Biomedical subjects

F Lampert

Publications and source records attributed to F Lampert.

At least 19 recordsLinked to original sources

PubMed2005

Engrafting periodontal fibroblasts with new 3-dimensional polylactide foams.

In clinical periodontics, "tissue engineering" techniques have been developed to guide the regenerative process following periodontal disease. We explored a new shaped biomaterial in order to promote cellular adhesion, proliferation and migration of periodontal cells. Granules of poly-D, L-lactide were foamed in specially designed moulds by a controlled gas-loading process. Explant cultures of periodontal tissue were seeded at different densities on the 3-dimensional scaffolds following analysis of cytotoxicity, cell proliferation and differentiation. The moulding procedure led to porous structures with predetermined tubes for cellular locomotion. Cells adhered to and populated the material's surface and inner cavities while retaining fibroblastic phenotype. An optimal ratio between cellular proliferation and mortality rate was achieved at a seeding density of > 10 6 cells/cm 3 scaffold. We designed modified polylactide scaffolds capable of acting as a stent and a cell carrier matrix. The foaming process is easy, cheap and suitable for commercial production.

Biocompatible Materials↗

PubMed2004

Bactericidal effect of a 980-nm diode laser in the root canal wall dentin of bovine teeth.

OBJECTIVE: The aim of this in vitro study was to investigate the antibacterial depth effect of continuous wave laser irradiation with a wavelength of 980 nm in the root canal wall dentin of bovine teeth. BACKGROUND DATA: The long-term success of an endodontic therapy often fails due to remaining bacteria in the root canal or dentin tubules, which cannot be sufficiently eliminated through the classical root canal preparation technique nor through rinsing solutions. MATERIALS AND METHODS: A total of 102 slices of bovine root dentin of different thicknesses (100, 300 and 500 micro m) were prepared. The samples were inoculated from one side with 5 micro L of an enterococcus faecalis suspension of defined concentration. Four slices per slice thickness served as a control group; the rest of the 30 slices per thickness were subjected to laser irradiation - 10 each of these slices were irradiated with distal outputs of 1.75, 2.3, and 2.8 Watts (W). After drying them for 30 sec, the back of the inoculated dentin slice was irradiated for 32 seconds with a 200- micro m fiber optical waveguide under constant movement of the fibers. The remaining bacteria were then detached in NaCl under vibration. The eluate produced by this was - taking account of the degree of dilution - plated out on sheep blood agar plates. After 24 h of incubation, the grown bacterial colonies were able to be counted out and evaluated. By doing so, they were compared with the non-irradiated, but otherwise identically treated control group. RESULTS: With a slice thickness of 100 micro m, the 980-nm diode laser achieved a maximum bacterial reduction of 95% at 1.75 W, 96% at 2.3 W, and 97% at 2.8 W. With a slice thickness of 300 micro m, a maximum of 77% of the bacteria was destroyed at 1.75 W, 87% at 2.3 W, and 89% at 2.8 W. The maximum bacterial reduction with a slice thickness of 500 micro m was 57% at 1.75 W, 66% at 2.3 W, and 86% at 2.8 W. CONCLUSION: The results of this research show that the 980-nm diode laser can eliminate bacteria that have immigrated deep into the dentin, thus being able to increase the success rate in endodontic therapy.

Animals↗

PubMed2001

Distal chromosome 17 gains in neuroblastomas detected by comparative genomic hybridization (CGH) are associated with a poor clinical outcome.

