PubMed Health⌕ Search

Biomedical subjects

M Latta

Publications and source records attributed to M Latta.

16 recordsLinked to original sources
PubMed2006

Effect of exponential polymerization on color stability of resin-based filling materials.

PURPOSE: To investigate the influence of a halogen light curing device used with constant or exponential polymerization mode on the color stability of contemporary resin-based filling materials. MATERIALS AND METHODS: Eight samples of Charisma (CH), Durafill (DU), Definite (DE), and Dyract AP (DY) each were light-cured with constant power or with soft-start mode (Translux Energy) for 20, 40 or 60s. The CIE-Lab-values (L(*), a(*), b(*)) were measured prior to and after performing dry aging, water aging or a Suntest (EN ISO 7491) and Delta L-, Delta a-, Delta b-, and Delta E-values were calculated. RESULTS: Statistical analysis (GLM and repetition of measures) showed significant changes (p<0.05) of the color values for each material's curing mode and time after each of the aging processes. Four-way ANOVA revealed significant differences (p<0.05) between Delta L-, Delta a-, Delta b-, and Delta E of the materials in dependence on the curing time and mode and aging condition. Exponentially-cured DU was the most color-unstable material after aging in water followed by the 20s exponentially cured DE and CH samples. After the Suntest, DY showed significant bleaching (negative Delta b) and the largest Delta E for all curing times and modes followed by the DE samples. DU and CH were the most color-stable materials in this test. SIGNIFICANCE: The extent of discoloration depends on the (a) curing time, (b) curing mode, (c) aging condition, and (d) material. For the constant curing mode, 40s curing time for the exponential 60s seems to be appropriate.

Analysis of Variance↗
PubMed2006

Spark erosion as a metal-resin bonding system.

PURPOSE: Resin to metal bond strength of two commercial and one experimental metal-resin bonding system for crown and bridge veneer composites was evaluated. MATERIALS AND METHODS: All specimens were prepared and tested according to ISO 10477 Amendment 1. A high precious alloy (HPA), a Co/Cr-alloy (Co/Cr) and pure titanium (Ti) were wet-ground to a final surface finish of 800 grit and air-dried. Twenty specimens of each metal were treated with Rocatec (RO), 20 with a Silano-Pen (SP), and 20 with the experimental spark-erosion (SE) bonding system. A light-curing opaque and a crown and bridge veneer composite were applied to each metal specimen. Ten specimens of each metal and bonding system were stored in water for 24h at 37 degrees C, 10 were thermocycled (TC) 5000 times in a water-bath between +5 and +55 degrees C prior to measuring shear bond strength (SBS). Scanning electron microscopy (SEM) was used to examine the treated metal surfaces. RESULTS: After 24h SE generated significantly higher SBS values on Ti than RO (p<0.05). SP revealed the highest SBS for Co/Cr. After TC significant decreases occurred for RO on HPA, for SP on Co/Cr and Ti, and for SP on Ti. SE revealed the highest mean bond values for all metals. These results and SEM proved that no sandblasting is required for SE to obtain good bond strength. SIGNIFICANCE: The experimental spark-erosion bonding system is an easy and very effective method for surface-treating alloys to obtain high SBS values for crown and bridge veneer composites.

Chromium Alloys↗
PubMed2006

The effects of thermocycling on the flexural strength and flexural modulus of modern resin-based filling materials.

OBJECTIVE: Evaluation of flexural strengths and flexural moduli of hybrids, packables, ormocers, compomers and flowables prior to, and after, thermocycling. MATERIALS AND METHODS: Twenty specimens, size (25 +/- 2) mm x (2 +/- 0.1) mm x (2+/-0.1) mm, of Herculite (HE), Point 4 (P4), TetricCeram (TC), Miris (MI), TetricCeram HB, (HB), Solitaire 2 (SO), Surefil A (SU), Definte (DE), Admira (AD), Dyract AP (DY), Compoglass (CO) and TetricFlow (TF) were made according to ISO 4049. A three-point-bending test was carried out for the first 10 specimens after 24 h water storage at 37 degrees C and for the second 10 specimens after 30-day water storage at 37 degrees C, followed by 5000 thermocycles between +5 and +55 degrees C. For each test series flexural strengths and flexural moduli were calculated. Statistical significance was p < 0.05. RESULTS: Flexural strengths of DY and CO missed the 80 MPa limit of ISO 4049 for occlusal fillings prior to and after thermocycling. AD missed it after thermocycling. HE, P4, TC, MI, HB, SO, SU, DE and TF did not significantly differ prior to and after thermocycling. Flexural strength of SO and AD significantly decreased after thermocycling. No change of the flexural moduli could be observed for any of the test materials. Only SU exceeded 10,000 MPa prior to and after thermocycling. SIGNIFICANCE: Only DY and CO missed the flexural strength limit of ISO 4049 for occlusal fillings. Only SU exceeded a flexural modulus of 10,000 MPa prior to and after thermocycling.

