• Korean Journal of Microbiology
• /
• v.48 no.1
• /
• pp.1-7
• /
• 2012

This study explores the interaction between the production of indole-3-acetic acid (IAA), a typical phytohormone auxin and the role of IAA biosynthetic pathways in each IAA producing rhizobacterial strain. The bacterial strains were isolated from rhizosphere of wild plants and identified as Acinetobacter guillouiae SW5, Bacillus thuringiensis SW17, Rhodococcus equi SW9, and Lysinibacillus fusiformis SW13. A. guillouiae SW5 exhibited the highest production of IAA using tryptophan-dependent pathways among the 4 strains. When indole-3-acetamide (IAM) was added, Rhodococcus equi SW9 showed the highest IAA production of $3824{\mu}g/mg$ protein using amidase activity. A. guillouiae SW5 also showed the highest production of IAA using two pathways with indole-3-acetonitrile (IAN), and its nitrile hydratase activity might be higher than nitrilase. B. thuringiensis SW17 showed the lowest IAA production, and most of IAA might be produced by the amidase activity, although the nitrilase activity was the highest among 4 strains. The roles of nitrile converting enzymes were relatively similar in IAA synthesis by Lysinibacillus fusiformis SW13. Tryptophan-independent pathway of IAA production was utilized by only A. guillouiae SW5.

• The Korean Journal of Physiology and Pharmacology
• /
• v.4 no.6
• /
• pp.515-523
• /
• 2000

Polymorphonuclear leukocytes (PMNs) play an important role in myocardial ischemia/reperfusion (MI/R) injury. Moreover, platelets are also important blood cells that can aggravate myocardial ischemic injury. This study was designed to test the effects of PMNs and platelets separately and together in provoking cardiac dysfunction in isolated perfused rat hearts following ischemia and reperfusion. Additional control rat hearts were perfused with $75{\times}10^6$ PMNs, with $75{\times}10^6$ platelets, or with $75{\times}10^6\;PMNs+75{\times}10^6$ platelets over a five minute perfusion followed by a 75 min observation period. No significant reduction in coronary flow (CF), left ventricular developed pressure (LVDP), or the first derivative of LVDP (dP/dt max) was observed at the end of the observation period in any non-ischemic group. Similarly, global ischemia (I) for 20 min followed by 45 minutes of reperfusion (R) produced no sustained effects on the final recovery of any of these parameters in any group of hearts perfused in the absence of blood cells. However, I/R hearts perfused with either PMNs or platelets alone exhibited decreases in these variables of $5{\sim}10%$ (p＜0.05 from control). Furthermore, I/R hearts perfused with both PMNs and platelets exhibited decreases of 50 to 60% in all measurements of cardiac function (p＜0.01). These dual cell perfused I/R hearts also exhibited marked increases in cardiac myeloperoxidase (MPO) activity indicating a significant PMN infiltration, and enhanced P-selectin expression on the coronary microvascular endothelium. All cardiaodynamic effects as well as PMN accumulation and P-selectin expression were markedly attenuated by a recombinant soluble PSGL-1 which inhibits selectin mediated cell adhesion. These results provide evidence that platelets and PMNs act synergistically in provoking post-reperfusion cardiac dysfunction, and that this may be largely due to cell to cell interactions mediated by P-selectin. These results also demonstrate that a recombinant soluble PSGL-1 reduces myocardial reperfusion injury by platelet and PMNs interaction.

• Journal of Animal Science and Technology
• /
• v.62 no.2
• /
• pp.159-173
• /
• 2020