PROCEDURE: To establish the significance of chromosome 17 aberrations in the biology of neuroblastomas, the fresh-frozen material of 53 primary neuroblastomas (average patient age: 20.8 months; stage 1 or 2: n = 10; stage 3: n = 10; stage 4: n = 10; stage 4s: n = 23) was studied by means of comparative genomic hybridization (CGH). Follow-up data were available for 52 of 53 cases studied (average follow-up period: 26.4 months). Except for one, all cases had previously been analyzed for MYCN status (semiquantitative Southern blot analysis). Studies of LOH 1p36 (VNTR-PCR) had been performed on 28 of 53 cases. RESULTS: Chromosome 17 gains were detected in 46 of 53 (86.8%) cases. Whole chromosome gains were mostly restricted to localized tumors (stage 1 or 2: 9 of 10 cases; stage 4s:19 of 23; stage 3: 2 of 10; stage 4:0 of 10 cases), whereas distal 17 gains were significantly associated with clinically advanced tumor stages and patients aged over 1 year at diagnosis. Univariate analyses revealed a statistically significant correlation of distal 17q gains with overall survival (P< 0.01, MYCN amplification: P< 0.01; 1p deletion: P< 0.01) and an elevated recurrency rate (17q: P= 0.02, MYCN amplification: P = 0.05; 1p deletion P= 0.3). There was a strong coincidence of distal 17q gains and 1p deletion or MYCN amplification (P < 0.01). CONCLUSION: Our data indicate that distal chromosome 17q gains are of major prognostic relevance for neuroblastoma patients. However, studies on a larger series of tumors have to be performed to assess whether or not these alterations are independent prognostic markers of a poor clinical outcome.

Child↗

PubMed2001

Multicentre analysis of patterns of DNA gains and losses in 204 neuroblastoma tumors: how many genetic subgroups are there?

PROCEDURE: Analysis of comparative genomic hybridization (CGH) data of 120 tumors from four different studies, and data of 84 previously unpublishied tumors, allowed delineation of at least six different genetic subsets of neuroblastomas. RESULTS AND CONCLUSIONS: A small number of tumors show no detectable imballances. A second group of tumors presents with gains and losses of whole chromosomes and is found predominantly in prognostically favorable stage 1 and 2 tumors. The remaining groups are characterized by the presence of partial chromosome imbalances, and are found mostly in stage 3, 4, and 4S tumors. The third group shows 17q gain without 11q loss, 1p loss, or MYCN amplification (MNA). The fourth group has 1p deletion or MNA, and finally, a fifth group shows 11q loss without 1p deletion or MNA, and is found mainly in stage 4 tumors. The latter group is significantly associated with losses of 3p, 4p, and 14q.

Chromosome Aberrations↗

PubMed2001

Microleakage of composite fillings in Er,Cr:YSGG laser-prepared class II cavities.

BACKGROUND AND OBJECTIVE: If there is insufficient bonding to the enamel, the shrinkage of composites that occurs during polymerization can result in a gap between the filling material and the cavity wall. This gap permits the passage of bacteria or their metabolic products and also of various molecules and ions. This leads to hypersensitivity or secondary caries and is thus one of the causes of the failures encountered in composite restorations. The aim of this study was to examine the quality of the margins of composite fillings in Er,Cr:YSGG laser-prepared cavities by means of dye penetration. The results were compared with those of restoration in conventionally prepared and conditioned cavities. STUDY DESIGN/MATERIALS AND METHODS: To this end, 45 class II cavities in extracted molars were prepared. The teeth were divided into three groups. The first group served as a control group. The cavities were prepared in the classical manner by using a diamond, beveled and subsequently conditioned by the etching method. In group 2, the cavities were prepared and conditioned exclusively with the Er,Cr:YSGG laser. In group 3, laser preparation was supplemented by conditioning of the cavity with phosphoric acid. RESULTS: No significant difference could be found between the classical preparation technique in combination with etching and the laser preparation method with supplementary etching (group 3). The degree of dye penetration was highest in the group undergoing laser-prepared restoration without additional etching (group 2) (Wilcoxon test, P < 0.017). CONCLUSIONS: Although it was found in previous studies that there is no significant difference between bond strength of acid etched enamel and Er,Cr:YSGG laser etched enamel, the dye penetration rate differs. On the basis of the results of our study, the additional use of etching after Er,Cr:YSGG laser preparation is recommended as it is used in the classical cavity preparation technique.

Coloring Agents↗

PubMed2001

Conditioning of enamel with Nd:YAG and CO2 dental laser systems and with phosphoric acid. An in-vitro comparison of the tensile bond strength and the morphology of the enamel surface.