Analysis of Variance↗
PubMed2004

Comparison of total nucleated cell measurements of UC blood samples using two hematology analyzers.

BACKGROUND: The total nucleated cell (TNC) content of umbilical cord blood (UCB) units currently serves as the most important measure for determining suitability for transplantation. Hence it is important that TNC measurements are performed in an accurate manner. TNC content is evaluated routinely by hematology analyzers (HA) as WBC counts. The objective of the study was to compare TNC content utilizing two different HA, one utilizing an impedance channel and optical channel, and the other using only an optical channel. METHODS: The HA utilized in this study used two different modes of operation for lysis, regular mode (RM) and extended lysis mode (ELM). Cell-Dyn 3200 (CD3.2) utilizes optical technology for WBC measurements, involving WBC optical count (WOC) and nuclear optical count (NOC), whereas the Cell-Dyn 3700 (CD3.7) utilizes both the impedance (WIC) and optical technology (WOC) for WBC measurements. TNC content was determined with 17 identical samples using CD3.2 in one laboratory and CD3.7 in the other laboratory. Cord blood samples processed to concentrate nucleated cells by either of the laboratories were sent by overnight courier and assays were performed on the same day by both laboratories. RESULTS: For CD3.7, the WOC values were consistently lower than the WIC using the regular mode, but showed no significant differences (P>0.05). The WIC and WOC values were comparable on using the ELM and RM. For CD3.2, WOC values using RM and NOC values using ELM showed no significant differences (P>0.05), even though the WOC measurement was lower than the NOC values for most samples. The best comparison of TNC measurement between the two HA could be achieved by comparing CD3.7-WIC with CD3.2-NOC values. The results were equivalent (P>0.05) and 12 of 17 samples had equal to or less than 10% difference (mean 9.5%). DISCUSSION: TNC measurements of UCB samples were essentially identical using the WIC channel of the Cell-Dyn 3700 and the NOC channel of the Cell-Dyn 3200.

Erythrocyte Count↗
PubMed2000

Metabolic depletion of ATP by fructose inversely controls CD95- and tumor necrosis factor receptor 1-mediated hepatic apoptosis.

Hepatocyte apoptosis is crucial in several forms of liver disease. Here, we examined in different models of murine liver injury whether and how metabolically induced alterations of hepatocyte ATP levels control receptor-mediated apoptosis. ATP was depleted either in primary hepatocytes or in vivo by various phosphate-trapping carbohydrates such as fructose. After the activation of the tumor necrosis factor (TNF) receptor or CD95, the extent of hepatocyte apoptosis and liver damage was quantified. TNF-induced cell death was completely blocked in ATP-depleted hepatocyte cultures, whereas apoptosis mediated by CD95 was enhanced. Similarly, acute TNF-induced liver injury in mice was entirely inhibited by ATP depletion with ketohexoses, whereas CD95-mediated hepatotoxicity was enhanced. ATP depletion prevented mitochondrial cytochrome c release, loss of mitochondrial membrane potential, activation of type II caspases, DNA fragmentation, and cell lysis after exposure to TNF. The extent of apoptosis inhibition correlated with the severity of ATP depletion, and TNF-induced apoptosis was restored when ATP was repleted by increasing the extracellular phosphate concentration. Our study demonstrates that TNF-induced hepatic apoptosis can be selectively and reversibly blocked upstream of mitochondrial dysfunction by ketohexose-mediated ATP depletion.

Adenosine Triphosphate↗
PubMed1997

Recombinational construction in Escherichia coli of infectious adenoviral genomes.

