PubMed · 15056078
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Benson T Massey. 2004. Whither the Bernstein test?. https://doi.org/10.1111/j.1572-0241.2004.04091.x
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PURPOSE: The complex superficial lipid layer of the tear film functions to prevent evaporation and maintain tear stability. Although classes of lipids found in the tear film have been reported, individual lipid species are currently being studied with more sophisticated. METHODS: The purpose of this work was to show the identification of fatty acids and the fatty acid amides in human meibomian gland secretions by using electrospray mass spectrometry. methods. Human meibomian gland secretions (meibum) were analyzed by electrospray quadrupole time-of-flight mass spectrometry (positive- and negative-ion mode). Accurate mass determination and collision-induced dissociation of meibum, and lipid standards were used to identify lipid species. RESULTS: Mass analysis of meibum in an acidic chloroform-methanol solution in positive-ion mode revealed a mass peak of m/z 282.3, which was identified as the protonated molecule of oleamide [C(18)H(35)NO+H](+). The high-resolution mass analysis of the m/z 282.2788 peak (oleamide) demonstrated a mass accuracy of 3.2 parts per million (ppm). Collision-induced dissociation of this species from meibum, compared with an oleamide standard, confirmed its identification. Myristic, palmitic, stearic, and oleic free fatty acids were identified in a similar manner, as were the other fatty acid amides (myristamide, palmitamide, stearamide, and erucamide). CONCLUSIONS: The findings indicate that oleamide (cis-9-octadecenamide), an endogenous fatty acid primary amide, is a predominant component of meibum when examined by electrospray mass spectrometry. The novel finding of oleamide and other members of the fatty acid amide family in the tear film could lead to additional insights into the role of fatty acid amide activity in human biological systems and may indicate a new function for this lipid class of molecules in ocular surface signaling and/or in the maintenance of the complex tear film.
AIM: To achieve secretory expression of mature sCD14 gene in Pichia pastoris and to analyze its LPS binding activity. METHODS: sCD14 gene was cloned into yeast expression vector pPIC9K. Recombinant plasmid was lineared by Sac I and transformed into Pichia pastoris GS115. Positive integrated clones were screened and expressed by methanol inducing. RESULTS: The mature sCD14 was expressed in BMMY with pH 6.5 after optimization of inducing expression condition. Western blot and ELISA data revealed recombinant products combined with specific antibody. The results from FACS showed recombinant product had biological activity of binding with LPS. CONCLUSION: The recombinant mature sCD14 molecules with favorable LPS-binding activity could be used for further study on its structure and functions.
OBJECTIVE: The purpose of this study was to evaluate the influence of symptomatic vulvovaginal candidiasis on the shedding of HIV-1 in cervicovaginal secretions of HIV-1-infected women. STUDY DESIGN: We obtained paired blood and cervicovaginal lavage samples from 66 HIV-infected women with symptomatic vulvovaginal candidiasis, and 249 HIV-infected control patients without genital infection. HIV-1 RNA in plasma, proviral HIV-1 DNA, HIV-1 RNA transcripts, and cell-free HIV-1 RNA in cervicovaginal secretions were quantitatively evaluated by competitive polymerase chain reaction (cPCR) and reverse transcriptase PCR (cRT-PCR). We used logistic regression on ordered data to assess the influence of vulvovaginal candidiasis on the HIV-1 load in cervicovaginal secretions adjusting for potential confounders. RESULTS: Overall, the amount of HIV-1 RNA in plasma was significantly correlated with HIV-1 DNA (Spearman rank 0.153 +/- 0.059, P = .006), HIV-1 RNA transcripts (Spearman rank 0.169 +/- 0.058, P = .003), and cell free HIV-1 RNA (Spearman rank 0.185 +/- 0.059, P = .001) load in cervicovaginal secretion. Forty-eight out of 182 (26.4%) patients who tested negative for HIV-1 RNA in plasma were positive for HIV-DNA in their cervicovaginal secretions. In logistic regression analysis vulvovaginal candidiasis was significantly associated with increasing loads of HIV-1 RNA transcripts (Odds ratio [OR] 1.97, 95% CI 1.09-3.57, P = .025) and cell free HIV-1 RNA (OR 2.03, 95% CI 1.10-3.73, P = .02) in cervicovaginal secretions. CONCLUSION: In HIV-infected women, vulvovaginal candidiasis is associated with an increased number of copies of cell-associated and cell-free HIV-1 RNA in cervicovaginal secretions.