The relationship between elastase and lactoferrin in healthy, gingivitis and periodontitis sites

OBJECTIVES: To compare the relative amounts of elastase (primary polymorphonuclear leucocyte granule constituent) and lactoferrin (secondary PMN granule constituent) in the gingival crevicular fluid (GCF) of healthy, gingivitis and periodontitis sites.
DESIGN: This cross-sectional study looked at the two GCF constituents in three categories of disease status within the same subject.
MATERIALS AND METHODS: Patients with chronic adult periodontitis were screened and those exhibiting all three types of sites ie periodontally healthy, gingivitis and periodontitis sites were recruited (n = 10) and had GCF collected from the three sites. Lactoferrin and elastase were measured in eluates of GCF by enzyme-linked immunosorbent assay.
RESULTS: The absolute amount of lactoferrin measured in ng per 30 s samples was significantly lower in healthy and gingivitis sites as compared to periodontitis sites however this difference failed to reach significance when the concentration of lactoferrin in GCF was used as the analytical unit. No significant differences were found for elastase levels at any sites when expressed as either absolute amounts or concentrations. Secondary granule release, as evidenced by lactoferrin levels, occurs during cell migration and the process is independent of primary granule release, which is thought to correlate with PMN activation. The relationship between granule constituents in the samples showed significant differences, the highest lactoferrinlelastase ratio being at periodontitis sites (P < 0.001).
CONCLUSIONS These findings imply a change in the relative amounts of elastase and lactoferrin released at different disease level sites, with an almost 10-fold increase in the proportion of lactoferrin to elastase in periodontitis sites over healthy and gingivitis sites. This variation in the release by PMNs of primary and secondary granule constituents may indicate alterations in PMN function in different disease environments.     

人乳中乳铁蛋白含量的初步检测分析

目的 了解人乳乳铁蛋白水平.方法 利用酶联免疫吸附双抗体夹心法(ELISA),分别测定36名产妇产后2、3 d的初乳,6、7 d的过渡乳及42 d的成熟乳标本中乳铁蛋白水平.结果 初乳的乳铁蛋白含量最高,过渡乳组和成熟乳组依次降低,分别为初乳组(2.63 ± 1.11) g/L、过渡乳组(1.96 ± 1.05) g/L、成熟乳组(1.42 ± 1.01) g/L,三组差异有统计学意义(P ＜ 0.01).剖宫产与顺产产妇的乳铁蛋白差异无统计学意义(P ＞ 0.05).产妇年龄＜ 28岁与≥ 28岁的乳铁蛋白含量差异无统计学意义(P ＞ 0.05).结论 人乳乳铁蛋白在初乳、过渡乳、成熟乳中的下降趋势与国外报道相符,但三组测定值均低于国外水平,尤其是初乳乳铁蛋白明显低于国外报道.     

Influence of aging on candidal growth and adhesion regulatory agents in saliva

Although oral candidiasis is frequently seen in the elderly, the factors determining candidal growth have insufficiently been explored. Hence, we examined the influence of aging on candidal adhesion and growth-inhibitory agents in saliva in 45 healthy volunteers and 60 patients with oral candidiasis. Both non-stimulated and stimulated salivary flow rates (SFRs) in the healthy controls decreased with aging. A gradual decrease of SFRs with aging was also observed in the patients, and the SFR levels were markedly lower than those in the controls. Although the salivary glucose levels were almost constant in all age groups, secretory immunoglobulin A and lactoferrin levels in saliva were significantly decreased statistically with age, and a marginal age-associated decrease in transferrin levels was also observed. In addition, the generation of superoxide from neutrophils in saliva and their Candida killing activity decreased with age, and these phenomena were more apparent in the patients. Furthermore, a larger number of Candida adhered to oral keratinocytes obtained from the elderly healthy controls than to those obtained from young controls. Correspondingly, keratinocytes from the aged controls showed more concanavalin-A binding sites than those from the young controls. However, oral Candida did not increase with increasing age in the controls, although an age-associated increase of oral Candida was observed in the patients. Taken together, these results indicate that the decreases of SFRs and salivary anti-candidal factors, suppression of salivary neutrophil function and the increase of candidal adhesion sites on keratinocytes predispose elderly individuals to oral candidiasis.     

