• Clinical and Experimental Pediatrics
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• v.46 no.6
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• pp.536-540
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• 2003

Purpose : Aspiration of foreign material into the lungs can cause acute or chronic pulmonary diseases. It is difficult to detect small amounts of aspiration due to the lack of safe, sensitive and specific diagnostic tests. Recently, in animal or human studies, it has been reported that immunochemistry for lactalbumin can be used to detect the minimal aspiration. So, the authors' investigation was designed to determine whether human milk phagocytized alveolar macrophages can be detected in human milk aspirated mice. Methods : Sixty four male mice, 6-8 weeks old and 30-40 gm weighing, were used for this study. About 0.05 mL of human milk or normal saline were given intranasally once per day for 1 day or 3 days. Under anesthesia with ketamine and xylazine, the trachea of each mouse was cannulated with an 18G Jelco needle and then, each mouse's lungs were lavaged three times with 0.5 mL of phosphate buffer solution at 2, 8, 24, and 48 hours after the last milk or normal saline instillation. Cells in bronchoalveolar lavage fluid were stained with Oil Red O and immunocytochemistry for alpha-lactalbumin. Results : Immunocytochemical reactivity for alpha-lactalbumin or lipid-laden alveolar macrophages were not observed in the normal saline aspirated groups. Immunocytochemical reactivity for alpha-lactalbumin were observed in the human milk aspirated groups. They showed a peak at 8 hours and decreased markedly at 24 hours but persisted even at 48 hours after aspiration. Immunocytochemical stain positive alveolar macrophages were noted similarly in number between single and multiple aspiration groups. Conclusion : These observations suggested that alveolar macrophages for lactalbumin could be more easily detected on immunocytochemistry than Oil Red O stain, and immunocytochemistry could be used as a sensitive and specific diagnostic test for the detection of human milk aspiration.

• Journal of Food Hygiene and Safety
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• v.5 no.4
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• pp.219-228
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• 1990

Whey proteins in milk were analyzed by polyacrylamide gel electrophoresis and compared with respect to electrophoregrams, densitograms and concentrations of whey proteins in raw and market milk classified according to 3 kinds of pasteurization by low temperature long time. high temperature short time and ultra-high temperature short time. Relative composition of major whey protein constituents such as bovine serum albumin, ${\alpha}\;-\;lactalbumin\;and\;{\beta}-lactoglobulin$ in raw milk were 3.71:11.44:84.85 and not affected by low temperature long time and high temperature short time pasteurization, even though there were the tendencies of some declining in the actual concentrations. But by ultra-high temperature short time pasteurization compositions of whey protein were changed to 0: 64.75: 35 in which reflected the disapprearance of bovine serum albumin and the extensive decrease of ${\beta}-lactoglobulin$. Storage of low temperature pasteurized milk at $5^{\circ}C$ resulted in a slight decrease of ${\alpha}\;-\;lactalbumin\;a\;{\beta}-lactoglobulin$, but storage at $25^{\circ}C$ did not make any changes until3rd days of storage. Most of whey proteins in high temperature short time pasteurized milk were not affected during storage at $5^{\circ}C\;and\;25^{\circ}C$, but bovine serum albumin and ${\alpha}\;-lactalbumin$ diminished in 2-3 days of storage. Whey proteins of milk treated with ultra-high temeperature were not affected during storage at $5^{\circ}C\;and\;25^{\circ}C$ except a slight decrease of ${\alpha}\;-lactalbumin$ in 2nd day of storage at $5^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.3
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• pp.305-308
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• 2006

A genetic study was conducted to elucidate the effect of alpha-Lactalbumin (${\alpha}$-LA) gene polymorphism on milk production traits involving total milk yield and daily milk yield during first lactation in two breeds of water buffaloes namely, Murrah and Bhadawari. Single strand conformation polymorphism (SSCP) was carried out to explore genetic polymorphism present at this locus. For this study, exon 1 region of ${\alpha}$-LA was analyzed. Finally, polymorphism data was associated with milk production traits by employing least square analysis. In Murrah buffalo, five genotypes such as AB, BB, BC, CC and CD and four alleles A, B, C and D were detected whereas in Bhadawari buffalo two genotypes namely, AB and BC and three alleles namely, A, B and C were found. Genotypes showed significant effects ($p{\leq}0.05$) on total milk yield and daily milk yield in Bhadawari buffalo but had non-significant effects on these traits in Murrah buffalo.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.7
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• pp.1041-1045
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• 2003

