant-like effect of CRS-induced depression by antineuroinflammatory and antioxidant (nuclear factor erythroid 2 related factor 2/heme oxygenase-1 activation) activities by inhibiting the hypothalamo-pituitary-adrenal axis mechanism. Further studies are needed to evaluate the potential of components of P. ginseng as an alternative treatment of depression, including clinical trial evaluation.
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