Oral mucosa model based on a collagen-elastin matrix.

BACKGROUND AND OBJECTIVE: The collagen-elastin matrix (Matriderm(®)) is used to treat deep and full-thickness burns and was recently described as a suitable scaffold for tissue engineering. The aim of the present study was to investigate the biocompatibility of Matriderm(®) for gingival use through creation of an oral mucosa model ex vivo. MATERIAL AND METHODS: Gingival fibroblasts and keratinocytes were cultured. A dermal area on the base of the collagen-elastin matrix was repopulated with fibroblasts. After 14 days, keratinocytes were seeded on this dermal area to engineer a multilayered mucosa. Analysis of the architecture was performed using light and electron microscopy. Immunohistochemical detection of collagen IV and cytokeratin was carried out. RESULTS: Based on this scaffold we generated a multilayered oral mucosa-like structure. Histological, immunohistochemical and electron microscopic analysis of the dermal/epidermal junction showed a typical basement membrane and hemidesmosomal structures. Neighboring keratinocytes formed desmosomes in the epidermal sections. Cytokeratin was detectable in all epidermal layers. These experiments revealed that the collagen-elastin matrix was highly biocompatible with gingival cells under ex vivo conditions. CONCLUSION: Employing tissue-engineering techniques with dermal and epidermal cells from the gingiva, a multilayered oral mucosa was generated and characterized with respect to biocompatibility for Matriderm(®). The results indicate that Matriderm(®) is suitable for the ex vivo growth of gingival tissue cells and is a useful scaffold with possible applications in periodontal therapy.

Enhanced hydrogen release by catalyzed hydrolysis of sodium borohydride-ammonia borane mixtures: a solution-state 11B NMR study.

Hydrolysis of mixtures consisting of sodium borohydride NaBH(4) (SB) and ammonia borane NH(3)BH(3) (AB) was studied in the absence/presence of a Co catalyst. The kinetics of the H(2) evolutions was measured. The reactions were followed in situ by solution-state (11)B NMR and the hydrolysis by-products characterized by NMR, XRD and IR. It is demonstrated that the combination of the two compounds gives a synergetic effect. SB rapidly reduces the Co catalyst precursor and the NH(4)(+) ions from AB contribute in the dispersion of the in situ formed Co nanoparticles. As a result, the kinetics of H(2) evolution is greatly improved. For instance, a hydrogen generation rate of 29.6 L min(-1) g(-1)(Co) was found for a mixture consisting of 81 wt% NH(3)BH(3), 9 wt% NaBH(4) and 10 wt% CoCl(2). By (11)B NMR, it was showed that the reaction mechanisms are quite trivial. As soon as the Co catalyst forms in situ, SB, rather than AB, hydrolyzes until it is totally converted. Then, the overall hydrolysis continues with that of AB. Both reactions follow a bimolecular Langmuir-Hinshelwood mechanism; no reaction intermediates were observed during the process. In fact, SB and AB convert directly into B(OH)(4)(-), which comes in equilibrium with a polyborate compound identified as B(3)O(3)(OH)(4)(-). All of these results are discussed herein.

Photocatalytic efficiencies of self-cleaning glasses. Influence of physical factors.

Two commercial types of self-cleaning glass (SCG) have been tested to confirm the real photocatalytic nature of their properties. This was done by using four photocatalytic tests: (i) in the gas phase with the total oxidation of acetylene; (ii) in water with the total degradation of malic acid, (iii) in water with the total degradation of methylene blue, and (iv) in the solid phase with the total oxidative degradation of a layer of stearic acid deposited on the self-cleaning glass surface, in contact with the superficial titania coating. The influence of various factors (temperature, humidity, wavelength, radiant flux, presence of inorganic particles stuck at the glass surface) was explained in line with the fundamentals of photocatalysis. The results helped to understand the behaviour of self-cleaning glass.

Environmental green chemistry as defined by photocatalysis.