In order to study the reaction behaviors of bovine $\alpha$-lactalbumin ($\alpha$-La), $\beta$-lactoglobulin ($\beta$-Lg), and their mixtures during heat treatment, samples were analyzed using native-polyacrylamide gel electrophoresis (Native-PAGE), sodium dodecylsulfate (SDS)-PAGE, and two-dimensional (2-D)-PAGE. The electrophoresis demonstrated that the loss of native-$\alpha$-La increased as temperature increased, and that the loss of apo-$\alpha$-La was slightly higher than that of holo-$\alpha$-La. The tests also showed that during heat treatment, a mixture of $\alpha$-La and $\beta$-Lg was less stable than $\alpha$-La alone. As such, it was assumed that $\beta$-Lg induced holo-$\alpha$-La to be less stable than apo-$\alpha$-La during heat treatment. The reaction behavior of $\alpha$-La (holo-, apo-form) during heat treatment showed similar patterns in the 2-D-PAGE electropherogram, but the mixture of $\alpha$-La and $\beta$-Lg created new bands. In particular, the results showed a greater loss of native $\alpha$-La in the holo-$\alpha$-La and $\beta$-Lg mixture than in the apo-$\alpha$-La and $\beta$-Lg mixture. Thus, it can be concluded that the holo-$\alpha$-La and $\beta$-Lg mixture was more intensively affected by heat treatment than other samples, and that free sulphydryl groups took part in the heat-induced denaturation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.3
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• pp.310-319
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• 2012

In order to investigate the biological effects of conformational changes in the folding state of bovine ${\alpha}$-lactalbumin (${\alpha}$-La), the protein was prepared and classified as apo form, microbial transglutaminase (MTGase) added form, or bovine ${\alpha}$-La made lethal to tumor cell (BAMLET) form. Apo ${\alpha}$-La form showed weaker cancer cell inhibitory activity (apoptosis) than native ${\alpha}$-La, which suggests that the metal ion-like $Ca^{2+}$ had a positive effect, whereas BAMLET form showed strong cancer cell apoptotic activity. The BAMLET form seemed to be a molten globule structure that increased hydrophobicity. MTGase added to apo ${\alpha}$-La polymer showed similar anti-cancer activity as native ${\alpha}$-La, and it was well hydrolyzed by digestive enzymes. NMR results showed that BAMLET interacted with oleic acid and produced a complex.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.9
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• pp.1327-1333
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• 2007

Lactose synthase catalyses the formation of lactose which is the major osmole of bovine milk and regulates the milk volume. Alpha-lactalbumin (${\alpha}$-LA) is involved in the synthesis of lactose synthase in the mammary gland. Therefore ${\alpha}$-LA is regarded as a plausible candidate gene for the milk yield trait. To determine whether ${\alpha}$-LA is associated with milk performance traits, 1,028 Chinese Holstein cows were used to detect polymorphisms in the ${\alpha}$-LA by means of single-strand conformation polymorphism (SSCP). Two nucleotide transitions were identified in the 5'flanking region and intron 3 of ${\alpha}$-LA. Associations of such polymorphisms with five milk performance traits were analyzed using a general linear model procedure. No significant associations were observed between these polymorphisms and the five milk performance traits (p>0.05). RH mapping placed ${\alpha}$-LA on BTA5q21, linked most closely to markers U63110, CC537786 and L10347 (LOD>8.3), which is far distant from the region of the quantitative trait locus (QTL) on bovine chromosome 5 for variation in the milk yield trait. In summary, based on our findings, we eliminated these SNPs from having an effect on milk performance traits.

• Journal of Animal Science and Technology
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• v.44 no.6
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• pp.801-808
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• 2002

Proteolytic activities of some commercial milk clotting enzymes(rennet, trypsin, pepsin, papain W-40, neutrase 1.5 and protease S) in bovine skim milk containing 0.02% $CaCl_2$ were determined by measuring DH(Degree of Hydrolysis), NPN(Non Protein Nitrogen) and by comparing patterns of SDS-PAGE(Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis). The DH of microbial enzymes(neutrase 1.5 and protease S) and trypsin in bovine skim milk were higher than those of pepsin and papain W-40. The amounts of NPN in the milk treated with trypsin and the other animal enzymes(rennet and pepsin) showed the highest and lowest degrees of proteolysis, respectively. SDS-PAGE showed that trypsin and protease S hydrolyzed $\alpha$-lactalbumin and papain W-40 hydrolyzed $\beta$-lactoglobulin slightly, while neutrase 1.5 hydrolyzed both $\alpha$-lactalbumin and $\beta$-lactoglobulin after treating for 90 min. Trypsin and protease S easily hydrolyzed ${\alpha}_s$-casein and $\beta$-casein, which were not hydrolyzed by rennet. Papain W-40 hydrolyzed $\kappa$-casein more than rennet as shown in SDS-PAGE. Based on the results of the experiments, the DH and NPN of trypsin, neutrase 1.5 and protease S were shown to be higher than those of the other enzymes. The SDS-PAGE patterns of papain W-40 and neutrase 1.5 were similar with that of rennet.