UNLABELLED: MATERIAL AND METHOD 1: The first stage of a three-stage in-vitro investigation used 146 maxillary incisors to identify optimum setting parameters for two CO2 and Nd:YAG standard laser systems applied in enamel conditioning for bracket bonding. RESULTS: 31 of the 75 parameter combinations investigated for the Nd:YAG laser and nine of the 71 investigated for the CO2 laser proved appropriate for further investigation of tensile strength. MATERIAL AND METHOD 2: In a second stage, tensile bond strength for the selected laser settings was determined in comparison to the conventional acid-etch technique for a further 210 teeth as the control group. RESULTS: The highest average tensile strength for the Nd:YAG laser was 4.1 MPa, with comparable values of 3.3 MPa for the CO2 Laser and 4.9 MPa for the acid-etch technique. MATERIAL AND METHOD 3: The third stage of the investigation involved comparative scanning electron microscopy of enamel surface morphology following laser application and acid-etching. RESULTS: The CO2 laser was found to produce craters of various dimensions, while the Nd:YAG laser produced honeycomb structures regionally similar to enamel samples from the acid-etch technique. CONCLUSION: The CO2 and Nd:YAG dental lasers tested produce enamel conditioning and tensile bond strength sufficient to meet the requirements of bracket bonding.

Acid Etching, Dental↗

PubMed2001

Assessment of acid production by various human oral micro-organisms when palatinose or leucrose is utilized.

One promising way of reducing caries is by using sucrose substitutes in food, e.g., palatinose or leucrose. Previous experiments addressing cariogenic potential of sucrose substitutes have focused mainly on Streptococcus mutans. However, given the many other micro-organisms in the oral cavity, this study compared the acid production of 100 bacterial strains representing 44 different species, by batch fermentation in a test tube containing, as a sole carbohydrate source, glucose, sucrose, palatinose, or leucrose. Selected strains were further analyzed in a fermenter. Additionally, 30 yeast strains were tested by an auxanographic sugar assimilation test. Only Lactobacillus spp., Stomatococcus mucilaginosus, Leuconostoc mesenteroides, and Weissella paramesenteroides, and some of the yeasts studied-i.e., Candida albicans, C. tropicalis, C. parapsilosis, and Saccharomyces cerevisiae-utilized leucrose and/or palatinose well. Strikingly, Stomatococcus mucilaginosus produced water-insoluble polysaccharides by fermentation of leucrose and palatinose. In the fermenter, the respective sucrose substitutes were not only cleaved but also utilized. Thus, extracellular cleavage by autochthonous micro-organisms may produce cariogenic cleavage products (glucose, fructose) that can be used by other well-characterized cariogenic bacteria found in the oral flora. Therefore, the anticariogenic potential of sucrose substitutes in food might be limited.

Bacteria↗

PubMed2000

Gene expression of neuronal nitric oxide synthase and adrenomedullin in human neuroblastoma using real-time PCR.

The objective of our study was to assess the gene expression of the antiproliferative systems neuronal nitric oxide synthase (nNOS) and adrenomedullin (AM) in human neuroblastoma. A novel real-time PCR method was evaluated using neuropeptide Y (NPY) for validation. Glyceraldehyd-3-phospate dehydrogenase (GAPDH) and NPY gene expression in neuroblastomas of 50 patients were measured in parallel by competitive quantitative and TaqMan real-time RT-PCR. AM and nNOS mRNA were determined by real-time PCR. Our results showed a linear relationship between competitive quantitative and real-time RT-PCR measurements of NPY and GAPDH (r = 0.87 and r = 0.92, respectively). AM and nNOS mRNA was found in all tumor samples. AM/GAPDH mRNA increased with higher differentiation according to Shimada (p = 0.013). There was no relation between MYCN amplification nor with the tumor stage (p = 0.78 and p = 0.30, respectively). AM/GAPDH did not relate to recurrence or death in a 5-year follow-up period. Neuronal NOS/GAPDH expression did not relate to any biological or clinical parameter of prognosis or differentiation. Similar results were obtained when the neuronal marker protein gene product 9.5 (PGP9.5) was used to normalize mRNA concentration. In conclusion, TaqMan real-time PCR appears to be a reliable method to quantify gene expression in neuroblastomas. Adrenomedullin mRNA in neuroblastoma is linked to tumor differentiation but not to prognostic markers.