Photocatalysis is efficient in several fields. Firstly, in selective mild oxidation: oxidation of gas and liquid hydrocarbons (alkanes, alkenes, cyclo-alkanes, aromatics) into aldehydes and ketons. Primary and secondary alcohols are also oxidized into their corresponding aldehydes or ketones. The high selectivity was ascribed to a photoactive neutral, atomic oxygen species. Once platinized (only 0.5wt.% Pt) titania may catalyze reactions involving hydrogen (deuterium-alkane isotopic exchange and alcohol dehydrogenation). For fine chemicals, high initial selectivities enable titania to address most of the twelve principles of "green chemistry", such as the synthesis of 4-tert-butyl-benzaldehyde, an important intermediate in perfume industry by direct selective oxidation of 4-tert-butyl-toluene with air. A new field recently appeared: thio-photocatalysis. Oxygen was replaced by sulfur, using H(2)S as a convenient and reactive source. For instance, the conversion of propene in 1-propanthiol was successfully obtained. The reaction was performed using either CdS or TiO(2). The latter was much more active than CdS. In environmental photocatalysis, titania becomes a total oxidation catalyst once in presence of water because of the photogeneration of OH radicals by neutralization of OH(-) surface groups by positive holes. Many toxic inorganic ions are oxidized in their harmless upper oxidized state. The total degradation of organic pollutants (pesticides, herbicides, insecticides, fungicides, dyes, etc. ...) is the main field of water photocatalytic decontamination. The UVA solar spectrum can de advantageously used as demonstrated by many campaigns performed in the solar pilot plant at the "Plataforma Solar de Almeria" (Spain).

Experimental microkinetic approach of the photocatalytic oxidation of isopropyl alcohol on TiO2. Part 1. Surface elementary steps involving gaseous and adsorbed C3H(x)O species.

The present study concerns an experimental microkinetic approach of the photocatalytic oxidation (PCO) of isopropyl alcohol (IPA) into acetone on a pure anatase TiO2 solid according to a procedure previously developed. Mainly, the kinetic parameters of each surface elementary step of a plausible kinetic model of PCO of IPA are experimentally determined: natures and amounts of the adsorbed species and rate constants (preexponential factor and activation energy). The kinetics parameters are obtained by using experiments in the transient regime with either a FTIR or a mass spectrometer as a detector. The deep oxidation (CO2 and H2O formation) of low concentrations of organic pollutants in air is one of the interests of the PCO. For IPA, literature data strongly suggest that acetone is the single route to CO2 and H2O and this explains that the present study is dedicated to the elementary steps involving gaseous and adsorbed C3H(x)O species. The microkinetic study shows that strongly adsorbed IPA species (two species denoted nd-IPA(sads) and d-IPA(sads) due to non- and dissociative chemisorption of IPA, respectively) are involved in the PCO of IPA. A strong competitive chemisorption between IPA(sads) and a strongly adsorbed acetone species controls the high selectivity in acetone of the PCO at a high coverage of the surface by IPA(sads). The kinetic parameters of the elementary steps determined in the present study are used in part 2 to provide a modeling of macroscopic kinetic data such as the turnover frequency (TOF in s(-1)) of the PCO using IPA/O2 gas mixtures.

Experimental microkinetic approach of the photocatalytic oxidation of isopropyl alcohol on TiO2. Part 2. from the surface elementary steps to the rates of oxidation of the C3H(x)O species.