A two-step gene replacement procedure was developed that generates infectious adenoviral genomes through homologous recombination in Escherichia coli. As a prerequisite, a human adenovirus serotype 5 (Ad5)-derived genome was first introduced as a PacI restriction fragment into an incP-derived replicon which, in contrast to ColE1-derivatives (e.g., pBR322 or pUC plasmids), is functional in a polA mutant of E. coli. Any modification can be introduced at will following two consecutive homologous recombinations between the incP/Ad5 replicon and the ColE1 plasmid. The overall procedure requires only the in vitro engineering of the ColE1-derivative by flanking the desired modification with small stretches of identical sequences. In the first step, a cointegrate between the tetracycline-resistant incP/Ad5 replicon and the kanamycin-resistant ColE1-derivative is selected by growing the polA host in the presence of both antibiotics. Resolution of this cointegrate is further selected in sucrose growth conditions due to the loss of a conditional suicide marker (the sacB gene of Bacillus subtilis) present in the ColE1 plasmid, leading to unmodified and modified incP/Ad5 replicons that can be differentiated upon restriction analysis. Consecutive rounds of this two-step cloning procedure allowed the introduction of multiple independent modifications within the virus genome, with no requirement for an intermediate virus. The potential of this procedure is demonstrated by the recovery of several E1E3E4-deleted adenoviruses following transfection of the corresponding E. coli-derived genomes in IGRP2 cells.

Adenovirus E1 Proteins↗
PubMed1997

Characterization of human apolipoprotein A-I expressed in Escherichia coli.

Human apolipoprotein A-I (apoA-I), with an additional N-terminal extension (Met-Arg-Gly-Ser-(His)6-Met) (His-apoA-I), has been produced in Escherichia coli with a final yield after purification of 10 mg protein/1 of culture medium. We have characterized the conformation and structural properties of His-apoA-I in lipid-free form, and in reconstituted lipoproteins containing two apoA-I per particle (Lp2A-I) by both immunochemical and physicochemical techniques. The lipid-free forms of the two proteins present very similar secondary structure and stability, and have also very similar kinetics of association with dimyristoyl phosphatidylcholine. His-apoA-I and native apoA-I can be complexed with 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC) to form similar, stable, either discoidal or spherical (sonicated) Lp2A-I particles. Lipid-bound native apoA-I and His-apoA-I showed very similar alpha-helical content (69% and 66%, respectively in discoidal Lp2A-I and 54% and 51%, respectively in spherical Lp2A-I). The conformation of His-apoA-I in lipid-free form and in discoidal or spherical Lp2A-I has also been shown to be similar to native apoA-I by immunochemical measurements using 13 monoclonal antibodies recognizing distinct apoA-I epitopes. In the free protein and in reconstituted Lp2A-I, the N-terminal has no effect on the affinity of any of the monoclonal antibodies and minimal effect on immunoreactivity values. Small differences in the exposure of some apoA-I epitopes are evident on discoidal particles, while no difference is apparent in the expression of any epitope of apoA-I on spherical Lp2A-I. The presence of the N-terminal extension also has no effect on the reaction of LCAT with the discoidal Lp2A-I or on the ability of complexes to promote cholesterol efflux from fibroblasts in culture. In conclusion, we show that His-apoA-I expressed in E. coli exhibits similar physicochemical properties to native apoA-I and is also identical to the native protein in its ability to interact with phospholipids and to promote cholesterol esterification and cellular cholesterol efflux.

Amino Acid Sequence↗
PubMed1996

Modulation of colonic aberrant crypt foci and proliferative indexes in colon and prostate glands of rats by vitamin E.

The effect of a high vitamin E diet on the early stages of colon carcinogenesis and on the proliferative indexes in the colon and in the prostate glands was investigated in rats. F344 male rats were injected with azoxymethane (AOM, 15 mg/kg sc). One week later, animals were randomly allocated into two dietary groups (n = 8 rats/group): normal vitamin E (50 IU/kg diet) and high vitamin E (200 IU/kg diet). The basal diet was the AIN-76 diet modified to contain high corn oil (23% wt/wt). After eight weeks of feeding, concentrations of vitamin E in plasma, liver, and prostate were analyzed. Enumeration of aberrant crypt foci (ACF) in colons and proliferative indexes of colons and prostate glands were determined. The total number of ACF and the average number of aberrant crypts (AC) per focus were similar in both dietary groups. ACF were classified as small (1-3 crypts/focus), medium (4-6 crypts/focus), or large (> or = 7 crypts/focus). Only the ACF in the small category showed a significant treatment effect, with values being lower in the high vitamin E group than in the control group (p < or = 0.05). No significant difference was observed in colonic proliferative indexes assessed by enumeration of metaphase cells, S phase cells, or cells exhibiting proliferative cell nuclear antigen (PCNA). The PCNA labeling index in the prostate glands and the activity of prostatic acid phosphatase in plasma were higher in high vitamin E-fed rats (p < or = 0.05) than in control animals. The present study demonstrates that additional vitamin E does not inhibit the induction and growth of ACF; also it enhances the proliferative status of the prostate glands.

