Impact of chemical peeling combined with negative pressure on human skin.

OBJECTIVE: In vivo changes in skin barrier function after chemical peeling with alpha hydroxyacids (AHAs) have been previously reported. However, the additional effects of physical treatment with chemical agents on skin barrier function have not been adequately studied. This study measured the degree of acute skin damage and the time required for skin barrier repair using non-invasive bioengineering methods in vivo with human skin to investigate the additional effect of a 4% AHA chemical jet accelerated at supersonic velocities. METHODS: Thirteen female subjects (average age: 29.54 ± 4.86 years) participated in this study. The faces of the subjects were divided into half according to the block randomization design and were then assigned to receive AHA peeling alone or AHA peeling combined with pneumatic pressure on each side of the face. Transepidermal water loss (TEWL), skin colour and skin blood flow were evaluated at baseline and at 30 min, 2, 5 and 7 days after treatment. RESULTS: The TEWL and skin blood flow were significantly increased after 30 min in chemodermabrasion compared with chemical peeling alone (P < 0.05). The TEWL and skin blood flow recovered to baseline after 2 days, and TEWL was significantly decreased at 7 days compared with chemical peeling alone (P < 0.05). CONCLUSIONS: Chemodermabrasion can temporarily impair skin barriers, but it is estimated that it can enhance the skin barrier function after 7 days compared to the use of a chemical agent alone. In addition, chemodermabrasion has a more effective impact in the dermis and relatively preserves the skin barrier.

Relationship between the HLA-G 14 bp insertion/deletion polymorphism and susceptibility to autoimmune disease: a meta-analysis.

Numerous studies have investigated the potential relationship between the human leukocyte antigen (HLA)-G 14-bp insertion/deletion (INS/DEL) polymorphisms and autoimmune disease (AID). However, published results are inconclusive. Our aim was to determine whether the 14-bp INS/DEL polymorphism in the HLA-G gene contributes to the risk of AID. A systemic literature search of the PubMed and EMBASE databases was conducted to identify eligible studies investigating the association of the HLA-G 14-bp INS/DEL polymorphism with AID. Our analysis included 11 publications involving a total of 6462 individuals. Overall, no significant association between the HLA-G 14-bp INS/DEL polymorphism and AID was detected in any comparison model. Further subgroup analyses based on AID types and ethnicity also revealed no significant associations. Our results suggest that the HLA-G 14-bp INS/DEL polymorphism is unrelated to the development of AID. Further studies including larger sample sizes are warranted to confirm these results.

N-glycosylation does not affect assembly and targeting of proglycinin in yeast.

Glycinin, a simple protein, and beta-conglycinin, a glycoprotein, are the dominant storage proteins of soybean and are suggested to be derived from a common ancestor. To investigate why glycinin does not require glycosylation for its maturation, we attempted N-glycosylation of proglycinin A1aB1b using site-directed mutagenesis and yeast expression system. An N-glycosylation consensus sequence Asn-X-Ser/Thr was created at positions 103, 183, 196, 284 and 457 in the variable regions being strongly hydrophilic revealed from the alignment of amino acid sequences of various glycinin-type proteins. Among five mutant proglycinins (Q103N, H183N, G198T, S284N, N459T), Q103N was fully glycosylated, and H183N and N459T were partly (around 20% of the expressed proteins), whereas others were barely or not glycosylated. The glycosylated proglycinin was susceptible to endo-beta-N-acetylglucosamidase and N-glycanase cleavages. N-glycosylation did not cause inconveniences to processing of signal peptide, assembly into trimers and targeting into the vacuoles. Thermal and trypsin sensitivity analyses of the glycosylated proglycinin suggested that N-linked glycan prevents protein-protein interaction but does not stabilize the protein conformation. The reason why glycinin does not require N-glycosylation for its maturation is discussed.

Real time measurement and control of thermodynamic water activities for enzymatic catalysis in hexane.