The present study concerns an experimental microkinetic approach of the photocatalytic oxidation (PCO) of isopropyl alcohol (IPA) into acetone on a pure anatase TiO2 solid according to a procedure previously developed. Mainly, the kinetic parameters of each surface elementary step of a plausible kinetic model of the PCO of IPA are experimentally determined: natures and amounts of the adsorbed species and rate constants (preexponential factors and activation energies). These kinetic parameters are used to evaluate a priori the catalytic activity (turnover frequency, TOF, in s(-1)) of the solid that is compared to the experimental value. The kinetics parameters are obtained by using experiments in the transient regime with either a FTIR or a mass spectrometer as a detector. The microkinetic study shows that only strongly adsorbed IPA species (two species denoted nd-IPA(sads) and d-IPA(sads) due to non- and dissociative chemisorption of IPA respectively) are involved in the PCO of IPA. A strong competitive chemisorption between IPA(sads) and a strongly adsorbed acetone species controls the high selectivity in acetone of the PCO at a high coverage of the surface by IPA(sads). The apparent rate constant (1.4 10(-3) s(-1)) of the Langmuir-Hinshelwood elementary step between IPA(sads) and the active oxygen containing species generated by the UV irradiation provides the TOF of the PCO for IPA/O2 gas mixtures. The kinetic parameters of the elementary steps determined by the experimental microkinetic approach allow us to provide a reasonable simulation of the experimental data (coverages of the adsorbed species and partial pressures of the gases of interest) recorded during a static PCO of IPA(sads) species.

Analysis of genetic polymorphisms at the interleukin-10 loci in aggressive and chronic periodontitis.

BACKGROUND/AIMS: Different cytokine genotypes have been described in periodontal disease. The aim of the present study was to investigate the genetic association of two previously described interleukin-10 (IL-10) polymorphisms in patients with aggressive (AP) and chronic periodontitis (CP) and to investigate possible associations with clinical manifestations. METHODS: Based on clinical parameters and radiographs, 23 patients with CP and 18 patients with AP were included in the study. Additionally, 21 age-matched healthy subjects served as a control group. Genomic DNA was isolated from whole blood samples and the IL-10 promoter sequences from positions - 597 to - 824 were amplified by polymerase chain reaction (PCR). Polymorphisms were detected by restriction-enzyme cleavage. The A and C alleles at the - 597 position were associated with the T and C alleles at the - 824 position, respectively. Fisher's exact test was used for the statistical analysis. RESULTS: No significant differences were observed in the allele frequencies between controls and AP patients (p = 0.70) or CP patients (p = 0.43), although the previously reported association between allele A at position - 597 and allele T at position - 824 was observed in our population. CONCLUSION: We conclude that the investigated polymorphisms are not associated with periodontal disease.

[Essential hypertension and stress. When do yoga, psychotherapy and autogenic training help?]. / Essenzielle Hypertonie und Stress. Wann helfen Yoga, Psychotherapie und autogenes Training?

Psychosocial factors play an important role in the development and course of essential hypertension, although "stress" can account for only 10% of blood pressure variance. A variety of psychotherapeutic interventions, such as relaxation techniques (autogenic training or progressive muscular relaxation), behavioral therapy or biofeedback techniques, can lower elevated blood pressure by an average of 10 mmHg (systolic) and 5 mmHg (diastolic). As a "secondary effect", such measures may also prompt the hypertensive to adopt a more health-conscious lifestyle.

Real-time optical coherence tomography for minimally invasive imaging of prostate ablation.

OBJECTIVE: Numerous ablation techniques have been developed to alleviate urethral obstruction and improve urodynamics in benign prostatic hyperplasia. Most techniques, however, rely on visual observation of surface changes for ablation end points. The feasibility of using real-time optical coherence tomography (OCT) for minimally invasive imaging to guide and monitor prostate resection is demonstrated with representative techniques of laser and radiofrequency ablation. Empiric comparisons of ablation dynamics are made, and the use of OCT as a high-resolution, subsurface modality for image guidance is evaluated. MATERIALS AND METHODS: Optical coherence tomography is a high-resolution, high-speed near-infrared imaging technique analogous to ultrasound imaging, except that reflections of light are detected rather than sound. High-speed OCT is used to image the dynamic process of laser and radiofrequency ablation of in vitro human prostate tissue. OCT images of ablation sites are compared with corresponding histology. RESULTS: Based on comparisons between OCT images and corresponding histology, OCT imaged transurethral prostate tissue morphology, including urethral sinuses and submucosal glands. Real-time OCT imaging provided rapid feedback and control of ablation dynamics. The compact and portable OCT technology is amenable to minimally invasive beam-delivery devices. CONCLUSIONS: Optical coherence tomography offers a minimally invasive means of assessing transurethral prostate morphology. Real-time OCT has the potential to provide image guidance of prostate resection for many of the existing surgical treatments directed at alleviating urethral obstruction associated with benign prostatic hyperplasia.

