Sarcolemmal membrane fraction from canine ventricle was isolated from the discarded pellet after the first homogenization in the isolation procedure of sarcoplasmic reticulum (Method 1) and the protein yield, purity, and sidedness of this preparation were compared to those of sarcolemmal fraction prepared by method of Lee et al. (Method 2) and a slight modification of original protocol of Jones et al. (Method 3). Method 1 differed from Method 2 essentially only in that vigorous homogenization was carried out by omnimixer and homogenization medium containing 30mM Tris-maleate was used in the first step. The sarcolemmal fraction was enriched from 45 to 50 and 29-fold in [$^3H$] ouabain, [$^3H$] DHA, [$^3H$] QNB binding and $Na^+$, $K^+$-ATPase activity, respectively, compared to homogenate. Total $Na^+$, $K^+$-ATPase activity of highly sarcolemma enriched fraction was 144.6$\pm$16.4$\mu\textrm{mol}$ Pi/mg protein/hr, which was about 85%, of total ATPase activity, and the yield of the preparation was 15.7 mg protein per 100g of starting ventricular tissue. The sarcolemmal preparation supported $^{45}Ca^{2+}$-uptake in the presence of ATP but this uptake was not dependent on oxalate. Sarcolemmal $Na^+$, $K^+$-ATPase activity and detectable [$^3H$] ouabain binding were increased about 32% and 35%, respectively, by pretreatment of sarcolemmal fraction with optimal concentration of sodium dodecylsulfate (0.3-0.4mg/mg protein), suggesting that this preparation contained about 24% of sealed rightside-out vesicles, 26% of sealed inside-out vesicles, and 5001o of freely permeable (leaky) form. This procedure showed the highest protein yield and leaky population, compared to Method 2 and 3. On the other hand, sarcolemmal fraction prepared by Method 2 and 3 showed low value in protein yield but comtained high population of inside-out (46%) and rightside-out (49%) vesicles, respectively, compared to present procedure (Method 1). The results indicate that vigorous homogenization decreases the population of sealed sarcolemmal vesicles but increases the sarcolemmal protein yield per gram tissue and that this procedure is available for further purification of sarcolemmal fraction and for the receptor binding study of sarcolemma.
Objective: This study was conducted to enhance our understanding of nitrogen (N) metabolism and mammary amino acid (AA) utilization in lactating cows with divergent phenotypes of residual feed intake (RFI). Methods: Fifty-three multiparous mid-lactation Holstein dairy cows were selected for RFI measurements over a 50-d experimental period. The 26 cows with the most extreme RFI values were classified into the high RFI (n = 13) and low RFI (n = 13) groups, respectively, for analysis of N metabolism and AA utilization. Results: Compared with the high RFI cows, the low RFI animals had lower dry matter intake (p<0.01) with no difference observed in milk yield between the two groups (p>0.10). However, higher ratios of milk yield to dry matter intake (p<0.01) were found in the low RFI cows than in the high RFI cows. The low RFI cows had significant greater ratios of milk protein to metabolizable protein (p = 0.02) and milk protein to crude protein intake than the high RFI cows (p = 0.01). The arterial concentration and mammary uptake of essential AA (p<0.10), branched-chain AA (p<0.10), and total AA (p<0.10) tended to be lower in the low RFI cows. Additionally, the low RFI cows tended to have a lower ratio of AA uptake to milk output for essential AA (p = 0.08), branched-chain AA (p = 0.07) and total AA (p = 0.09) than the high RFI cows. Conclusion: In summary, both utilization of metabolizable protein for milk protein and mammary AA utilization are more efficient in cows with lower RFI than in the high RFI cows. Our results provide new insight into the protein metabolic processes (related to N and AA) involved in feed efficiency.
Isoflavone was extracted with various concentration of aqueous methanol using whole hypocotyls as the starting material. Whole hypocotyls were preferred as the raw material because the residue could be easily removed from the solvent after the extraction process. Extraction yield was almost constant at the methanol concentration of $20-80\%$. Most of the isoflavone was extracted within 1 hr, and the extraction yield remained almost constant thereafter. When the concentration of methanol was $80\%$, the content of total solid was reduced due to the reduced extraction of contaminating protein as the result of protein insolubilization. Among resins tested, Diaion HP-20, Amberlite XAD-16, and Amberlite IRC-50 showed the highest capacity to absorb the compound. Open column chromatography with Diaion HP-20 showed that $80\%$ aqueous ethanol was most efficient as the eluting solvent with final recovery of the phytochemical being more than $95\%$. Maximum adsorption of the phytochemical occurred at the acidic pH 2-4. When the spatial velocity was increased to 15 and more, the degree of adsorption was decreased, whereas below spatial velocity of 15, the adsorption capacity of isoflavone to the resin was almost constant. The purity of the isoflavone purified by column chromatography was $78\%$.