Acid Phosphatase↗
PubMed1994

Identification of specific amphipathic alpha-helical sequence of human apolipoprotein A-IV involved in lecithin:cholesterol acyltransferase activation.

To investigate the structure-function relationship of human apolipoprotein A-IV (apoA-IV), several deletion mutants of this protein were constructed by sequentially removing pairs of 22-residue repeats, potentially having an amphipathic alpha-helical conformation. The mutants, produced as recombinant poly-histidine-tagged apolipoproteins (t-apo) in Escherichia coli, assembled with phosphatidylcholine (i.e. dimyristoylphosphatidylcholine, palmitoyloleoylphosphatidylcholine, or egg lecithin) as did native apoA-IV. Lecithin:cholesterol acyltransferase (LCAT) cofactor function, measured as cholesterol esterification occurring when t-apo-phosphatidylcholine-cholesterol complexes were incubated with purified enzyme, decreased significantly when pairs of repeats between residues 117 and 248 were deleted and most markedly when residues 117-160 were deleted. LCAT cofactor activity decreased by 90 and 75%, respectively, when egg lecithin or palmitoyloleoylphosphatidylcholine was used to form the particles with the delta aa 117-160 mutant. Thus, on the basis of deletion scanning of t-apo, residues 117-160 seem to be involved in the LCAT cofactor function of apoA-IV.

Amino Acid Sequence↗
PubMed1994

Purification and characterization of recombinant human apolipoprotein A-II expressed in Escherichia coli.

We have expressed recombinant human apolipoprotein A-II (apoA-II) in Escherichia coli, as a fusion protein with Schistosoma japonicum glutathione-S-transferase (GST). The GST-AII fusion protein was recovered by affinity chromatography using glutathione as a ligand. After thrombin cleavage and removal of the GST carrier, recombinant apoA-II was obtained in a highly purified form and was exclusively composed of dimeric apoA-II. Kinetics of association to dimyristoylglycerophosphocholine (Myr2GroPCho) vesicles showed that recombinant apoA-II exhibited the same pattern of association as human plasma apoA-II. Electron microscopic analysis of the complexes showed a typical pattern of rouleaux, characteristic of stacked discs, with a diameter similar to that determined by gradient-gel electrophoresis. Circular dichroism measurements showed that the alpha-helical content of both plasma and recombinant apoA-II increased similarly when the proteins associated with Myr2GroPCho vesicles, at the expense of a random-coil structure. Lipid-bound apoA-II consisted of 70-72% alpha helices, suggesting the presence of three 18-residue alpha helices/apoA-II monomer. Cross-linking experiments indicated that Myr2GroPCho complexes contained two molecules dimeric apoA-II/vesicle. Recombinant apoA-II was as efficient as plasma apoA-II in associating with HDL subclasses, and in displacing apoA-I from dipalmitoylglycerophosphocholine/cholesterol/apoA-I complexes, most likely due to its highly ordered secondary structure when associated with Myr2GroPCho vesicles. These findings demonstrate that recombinant apoA-II exhibits the same structural and functional properties as human plasma apoA-II. Thus, the expression system utilized is appropriate to produce mutagenized forms to further structure/function analysis.

Amino Acid Sequence↗
PubMed1991

Functional characterization of human recombinant apolipoprotein AIV produced in Escherichia coli.

Apolipoprotein AIV (apoAIV), a protein which is known to activate the enzyme lecithin: cholesterol acyltransferase, to bind to apoAI/AII receptor sites and also to promote cholesterol efflux from adipose cells, may play an important role in reverse cholesterol transport. In this report, the high-level production of soluble recombinant mature human apoAIV (isoform 1) in Escherichia coli is described. The recombinant protein was purified by avoiding lipid extraction or denaturation. The apoAIV preparation was analysed by its reactivity with antibodies raised against human apoAIV, SDS-gel electrophoresis, isoelectric focusing and N-terminal sequencing. The purified recombinant protein retains an extra methionine at the N-terminus. Purified recombinant and natural apoAIV proteins were indistinguishable with regard to their denaturation properties, thermo-stability or their fluorescence emission properties in the presence of various quantities of a quenching agent. Complexes of ApoAIV with L-alpha-dimyristoyl-glycerophosphocholine (Myr2GroPCho), glycerophosphocholine (GroPCho), or L-alpha-1-palmitoyl-2-oleoylglycerophosphocholine (PamOleGroPCho) prepared from plasmatic and from recombinant apoAIV proteins have similar densities as revealed by analytical centrifugation. They also share the same cofactor properties for the lecithin:cholesterol acyltransferase reaction. Recombinant apoAIV complex with Myr2GroPCho was also able to bind to the same apoAI/AII receptor sites and to promote cholesterol efflux to an equal extent from adipose cells. It is concluded that the recombinant protein is functionally identical to the plasmatic apoAIV and may therefore be very useful in helping to elucidate the physiological role of apoAIV.

