In order to assay the efficacy of newly synthesized antiviral compounds, in vitro studies of their active intracellular phosphorylated metabolites were established as compared with Zidovudine (ZDV). Antiviral base analogs require intracellular phosphorylation prior to the inhibition of HIV replication. Therefore, antiviral drugs concentrations in plasma have not reflected any direct relationship with activity or toxicity. A method has been developed to measure the concentration of total phosphorylated metabolites inside peripheral blood mononuclear cells using modified commercial radioimmunoassay (RIA). ZDV 5'-monophosphate was synthesized and used as a procedural control for RIA modification. PBMCs were isolated from whole blood and incubated with ZDV for 20 h to allow metabolic phosphorylation. Viable cells were extracted overnight with 60% methanol. After evaporation, the extract was reconstituted in Tris buffer. Samples were split into two fractions, one of which was treated with alkaline phosphatase (AP) to liberate phosphate groups. Concentrations of phosphorylated metabolites were determined by subtracting thε concentration of non-AP-treated fraction from that of the treated fraction. Recovery of phosphorylated ZDV from cell extracts was approximately 90%, and reproducibility was acceptable (coefficients of variation <15% for concentrations${\geq}$0.25 ng/mL). Intracellular concentrations $(0.135{\sim}5.019\;nmole/10^6\;cells)$ followed a nonlinear dose-response relationship over the range $0.015{\sim}2.996mM$ extracellular ZDV, with concentration-dependant saturation.
Hyun Sang Kim;Jun Sik Eom;Shin Ja Lee;Youyoung Choi;Seong Uk Jo;Sang Suk Lee;Eun Tae Kim;Sung Sill Lee
Animal Bioscience
/
v.36
no.8
/
pp.1199-1208
/
2023
Objective: The purpose of this study was to perform a comparative analysis of metabolite levels in serum and milk obtained from cows fed on different concentrate to forage feed ratios. Methods: Eight lactating Holstein cows were divided into two groups: a high forage ratio diet (HF; 80% Italian ryegrass and 20% concentrate of daily intake of dry matter) group and a high concentrate diet (HC; 20% Italian ryegrass and 80% concentrate) group. Blood was collected from the jugular vein, and milk was sampled using a milking machine. Metabolite levels in serum and milk were estimated using proton nuclear magnetic resonance and subjected to qualitative and quantitative analyses performed using Chenomx 8.4. For statistical analysis, Student's t-test and multivariate analysis were performed using Metaboanalyst 4.0. Results: In the principal component analysis, a clear distinction between the two groups regarding milk metabolites while serum metabolites were shown in similar. In serum, 95 metabolites were identified, and 13 metabolites (include leucine, lactulose, glucose, betaine, etc.) showed significant differences between the two groups. In milk, 122 metabolites were identified, and 20 metabolites (include urea, carnitine, acetate, butyrate, arabinitol, etc.) showed significant differences. Conclusion: Our results show that different concentrate to forage feed ratios impact the metabolite levels in the serum and milk of lactating Holstein cows. A higher number of metabolites in milk, including those associated with milk fat synthesis and the presence of Escherichia coli in the rumen, differed between the two groups compared to that in the serum. The results of this study provide a useful insight into the metabolites associated with different concentrate to forge feed ratios in cows and may aid in the search for potential biomarkers for subacute ruminal acidosis.
