• Animal Bioscience
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• v.35 no.9
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• pp.1351-1359
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• 2022

Objective: The objective of this study was to analyse the differentially abundant proteins caused by freeze-thawing of ram sperm and explore candidate proteins of interest for their ability to improve ram sperm cryopreservation outcomes in vitro. Methods: Sperm were from three mature Dorper. Fresh and frozen sperm proteins were extracted, and the differentially abundant proteins were analysed by mass spectrometry. Among these proteins, lactoferrin (LTF) was selected to be added before cryopreservation. Next, sperm samples were diluted in Tris extender, with the addition of 0, 10, 100, 500, and 1,000 ㎍/mL of LTF. After thawing, sperm quality was evaluated by motility, plasma membrane integrity, mitochondrial activity and reactive oxygen species (ROS). Results: Cryopreservation significantly altered the abundance of 40 proteins; the abundance of 16 proteins was increased, while that of 24 proteins was decreased. Next, LTF was added to Tris extender applied to ram sperm. The results showed that sperm motility and plasma membrane integrity were significantly improved (p<0.05) by supplementation with 10 ㎍/mL LTF compared to those in the control group. There was no significant difference in mitochondrial activity between the 0 ㎍/mL group and other groups (p>0.05). Supplementation of the cryoprotective extender with 10 ㎍/mL LTF led to decreased ROS levels compared with those in the control and other groups (p<0.05). Conclusion: The LTF is an important protein during cryopreservation, and the addition of 10 ㎍/mL LTF to a cryoprotective extender can significantly improve the function of frozen ram sperm.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.3
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• pp.348-352
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• 2003

An investigation was made into the protein profile of colostrum/milk of ten Murrah buffaloes and of their ten buffalo calves during their first week of neonatal life to study the materno-neonatal transfer of immunoglobulins (Ig). Calves were pail fed 3.5 liter of colostrum and/or milk per calf/day exclusively from their dam. First blood sample from newborn calves was collected before colostrum feeding on the day of birth (day zero) and the sampling continued daily for seven days after colostrum/milk feeding. Colostrum/milk Ig and IgG values were $4.82{\pm}2.60$, $2.19{\pm}1.90$, $1.12{\pm}0.82$, $0.69{\pm}0.44$, $0.59{\pm}0.31$, $0.47{\pm}0.20$, $0.40{\pm}0.22$, $0.40{\pm}0.25$ and $3.58{\pm}1.90$, $1.08{\pm}0.92$, $0.52{\pm}0.40$, $0.31{\pm}0.20$, $0.27{\pm}0.14$, $0.22{\pm}0.08$, $0.18{\pm}0.09$, $0.14{\pm}0.08$ respectively during 0-7 days post partum. The concentration of total colostrum/milk proteins, Ig, IgG and albumin were highest within 12 h post-partum. Thereafter, the concentrations followed a declining trend which may be attributed to the reduced transfer of proteins from the maternal blood, declining synthesis by the mammary glands and/or depletion of stored proteins. The concentrations of plasma Ig and IgG before colostrum feeding on day zero were $0.42{\pm}0.09$ and $0.08{\pm}0.03$ respectively. The levels of plasma Ig were $1.90{\pm}0.37$, $1.80{\pm}0.31$, $1.80{\pm}0.26$, $1.81{\pm}0.28$, $1.78{\pm}0.31$, $1.79{\pm}0.21$, $1.80{\pm}0.32$ and of IgG were $1.57{\pm}0.41$, $1.30{\pm}0.29$, $1.31{\pm}0.21$, $1.27{\pm}0.18$, $1.23{\pm}0.21$, $1.23{\pm}0.16$, $1.26{\pm}0.21$ on days 1-7 after birth after colostrum/milk feeding. The concentrations of total plasma proteins, Ig, IgG were lowest before colostrum feeding and increased significantly (p<0.05) after colostrum feeding in buffalo neonates. The results suggest that the highest amounts of colostral Ig and IgG were available on the day of parturition and thus the calves should receive colostrum as early after birth as possible. Colostrum Ig and IgG concentrations were not correlated to plasma Ig and IgG concentrations in the post-suckle buffalo calves and therefore, colostrum Ig and IgG concentrations were probably not the principle determinants of calf post-suckle plasma Ig and IgG concentrations.