The esterification reaction of geraniol with acetic acid catalyzed by immobilized Candida antarctica lipase B was studied in hexane using a pervaporation-assisted batch reactor. The effect of thermodynamic water activity (a(w)) on the initial reaction rate was investigated at a(w) ranging from 0.02 to 1.0. The a(w) was monitored on-line in real time. a(w) was actively controlled throughout the reaction by using highly water-selective membrane pervaporation. This novel combination of a(w) sensing and control eliminates changes in a(w) during the reaction even in the initial phase of relatively rapid water release during an esterification. No chemicals are introduced for a(w) control, and no purge gases or liquids are needed. A maximum in the initial reaction rate was found approximately at a(w)=0.1. The initial reaction rate declined quickly at higher a(w), and dropped precipitously at lower a(w).

Essential role of Rac GTPase in hydrogen peroxide-induced activation of c-fos serum response element.

In the present study, we investigated whether hydrogen peroxide activates c-fos serum response element (SRE) in Rat-2 fibroblast cells. By transient transfection analysis, exogenous H2O2 stimulated SRE-dependent reporter gene activity in a dose and time-dependent manner. Also, we examined the role of Rac GTPase and phospholipase A2 (PLA2) in the H2O2-induced SRE activation. Either transfection of a dominant negative Rac mutant, RacN17, plasmid or pretreatment of mepacrine, a potent inhibitor of PLA2, blocked H2O2-induced SRE activation dramatically. Together, these findings suggest a critical role of 'Rac and subsequent activation of phospholipase A2' in the signaling pathway of H2O2 to SRE.

Effects of gamma radiation on the conformational and antigenic properties of a heat-stable major allergen in brown shrimp.

This study was performed to evaluate the application of food irradiation technology as a method for reducing shrimp allergy without adverse effects. Shrimp heat-stable protein (HSP) was isolated and gamma irradiated at 0, 1, 3, 5, 7, or 10 kGy in the condition of solution (1 mg/ml), and fresh shrimp was also irradiated. Conformational change of irradiated HSP was monitored by means of spectrometric measures, enzyme-linked immunosorbent assay with mouse monoclonal antibody, or human patients' sera and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The ability of the immunoglobulin E of patients allergic to shrimp to bind to irradiated HSP was dose dependently reduced. The amount of intact HSP in an irradiated solution was reduced by gamma irradiation, depending on the dose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the main band disappeared and the traces induced from coagulation appeared at a higher molecular weight zone. The binding ability of immunoglobulin E to allergens in the extracts from irradiated shrimp decreased, depending on the dose. The results provide a new method so that food irradiation technology can be applied to reduce allergenicity of shrimp.

Potential and limitations of alum or zeolite addition to improve the performance of a submerged membrane bioreactor.

In this study, alum and natural zeolite were added to a submerged membrane bioreactor (MBR) not only to reduce membrane fouling but also to increase the removal of nitrogen and phosphorus. Alum addition reduced significantly the rising rate of suction pressure and also resulted in stable and better COD removal. Although phosphorus removal was more than 90% by chemical precipitation, nitrification inhibition was observed. With the addition of natural zeolite, membrane permeability was greatly enhanced by the formation of rigid floc that had lower specific resistance than that of the control activated sludge floc. In particular, the nitrification efficiency was over 95% even at N-shock loading due to the ion-exchange capacity of zeolite. The mechanisms for improved membrane permeability through alum or zeolite addition were discussed in detail.

Pituitary adenylate cyclase-activating polypeptide-immunoreactive cells in the ageing gerbil hippocampus.

In the present study, we investigated age-related changes in pituitary adenylate cyclase-activating polypeptide (PACAP) immunoreactivity and its protein levels in the gerbil hippocampus at various ages using immunohistochemistry and western blot analysis. In the post-natal month 1 (PM 1) group, PACAP-immunoreactive cells were found in all hippocampal subregions. The number of PACAP-immunoreactive cells was decreased in the PM 3 group and was still more decreased in the PM 6 and 12 groups. Thereafter, in the PM 18 and 24 groups, PACAP-immunoreactive cells were significantly increased again. However, in the mossy fibre zone, PACAP immunostaining was very strong in the adult group, especially in the PM 6 group. In addition, PACAP protein level was highest at PM 6, showing a slight decrease at PM 24. These results indicate that PACAP-immunoreactive cells are lowest in the adult stage and highest in the aged stage. However, PACAP immunoreactivity in the mossy fibre zone and PACAP protein level in the hippocampus are highest in the adult stage.

Intramitochondrial pyruvate attenuates hydrogen peroxide-induced apoptosis in bovine pulmonary artery endothelium.