A 4×2 factorial feeding trial was designed to investigate the effect of replacing soybean meal (SBM) with cottonseed meal (CSM) in wheat/sorghum/SBM-based diets fed with or without microbial enzymes in diets on the performance, visceral organ development and digestibility of nutrients of broiler chickens. Four graded levels of CSM - none (0%), low (4%, 8%, and 12%), medium (5%, 10%, and 15%), and high (6%, 12%, and 18%) of complete diets in starter, grower and finisher, respectively were fed with or without 100 mg/kg of xylanase and β-glucanase blend. Eight isocaloric and isonitrogenous diets were formulated using least-cost method to meet the nutrient specifications of Ross 308 male broilers. Each treatment was randomly assigned to 6 replicates (10 birds per replicate). There were CSM-enzyme interactions (p < 0.05) on feed intake (FI) and weight gain (WG) in the starter phase. Enzyme supplementation improved (p < 0.05) feed conversion ratio (FCR) in the grower and finisher phases, and increased WG in growing and finishing birds. CSM inclusion reduced (p < 0.05) the weight of gizzard and proventriculus in starter chicks, while these organs were bigger (p < 0.05) in the grower phase. The test ingredient decreased (p < 0.05) small intestinal weight in starter and grower birds. The CSM increased the absolute weight of thighs (p < 0.05) while breast meat was increased (p < 0.01) by enzyme addition. Starch digestibility was improved (p < 0.01) by enzyme inclusion and decreased (p < 0.01) by CSM. Enzyme supplementation improved (p < 0.05) the ileal digestibility of gross energy and protein. The results demonstrate that CSM can substitute up to 90% SBM in broiler chicken diets without compromising performance, and the nutritive value of CSM-containing diets can effectively be improved by enzyme supplementation.

• Korean Journal of Food Science and Technology
• /
• v.47 no.3
• /
• pp.299-305
• /
• 2015

The Folin-Denis assay using the Folin-Ciocalteu (F-C) reagent has been commonly used for analyzing the total phenolic compound content in various food products. In the present study, the effects of proteins on the reactivity of the F-C reagent with different phenolic compounds were investigated. Bovine serum albumin (BSA) or skim milk proteins showed a concentration-dependent increase in color response in the Folin-Denis assay; these proteins decreased the color response of most phenolic compounds tested. The reactivity of phenolic compounds was significantly less pronounced in the presence of BSA and this interference was greater at higher concentrations of phenolic compounds. The reactivity of phenolic compounds with the F-C reagent was reduced significantly by their oxidation; the reaction of the oxidized products with the F-C reagent was more severely affected by BSA. The interfering effects in the Folin-Denis assay might be attributable to binding interactions of phenolic compounds with proteins.

• Bulletin of the Korean Chemical Society
• /
• v.31 no.1
• /
• pp.52-58
• /
• 2010

Human histamine H1 receptor (HHR1) is a G protein-coupled receptor and a primary target for antiallergic therapy. Here, the ligand-based three-dimensional pharmacophore models were built from a set of known HHR1 inverse agonists using HypoGen module of CATALYST software. All ten generated pharmacophore models consist of five essential features: hydrogen bond acceptor, ring aromatic, positive ionizable and two hydrophobic functions. Best model had a correlation coefficient of 0.854 for training set compounds and it was validated with an external test set with a high correlation value of 0.925. Using this model Maybridge database containing 60,000 compounds was screened for potential leads. A rigorous screening for drug-like compounds unveiled RH01692 and SPB00834, two novel molecules for HHR1 with good CATALYST fit and estimated activity values. The new lead molecules were docked into the active site of constructed HHR1 homology model based on recently crystallized squid rhodopsin as template. Both the hit compounds were found to have critical interactions with Glu177, Phe432 and other important amino acids. The interpretations of this study may effectively be deployed in designing of novel HHR1 inverse agonists.

• KSBB Journal
• /
• v.15 no.6
• /
• pp.649-657
• /
• 2000

The factors affecting the back-extraction efficiency of Bovine Serum Albumin (BSA, Mw. 65kDa, pl 4.9) solubilized in an AOT reverse micellar solution, prepared by the injection method, to an excess aqueous phase were investigated. In particular, the effects of pH, the types of salts, alcohols added as cosurfactants, and their concentrations in the aqueous phase were examined. Furthermore, by comparing the CD spectra of the back-extracted BSA and the feed BSA, the structural changes of BSA during the extraction process were determined. The addition of 1:1 salt such as KCl or NaCl to the aqueous phase resulted in almost a 100% extraction to the aqueous phase at a pH higher than its isoelectric point pl. This high efficiency of back-extraction might be due to the change in the interactions between the protein and micellar aggregates driven by the added salt. For 1:2 salts like $CaCl_2$ and $MgCl_2$, BSA was back-extracted with lower than 20% extraction efficiency. Maximum extraction efficiencies were attained at about pH=7 and pH=8 for monovalent and divalent salts, respectively. The addition of alcohols as cosurfactants led to an improvement in monovalent and divalent salts, respectively. From the CD spectra of BSA extracted to the aqueous phase, it was observed that denaturation of BSA was not significant. In certain back-extraction conditions, the extracted BSA showed even higher activity than the feed BSA.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.11
• /
• pp.1705-1714
• /
• 2019

