• Asian-Australasian Journal of Animal Sciences
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• v.22 no.9
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• pp.1267-1278
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• 2009

Three beef steers fitted with permanent cannulae in the rumen and duodenum were used to determine the effects of protein supply from soyhulls (SH) and wheat bran (WB) on ruminal metabolism, blood metabolites, nitrogen metabolism, nutrient digestion and concentrations of soluble non-ammonia nitrogen (SNAN) in ruminal (RD) and omasal digesta (OD). In a 3${\times}$3 Latin square design, steers were offered rice straw and concentrates formulated either without (control) or with two brans to increase crude protein (CP) level (9 vs. 11% dietary DM for control and bran-based diets, respectively). The brans used were SH and WB that had similar CP contents but different ruminal CP degradability (52 vs. 80% CP for SH and WB, respectively) for evaluating the effects of protein degradability. Ruminal ammonia concentrations were higher for bran diets (p<0.01) than for the control, and for WB (p<0.001) compared to the SH diet. Similarly, microbial nitrogen and blood urea nitrogen were significantly increased (p<0.05) by bran and WB diets, respectively. Retained nitrogen tended (p<0.082) to be increased by SH compared with the WB diet. Intestinal and total tract CP digestion was enhanced by bran diets. In addition, bran diets tended (p<0.085) to increase intestinal starch digestion. Concentrations of SNAN fractions in RD and OD were higher (p<0.05) for bran diets than for the control, and for WB than for the SH diet. More rumendegraded protein supply resulting from a higher level and degradability of CP released from SH and WB enhanced ruminal microbial nitrogen synthesis and ruminal protein degradation. Thus, free amino acids, peptides and soluble proteins from microbial cells as well as degraded dietary protein may have contributed to increased SNAN concentrations in the rumen and, consequently, the omasum. These results indicate that protein supply from SH and WB, having a low level of protein (13 and 16%, respectively), could affect ruminal metabolism and nutrient digestion if inclusion level is relatively high (>20%).

• Korean Journal of Plant Tissue Culture
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• v.24 no.2
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• pp.119-124
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• 1997

Plantlets were regenerated from various explants (shoot tip, leaf blade, petiole and root segments) via organogenesis and/or somatic embryogenesis from Armoracia rusticana(Lam) Gaerth., Mey, et Scherb.. Shoot regeneration rate from callus was highest on the MS mediums supplemented with 0.5 ㎎/L IAA, 5.0㎎/L BA and 10.0㎎/L spermine. A Low frequency of regeneration occurred on hormone-free MS medium. Multiple shooks were regenerated at a pH of 4.0 to 8.0 on MS medium supplemented with 1.0 ㎎/L BA and 0.1 ㎎/L NAA. Polyamines promoted shoot- and root-formation by 2 to 4 times normal, Specific proteins associated with organogenesis were identified. Somatic embryogenesis occurred directly from the leaf blade, petiole and root segments cultured on MS medium with 2.0 ㎎/L BA and 2.0 ㎎/L BA and 2.0 ㎎/L NAA. Three types of regeneration in A, rusticana were clearly established, which could be applied to the study of morphogenesis and genetics at cell, tissue and organ levels.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.2
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• pp.224-233
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• 2012

The limiting sequence and relative ratio of lysine (Lys), methionine (Met), and threonine (Thr) for calves about 2 mo of age fed milk replacers (MR) containing soy protein are not clearly defined. The objective of the study was to investigate the effect of supplementing MR containing 22% CP, half from soy protein concentrate (SPC, 40.56% CP, flour) and half from whey proteins, with Lys, Met, and Thr to estimate amino acid (AA) sequence and their relative ratio for calves about 2 mo of age. A method of partial deduction of AA was adopted. Twenty-four newborn calves (half males and half females, $40.7{\pm}0.9$ kg of BW) were fed 1 of 4 MR diets for 56 d (n = 6/diet). The diets were supplemented with all (positive control) or with 2 of the 3 AAs: Lys, Met and Thr, (i.e., PC (22% CP, 2.34% Lys, 0.72% Met and 1.80% Thr), PC-Lys (22% CP, 1.64% Lys, 0.72% Met and 1.80% Thr), PC-Met (22% CP, 2.34% Lys, 0.50% Met and 1.80% Thr), and PC-Thr (22% CP, 2.34% Lys, 0.72% Met and 1.26% Thr)). Calves were fed thrice daily; starter (20% CP, 1.03% Lys, 0.30% Met and 0.69% Thr), hay (3.23% CP, 0.29% Lys, 0.12% Met and 0.23% Thr) and water were offered free choice. Starter and hay were only offered beginning on d 36 (after 5 wk) and d 43 (after 6 wk), respectively. BW, body size and blood samples measures were taken every two weeks. Three-day total collection of feed refusals, feces, and urine were recorded starting at d 33 and d 54 of age, respectively. From the results, the limiting sequence and relative ratio between the 3 AAs in calves with different diet structures were calculated. The limiting sequence of the 3 AAs were ranked as Lys, Met and Thr; the proper ratio was 100:29:70 for MR-only diet and 100:30:60 for diets consisted of MR, starter and hay. Nitrogen digestion and utilization and nutrient digestibility were negatively affected by AA deletion treatments. From the evidence of this experiment, it did not appear that the AA limiting sequence was selectively altered by differences in diet structures such as would be encountered in practice. The relative ratio between the 3 AAs varied with the offer of starter and hay to calves, and the average ratio was 100:29.5:65 for calves during 2 to 10 wk of age.