This study was performed to determine whether the infusion of nucleosides and a nucleotide mixture directly ito intestinal lumen can induce a regenerative effect on impaired intestinal mucosa. The effects of massive small bowel resention and also total parenteral nutrition were induced by surgical creation of Thirty-Vella fistual in male Sprague-Dawley rats. The rats received saline solution (Control group) or nucleosides and a nucleotide mixture(lower concentration group(Nucl) or higher concentration group (Nuc2) every two days into the fistula. Mucosal protein, DNA , ornithine decarboxylase(ODC) activity, and morphometry were evaluated at 9 or 21 days postoperation in the fistual and also in the residual ileal segment. On the 9th day, mucosal protein, DNA content, and villous surface area in the fistula and also in the residual ileum increased in rats that received nucleosides and a nucleotide mixture of lower concentration (Nuc 1). On the 21 th day, there were no significant differences in intestinal mucosa between the control group and the lower level nucleoside nucleotide mixture-treated group. The fistula villous height of the higher nucleosides and a nucleotide mixture group was higher than in the control rats. Fistula mucosal ODC activities were not significantly different between groups although the mucosal ODC activity of the residual ileal segment was increased on the 9th day. Our data suggests that this animal model is suitable for studying the effect of dietary factors on intestinal mucosal growth and regeneration after villous stropy , differentiating direct effects of diet on the intestine from systemic effects. It is also suggested that external nucleosides and nucleotides have supportive effects on intestinal mucosal regeneration.
The experiment was designed on 42 non pregnant Black Bengal goat. Out of which 18 were subjected to a superovulatory treatment comprising of eCG and hCG for embryo transfer study. The remaining 24 goats received no treatment and served as control for parameter studied as well as recipient for embryo transfer studies. Important biochemical constituents such as acid and alkaline phosphatase, total protein and cholesterol and inorganic phosphorus were estimated in the follicular fluid of control and treated group and the values were separately recorded for small medium and large size follicle. The results indicated a significant effect on acid phosphotase activity due to size of follicle. The value increased progressively from small to medium and from medium to large follicles. Alkaline phosphotase activity showed reverse trend. Alkaline phosphotase decreased progressively as size increased. The concentration of inorganic phosphorus did not reveal any significant difference between the control and treatment groups and also between the different size follicles. The concentration of protein decreased significantly from small to medium and from medium to large, although no difference was observed between the control and treatment groups. The concentration of Cholesterol in the follicular fluid indicated a significant increase from small to medium and to large follicle. Here also no difference was observed due to treatment. Similar in the composition of follicular fluid in the respect of above mentioned constituents indicated no of super ovulatory treatment on follicular fluid composition.
The climate and marketing system of raw milk in Taiwan create problems in balance feeding of protein and energy in lactating cows in Taiwan. Level of urea nitrogen both in bulk milk and serum reflects ruminal protein degradation and post-ruminal protein provision, whereas milk protein concentration responds to dietary energy intake and bacterial protein production in the rumen. Establishment of a range of reference standards in milk protein and urea nitrogen levels can be applied as a noninvasive economical feeding guide to monitor the balance of protein and energy intake. Standard reference levels of 3.0% milk protein and 11-17 mg/dL milk urea nitrogen (MUN) were established. Level of milk protein below 3.0% is regarded as indicating inadequate dietary energy whereas MUN below or above the range is regarded as a deficiency or surplus in dietary protein. Results from analysis of bulk a milk samples collected from 174 dairy herds over Taiwan showed that only one quarter (25.29%) of the herds received a balanced intake of protein and energy, 33.33% adequate protein with energy inadequate, 22.99% herds in protein surplus with energy inadequate, 10.35% herds in protein surplus with energy adequate, 4.6% protein deficiency with energy adequate, and 3.45% herds with both protein and energy inadequate. Energy inadequate herds accounted for 60% of the total dairy herds in Taiwan with 56% adequate, 38% surplus and 6% inadequate in protein. In comparing milk sampled from bulk milk on different seasons from Lee-Kang area in the southern Taiwan, the concentrations of milk fat and milk protein were significantly higher in the cool season (February) than in the warm season (August) (p<0.05), whereas the urea nitrogen in the milk was significantly lower in the cool season than in the warm season (p<0.05). This indicated that lactating cows had excess protein and/or inadequate energy intake in the warm season in this area. It appears that the major problem feeding in lactating cows is energy intake shortage, especially during the warm season in Taiwan.
Journal of Physiology & Pathology in Korean Medicine
/
v.25
no.6
/
pp.1020-1025
/
2011
Effects of Hwangryeun(Copitidis rhizoma) extract on recovery of liver function were researched in $CCl_4$ intoxicated rats. Concentration ${\alpha}$-fetoprotein, total protein in plasma showed a tendency to decrease in Hwangryeun(Copitidis rhizoma) extract applied groups while that of albumin showed a increase. Activities of plasma aspartate aminotransferase and alanine aminotransferase in Hwangryeun(Copitidis rhizoma) extract applied groups showed a lower value than those of controlled groups. Alkaline phosphatase, lactate dehydrogenase and ${\gamma}$-glutamyl transpeptidase activities showed a tendency to decrease in Hwangryeun(Copitidis rhizoma) extract applied groups. Concentration of plasma triglyceride and total cholesterol showed a lower values than that of control group.