Objective: Reducing roughage feeding without negatively affecting rumen health is of interest in ruminant nutrition. We investigated the effects of roughage sources and concentrate types on growth performance, ruminal fermentation, and blood metabolite levels in growing cattle. Methods: In this 24-week trial, 24 Hanwoo cattle ($224{\pm}24.7kg$) were fed similar nitrous and energy levels of total mixed ration formulated using two kinds of roughage (timothy hay and ryegrass straw) and two types of concentrate mixes (high starch [HS] and high fiber [HF]). The treatments were arranged in a $2{\times}2$ factorial, consisting of 32% timothy-68% HS, 24% timothy-76% HF, 24% ryegrass-76% HS, and 17% ryegrass-83% HF. Daily feed intakes were measured. Every four weeks, blood were sampled, and body weight was measured before morning feeding. Every eight weeks, rumen fluid was collected using a stomach tube over five consecutive days. Results: The mean dry matter intake (7.33 kg) and average daily gain (1,033 g) did not differ among treatments. However, significant interactions between roughage source and concentrate type were observed for the rumen and blood parameters (p<0.05). Total volatile fatty acid concentration was highest (p<0.05) in timothy-HF-fed calves. With ryegrass as the roughage source, decreasing the roughage inclusion rate increased the molar proportion of propionate and decreased the acetate-to-propionate ratio; the opposite was observed with timothy as the roughage source. Similarly, the effects of concentrate types on plasma total protein, alanine transaminase, Ca, inorganic P, total cholesterol, triglycerides, and creatinine concentrations differed with roughage source (p<0.05). Conclusion: Decreasing the dietary roughage inclusion rate by replacing forage neutral detergent fiber with that from non-roughage fiber source might be a feasible feeding practice in growing cattle. A combination of low-quality roughage with a high fiber concentrate might be economically beneficial.

• Animal Bioscience
• /
• v.34 no.1
• /
• pp.74-84
• /
• 2021

Objective: Feed additives that modify rumen fermentation can be used to prevent metabolic disturbances such as acidosis and optimize beef cattle production. The study evaluated the effects of liquid and powdered forms of polyclonal antibody preparation (PAP) against Streptococcus bovis and Fusobacterium necrophorum on rumen fermentation parameters in ruminally cannulated non-lactating dairy cows that were adapted or unadapted to a high concentrate diet. Methods: A double 3×3 Latin square design was used with three PAP treatments (control, powdered, and liquid PAP) and two adaptation protocols (adapted, unadapted; applied to the square). Adapted animals were transitioned for 2 weeks from an all-forage to an 80% concentrate diet, while unadapted animals were switched abruptly. Results: Interactions between sampling time and adaptation were observed; 12 h after feeding, the adapted group had lower ruminal pH and greater total short chain fatty acid concentrations than the unadapted group, while the opposite was observed after 24 h. Acetate:propionate ratio, molar proportion of butyrate and ammonia nitrogen concentration were generally greater in adapted than unadapted cattle up to 36 h after feeding. Adaptation promoted 3.5 times the number of Entodinium protozoa but copy numbers of Streptococcus bovis and Fibrobacter succinogens genes in rumen fluid were not affected. However, neither liquid nor powdered forms of PAP altered rumen acidosis variables in adapted or unadapted animals. Conclusion: Adaptation of cattle to highly fermentable carbohydrate diets promoted a more stable ruminal environment, but PAP was not effective in this study in which no animal experienced acute or sub-acute rumen acidosis.