In the hydrogen peroxide (H2O2) apoptosis model of the murine thymocyte, redox reactant and antioxidant pyruvate prevents programmed cell death. We tested the hypothesis that such protection was mediated, at least in part, via pyruvate handling by mitochondrial metabolism. Cultured bovine pulmonary artery endothelial cells were incubated for 30 min with 0.5 mM H2O2 in the absence and presence of 0.5 mM alpha-cyano-3-hydroxycinnamate, as a selective inhibitor of the mitochondrial pyruvate transporter. In controls H2O2 decreased cell viability by 30% within 24 h; this was associated with apoptosis-like bodies, nuclear condensation, and biochemical DNA damage consistent with programmed cell death. Pyruvate (0.1-20 mM) enhanced cell viability in a dose-dependent manner, with > or = 85% viable cells at > or = 3 mM and no DNA laddering, no positive nick-end labeling (TUNEL), and no detectable Annexin V or propidium iodide staining. In contrast, using > or = 5 mM L-lactate as a cytosolic reductant or acetate as a redox-neutral substrate, cell death increased to approximately 40%, which was associated with intense DNA laddering, positive TUNEL and Hoechst 33258 assays. Alpha-cyano-3-hydroxycinnamate alone did not significantly decrease endothelial viability but reduced viability from 85+/-3 to 71+/-4% (p = 0.023) in presence of 3 mM pyruvate plus H2O2; pathological cell morphology and DNA laddering under the same conditions suggested loss of pyruvate protection against apoptosis. Since alpha-cyano-3-hydroxycinnamate re-distributed medium pyruvate and L-lactate consistent with selective blockade of pyruvate uptake into the mitochondria, the findings support the hypothesis that pyruvate protection against H2O2 apoptosis is mediated in part via the mitochondrial matrix compartment. Possible mediators include anti-apoptotic bcl-2 and/or products of mitochondrial pyruvate metabolism such as citrate that affect metabolic regulation and anti-oxidant status in the cytoplasm.

Growth inhibition and differentiation of the human colon carcinoma cell line, Caco-2, by constitutive expression of insulin-like growth factor binding protein-3.

The human colon carcinoma cell line, Caco-2, produces insulin-like growth factor binding protein-3 (IGFBP-3), the secretion of which correlates with markers of enterocyte differentiation. To investigate whether IGFBP-3 inhibits proliferation or induces differentiation, Caco-2 cells were stably transfected with an IGFBP-3 cDNA expression construct or pcDNA3 vector as a control. Accumulation of IGFBP-3 mRNA and secretion of the protein into conditioned medium 9 days after plating were readily detected in the transfected cells, whereas these parameters were undetectable in pcDNA3-transfected cells. Insulin-like growth factor binding protein-3-expressing cells grew at a rate similar to the controls for 6 days after plating, but achieved a much lower final density between days 10 and 12. By day 9 of culture, accumulation of sucrase-isomaltase mRNA, a marker of enterocytic differentiation of Caco-2 cells, was evident in the IGFBP-3-expressing cells, but was undetectable in the controls. These results indicate that IGFBP-3 may inhibit proliferation and induce early differentiation of Caco-2 cells.

Rac and p38 kinase mediate 5-lipoxygenase translocation and cell death.

5-Lipoxygenase (5-LO) is a key enzyme involved in the synthesis of leukotrienes from arachidonic acid, and its activation is usually followed by translocation to the nuclear envelope. The details of mechanisms involved in the translocation of 5-LO are not well understood, though Ca(2+) is known to be essential. Here we show that ionomycin, a Ca(2+) ionophore, induces 5-LO translocation and necrotic cell death in Rat-2 fibroblasts, suggesting a potential relationship between activation of 5-LO and cell death. These effects were markedly attenuated in Rat2-Rac(N17) cells expressing a dominant negative Rac1 mutant. Pretreatment with SB203580, a specific inhibitor of p38 MAP kinase, or EGTA, a Ca(2+) chelator, likewise diminished ionomycin-induced 5-LO translocation and cell death, but PD98059, a MEK inhibitor, did not. Thus, Rac and p38 MAP kinase appear to be components in a Ca(2+)-dependent pathway leading to 5-LO translocation and necrotic cell death in Rat-2 fibroblasts.