Nejad, Jalil Ghassemi;Lee, Bae-Hun;Kim, Byong-Wan;Ohh, Sang-Jip;Sung, Kyung Il
Asian-Australasian Journal of Animal Sciences
/
v.29
no.3
/
pp.378-383
/
2016
The objective of this study was to determine the effects of chromium methionine (Cr-Met) chelate supplementation on blood metabolites and fatty acid profile of beef from Holstein steers during late fattening period. Fifteen Holstein steers were allotted randomly into two groups including the control (non Cr-Met feeding, NCM, ave. body weight [BW] = $483{\pm}25.7kg$) and the treatment (Cr-Met feeding for 4 months, 4CM, ave. $BW=486{\pm}27.5kg$) group. The feeding amount of Cr-Met to animals was limited to 400 ppb/cow/d and was supplemented to total mixed ration. No difference in blood albumin, alkaline phosphatase, urea-nitrogen, calcium, creatine, glucose, total protein, triglyceride, and cholesterol were observed between the treatment groups (p>0.05). The level of high density lipoprotein was higher in the 4CM group than the NCM group, whereas low density lipoprotein was lower in the 4CM group (p<0.05). The fatty acid composition (caprate, laurate, myristate, pentadecanoate, palmitate, palmitoleate, margarate, cis-11 heptadodecanoate, stearate, oleate, trans-vaccenate, linoleate, cis-11 eicosenoate, docosa hexaenoic acid, and docosa pentaenoic acid) of the beef showed no difference between the two groups (p>0.05). The arachidonic acid level tended to be higher in the 4CM than the NCM group (p = 0.07). Cr-Met had no influence (p>0.05) on the ratio of saturated, unsaturated, unsaturated/saturated, monounsaturated/saturated and polyunsaturated/saturated fatty acids whereas the ratio of polyunsaturated fatty acids (PUFA) in the 4CM group was comparatively higher than the NCM group (p<0.05). This study concluded that feeding Cr-Met supplementation in 400 ppb/d to Holstein steers for 4 months during late fattening period can improve some blood metabolites and beef quality by increasing PUFA and gamma-linoleate compositions of beef.
The effect of cimetidine administration on the pharmacokinetic parameters of cyclosporine were determined in healthy voluteers. This study was performed in 10 volunteers of age ranged 22-48 years and body weight 48-62 kg. This study was performed with cross-over design. Mono cyclosporine and cyclosporine metabolites was extracted from whole blood analysed by fluororescence polarization immune assay (TDX-FLX, Abbott). After coadministration of cimetidine (300 mg) with cyclosporine (300 mg) orally, maximum concentration of mono cyclosporine was significantly increased $1221{\pm}143\;ng/ml\;to\;1562{\pm}184\;ng/ml$ (P<0.05), area under the time curve of cyclosporine (12 hr) also was significantly increased $7478{\pm}829\;ng/ml{\cdot}hr\;to\;9721{\pm}879\;ng/ml{\cdot}hr$ (P<0.05) and absolute baioavailability of cyclosporine was increased $50\pm5.6\%\;to\;57.6\pm6.1\%\;(P<0.05)$ compared to control group. The blood concentrations of cyclopsorine metabolites were significantly decrased (P<0.05) after coadministration of cimetidine. In cimetidine pretreated group, blood mono cyclosporine concentrations were increased significan시y $1220.0\pm203.00\;ng/ml\;to\;1510.0\pm204.00\;ng/ml$ compared with control group (P<0.05). In the mono cyclosporine pharmacokinetic parameter after oral administration absorption rate and maximum concentration were significantly higher in cimetidine coadministered and pretreated group than control group (P<0.05). The ratio of metabolites and mono cyclosporine concentrations was decreased significantly from $70.8\%\;in\;control\;to\;34.8\%$ in coadministration of cimetidine orally. As matter of facts these reults are considered to inhibition of cyclosporine hepatic metabolism and increasing of cyclosporine absorption rate in gastrointestinal tract because of maintaining cyclosporine stability in elevated gastric pH by cimetidine. We considered, it appeares that cimetidine increase bioavailability of cyclosporine by increasing oral absorption and by decreasing hepatic clearance. But the absorption and clearance of cyclosporine was highly variable individually, and therefore we consider that cyclosporine blood level monitoring would be essential in patients with cimetidine co-administration.
