A New Low Cost, Fully Automated Amino Acid Analyzer Using a Gradient HPLC

Klapper, David G.

doi:10.1007/978-1-4612-5832-2_45

Cited by 25 publications

(15 citation statements)

References 5 publications

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“…The analysis procedure was similar to that of Klapper [33] and Cohen and Strydom [34]. The samples were vacuum-dried and placed in a hydrolysis vessel containing some constant boiling HCl and 1% (v/v) phenol.…”

Section: Methodsmentioning

confidence: 99%

The role of crude human saliva and purified salivary MUC5B and MUC7 mucins in the inhibition of Human Immunodeficiency Virus type 1 in an inhibition assay

Habte

Mall

Beer

et al. 2006

Virol J

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Background: Despite the continuous shedding of HIV infected blood into the oral cavity and the detectable presence of the AIDS virus at a high frequency, human saliva is reported to inhibit oral transmission of HIV through kissing, dental treatment, biting, and aerosolization. The purpose of this study was to purify salivary MUC5B and MUC7 mucins from crude saliva and determine their anti-HIV-1 activities.

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Section: Methodsmentioning

confidence: 99%

The role of crude human saliva and purified salivary MUC5B and MUC7 mucins in the inhibition of Human Immunodeficiency Virus type 1 in an inhibition assay

Habte

Mall

Beer

et al. 2006

Virol J

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“…Briefly, mucin was prepared according to the method of Schroten et al [19] , semipurified by gel filtration on Sepharose CL-4B, and subsequently purified by caesium chloride isopycnic density gradient ultracentrifugation [20] . Mucin was further analyzed by SDS-PAGE [21] and identified by Western blotting [22] and amino acid analysis [23,24] . Glycoprotein was estimated by the periodic acidSchiff (PAS) procedure of Mantle and Allen [25] and protein according to the method of Lowry et al [26] .…”

Section: Mucin Preparation Purification Identification and Analysismentioning

confidence: 99%

Inhibition of Human Immunodeficiency Virus Type 1 Activity by Purified Human Breast Milk Mucin (MUC1) in an Inhibition Assay

et al. 2007

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It has been reported that breast-feeding is responsible for approximately 40% of the HIV transmissions from HIV-positive mothers to children. Human breast milk, however, is known to contain numerous biologically active components which protect breast-fed infants against bacteria, viruses, and toxins. The purpose of this study was to purify and characterize breast milk mucin and to determine its anti-HIV-1 activity in an HIV inhibition assay. Sepharose CL-4B column chromatography and caesium chloride isopycnic density gradient purification were used to isolate and purify the mucin. Following Western blotting and amino acid analysis, an HIV-1 inhibition assay was carried out to determine the anti-HIV-1 activity of crude breast milk and purified milk mucin (MUC1) by incubating them with HIV-1 prior to infection of the human T lymphoblastoid cell line (CEM SS cells). SDS-PAGE analysis of the mucin, together with its amino acid composition and Western blotting, suggested that this purified mucin from human breast milk was MUC1. The HIV inhibition assay revealed that while the purified milk mucin (MUC1) inhibited the HIV-1 activity by approximately 97%, there was no inhibition of the HIV-1 activity by crude breast milk. Although the reason for this is not clear, it is likely that because the MUC1 in crude milk is enclosed by fat globules, there may not be any physical contact between the mucin and the virus in the crude breast milk. Thus, there is a need to free the mucin from the fat globules for it to be effective against the virus.

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“…1-mg samples of lyophilized tissue were hydrolysed (110 • C, 22 h) under an inert atmosphere in a sealed glass tube using 0.5 ml of HCl (5.7 M) containing 0.25 % (v/v) phenol. After cooling, opening of the tubes and drying (in vacuo), the samples were redissolved in 1 ml of citrate buffer (pH 2.2) and applied to an HPLC cation-exchange column (Waters) maintained at 62 • C. Amino acids were separated using a pH gradient from 3.05 to 9.45, with detection relying on post-column fluorescence using orthophthalaldehyde [18], and reported as nmol/mg of dry tissue.…”

Section: Tissue Preparation and Fixationmentioning

confidence: 99%

The effects of cross‐link density and chemistry on the calcification potential of diamine‐extended glutaraldehyde‐fixed bioprosthetic heart‐valve materials

Bezuidenhout

Oosthuysen

Human

et al. 2009

Biotech and App Biochem

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Despite indications that GA (glutaraldehyde)-crosslinked tissues remain prone to long-term degradation and calcification, it is still the reagent of choice in the fixation of bioprosthetic heart valves. We have shown previously that increased GA concentrations and diamine extension of cross-links with lysine incorporation lead to mitigated in vivo calcification, mainly of porcine aortic-wall tissue. The present study was performed to assess the correlation between the cross-link density of all three commonly used tissue types [PW (porcine aortic wall), PL (porcine aortic leaflet) and BP (bovine pericardium)] and tissue calcification in the subcutaneous rat model after GA treatment with or without lysine. The effect of lysine enhancement, and increased GA concentration in the presence of lysine, resulted in significant increases in tissue cross-linking in all three tissue types. Although increased GA concentration on its own resulted in decreased calcification without an increase in cross-link density, overall positive correlations were found between denaturation temperature and RPD (resistance towards protease degradation) [correlation coefficient (rho) values: rhoPW =0.922, rhoPL =0.783 and rhoBP =0.955], whereas negative correlations existed between RPD and calcification (rhoPW=-0.836, rhoPL=-0.929 and rhoBP=-0.579). The combination of lysine enhancement and an increase in GA concentration from 0.2 to 3% resulted in 79, 44 and 56% decreases in calcification in PW, PL and BP. In the case of BP, a decrease in calcification of 81% could be achieved merely by adding lysine extension to low-concentration (0.2 %) GA cross-linking. Thus it is concluded that the increase in cross-link density achieved by lysine incorporation, and by increased GA concentration in the presence of lysine, results in significant and marked decreases in calcification of all three types of tissues commonly used in bioprosthetic heart valves.

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A New Low Cost, Fully Automated Amino Acid Analyzer Using a Gradient HPLC

Cited by 25 publications

References 5 publications

The role of crude human saliva and purified salivary MUC5B and MUC7 mucins in the inhibition of Human Immunodeficiency Virus type 1 in an inhibition assay

The role of crude human saliva and purified salivary MUC5B and MUC7 mucins in the inhibition of Human Immunodeficiency Virus type 1 in an inhibition assay

Inhibition of Human Immunodeficiency Virus Type 1 Activity by Purified Human Breast Milk Mucin (MUC1) in an Inhibition Assay

The effects of cross‐link density and chemistry on the calcification potential of diamine‐extended glutaraldehyde‐fixed bioprosthetic heart‐valve materials

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