phosphatase (ALP), a membrane-bound gly- ..... liberation of the cytoplasmic or outer membrane-bound enzymes from injured mammary gland epithelial cells ...
Research Article
Alkaline phosphatase activity in human colostrums as a valuable predictive biomarker for lactational mastitis in nursing mothers
Keywords: alkaline phosphatase n lactational mastitis
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Methods At the beginning of the investigation our study included 87 nursing mothers with normal vaginal delivery, who gave birth to healthy, single, term infants, Of them, 27 were excluded from the study due to their inability (lack of milk, social reasons.) to breastfeed their babies for more than 3 months, what is conventionally accepted period by the authors of the manuscript as a sufficient time for developing mastitis. The study group finally consisted of 60 healthy nursing mothers, admitted for delivery at the Clinic of Obstetrics and Gynecology in Clinical Center, Nis, Serbia, between May and July 2010. A total amount of 2 ml of colostrums was obtained from once from each breast with a manual breast pump (Ginevri, Milan, Italy). The samples were taken either the first or second lactation day in the morning, depending on the first beginning of colostrum arrival. In order to prevent separation of fat and lost of membrane enzymes which are stored in milk fat globules the samples were diluted three-fold by adding in sterile physiological NaCl, and stored at -20°C until they were analyzed. During the analysis, samples were diluted again (1:10) with 10% SDS (sodium dodecyl-sulphate) to eliminate the opalescence, which comes from the large amounts of proteins and lipids. The alkaline phosphatase activity was measured at pH 10.3 using p-nitrophenyl phosphate as substrate by using commercial Elitech- France kit, ALP-DEA. All nursing mothers were prospectively followed from day 1 postpartum to the end of their
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Milk synthesis starts in the lactocytes of the mammary gland at the end of pregnancy to support the nutrition and to promote the health of newborn babies [1] . The occurrence of the enzyme alkaline phosphatase (ALP), a membrane-bound glycoprotein in milk, was first recognized in 1925 and subsequently characterized as an indigenous enzyme to milk [2] . Biochemical investigations have shown that ALP, which is also detectable in the lactocytes, plays a very important diagnostic role in clinical medicine, since its activity varies in different tissues and serves as a specific indicator of disease states. Recently, in 2006, Fox and Kelly suggested an important role of mammary gland ALP in milk secretion [3] . Although the exact mechanisms by which the lactocytes are damaged during lactational mastitis aren’t still fully understood, liberation of the cytoplasmic or outer membrane-bound enzymes from injured mammary gland epithelial cells into human milk is one of the common consequences of mastitis [3] . Currently, measuring the bovine milk enzymes that leak out into milk as a result of mammary gland cell damage has been shown to be a suitable diagnostic method for identifying subclinical mastitis in dairy cows [4–6] . The aim of this study was to determine ALP activity in human colostrum as a possible predictive biomarker for lactational mastitis in nursing mothers.
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Aims: Biochemical investigations have shown that an indigenous milk enzyme, alkaline phosphatase (ALP) ,which is detectable in the lactocytes, plays a very important diagnostic role in clinical medicine, since its activity varies in different tissues and serves as a specific indicator of disease states. The purpose of this study was to evaluate ALP activity in human colostrum as a possible early predictive biomarker for lactational mastitis in nursing mothers. Patients & methods: During a period from May to July 2010, a total of 60 healthy nursing mothers were recruited for this study. Results: The mean level of colostrum ALP activity from the affected breasts was significantly higher when compared with ALP activity from the contralateral asymptomatic as well as ‘healthy’ breasts (p
