Advanced quantitative proteomics to evaluate molecular effects of low-molecular-weight hyaluronic acid in human dermal fibroblasts

Hyaluronic acid (HA) is physiologically synthesized by several human cells types but it is also a widespread ingredient of commercial products, from pharmaceuticals to cosmetics. Despite its extended use, the precise intra- and extra-cellular effects of HA at low-molecular-weight (LWM-HA) are currently unclear. At this regard, the aim of this study is to in-depth identify and quantify proteome's changes in normal human dermal fibroblasts after 24 h treatment with 0.125, 0.25 and 0.50 % LMW-HA (20 1250 kDa) respectively, vs controls. To do this, a label-free quantitative proteomic approach based on high-resolution mass spectrometry was used. Overall, 2328 proteins were identified of which 39 significantly altered by 0.125 %, 149 by 0.25 % and 496 by 0.50 % LMW-HA. Protein networking studies indicated that the biological effects involve the enhancement of intracellular activity at all concentrations, as well as the extracellular matrix reorganization, proteoglycans and collagen biosynthesis. Moreover, the cell's wellness was confirmed, although mild inflammatory and immune responses were induced at the highest concentration. The more complete comprehension of intra- and extra-cellular effects of LMW-HA here provided by an advanced analytical approach and protein networking will be useful to further exploit its features and improve current formulations

T cell-mediated hypersensitivity to quinolones: mechanisms and cross-reactivity

BACKGROUND: Quinolones are widely used, broad spectrum antibiotics that can induce immediate- and delayed-type hypersensitivity reactions, presumably either IgE or T cell mediated, in about 2-3% of treated patients. OBJECTIVE: To better understand how T cells interact with quinolones, we analysed six patients with delayed hypersensitivity reactions to ciprofloxacin (CPFX), norfloxacin (NRFX) or moxifloxacin (MXFX). METHODS: We confirmed the involvement of T cells in vivo by patch test and in vitro by means of the lymphocyte proliferation test (LTT). The nature of the drug-T cell interaction as well as the cross-reactivity with other quinolones were investigated through the generation and analysis (flow cytometry and proliferation assays) of quinolone-specific T cell clones (TCC). RESULTS: The LTT confirmed the involvement of T cells because peripheral blood mononuclear cells (PBMC) mounted an enhanced in vitro proliferative response to CPFX and/or NRFX or MXFX in all patients. Patch tests were positive after 24 and 48 h in three out of the six patients. From two patients, CPFX- and MXFX-specific CD4(+)/CD8(+) T cell receptor (TCR) alphabeta(+) TCC were generated to investigate the nature of the drug-T cell interaction as well as the cross-reactivity with other quinolones. The use of eight different quinolones as antigens (Ag) revealed three patterns of cross-reactivity: clones exclusively reacting with the eliciting drug, clones with a limited cross-reactivity and clones showing a broad cross-reactivity. The TCC recognized quinolones directly without need of processing and without covalent association with the major histocompatability complex (MHC)-peptide complex, as glutaraldehyde-fixed Ag-presenting cells (APC) could present the drug and washing quinolone-pulsed APC removed the drug, abrogating the reactivity of quinolone-specific TCC. CONCLUSION: Our data show that T cells are involved in delayed immune reactions to quinolones and that cross-reactivity among the different quinolones is frequent

Pembuatan Bioetanol dari Umbi Singkong Karet dengan Metode Hidrolisis Asam dan Enzim

Bioetanol merupakan salah satu bahan bakar alternatif terbarukan yang dapat menggatikan bahan bakar fosil . Tujuan penelitian ini adalah untuk membandingkan bioetanol yang dihasilkan dari tepung( ampas singkong karet, singkong karet parut, dan pati singkong karet) dengan metode hidrolisa asam klorida dan Hidrolisa Enzim α – amilase, Enzim glukoamilase,terhadap bioethanol yang dihasilkan dengan berat tepung sebanyak 300 gram. Hasil penelitian menunjukkan bahwa Bioetanol dari tepung singkong karet parut yang dihidrolisis dengan enzim menghasilkan volume sebanyak 2,7 ml memiliki kadar etanol tertinggi, yaitu 84,25% dan indeks biasnya adalah 1,361 mendekati indek bias etanol murni yaitu 1,3633, sedangkan singkong karet parut yang dihidrolisis asam menghasilkan bioetanol dengan volume sebanyak 1,6 ml dan indek bias 1,332 dan kadar alkohol yaitu 24,25 % Hasil penelitian yang menunjukan bahwa kadar gula tertinggi diperoleh dari singkong karet parut, yaitu 17,748 ppm dari hasil hidrolisis enzim dan 18,744 ppm dari hasil hidrolisis asam. Bioetanol dari singkong karet parut yang dihidrolisis dengan enzim menghasilkan volume sebanyak 2,7 ml memiliki kadar etanol tertinggi, yaitu 84,25% dari hasil perhitungan data indeks bias dan 38,571% dari hasil analisis gas chromatography, sedangkan singkong karet parut yang dihidrolisis asam menghasilkan bioetanol dengan volume sebanyak 1,6 ml memiliki kadar yang lebih rendah, yaitu 24,25% dari hasil perhitungan data indeks bias dan 15,640% dari hasil analisa menggunakan gas chromatography

JESSI project: advanced technolgoy for 0.25 #mu#m CMOS and below. Subproject: process modules evaluation and integration. Theme 5 Multilevel metallization. Final report

Titanium- and cobaltdisilicide were investigated as material for shallow, low-resistive contacts in a CMOS-metallization system. ITM and DDS were used as self-aligned manufacturing processes. The TiSi_2- and CoSi_2-contact-systems were compared concerning manufacturing process and electrical and physical properties. The CoSi_2-contact-system shows the best properties. CVD-W was used to fill up vias and interconnects. To this a selective tungsten CVD process on CoSi_2 and a blanket deposition process were developed. Due to the excellent selectivity concerning the contact filling, a planarization effect was achieved. Accelerated tests show a high current-carrying capacity for the interconnects. Electromigration was identified as degradation mechanism. PECVD- and TEOS-PECVD-processes were investigated for the deposition of intermetal-dielectrics. An electrical and physical characterization of the layers have been performed. Processes with good step coverage and processes with a planarization effect were specified. Etch-back procedures using photo-resist and spin-on-techniques with various polyimides and a spin-on-glass were investigated in view of a planarization of the chip surface. The best results were achieved using the spin-on-technique with polyimide. (orig.)SIGLEAvailable from TIB Hannover: F94B1507+a+b / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Forschung und Technologie (BMFT), Bonn (Germany)DEGerman