Interleukin-4 polymorphisms in early onset periodontitis.

BACKGROUND, AIMS: Periodontitis is the result of a complex interplay between oral bacteria and the host response, often modulated by behavioral factors. Early-onset periodontitis (EOP) is defined by the age of onset, the distribution of lesions and specific microbial pathogens. METHOD: Studies in twins suggested a genetic contribution to the pathogenesis of periodontitis. EOP heritable factors may be related to immune mechanisms which could enhance the pathogenic potential of plaque bacteria in susceptible individuals. Among others, Interleukin 4 (IL-4) is a potent cytokine in the immune response and is a potent down regulator of macrophage function. In the present study, we report a specific genotype of the IL-4 gene, which was detected by specific primers and PCR analysis. RESULTS: In the EOP-group 27.8% were IL-4 promotor- and intron polymorphism positive (PP+ and IP+). None of the age-matched healthy controls or patients with adult periodontitis (n=25) carried the markers. Moreover, serum IL-4 levels of PP+ and IP+ patients were below the detection limit and significantly different (p<0.01) from the IL-4 concentrations of healthy controls and PP- and IP- patients.

Concentration of interleukin-1beta and neutrophil elastase activity in gingival crevicular fluid during experimental gingivitis.

BACKGROUND/AIM: The aim of the present study was to measure interleukin-1beta concentrations and neutrophil elastase activity in gingival crevicular fluid (GCF) during experimental gingivitis in humans. MATERIAL AND METHODS: 12 healthy young men participated. After prophylaxis, they performed optimal hygiene to reach plaque and gingivitis indices of or approaching zero. All oral hygiene measures were then ceased for a period of 18 days. The Quigley-Hein plaque index (PLI) and Saxer & Mühlemann papillary bleeding index (PBI) were assessed. GCF samples were taken from the mesiobuccal site of two contralateral teeth in the upper jaw by means of periopapers at baseline and on days 3, 7, 14 and 18. After measuring the gingival crevicular fluid volume (GCFV) with the Periotron 8000, the samples were analyzed in our laboratory for the detection of IL-1beta concentration by ELISA. RESULTS: PLI and PBI showed a reduction prior to baseline reaching almost zero, both increasing from day 0 to day 18 (PLI=from 0.1 to 2.9, PBI=from 0 to 2.0). IL-1beta concentration increased from 229.25 ng/ml (day 0) to 526.13 ng/ml (day 18). Clinical data and IL-1beta concentrations were correlated with elastase activity (EA). No significant correlation could be demonstrated between the clinical parameters assessed and IL-1beta or EA (Spearman rank correlation coefficient). A correlation between GCFV and PBI from day 0 to day 18 could be demonstrated. CONCLUSION: Overall, both IL-1beta and EA showed an increase from baseline throughout the whole study.

Analysis of protein-protein interactions in mitochondria by coimmunoprecipitation and chemical cross-linking.

Many different techniques have been employed to analyze protein-protein interactions. Coimmunoprecipitation and chemical cross-linking have been used extensively to study mitochondrial biogenesis. Both techniques have proven to be powerful methods to investigate the sequential interactions of precursor proteins with the various components of the translocation machineries in the mitochondrial membranes. Similarly, protein-protein interactions during processes such as protein synthesis, folding, and degradation can be studied. Moreover, the composition of the oligomeric protein complexes of mitochondria, such as respiratory chain complexes or protein translocation machineries, can be determined. The general principles and protocols of these methods are described and illustrated with typical examples.