This study was designed to investigate the effects of supplementation of spent mushroom substrates (SMS) on rumen fermentation and blood metabolites in Hanwoo steers. The experiment was conducted as a duplicated Latin square design with six Hanwoo steers ($600{\pm}47\;kg$), each permanently fitted with a ruminal cannula. There were three treatments; i) control (concentrates 4.8 kg +rice straw 1.2 kg/d), ii) Pleurotus eryngiia (PE) treatment (concentrates 4.8 kg+rice straw 0.73 kg+Pleurotus eryngiia 1.20 kg/d) and iii) Pleurotus osteratus (PO) treatment (concentrates 4.8 kg+rice straw 0.73 kg+Pleurotus osteratus 1.20 kg/d). There were no major effects of different dietary treatments on rumen parameters such as pH, ammonia-N, individual and total VFA production. Parameters of N utilization, including blood urea nitrogen (BUN), total protein and albumin levels, were not significantly different among the treatments, except for creatinine. Thus, the present results indicated that protein utilization was mostly unaffected by SMS treatments such as PE and PO, even though creatinine concentration was lower in PE compared with control and PO treatments (p<0.05). The present results indicate that Pleurotus eryngii and Pleurotus osteratus could be used as a forage source to replace 40% of rice straw without any negative effects on rumen fermentation and blood metabolites in Hanwoo steers.
Kim, Min Hee;Lee, Jongsung;Jung, Sehyun;Kim, Joo Wan;Shin, Jae-Ho;Lee, Hae-Jeung
Journal of Ginseng Research
/
v.41
no.2
/
pp.120-126
/
2017
Background: The present study investigated the effect of ginseng berry hot water extract (GBx) on blood flow via the regulation of lipid metabolites and blood coagulation in rats fed a high-fat diet (HFD). Methods: Sixty rats were divided into five groups in descending order of body weight. Except for the control group, the other four groups were fed a HFD containing 45% kcal from fat for 11 wk without GBx. GBx groups were then additionally treated by gastric gavage with GBx dissolved in distilled water at 50 (GBx 50) mg/kg, 100 (GBx 100) mg/kg, or 150 (GBx 150) mg/kg body weight for 6 wk along with the HFD. To investigate the effects of GBx on rats fed a HFD, biochemical metabolite, blood coagulation assay, and histological analysis were performed. Results: In the experiments to measure the serum levels of leptin and apolipoprotein B/A, GBx treatment attenuated the HFD-induced increases in these metabolites (p < 0.05). Adiponectin and apolipoprotein E levels in GBx-treated groups were significantly higher than the HFD group. Prothrombin time and activated partial thromboplastin time were increased in all GBx-treated groups. In the GBx-treated groups, the serum levels of thromboxane $A_2$ and serotonin were decreased and concentrations of serum fibrinogen degradation products were increased (p < 0.05). Moreover, histomorphometric dyslipidemia-related atherosclerotic changes were significantly improved by treatment with GBx. Conclusion: These results suggest the possibility that GBx can ameliorate blood flow by decreasing intima-media thickness via the regulation of blood coagulation factors related to lipid metabolites in rats fed a HFD.
Kim, Hyun-Jin;Cho, Chang-Won;Hwang, Jin-Taek;Son, Nari;Choi, Ji Hea;Shim, Gun-Sub;Han, Chan-Kyu
Journal of Ginseng Research
/
v.37
no.3
/
pp.371-378
/
2013
Serum and liver metabolites in rats fed red ginseng (RG) were analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. The mass data were analyzed by partial least squares-discriminant analysis (PLS-DA) to discriminate between control and RG groups and identify metabolites contributing to this discrimination. The RG group was clearly separated from the control group on PLS-DA scores plot for serum samples, but not liver samples. The major metabolites contributing to the discrimination included lipid metabolites (lysophosphatidylcholine, acyl-carnitine, and sphingosine), isoleucine, nicotinamide, and corticosterone in the serum; the blood levels of all but isoleucine were reduced by RG administration. Not all metabolites were positively correlated with the health benefits of RG. However, the blood levels of lysophosphatidylcholine, which stimulate various diseases, and long-chain acylcarnitines and corticosterone, which activate the stress response, were reduced by RG, suggesting long-term RG might relieve stress and prevent physiological and biological problems.