• Proceedings of the Membrane Society of Korea Conference
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• 1994.10a
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• pp.72-73
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• 1994

Whey는 일명 lactoserum 이라고도 하며 치즈제조시 우유를 응고시키는 과정에서 Casein과 지방으로부터 분리되어 나오는 액상의 부산물로 본래 우유 부피의 약 90%를 차지하며, 용해성 단백질, 유당, 비타민과 무기질 등을 함유하고 있다. 유청에 함유되어 있는 단백질은 건조고형분의 약 13%가 되는데, 주요 단백질은 $\beta$-lactoglobulin(50%), $\alpha$-lactalbumin(22%), Serum albumin(5%), Immunoglobulin(12%), Proteose-peptone(10%) 등이 있다. 유청단백질중 가장 많이 함유되어 있는 $\beta$-lactoglobulin은 구형의 단백질로 단량체의 분자량은 약 18,400이며, pH 3.5~7 범위내에서는 해리되지 않는 이량체(dimer)를 형성한다. pH 3.5 이하에서 이량체는 해리되고 다량체의 형성으로 재평성한다. pH 7.0 이상의 알칼리 영역에서는 Conformational Change가 일어나는 것으로 알려져 있으며, 등전점(isoelectric point)은 pH 5.2이다. $\alpha$-lactalbumin은 14,200의 분자량을 가지는 구형의 단백질로 등전점은 pH 4.8이다.

• Food Science of Animal Resources
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• v.22 no.1
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• pp.59-65
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• 2002

This study was carried out to evaluate the effect of whey protein concentrate-80%(WPC-80) and whey protein isolate(WPI) on the growth of B. bifidum Bb-11. Whey proteins($\alpha$-lactalbumin, $\beta$-lactoglobulin) were digested with trypsin, then their hydrolyzates were separated into three fractions (>10,000Da, 3,000∼10,000Da, <3,000Da) by two-step ultrafiltration process with Centriprep 10 and Centricon-30. These three fractions by molecular weight were evaluate growth-promoting effects for the B. bifidum Bb-11. The results obtained were summarized as follows; The growth rate of B. bifidum Bb-11 tended to increase by supplementation of WPC-80 to basal medium, but decreased by supplementation of WPI. Two whey proteins were hydrolyzed by trypsin at 40$\^{C}$ for 6 hrs, and three fractions were collected by UF treatment and concentrated by Centricon-30. Collected concentrations of protein of F-I and F-II and F-III from $\alpha$-lactalbumin were 11.53mg, 7.79mg, and 5.21 mg and those of protein from $\beta$-lactoglobulin were 4.13mg, 5.30mg, and 9.351mg, respectively. Three fractions of $\alpha$-lactalbumin hydrolyzates promoted the growth rate of B. dbifidum Bb-11. Growth promoting activities of hydrolyzates(F-I and F-II) with molecular weight below 10,000Da were stronger than that of hydrolyzate(F-III) above 10,000Da. However, there was no significant difference between the hydrolyzate F-I and F-II. Three fractions of $\beta$-lactoglobulin hydrolyzates improves the growth rate of B.bifidum Bb-ll. The growth of B.bifidum Bb-ll was decreased after 24 hr incubation by supplementation of either F-II or F-III fraction compared to basal Whey medium, but maintained the enhancement by supplementation of F-I.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.6
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• pp.922-927
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• 1996

In order to investigate heat-induced gelation properties of $\alpha-lactalbumin(\alpha-La),$ total free SH groups, half-cystine, and disulfide bond contents of $\alpha-La$ gels prepared in 0.1M Tris-HCI buffer(pH 8.0) were measured. The samples were heated at $90^{\circ}C$ for 40 minutes under different PH and concentrations of NaCl, $CaCl_2,$ $\alpha-La,$ N-ethylrnaleirnide(NEM), and dithiothreitol(DTT). Total free SH groups were low at high concentrations of $\alpha-La$ and at pH 6.5~8.5, and were $14.72~18.58\mu\textrm{m}ol/g$ and $14.17~16.11\mu\textrm{m}ol/g,$ respectively. Half-cystine contents of NEM-induced gels decreased with increasing concentration of NEM, and were $1.03~39.17\mu\textrm{m}ol/g.$ Disulfide bonds of DTT-induced gels increased with increasing concentration of DTT, and were $70.04~71.80\mu\textrm{m}ol/g$.