Mba1, a novel component of the mitochondrial protein export machinery of the yeast Saccharomyces cerevisiae.

The biogenesis of mitochondria requires the integration of many proteins into the inner membrane from the matrix side. The inner membrane protein Oxa1 plays an important role in this process. We identified Mba1 as a second mitochondrial component that is required for efficient protein insertion. Like Oxa1, Mba1 specifically interacts both with mitochondrial translation products and with conservatively sorted, nuclear-encoded proteins during their integration into the inner membrane. Oxa1 and Mba1 overlap in function and substrate specificity, but both can act independently of each other. We conclude that Mba1 is part of the mitochondrial protein export machinery and represents the first component of a novel Oxa1-independent insertion pathway into the mitochondrial inner membrane.

The ADP ribosylation factor-nucleotide exchange factors Gea1p and Gea2p have overlapping, but not redundant functions in retrograde transport from the Golgi to the endoplasmic reticulum.

The activation of the small ras-like GTPase Arf1p requires the action of guanine nucleotide exchange factors. Four Arf1p guanine nucleotide exchange factors have been identified in yeast: Sec7p, Syt1p, Gea1p, and its homologue Gea2p. We identified GEA2 as a multicopy suppressor of a sec21-3 temperature-sensitive mutant. SEC21 encodes the gamma-subunit of coatomer, a heptameric protein complex that together with Arf1p forms the COPI coat. GEA1 and GEA2 have at least partially overlapping functions, because deletion of either gene results in no obvious phenotype, whereas the double null mutant is inviable. Conditional mutants defective in both GEA1 and GEA2 accumulate endoplasmic reticulum and Golgi membranes under restrictive conditions. The two genes do not serve completely overlapping functions because a Deltagea1 Deltaarf1 mutant is not more sickly than a Deltaarf1 strain, whereas Deltagea2 Deltaarf1 is inviable. Biochemical experiments revealed similar distributions and activities for the two proteins. Gea1p and Gea2p exist both in membrane-bound and in soluble forms. The membrane-bound forms, at least one of which, Gea2p, can be visualized on Golgi structures, are both required for vesicle budding and protein transport from the Golgi to the endoplasmic reticulum. In contrast, Sec7p, which is required for protein transport within the Golgi, is not required for retrograde protein trafficking.

The mitochondrial proteins Ssq1 and Jac1 are required for the assembly of iron sulfur clusters in mitochondria.

Mitochondria of the yeast Saccharomyces cerevisiae contain three different Hsp70 chaperones, Ssc1, Ecm10 and Ssq1. Ssc1 is an essential protein that mediates the import of nuclear-encoded proteins into the organelle and their subsequent folding. The nucleotide state of Ssc1 is thereby regulated by the nucleotide exchange factor Mge1. Here, we show that Mge1 interacts with Ssq1 in an ATP-dependent manner, suggesting that Mge1 also regulates Ssq1 function. In contrast to Ssc1, Ssq1 does not associate with the Tim44 subunit of the protein translocating complex, indicating a different function of both chaperones. Mutants in Ssq1 were reported to have low levels of iron sulfur (FeS) cluster-containing enzymes. Employing an assay that allowed us to monitor the conversion of the apoform of mitochondrial ferredoxin into its FeS-containing holoform, Ssq1 was demonstrated to be required for the FeS cluster assembly in mitochondria. The mitochondrial DnaJ homolog Jac1 is crucial for this process, whereas Mdj1 function is dispensable. Furthermore, the presence of frataxin is necessary for FeS cluster assembly into ferredoxin suggesting a role for frataxin at the level of the formation of holo-ferredoxin.

Microassay for the detection of elastase activity in the gingival crevice.