The effect of dietary supplementation of sodium salt of isobutyric acid in low protein (10% CP) wheat straw based diet on nutrient utilization and rumen fermentation was studied in ruminally fistulated male crossbred cattle. The study included a 7 day metabolism and a 3 day rumen fermentation trials. The cattle were distributed into two equal groups of 4 each. The animals of control group were fed a basal diet consisting of wheat straw, concentrate mixture and green maize fodder in 40:40:20 proportion whereas branched chain volatile fatty acid (BCFA) supplemented group received a basal diet + isobutyric acid at 0.75 percent of basal diet. The duration of study was 36 days. The feed intake between experimental groups did not differ significantly and the average total DMI (% BW) was 2.01 and $2.28kg\;day^{-1}$ in control and BCFA supplemented diets. The dietary supplementation of BCFA improved (p<0.05) the DM, OM, NDF and cellulose digestibility by 4.46, 6.63, 10.57 and 11.31 per cent over those fed control diet. The total N retention on BCFA supplementation was improved (p<0.01) due to decreased (p<0.05) urinary N excretion. The concentrations of ruminal total N was 37.07 and $34.77mg\;100ml^{-1}$ in control and BCFA fed groups, respectively. Dietary supplementation BCFA significantly (p<0.01) reduced the ruminal ammonia N concentration as compared to control and the mean values ($mg\;100ml^{-1}$) were 13.18 and 9.42 in control and BCFA fed groups. The total VFA concentration was higher (p<0.01) in BCFA supplemented group (101.14 mM) than the control (93.05 mM). Among the VFAs, the molar proportion of acetate was higher (p<0.01) in BCFA supplemented group (71.07 mM) as compared to control (64.98 mM). However, the concentration of propionate and butyrate remained unchanged. Amino acids composition of bacterial hydrolysates was similar in both the groups. Ruminal outflow rate of liquid digesta was higher (p<0.01) in BCFA fed group ($67.56l\;day^{-1}$) than control ($52.73l\;day^{-1}$). It is concluded that the dietary supplementation of Na-salt of isobutyric acid in low protein diet improved the nutrient utilization and ruminal fermentation characteristics.
Huang, Chengfei;Li, Ping;Ma, Xiaokang;Jaworski, Neil William;Stein, Hans-Henrik;Lai, Changhua;Zhao, Jinbiao;Zhang, Shuai
Asian-Australasian Journal of Animal Sciences
/
v.31
no.8
/
pp.1315-1324
/
2018
Objective: An experiment was conducted to investigate the effects of different diet formulations: F1 (Two complicated basal diets containing different crude protein levels plus tested feedstuff) vs F2 (A simple corn soybean meal [SBM] basal diet plus tested feedstuff) combined with total collection (TC) or chromic oxide ($Cr_2O_3$) marker or acid-insoluble ash (AIA) marker method, and freeze-dry or oven-dry (OD) technique on estimation of nutrient digestibility in diets fed to growing pigs. Methods: In F1, twelve barrows were allocated to two $6{\times}4$ Youden Squares. The treatment diets included a high protein basal (HPB) diet, a low protein basal (LPB) diet, a corn diet and a wheat bran (WB) diet formulated based on the HPB diet, and a SBM diet and a rapeseed meal (RSM) diet formulated based on the LPB diet. In F2, eight barrows were allocated to two $4{\times}4$ Latin Squares. The treatment diets included a corn basal diet, a SBM basal diet formulated based on the corn diet, and a WB diet and a RSM diet formulated based on the SBM diet. Results: Concentration of digestible (DE) and metabolizable energy (ME), and the apparent total tract digestibility of gross energy, ash, neutral detergent fibre, and acid detergent fibre determined by $Cr_2O_3$ marker method were greater than those determined by TC and AIA marker methods in HPB, LPB, and RSM diets formulated by F1 and in corn diet formulated by F2 (p<0.05). The DE values in WB and both DE and ME values in SBM and RSM estimated using F1 were greater than those estimated using F2 (p<0.05). Conclusion: From the accuracy aspect, the AIA marker or TC method combined with OD technique is recommended for determining the energy concentration and nutrient digestibility of components in diets fed to growing pigs.
Physicochemical changes of milt during the spermiation period were investigated in black seabream (Acanthopagrus schlegeli) reared in recirculating seawater system. The spermiation period for the milt collection in cultured brood stock was from 11 April to 4 June. During the spermiation period, average milt volume (ml/100 g body weight) was $0.70{\pm}0.33\;ml$ and maintained high level from 2 May to 4 June. The total number of stripping spermatozoa per 100 g body weight reached the maximum value $(3.32{\times}10^{10})$ in 9 May, then decreased rapidly thereafter. Spermatozoa concentration per ml reached the minimum value in 2 May. There was no change in spermatocrit for the spermiation period. Total protein, total lipid, glucose and Na concentration in spermatozoa and seminal fluid were lower than those in plasma. Total protein, total lipid and K concentration in spermatozoa were higher than those in seminal fluid. The glucose concentration in spermatozoa and seminal fluid in April and May were significantly higher than those in June.
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