• Journal of Veterinary Clinics
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• v.32 no.1
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• pp.62-68
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• 2015

There is no reliable indicator available for the diagnosis of horse laminitis, although the disease is common and costly. This study was performed to develop a specific diagnostic biomarker for laminitis. We have identified 33 differentially expressed proteins in plasma of a horse suffering laminitis that is experimentally induced by an overdose of oligofructose, in comparison with normal horse plasma. Among the proteins, myosin-9 mRNA was found in RNA sequencing analysis to be expressed specifically in laminitis tissues compared to other horse tissues. It is thus suggested that expression of plasma myosin-9 may be used for the diagnosis of equine laminitis.

• Journal of Pharmaceutical Investigation
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• v.20 no.4
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• pp.179-191
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• 1990

A central dogma of pharmacology is that only unbound drug is capable of translocation across biological membrane. Thus, hepatic uptake is assumed to be solely determined by the unbound concentration of the diffusible moiety at the surface of the liver cell. However, an increasing number of experimental observations with xenobiotics that are normally very extensively bound to plasma proteins (>99%) appear to be inconsistent with these assumptions. This suggested that in addition to progressive spontaneous dissociation within the liver sinusoids and space of Disse, direct interactions of the albumin-drug complex at the plasma membrane may facilitate dissociation of the complex. To explain this phenomena. called albumin-mediated uptake, 4 mechanisms have been suggested. The validity of such hypotheses needs to be examined by the further study. Because albumin-mediated uptake has also been observed to occur in other plasma proteins, protein-mediated uptake rather than albumin-mediated uptake seems to be acceptable.

• Journal of Pharmaceutical Investigation
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• v.30 no.2
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• pp.73-83
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• 2000

Polyethylene glycol (PEG) is a water soluble, biocompatible, non-toxic polymer and PEGylation is a well established technique for the modification of therapeutic proteins and peptides. PEG-protein drugs have been extensively studies in relation to therapies for various diseases: cancer, inflammation and others. The covalent attachment of PEG to proteins and peptides prolonged plasma half-life, reduced antigenicity and immunogenicity, increased thermal and mechanical stability, and prevented degradation by enzymes. Several chemical groups for general and site specific conjugation have been exploited to activate PEG for amino group, carboxyl group, and cysteine groups. PEGylation of many proteins and peptides have been studied to enhance their properties for the potential uses. Also, the different positional isomers in several PEG-proteins have shown the difference in vivo stability and biological indicating that the site of PEG molecule attachment is one of the important factor to develop PEG-proteins as potential therapeutic agents.

• The Plant Pathology Journal
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• v.31 no.4
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• pp.323-333
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• 2015

As sessile organisms, plants are exposed to persistently changing stresses and have to be able to interpret and respond to them. The stresses, drought, salinity, chemicals, cold and hot temperatures, and various pathogen attacks have interconnected effects on plants, resulting in the disruption of protein homeostasis. Maintenance of proteins in their functional native conformations and preventing aggregation of non-native proteins are important for cell survival under stress. Heat shock proteins (HSPs) functioning as molecular chaperones are the key components responsible for protein folding, assembly, translocation, and degradation under stress conditions and in many normal cellular processes. Plants respond to pathogen invasion using two different innate immune responses mediated by pattern recognition receptors (PRRs) or resistance (R) proteins. HSPs play an indispensable role as molecular chaperones in the quality control of plasma membrane-resident PRRs and intracellular R proteins against potential invaders. Here, we specifically discuss the functional involvement of cytosolic and endoplasmic reticulum (ER) HSPs/chaperones in plant immunity to obtain an integrated understanding of the immune responses in plant cells.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.3
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• pp.358-364
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• 2001