Adolescent↗

PubMed2000

The human GRAF gene is fused to MLL in a unique t(5;11)(q31;q23) and both alleles are disrupted in three cases of myelodysplastic syndrome/acute myeloid leukemia with a deletion 5q.

We have isolated the human GRAF gene (for GTPase regulator associated with the focal adhesion kinase pp125(FAK)). This gene was fused with MLL in a unique t(5;11)(q31;q23) that occurred in an infant with juvenile myelomonocytic leukemia. GRAF encodes a member of the Rho family of the GTPase-activating protein (GAP) family. On the protein level, it is 90% homologous to the recently described chicken GRAF gene that functions as a GAP of RhoA in vivo and is thus a critical component of the integrin signaling transduction pathway. The particular position of the human GRAF gene at 5q31 and the proposed antiproliferative and tumor suppressor properties of its avian homologue suggest that it also might be pathogenetically relevant for hematologic malignancies with deletions of 5q. To investigate this possibility, we sequenced 4-5 individual cDNA clones from 13 cases in which one allele of GRAF was deleted. We found point mutations within the GAP domain of the second GRAF allele in one patient. In two additional patients we found an insertion of 52 or 74 bp within the GRAF cDNA that generates a reading frame shift followed by a premature stop codon. GRAF maps outside the previously defined commonly deleted 5q31 region. Nevertheless, inactivation of both alleles in at least some cases suggests that deletions and mutations of the GRAF gene may be instrumental in the development and progression of hematopoeitic disorders with a del(5q).

Artificial Gene Fusion↗

PubMed2000

Microhardness changes in surface enamel after application of bioadhesive fluoride tablets in situ.

The effect of fluoride released by bioadhesive tablets was evaluated in an in situ model in human volunteers. Eight volunteers carried four to six polished human enamel samples in a lower lingual device for a period of 5 days without fluoride supplements (control) and 5 days using one bioadhesive tablet (0.5 mg F) per day, placed in the lower buccal sulcus. Changes in mineral content were measured in terms of surface microhardness indentation length (load 0.98 N, Knoop diamond indenter). Some enamel samples were analyzed for their fluoride content by means of a surface etch biopsy. The indentation length increased significantly in both experiments, but demineralization was less pronounced with fluoride (indentation length increased from 44 +/- 7 to 48 +/- 13 microns) than in the control (44 +/- 7 to 58 +/- 13 microns). The fluoride concentration and the amount of calcium in the acid etch biopsy specimens of the enamel samples did not differ significantly between the two groups. Bioadhesive, fluoride-releasing tablets have therefore demonstrated the potential to reduce demineralization by elevating salivary and plaque fluoride concentration for a period of several hours. This self-administered device can be a supplementary means for prevention of caries.

Adhesives↗

PubMed2000

Quantitative DNA fragment analysis for detecting low amounts of hepatitis B virus deletion mutants in highly viremic carriers.

Many variants of hepatitis B virus (HBV) with deletions in the viral genome have been identified. Some of these variants are indicator or even effector of a more severe course of hepatitis. These deletion mutants contribute a variable and sometimes very low proportion to the viral population. For early detection of small amounts of deletion mutants among a large number of wild-type genomes, we applied a new screening method designated quantitative fragment analysis (QFA). By QFA the whole viral genome can be scanned for the presence of deletions or insertions of >/=3 nucleotides representing more than 2% of the viral population. Using QFA we showed that an often described deletion of 8 nucleotides is packaged in viral capsids and not a polymerase chain reaction (PCR) artifact. QFA was applied to study the emergence of deletion mutants in a group of 18 pediatric patients who had been infected from a common source while being under multidrug cancer chemotherapy. All patients had developed a highly viremic asymptomatic HBV carrier state. In 3 of these patients 3 different kinds of HBV deletion mutants were found by QFA: 8 bp deletions within the core promoter, core gene deletions from 8 to 86 bp, and large deletions of up to 1,989 bp spanning the precore/core and the preS/S reading frames. PCR primers that specifically amplify deletion variants enabled the detection of additional patients harboring the investigated variant.