Ascitic fluid analysis for diagnosis and monitoring of spontaneous bacterial peritonitis

Polymorphonuclear （PMN） cell count in the ascitic fluid is essential for the diagnosis and management of spontaneous bacterial peritonitis （SBP）. To date, PMN cell count is routinely performed by traditional manual counting. However, this method is time-consuming, costly, and not always timely available. Therefore, considerable efforts have been made in recent years to develop an alternative test for a more rapid diagnosis and monitoring of SBP. The use of urinary reagent strips was proposed to achieve an ＂instant＂ bedside diagnosis of SBP. A series of reports evaluated the urine strip test for SBP diagnosis and reported promising results. However, a recent large multicenter study revealed a surprising lack of diagnostic efficacy of the urine screening test for SBP diagnosis. Another method, more recently proposed as an alternative to the manual PMN count, is the measurement of lactoferrin in ascitic fluid, but the data available on the diagnostic value of this test are limited to a single study. However, both urinary reagent strips and ascitic lactoferrin tests are qualitative methods and need, therefore, to be fur- ther confirmed by standard cytology of the ascitic fluid. To date, the only quantitative method proposed as a valid alternative to manual PMN counting is automated blood cell counters, commonly used in all laboratories for blood cell counting. Data available in the literature on the diagnostic performance of this method are limited but very promising, and this tool seems to have the potential to replace the manual counting method.     

Sialochemical markers of salivary gland involvement with Sjögren's syndrome secondary to rheumatoid arthritis and primary biliary cirrhosis

Abstract: Sjögren’s syndrome is an autoimmune condition affecting the lacrimal and salivary glands and can be associated with rheumatoid arthritis and primary biliary cirrhosis. Parotid salivas collected from patients and normal controls were analysed for lactoferrin, IgA and beta₂‐microglobulin (measured by ELISA), and cystatin (measured by a enzyme inhibition assay). Output data provided less variable means, whilst expressing results as a proportion of the total protein provided greater specificity as markers for Sjögren’s syndrome. Levels of specificity for IgA, lactoferrin and beta₂‐microglobulin were all high (100, 95 and 100%, respectively). Sensitivity levels of these markers (but not cystatin) tended to be similar for Sjögren’s syndrome secondary to primary biliary cirrhosis (IgA, 25%; lactoferrin, 63%; and beta₂‐microglobulin, 50%), compared to Sjögren’s syndrome secondary to connective tissue diseases such as rheumatoid arthritis (IgA, 50%; lactoferrin, 86%; and beta₂‐microglobulin; 38%).     

Enzyme immunoassay of specific human IgE to purified cows’ milk allergens

An immunometric enzyme immunoassay for specific immunoglobulin E (IgE) against five purified cows’ milk allergens, β‐lactoglobuHn, α‐lactalbumin, bovine serum albumin, lactoferrin and whole casein fraction, has been developed. Allergens were immobilized on microtitration plates. After incubations with sera from allergic patients, specific IgE bound to the plastic were detected using a monoclonal anti‐human IgE antibody labelled with acetylcholinesterase. Quantitative determinations were made by comparison with a dose‐response curve obtained under the same conditions with standard total IgE. A quantification limit of 0.08 IU ml^‐1 can thus be obtained with a coefficient of variation of lower than 5%. This allows specific IgE determinations of clinical significance in sera from allergic patients at a dilution of at least 1/10 (i.e. in a few μl of serum) with good precision and reproducibility. The determinations of specific IgE in sera from 11 patients allergic to cows’ milk showed an excellent correlation of this assay with clinical data.     

Lactoferrin in human amniotic fluid

The concentration of human lactoferrin (LF) was measured byradioimmunoassay or non-competitive avidin-biotin assay in amnioticfluid, cord blood and in the decidua, trophoblast, fetal membranesand umbilical cord. Amniotic fluid was obtained by amniocentesis,and cord blood and tissue samples were taken after deliveryor elective Caesarean section. No detectable concentration ofLF was found in amniotic fluid before week 20 of pregnancy.A significant increase in the LF concentration was observedaround week 30 and it remained high until term. In cord blood,an undetectable or low concentration of LF was measured. Intissue specimens the amount of LF was highest in the decidua(9–95µ/g a moderate concentration was assayed inthe amniotic, (2–37µg/g) and chorion (2–26µg/g) membrane and in the trophoblast (5–35 µg/g).In the umbilical cord, the concentration was > 1µg/g.These results suggest a decidual origin of LF. The role of LFduring pregnancy is discussed