The physiochemical properties and interactions between porcine blood and waxy rice were determined. Addition of calcium chloride (0.15%) improved acceptability of blood cake and increased the gelatinization degree of waxy rice. The water-holding capacity of porcine blood gel (blood/water=60/40, v/v), extent of absorption and gelatinization of waxy rice, and scanning electron microscopy showed that blood protein matrix and waxy rice are competitors for holding water in the cooking procedure. Non-haem iron content increased linearly (R=0.95) when heating temperature rose. The presence of blood proteins caused increasing of peak temperature (Tp) of gelatinization in differential scanning calorimetric thermal gram, The microstnlcture of plasma proteins and haemoglobin appeared continuous changes, and interacted with surface of waxy rice flour in terms of network and mosaic form, respectively. The electrophoretic patterns revealed an interaction between plasma proteins and waxy rice glutelin and haemoglobin when heated could be found at temperatures above $60^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1594-1599
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• 2007

Plasma membrane proteins from pig adipocytes, brain, heart, kidney, liver and spleen were isolated using a 32% sucrose gradient. An adult male sheep was immunized three times at 3-wk intervals with the purified pig adipocyte plasma membrane (APM) proteins. Blood samples were taken from the immunized sheep 12 d after the third immunization. Antiserum showed strong reactivity with APM proteins determined by ELISA, and the reactivity could be detected at dilutions in excess of 1:128,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver or spleen. Ninety weanling pigs were allocated randomly to three treatment groups and were injected i.p. with 40 ml of antiserum (n = 30) or 20 ml of lyophilized antiserum (21.5 mg/ml; n = 30). A control group (n = 30) received 40 ml of saline, and all pigs were slaughtered at 24 wk of age. The polyclonal antiserum did not change BW or ADG. Carcass percentage of pigs was numerically increased by the antiserum treatment compared with control. Both antiserum treatments did not significantly (p>0.05) affect body composition, including body fat content, relative to the control group.

• Food Science of Animal Resources
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• v.40 no.2
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• pp.172-182
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• 2020

In the present study, we determined the degree of hydrolysis (DH) of porcine blood plasma proteins, albumin, and globulin hydrolyzed by six proteases (alcalase, neutrase, flavourzyme, protamex, trypsin, and papain) for various reaction times. Moreover, antimicrobial activities of hydrolysates against five pathogenic microorganisms (Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium, Escherichia coli, and Shigella flexneri) were investigated. Alcalase, trypsin, and papain hydrolysates of the three porcine blood proteins showed higher DH values than hydrolysates produced by the other three proteases. DH of the three porcine blood proteins hydrolyzed by the six proteases failed to increase after 2 h of hydrolysis. In antimicrobial tests, hydrolysates (hydrolysis time of 2 h) showed antibacterial activity only against B. cereus. Albumin hydrolysates showed higher antimicrobial activity than globulin and plasma hydrolysates. Albumin hydrolysates obtained with flavourzyme, protamex, and trypsin showed higher antimicrobial activity than those obtained with the other three proteases.

• Korean Journal of Food Science and Technology
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• v.23 no.4
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• pp.428-432
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• 1991

This study was carried out to investigate the effect of NaCl, pH and phosphate on the functional properties of a mixed system of plasma protein and myofibrillar proteins. The solubility of plasma protein, myofibrillar protein and the mixture (plasma+myofibrillar protein) increased according to the increase of NaCl concentration ($0{\sim}4%$) and pH $pH4{\sim}8$). The solubility, emulsifying activity and capacity of the mixture were lower than those of plasma protein, whereas higher than those of myofibrillar protein. The gel strength of the mixture and myofibrillar protein showed a significant increase when NaCl concentration was increased from 2 to 3%. The gel strength of myofibrillar protein increased about four times when 0.3% polyphosphate added to the sample containing 2% NaCl, whereas the moisture loss of the mixture and myofibrillar protein decreased significantly. The gel strength of plasma protein, myofibrillar protein and the mixture increased slightly at $3{\sim}5%$ protein concentration, whereas the gel strength of those increased significantly as the protein concentration increased from 5 to 9%.