Adolescent↗

PubMed2000

Diode laser radiation and its bactericidal effect in root canal wall dentin.

OBJECTIVES: The aim of this study was to investigate the antibacterial effect of a diode laser in deep root canal dentin. BACKGROUND DATA: The microbial colonization of root canal dentin can lead to failures in conventional endodontic treatment if an inadequate bacterial reduction only is achieved through canal treatment and chemical disinfection. METHODS: 100 microm, 300 microm and 500 microm bovine dentin slices obtained by longitudinal sections were sterilized and inoculated on one side with an Enterococcus faecalis suspension. Laser radiation was performed on the opposite side with the diode laser (810 nm) at a setting of 3 W in continuous mode (CW). Radiation was performed using a 400-microm tapered fiber tip at an angle of approximately 5 degrees to the surface over a period of 30 seconds. The output power at the distal end of the tip was 0.6 W. The bacteria were then eluted through vibration and cultured on blood agar plates. The colony count reflected the antibacterial effect of laser radiation as a function of the layer thickness. RESULTS: A mean bacterial reduction of 74% was achieved even with a 500-microm thick slice. CONCLUSION: This investigation indicates that the diode laser radiation reduces the number of bacteria in deep layers of infected root canal wall dentin.

Animals↗

PubMed2000

The bactericidal effect of Ho:YAG laser irradiation within contaminated root dentinal samples.

OBJECTIVE: This in vitro study investigates the bactericidal effect of pulsed Ho:YAG laser irradiation in the depth of contaminated dentin specimens. BACKGROUND DATA: Previous studies have shown the effectiveness of laser irradiation in bacterial reduction of infected root canal. METHODS: Root dentin of bovine teeth were sliced longitudinally in 180 samples of 100 microm, 300 microm, and 500 microm thickness, sterilized, dried, and inoculated on one side, with 1 microL of Enterococcus faecalis suspension. The opposite side's were irradiated four times for 5 seconds each with Ho:YAG laser irradiation, a wavelength of 2.10 microm, using four different energy settings: 1 W/5 Hz; 1 W/10 Hz; 1.5 W/5 Hz, and 2.0 W/5 Hz through a 320-microm quartz fiber at an angle of approximately 5 degrees. In addition, two control groups were investigated, the first was inoculated and not submitted to any treatment, the second was inoculated and treated with NaOCl and H2O2. The remaining bacteria from each dentin sample in a transport media were removed by vibration, serially diluted, and plated out on culture dishes selective for Enterococcus faecalis. RESULTS: When compared with the untreated control group or even with the group treated with NaOCl plus H2O2, counting of colonies forming units (CFU) from the laser-treated samples revealed a high significant bacterial elimination with a maximum of 98.46% and a minimum of 83.65%. CONCLUSIONS: Our findings demonstrate a significant decrease of the bacterial population in depth, suggesting that the Ho:YAG laser irradiation could be effective to eliminate the microorganisms harbored within dentin or contaminated canals.

Animals↗

PubMed1999

A DNA damage repair mechanism is involved in the origin of chromosomal translocations t(4;11) in primary leukemic cells.

Some chromosomal translocations involved in the origin of leukemias and lymphomas are due to malfunctions of the recombinatorial machinery of immunoglobulin and T-cell receptor-genes. This mechanism has also been proposed for translocations t(4;11)(q21;q23), which are regularly associated with acute pro-B cell leukemias in early childhood. Here, reciprocal chromosomal breakpoints in primary biopsy material of fourteen t(4;11)-leukemia patients were analysed. In all cases, duplications, deletions and inversions of less than a few hundred nucleotides indicative of malfunctioning DNA repair mechanisms were observed. We concluded that these translocation events were initiated by several DNA strand breaks on both participating chromosomes and subsequent DNA repair by 'error-prone-repair' mechanisms, but not by the action of recombinases of the immune system.

Adolescent↗

PubMed1999

Multiplex PCR--a rapid screening method for detection of gene rearrangements in childhood acute lymphoblastic leukemia.