BACKGROUND, AIMS: A new microassay for the detection of elastase activity (EA) in gingival crevicular fluid (GCF) has been established. GCF was collected with Periopaper strips and quantified in a Periotron. METHODS: Enzyme activity was measured in a microtiter plate reader, using a fluorometric assay. To ensure quality and precision of the assay, recovery rates were determined at different activities with a recovery of >90%. In a 2nd step, stability of the enzyme was investigated during storage at room temperature, +4 degrees C, -22 degrees C, -88 degrees C. GCF samples retained elastase activity of almost 100% after a storage of 3 days at -22 degrees C. In a group of 12 healthy volunteers, elastase activity was assayed throughout an 18 day experimental gingivitis protocol. RESULTS: Median activity increased from 481 microU/microl at baseline to 1444 microU/microl at day 18, which was accompanied by the development of the signs of gingivitis. The increase of EA during the experimental phase of the study was highly significant (p<0.001) and correlated well with the increasing severity of gingivitis. CONCLUSION: The data suggest that elastase activity in GCF is an excellent quantitative measure of gingival inflammation.

[The Roemer Award of the German College of Psychosomatic Medicine (1976-1998): quantitative data and contents]. / Der Roemer-Preis des DKPM (1976-1998): Quantitative und inhaltliche Aspekte.

The Roemer-Award of the German College of Psychosomatic Medicine might be the most attractive research award in the German Psychosomatic field. This review starts with an appreciation of its namer, Hans Roemer. Then the award winners between 1976 and 1998 are summarized. Quantitative analyses of the material reveal interesting data about the number of applications during these years and about how the applications and the awarded work are distributed over the different issues of the research field. It is shown that studies dealing with psychosomatic aspects in internal medicine were submitted and awarded most frequently.

The apoptosis mediator mDAP-3 is a novel member of a conserved family of mitochondrial proteins.

Programmed cell death is essential for organ development and regeneration. To identify molecules relevant for this process, full length cDNA cloning of a short, developmentally regulated murine cDNA fragment, MERM-3, was performed and showed a 1.7 kb mRNA encoding a 45 kDa protein with an ATP/GTP binding motive (P-loop). Sequence analysis revealed an 82% amino acid identity to the human death associated protein 3 (hDAP-3), a positive mediator of apoptosis. The full length sequence being the murine orthologue of hDAP-3 is therefore referred to as mDAP-3. In situ hybridization and northern blot analysis showed an abundant mRNA expression with a pronounced expression in highly proliferative epithelial compartments. For mDAP-3, cytochrome c release and induction of cell death could be demonstrated by overexpression of a mDAP-3/EGFP fusion protein. DAP-3 mediated apoptosis was shown to depend on a functional P-loop. Intracellular localization studies using the mDAP-3/EGFP fusion protein, cell fractionation and protease protection experiments localized mDAP-3 to the mitochondrial matrix. DAP-3, in contrast to cytochrome c, retained its mitochondrial localization during apoptosis induction. A mutant of a putative yeast orthologue of mDAP-3, YGL129c, here referred to as yDAP-3, has been shown to exhibit disrupted mitochondrial function. yDAP-3 deficient mutants could be shown to progressively loose mitochondrial DNA. Loss of mitochondrial DNA in yDAP-3 was partially prevented by transfection of the yDAP-3 deficient mutant with mDAP-3, indicating functional complementation by murine DAP-3 in the yeast system. These data identify mDAP-3 as one of the first proapoptotic factors in the mitochondrial matrix and provide evidence for a critical, evolutionary conserved role of members of the DAP-3 protein family for mitochondrial biogenesis.

What fuels polypeptide translocation? An energetical view on mitochondrial protein sorting.

Protein sorting into mitochondria is achieved by the concerted action of at least four translocation complexes. Vectorial transport of polypeptide chains by these complexes requires different driving forces. In particular, Deltapsi, matrix adenosine triphosphate and the free energy of the binding to other protein components are used in series to achieve sorting of proteins to the various mitochondrial subcompartments. The processes providing the translocation energy are presented in this review and their impact for protein sorting into and within mitochondria is discussed.