• Yonsei Medical Journal
• /
• v.59 no.9
• /
• pp.1096-1106
• /
• 2018

Purpose: Alzheimer's disease (AD) is the sixth most common cause of death in the United States. MicroRNAs have been identified as vital players in neurodegenerative diseases, including AD. microRNA-128 (miR-128) has been shown to be dysregulated in AD. This study aimed to explore the roles and molecular mechanisms of miR-128 in AD progression. Materials and Methods: Expression patterns of miR-128 and peroxisome proliferator-activated receptor gamma ($PPAR-{\gamma}$) messenger RNA in clinical samples and cells were measured using RT-qPCR assay. $PPAR-{\gamma}$ protein levels were determined by Western blot assay. Cell viability was determined by MTT assay. Cell apoptotic rate was detected by flow cytometry via double-staining of Annexin V-FITC/PI. Caspase 3 and $NF-{\kappa}B$ activity was determined by a Caspase 3 Activity Assay Kit or $NF-{\kappa}B$ p65 Transcription Factor Assay Kit, respectively. Bioinformatics prediction and luciferase reporter assay were used to investigate interactions between miR-128 and $PPAR-{\gamma}$ 3'UTR. Results: MiR-128 expression was upregulated and $PPAR-{\gamma}$ expression was downregulated in plasma from AD patients and $amyloid-{\beta}$ $(A{\beta})-treated$ primary mouse cortical neurons (MCN) and Neuro2a (N2a) cells. Inhibition of miR-128 decreased $A{\beta}-mediated$ cytotoxicity through inactivation of $NF-{\kappa}B$ in MCN and N2a cells. Moreover, $PPAR-{\gamma}$ was a target of miR-128. $PPAR-{\gamma}$ upregulation attenuated $A{\beta}-mediated$ cytotoxicity by inactivating $NF-{\kappa}B$ in MCN and N2a cells. Furthermore, $PPAR-{\gamma}$ downregulation was able to abolish the effect of anti-miR-128 on cytotoxicity and $NF-{\kappa}B$ activity in MCN and N2a cells. Conclusion: MiR-128 inhibitor decreased $A{\beta}-mediated$ cytotoxicity by upregulating $PPAR-{\gamma}$ via inactivation of $NF-{\kappa}B$ in MCN and N2a cells, providing a new potential target in AD treatment.

• Biomolecules & Therapeutics
• /
• v.30 no.6
• /
• pp.553-561
• /
• 2022

Chronic myeloid leukemia (CML) is a slowly progressing hematopoietic cell disorder. Sphingosine kinase 1 (SPHK1) plays established roles in tumor initiation, progression, and chemotherapy resistance in a wide range of cancers, including leukemia. However, small-molecule inhibitors targeting SPHK1 in CML still need to be developed. This study revealed the role of SPHK1 in CML and investigated the potential anti-leukemic activity of hirsuteine (HST), an indole alkaloid obtained from the oriental plant Uncaria rhynchophylla, in CML cells. These results suggest that SPHK1 is highly expressed in CML cells and that overexpression of SPHK1 represents poor clinical outcomes in CML patients. HST exposure led to G2/M phase arrest, cellular apoptosis, and downregulation of Cyclin B1 and CDC2 and cleavage of Caspase 3 and PARP in CML cells. HST shifted sphingolipid rheostat from sphingosine 1-phosphate (S1P) towards the ceramide coupled with a marked inhibition of SPHK1. Mechanistically, HST significantly blocked SPHK1/S1P/S1PR1 and BCR-ABL/PI3K/Akt pathways. In addition, HST can be docked with residues of SPHK1 and shifts the SPHK1 melting curve, indicating the potential protein-ligand interactions between SPHK1 and HST in both CML cells. SPHK1 overexpression impaired apoptosis and proliferation of CML cells induced by HST alone. These results suggest that HST, which may serve as a novel and specific SPHK1 inhibitor, exerts anti-leukemic activity by inhibiting the SPHK1/S1P/S1PR1 and BCR-ABL/PI3K/Akt pathways in CML cells, thus conferring HST as a promising anti-leukemic drug for CML therapy in the future.