Chromosomal rearrangements in childhood acute lymphoblastic leukemia (ALL) play an important role in the identification of clinical relevant subgroups. For rapid and easy detection of the clinically most important gene rearrangements, a nested multiplex reverse transcriptase polymerase chain reaction (multiplex PCR) was developed. This multiplex PCR enables the detection of M-BCR/ABL, m-BCR/ABL, TEL/ AML1, and MLL/AF4 fusion transcripts in one PCR reaction. However, the existence of splicing variants and different breakpoints on the DNA level hampers the discrimination of the rearrangements by their fragment size on an agarose gel. Therefore, one of the internal primers of each translocation (ABL-2, TEL-2, AF4-2) was labeled with a characteristic fluorescent dye, and an automatic fluorescence-based DNA fragment analysis was performed. The sensitivity of this multiplex PCR is in the same range as that of the corresponding single PCR reaction and allows a fast screening for the detection of therapy-relevant rearrangements, with a high turnover of samples.

Base Sequence↗

PubMed1999

Biology and clinical significance of the TEL/AML1 rearrangement.

The accurate identification of chromosomal abnormalities in patients with leukemia is essential for diagnosis and treatment assignment. Recent technical improvements in the detection of such aberrations have demonstrated that the previously unrecognized chromosomal translocation t(12;21) is the most prevalent structural aberration in childhood acute lymphoblastic leukemia. Both genes involved, translocation ets like gene (or ETV6) on chromosome 12 and acute myeloid leukemia 1 gene (or CBF alpha) on chromosome 21 had been identified for several years previously, which facilitated the rapid development of molecular diagnostic assays and their implementation in therapy trials. Although first described less than four years ago, the TEL/AML1 story is an excellent example of how close collaboration between physicians and molecular biologists is mandatory for achieving general insights into the molecular pathogenesis of leukemia and for further improvements in diagnosis and in monitoring response to chemotherapy.

Child↗

PubMed1999

Flow cytometry to monitor phagocytosis and oxidative burst of anaerobic periodontopathogenic bacteria by human polymorphonuclear leukocytes.

The reduced susceptibility to phagocytosis found among some periodontopathogenic anaerobes may account for the differences between invasive and non-invasive strains. We applied flow cytometry as a powerful tool to analyze and quantify phagocytosis using standardized cultures of oral anaerobes (Porphyromonas gingivalis, Prevotella intermedia, P. nigrescens, Capnocytophaga gingivalis, C. ochracea, C. sputigena, Fusobacterium nucleatum and Peptostreptococcus micros) and heparinized whole blood. Bacteria were labeled by a fluorescein-methylester and their esterase activity, resulting in green fluorescence. Ingested bacteria could be detected easily and quantified by a shift towards green fluorescence in the PMNL population involved and a concomitant decrease in the bacterial population. Furthermore, the oxidative burst of PMNLs was detected in parallel assays using the dye DHR123 which becomes fluorescent upon oxidation during the oxidative burst process. We found a great diversity in phagocytosis susceptibility determined by estimating the portion of phagocytosing PMNLs, ranging from 10.6% (strain W83) to > 99.4% (e.g. ATCC 33277T) in P. gingivalis and from 15.9% (strain MH5) to > 95% (ATCC 33563T) in P. nigrescens. In contrast, almost all P. intermedia strains as well as the representatives of the other anaerobic, putative periodontopathic species tested showed no or only moderate resistance in the phagocytosis assay. Comparison of clinical data of patients and the extent of phagocytosis resistance of the corresponding P. gingivalis strains suggests that this virulence factor may contribute to the clinical outcome.

Bacteria, Anaerobic↗

PubMed1999

Simultaneous detection of Bacteroides forsythus and Prevotella intermedia by 16S rRNA gene-directed multiplex PCR.

In a 16S rRNA gene-directed multiplex PCR, Prevotella intermedia- and Bacteroides forsythus-specific reverse primers were combined with a single conserved forward primer. A 660-bp fragment and an 840-bp fragment that were specific for both species could be amplified simultaneously. A total of 152 clinical samples, subgingival plaque and swabs of three different oral mucosae, from 38 periodontitis patients were used for the evaluation.

Adult↗