The objective of the study was to determine the relationship between faecal egg counts and nutritionally-related blood metabolites in Nguni goats of South Africa. Body weights, body condition scores (BCS), FAMACHA scores, faecal and blood samples were collected from 96 Nguni castrates. Faecal samples were analysed using the modified McMaster technique for nematodes and the sedimentation method for trematodes. Blood was analysed for packed cell volume (PCV), glucose, cholesterol, total protein, albumin, urea and creatinine. Season had an effect on glucose, globulin, total protein, creatinine, PCV and faecal egg counts (FEC). Globulin, PCV, creatinine and FEC were significantly higher in the wet season compared to the dry season. A quadratic relationship existed between faecal egg count loads and BCS whilst negative linear relationships were observed between faecal egg counts and creatinine, albumin and cholesterol levels of Nguni goats.
Vakili, A.R.;Khorrami, Behzad;Mesgaran, M. Danesh;Parand, E.
Asian-Australasian Journal of Animal Sciences
/
v.26
no.7
/
pp.935-944
/
2013
Essential oils have been shown to favorably effect in vitro ruminal fermentation, but there are few in vivo studies that have examined animal responses. The objective of this study was to evaluate the effects of thyme (THY) and cinnamon (CIN) essential oils on feed intake, growth performance, ruminal fermentation and blood metabolites in feedlot calves fed high-concentrate diets. Twelve growing Holstein calves ($213{\pm}17kg$ initial BW) were used in a completely randomized design and received their respective dietary treatments for 45 d. Treatments were: 1-control (no additive), 2-THY (5 g/d/calf) and 3-CIN (5 g/d/calf). Calves were fed ad libitum diets consisting of 15% forage and 85% concentrate, and adapted to the finishing diet by gradually increasing the concentrate ratio with feeding a series of transition diets 5 wk before the experiment started. Supplementation of THY or CIN did not affect DMI and ADG, and feed efficiency was similar between treatment groups. There were no effects of additives on ruminal pH and rumen concentrations of ammonia nitrogen and total VFA; whereas molar proportion of acetate and ratio of acetate to propionate decreased, and the molar proportion of propionate increased with THY and CIN supplementation. Rumen molar concentration of butyrate was significantly increased by adding CIN compared to control; but no change was observed with THY compared with control group. No effects of THY, or CIN were observed on valerate, isobutyrate or isovalerate proportions. Plasma concentrations of glucose, cholesterol, triglyceride, urea-N, ${\beta}$-hydroxybutyrate, alanine aminotransferase and aspartate aminotransferase were not changed by feeding THY or CIN. Results from this study suggest that supplementing a feedlot finishing diet with THY or CIN essential oil might be useful as ruminal fermentation modifiers in beef production systems, but has minor impacts on blood metabolites.
Jin, Yan Mei;Zhang, Xiao Qing;Badgery, Warwick B;Li, Peng;Wu, Jun Xi
Asian-Australasian Journal of Animal Sciences
/
v.32
no.10
/
pp.1630-1639
/
2019
Objective: Sixty Pengbo semi-wool sheep ewes (approximately 1.5-years-old; $31.33{\pm}0.43kg$) were randomly assigned to two groups, either grazing (G) or dry lot feeding (D), to examine the effects of traditional daily grazing and dry lot feeding on performance and blood metabolites during the cold season in Tibetan Plateau. Methods: The ewes in the G group were grazed continuously each day and housed in one shed each evening, while the ewes in the D group were housed in another shed all day. All animals were fed 400 g/d of commercial concentrate, and grass hay was available freely throughout the experimental period. Results: Compared with the G group, the ewes in the D group had higher (p<0.05) live weight and weight gain. The D group ewes had greater (p<0.05) numbers of white blood cells and platelets, while they had lower (p<0.05) platelet-large cell ratios, cholesterol, high-density lipoprotein cholesterol and glutathione peroxidase, as compared with the G group ewes. Additionally, three serum metabolites, abscisic acid, xanthoxin and 3,4-dihydroxy-5-polypren, were upregulated (p<0.05) in the G group in comparison with the D group. Conclusion: In conclusion, a dry lot feeding regime during the winter and spring period will increase the productivity of sheep and improve blood physiological and biochemical profiles.
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