Sphingolipid identification and skin barrier recovery capacity of a milk sphingolipid-enriched fraction (MSEF) from buttermilk powder.

OBJECTIVE: In this study, we isolated the milk sphingolipid-enriched fraction (MSEF) of sweet buttermilk powder and conducted a clinical trial for evaluating its efficacy in skin barrier recovery. METHODS: Milk sphingolipid-enriched fraction was isolated via solvent extraction of buttermilk powder, and further concentrated by removing the phospholipids and neutral lipids. A cream containing 1% MSEF was used during clinical trials to assess for water holding and skin barrier recovery capacities. RESULTS: The main components of the MSEF were sphingomyelin, glucosylceramide and lactosylceramide, confirmed by TLC, HPLC, MS and NMR. The MSEF cream-treated group had a significantly higher (P < 0.05) water holding capacity, compared with the base cream (vehicle) group. Compared with that in the base cream group, transepidermal water loss (TEWL) recovery increased in the presence of the sphingolipid-containing MSEF cream (MSEF group), with a significant difference (P < 0.05) recorded on day 14. CONCLUSION: The MSEF cream contributed to improving the water holding capacity and skin barrier recovery of damaged skin. Therefore, sphingolipid-containing MSEF can be useful for strengthening or repairing skin barrier function.

OBJECTIF: Dans la présente étude, nous avons isolé la fraction enrichie en sphingolipides du lait (milk sphingolipid-enriched fraction, MSEF) du lait de baratte doux en poudre et mené un essai clinique pour évaluer l'efficacité de la récupération de la barrière cutanée. MÉTHODES: La fraction enrichie en sphingolipides du lait est isolée par l'extraction de solvant du lait de baratte en poudre, et ensuite concentrée en enlevant les phospholipides et les lipides neutres. Une crème contenant un pourcent (1%) de la fraction enrichie en sphingolipides du lait (MSEF) est utilisée pendant des essais cliniques visant à évaluer les capacités de la rétention d'eau et de la récupération de la barrière cutanée. RÉSULTATS: Les principales composantes de la fraction enrichie en sphingolipides du lait (MSEF) étaient la sphingomyéline, la glucosylcéramide, et le lactosylcéramide, confirmées par la chromatographie sur couche mince (TLC), la chromatographie liquide à haute performance (CLHP), la spectrométrie de masse (MS) et la RMN(NMR). La groupe traitée par la crème enrichie en sphingolipides du lait (MSEF) avait une capacité de la rétention d'eau beaucoup plus élevée (P < 0.05), par rapport au groupe utilisant la crème de base (véhicule). Comparé au groupe utilisant la crème de base, la récupération de la perte d'eau transépidermique (TEWL) a augmenté en présence de la fraction enrichie en sphingolipides du lait (MSEF), avec une différence significative (P < 0.05) enregistrée au 14ème jour. CONCLUSION: La crème enrichie en sphingolipides du lait (MSEF) a contribué à améliorer les capacités de la rétention d'eau et de la récupération de la barrière cutanée de la peau endommagée. Par conséquent, la fraction enrichie en sphingolipides du lait (MSEF) peut être utile pour renforcer ou réparer la fonction de barrière cutanée.

RNA interference validation of detoxification genes involved in ivermectin tolerance in Drosophila melanogaster.

Previously, we observed increased transcription levels of specific cytochrome P450 monooxygenase (P450) and adenosine triphosphate binding cassette (ABC) transporter genes in human body lice, Pediculus humanus humanus, following exposure to ivermectin using the non-invasive induction assay, which resulted in tolerance. To confirm the roles of these genes in induction and tolerance, the robust genetic model insect Drosophila melanogaster was chosen. Orthologous genes corresponding to the body louse P450 (Cyp9f2, Cyp6g2 and Cyp9h1) and ABC transporter (Mrp1, GC1824 as an ABCB type and CG3327 as an ABCG type) genes were selected for in vivo bioassay. Following a brief treatment with a sublethal dose of ivermectin, the mortality response was significantly slower, indicating the presence of tolerance. Concurrently, the transcription levels of Cyp9f2 and Mrp1 at 3 h and those of Cyp6g2, Cyp9h1, Mrp1, CG1824 and CG3327 at 6 h post-treatment were upregulated, indicating gene induction. In behavioural bioassay using GAL4/UAS-RNA interference transgenic fly lines, increased susceptibility to ivermectin was observed following heat shock in the Cyp9f2 , Cyp6g2 , Cyp9h1 , Mrp1 or CG3327-knockdown flies. Considering that these five genes are orthologous to those which had the largest over-expression level following ivermectin-induced tolerance in the body louse, the current results suggest that they are also associated with ivermectin detoxification in D. melanogaster and that body lice and D. melanogaster are likely to share, in part, similar mechanisms of tolerance to ivermectin.

A panel of antibodies for the immunostaining of Bouin's fixed bone marrow trephine biopsies.

AIMS: To assess a panel of antibodies on Bouin's fixed bone marrow trephine (BMT) biopsies. These biopsies are widely used in routine diagnosis of various haematological malignancies and may be the sole material available in many centres; however, information regarding the immunostaining of this material is lacking. METHODS: Biopsies were taken from 72 patients presenting with various haematological malignancies (leukaemia, 38; lymphoma, 14; multiple myeloma, 20). A panel of antibodies was assessed on Bouin's fixed BMT biopsies by the alkaline phosphatase-antialkaline phosphatase method. RESULTS: Three B (MB2, LN-2, Ki-B5) and two T cell lineage antibodies (UCHL-1, CD3-r) reliably identified lymphoid cells, while MPO-r, Leu-M1/CD15, and KP-1/CD68 recognised cells from the myeloid or histiocytic/macrophage series. Reed-Sternberg cells were stained by LN-2, Leu-M1, and CD30. Antibodies specific for plasma cells (VS38) and hairy cells (DBA.44) gave a variable pattern of staining. Among the proliferation markers, proliferative cell nuclear antigen but not Ki-67 related antibodies were effective. CONCLUSION: This study presents a panel of antibodies with reactivity not restricted to common fixatives that are also suitable for Bouin's fixed BMT biopsies.

Application of laser-induced fluorescence for determination of trace uranium, europium and samarium.

Trace amounts of uranium, europium and samarium can be determined by means of laser-induced fluorescence. The fluorescence emission and excitation spectra were observed by adding fluorescence enhancing reagents. The fluorescence lifetimes of these elements were also measured. The intensities of emission were found to be linear with respect to the concentration of the element over a wide range. The detection limits for U(VI), Eu(III), and Sm(III) were established as 0.05, 0.1, and 10 ng/ml, respectively. The study suggested that this is a suitable technique for the trace determination of uranium and rare earth elements.