• Journal of Life Science
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• v.27 no.9
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• pp.1003-1010
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• 2017

An open reading frame coding for mannanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned into Escherichia coli by polymerase chain reaction amplification, and completely sequenced. This mannanase gene, designated man26AT, consisted of 3,162 nucleotides encoding a polypeptide of 1,053 amino acid residues. Based on the deduced amino acid sequence, Man26AT was identified as a modular enzyme, which included a catalytic domain belonging to the glycosyl hydrolase family 26 and two carbohydrate-binding modules, CBM27 and CBM11. The amino acid sequence of Man26AT was homologous to that of several putative mannanases, with identity of 81% for P. ihumii and identity of less than 57% for other strains of Paenibacillus. A cell-free extract of recombinant E. coli carrying the man26AT gene showed maximal mannanase activity at $55^{\circ}C$ and pH 5.5. The enzyme retained above 80% of maximal activity after preincubation for 1 h at $50^{\circ}C$. Man26AT was comparably active on locust bean gum (LBG), galactomanan, and kojac glucomannan, whereas it did not exhibit activity on carboxymethylcellulose, xylan, or para-nitrophenyl-${\beta}$-mannopyranoside. The common end products liberated from mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, or LBG by Man26AT were mannose, mannobiose, and mannotriose. Mannooligosacchrides larger than mannotriose were found in enzymatic hydrolyzates of LBG and guar gum, respectively. However, Man26AT was unable to hydrolyze mannobiose. Man26AT was intracellularly degraded into at least three active proteins with different molecular masses by zymogram.

• Applied Chemistry for Engineering
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• v.31 no.2
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• pp.220-225
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• 2020

Hair is made of proteins containing various amino acids. Ultraviolet (UV) radiation is believed to be responsible for the most damaging effects of sunlight, and also plays an important role in hair aging. The purpose of this study was to investigate the changes in morphological and chemical structures after ultraviolet B (UVB) irradiation of human hair. The UVB-irradiated hair showed characteristic morphological and structural changes, compared to those of the normal hair. The result from a scanning electron microscope (SEM) equipped with an energy dispersive X-ray diffractometer (EDX) showed that the scale of UV-irradiated hair appeared to be rough and the amount of oxygen element was higher than that of the normal hair. Fluorescence and three dimensional (3D) topographical images were obtained by a confocal laser scanning microscope (CLSM). In 3D images, the green emission intensity of normal hair was much higher than that of fluorescing UVB-irradiated hair. The intensity of green emission reflects the intrinsic fluorescence of hair protein. Also, a fluorescent imaging method using fluorescamine reagent was used to identify the free amino groups resulting from a peptide bond breakage in UVB-irradiated hair. Strong blue fluorescence of UVB-irradiated hair, which indicates a very high level of amino groups, was observed by CLSM. Therefore, the fluorescamine as an extrinsic fluorescence could provide a useful tool to identify the peptide bond breakage in UVB-irradiated hair. Infrared image mapping was also employed to assess the cross-sections of normal and UVB-irradiated specimens to examine the oxidation of disulfide bonds. The degree of peak areas with strong absorbance for the disulfide mono-oxide was spread from the outside to the inside of hair. The spectroscopic techniques used alone, or in combination, launch new possibilities in the field of hair cosmetics.

