Proteomic Profiling of Human Keratinocytes Undergoing UVB-Induced Alternative Differentiation Reveals TRIpartite Motif Protein 29 as a Survival Factor

BACKGROUND: Repeated exposures to UVB of human keratinocytes lacking functional p16(INK-4a) and able to differentiate induce an alternative state of differentiation rather than stress-induced premature senescence. METHODOLOGY/PRINCIPAL FINDINGS: A 2D-DIGE proteomic profiling of this alternative state of differentiation was performed herein at various times after the exposures to UVB. Sixty-nine differentially abundant protein species were identified by mass spectrometry, many of which are involved in keratinocyte differentiation and survival. Among these protein species was TRIpartite Motif Protein 29 (TRIM29). Increased abundance of TRIM29 following UVB exposures was validated by Western blot using specific antibody and was also further analysed by immunochemistry and by RT-PCR. TRIM29 was found very abundant in keratinocytes and reconstructed epidermis. Knocking down the expression of TRIM29 by short-hairpin RNA interference decreased the viability of keratinocytes after UVB exposure. The abundance of involucrin mRNA, a marker of late differentiation, increased concomitantly. In TRIM29-knocked down reconstructed epidermis, the presence of picnotic cells revealed cell injury. Increased abundance of TRIM29 was also observed upon exposure to DNA damaging agents and PKC activation. The UVB-induced increase of TRIM29 abundance was dependent on a PKC signaling pathway, likely PKCdelta. CONCLUSIONS/SIGNIFICANCE: These findings suggest that TRIM29 allows keratinocytes to enter a protective alternative differentiation process rather than die massively after stress

Caractérisation du consortium microbien d'un grain de kéfir

Les grains de kéfir sont des ferments lactiques constitués d'une microflore complexe et diversifiée. Celle d'un grain de kéfir (KJ) a été caractérisée par une approche méthodologique classique d'isolements microbiens sur des milieux de culture sélectifs et, pour les bactéries lactiques, par une approche indépendante d'isolements bactériens par culture. Cette dernière consistait en l'analyse de la séquence des régions V1 et V2 de l'ADNr 16S, amplifiées à partir de l'ADN extrait du grain. La sensibilité de la méthode a été augmentée à cette occasion en dissociant les amplifications géniques des populations lactiques minoritaires et majoritaires du grain. La flore identifiée dans le grain KJ comprenait Acetobacter sp., Kazachstania exigua, Lactobacillus kefiranofaciens subsp. kefirgranum, Lb. kefiri, Lb. parakefiri, Lactococcus lactis subsp. lactis et Leuconostoc mesenteroides, et était dépourvue de microcoques, de pédiocoques, de Weissella viridescens, de bactéries indicatrices de défauts d'hygiène, de Salmonella sp. et de Listeria monocytogenes. Dans le but de vérifier le caractère complet des espèces mises à jour, une reconstitution du grain à partir des micro-organismes qui en ont été isolés a été envisagée. Des conditions expérimentales favorables à la formation de grains dans un substrat lacté ont été recherchées à partir d'extraits du grain KJ renfermant un consortium microbien a priori complet. Ces essais de reconstitution n'ont pas conduit à la formation de grains de kéfir mais l'un d'eux a conduit à la formation de biofilms. Cet évènement a été reproduit dans du lait de façon répétable avec des consortiums reconstitués à partir des micro-organismes individuels isolés du grain KJ

Impact of skimming and microfiltration processes on equol concentration in milk

Equol is a microbial metabolite of isoflavones that could be used as therapeutic agent against several diseases and cancers. Cow's milk could be a potential source of equol in the human diet, but more studies are needed on the changes in equol concentration during the technological processing of milk and dairy products. This first exploratory study sought to assess the impact of industrial skimming and microfiltration processes on equol concentration in conventional cow's milk. The milk samples used in this study came from bulk cow’s milk collected in Wallonia (Belgium), by a local dairy and processed in a local cheese factory. Six random sampling were conducted during spring and, for each of them, samples from raw, skimmed and microfiltered milk were analyzed. Equol was present in all samples at a concentration of between 3.2 and 10.3 µg.L-1. A Wilcoxon's signed rank test was then performed on the difference of data, having raw milk as reference, irrespective of the collection date. The results showed that the skimming process slightly increased the equol concentration in milk and therefore that equol had no or little affinity with the lipid milk fraction. The results also showed that, with the microfiltration process, a small proportion of equol was retained. Equol concentration has returned to the same level as that found in raw milk. This might be because of a chemical affinity either with bigger molecules that are physically retained or through direct interaction with the membrane. This scoping study paves the way for more extensive studies on the interaction between equol and other components of the milk.PhytoHealt

Forage legumes: the preferred choice for increasing equol levels and improving fatty acid composition in milk

The type of grass made available to cows and, more specifically, the proportion of legumes influences the quality of dairy products, including milk. Here, we carried out 6 experiments whose results underscore how forage legumes affect milk quality, namely fatty acid composition and equol levels. When animals are given silage, legumes are better than grasses for improving milk fatty acid composition. Similarly, whether animals are grazing or green fed, forage legumes, and especially red clover, increase milk equol levels. When milk is processed to create dairy products, equol levels remain consistent. With the exception of organically produced milk, commercially available milk has much lower levels of equol than did the milk produced by our experimental animals given access to legumes.PhytoHealth - GrassMil

Sweet's syndrome with arthritis in an 8-month-old boy

Sweet's syndrome was diagnosed in a 4-month-old boy. He was successfully treated with systemic corticosteroids. At the age of 8 months, he developed acute arthritis in his right knee. The synovial fluid was analyzed and revealed a very high neutrophil count and neutrophil activation with a detectable level of intraarticular granulocyte-monocyte colony stimulating factor (GM-CSF). Prednisone injection into the knee led to dramatic improvement. No recurrence occurred. Although arthritis and/or arthralgia are common features in adult patients with Sweet's syndrome, this is the first reported case of Sweet's arthritis in a child

Transplantation of umbilical cord blood in a refractory lymphoma.

A successful cord blood transplantation combined with hematopoietic growth factor was performed in a boy presenting with refractory mediastinal T-cell lymphoma. Cord blood cells were collected from an HLA-identical sibling at the time of delivery. A transient and corticosensitive acute grade II graft versus host disease was observed. One year after transplantation, the child is still in remission with complete engraftment. This is the first report of cord blood transplantation in a patient with refractory lymphoma

The effect of commonly used dairy processing techniques and unit operations on the equol content of dairy products

Since a large portion of the milk consumed by human is processed, the aim of this study was to determine possible changes in equol content upon processing milk. Individual milks with different levels of equol content were collected from the morning milking to make skimmed milk and cream, as well as pasteurised and sterilised full fat milks. Pasteurised milk was further processed into yoghurt, kefir, cottage cheese and whey. Yoghurt and kefir were also produced from sterilised milk. Equol content was not affected by either pasteurisation or sterilisation. Compared with raw milk, a higher concentration of equol was measured in skimmed milk, but equol content in cream was lower. The concentration of equol remained unchanged after yoghurt production but was reduced by more than 50% after kefir production. The equol content in whey was low compared with that of raw milk, while it increased sharply in cottage cheese