• Korean Chemical Engineering Research
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• v.60 no.2
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• pp.300-307
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• 2022

Time-resolved serial femtosecond crystallography (TR-SFX) is a powerful technique for determining temporal variations in the structural properties of biomacromolecules on ultra-short time scales without causing structure damage by employing femtosecond X-ray laser pulses generated by an X-ray free electron laser (XFEL). The mixing rate of reactants and biomolecule samples, as well as the hit rate between crystal samples and x-ray pulses, are critical factors determining TR-SFX performance, such as accurate image acquisition and efficient sample consumption. We here develop two distinct sample delivery systems that enable ultra-fast mixing and on-demand droplet injecting via pneumatic application with a square pulse signal. The first strategy relies on inertial mixing, which is caused by the high-speed collision and subsequent coalescence of droplets ejected through a double nozzle, while the second relies on on-demand pneumatic jetting embedded with a 3D-printed micromixer. First, the colliding behaviors of the droplets ejected through the double nozzle, as well as the inertial mixing within the coalesced droplets, are investigated experimentally and numerically. The mixing performance of the pneumatic jetting system with an integrated micromixer is then evaluated by using similar approaches. The sample delivery system devised in this work is very valuable for three-dimensional biomolecular structure analysis, which is critical for elucidating the mechanisms by which certain proteins cause disease, as well as searching for antibody drugs and new drug candidates.

• Korean Journal of Veterinary Research
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• v.5 no.1
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• pp.1-16
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• 1965

I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

• Applied Microscopy
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• v.32 no.2
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• pp.91-105
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• 2002

Urea transport in the kidney is mediated by a family of transporter proteins that includes renal urea transporters (UT-A) and erythrocyte urea transporters (UT-B). The cDNA of five isoforms of rat UT-A, UTA1, UT-A2, UT-A3, UT-A4, and UT-A5 have been cloned. The purpose of this study was to examine the expression of UT-A (L194), which marked UT-A1, UT-A2 and UT-A4. Male Sprague-Dawley rats, weighing approximately 200 g, were divided into three group: control rats had free access to water, dehydrated rats were deprived of water for 3 d, and water loaded rats had free access to 3% sucrose water for 3 d before being killed. The kidneys were preserved by in vivo perfusion through the abdominal aorta with the 2% paraformaldehyde-lysine- periodate (PLP) or 8% paraformaldehyde solution for 10 min. The sections were processed for immunohistochemical studies using pre-embedding immunoperoxidase method and immunogold method. In the normal rat kidney, UT-A1 was expressed intensely in the cytoplasm of the inner medullary collecting duct (IMCD) cell and UT-A2 was expressed on the plasma membrane of the terminal portion of the shortloop descending thin limb (DTL) cells (type I epithelium) and of the long-loop DTL cells (type II epithelium) in the initial part of the inner medulla. Immunoreactivity for UT-A1 in the IMCD cells, was decreased in dehydrated animals whereas strongly increased in water loaded animals compared with control animals. In the short-loop DTL, immunoreactivity for UT-A2 was increased in intensity in both dehydrated and water loaded groups. However, in the long-loop DTL of the outer part of the inner medulla, immunoreactivity for UT-A2 was markedly increase in intensity in dehydrated group, but not in water loaded group. In conclusion, in the rat kidney, UT-A1 is located in the cytoplasm of IMCD cells, whereas UT-A2 is located in the plasma membrane of both the short-and long-loop DTL cells. Immunohistochemistry studies revealed that UT-A1 and UT-A2 may have a different role in urea transport and are regulated by different mechanisms.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.5
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• pp.408-418
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• 1987

The quality of different grades of dried lavers obtained from three culture areas was evaluated and its changes during the storage at different levels of water activity were measured. Not much differences in general chemical composition between the locality was detected except some in the content of lipid and pigments. But the quality grades of dried lavers were mainly depended upon the content of protein and pigments including chlorophyll a, carotenoids, and biliproteins although there was little difference in amino acid composition of the proteins, and glutamic acid, aspartic acid and alanine were high in general. The lipid of dried lavers was composed of a high level of polyunsaturated fatty acids, particularly, of eicosapentaenoic acid which amounted to as much as a half of the total lipid, and of palmitic acid that reached a quarter depending on grades. The quality of dried layers was significantly changed by equilibrium moisture level when stored for three months at different water activities in range of 0.1 to 0.6. The loss of chlorophyll a, carotenoid, biliproteins, ascorbic acid, and browning were markedly retarded at aw 0.1 to 0.2. Oxidation of polyunsaturated fatty acids and the loss of free amino acids were also minimized at aw 0.2. Glutamic acid and methionine were reduced very fast during the storage.