Apolipoproteins↗
PubMed1991

Phosphorylation of human recombinant tyrosine hydroxylase isoforms 1 and 2: an additional phosphorylated residue in isoform 2, generated through alternative splicing.

The single human tyrosine hydroxylase (TH) gene generates four different mRNA species through alternative splicing events. TH-1 and TH-2 mRNAs are expressed mostly in the brain. We have produced large amounts of the corresponding proteins in Escherichia coli to analyze their respective molecular characteristics. The polypeptides have molecular weights similar to those of TH expressed in Xenopus oocytes and react with antibodies to TH. The two isoforms were purified with a purity of 90% using a three-step procedure. The phosphorylation sites have been determined in the two isoforms after labeling with [gamma-32P]ATP in the presence of cAMP-dependent protein kinase (PKA) or calmodulin-dependent protein kinase II (CaM-PK II). In both isoforms, Ser-40 was found to be phosphorylated by PKA, and Ser-19 and Ser-40 were found to be phosphorylated by CaM-PK II. The putative phosphorylation site generated by alternative splicing (Ser-31) was phosphorylated specifically by CaM-PK II in TH-2 only. The kinetic properties of the two isoforms in the presence of various concentrations of the substrate (tyrosine) and of the natural cofactor [6R)-tetrahydrobiopterin) were also analyzed. TH produced in E. coli is unphosphorylated but nevertheless active. At 50 microM tyrosine and 300 microM (6R)-tetrahydrobiopterin, the specific activities of TH-1 and TH-2 are 1300 and 620 nmol of dihydroxyphenylalanine/min/mg, respectively. Phosphorylation of TH-1 and TH-2 by PKA activates both isoenzymes as shown by the increase in the affinity for the cofactor. No changes in kinetic parameters of the isoenzymes were observed after phosphorylation by CaM-PK II. Dopamine was found to inhibit both TH isoenzymes to the same extent as shown by their similar Ki values for dopamine. These values were increased after phosphorylation of each enzyme by PKA. Unlike TH-1, phosphorylation of TH-2 by CaM-PK II resulted in an increase of the Ki value for dopamine. This property may be related to the presence of the additional phosphorylated residue in TH-2 isoform.

Base Sequence↗
PubMed1990

Tryptophan promoter derivatives on multicopy plasmids: a comparative analysis of expression potentials in Escherichia coli.

A collection of variant plasmids expressing either Escherichia coli galactokinase or human serum albumin under the control of several E. coli trp promoter derivatives were constructed and studied for both efficiency of expression and regulation by tryptophan. Several variables, including the length of the upstream region, tandem duplications of a core promoter, and the insertion of the trp repressor trpR gene onto the expression vector, were studied. It is shown that derivatives containing sequences upstream from the -35 region or multiple copies of the trp promoter produce twofold higher levels of protein than plasmids with a minimal trp promoter truncated at -40. We show that the expression of a heterologous protein such as albumin can be significantly improved (13% vs. 7% of total proteins) if both the upstream trp promoter region, which enhances promoter strength, and an intact trpR gene, are included on the plasmids.

Base Sequence↗
PubMed1989

Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1 beta.

Expression plasmids carrying the coding sequence of mature human interleukin 1 beta (IL 1 beta) linked either to a Met start codon, or fused to different efficient Escherichia coli secretion signal sequences, have been constructed. In the latter case, we used signal peptides derived either from an outer membrane protein (OmpA) or from a periplasmic protein (PhoA). The synthesis of IL1 beta from these fusions was investigated in an otherwise strictly isogenic context using identical conditions of derepression and culture media. The Met-IL1 beta fusion produced a soluble cytoplasmic protein which could be released from the cells by osmotic shock whereas the OmpA and PhoA fusions were always insoluble. The extent of sOmpA-IL1 beta maturation was found to vary from 50 to 100%, mainly depending on the medium used, whereas no significant maturation of the signal peptide could be detected in the case of the sPhoA-IL1 beta fusion. Immuno-electron microscopy revealed that the sOmpA-IL1 beta fusion was targeted to the inner membrane, whereas the sPhoA-IL1 beta fusion remained within the cytoplasm and thus did not appear to enter the secretion pathway. Amplifying the E. coli signal peptidase lep gene on a multicopy plasmid did not improve signal peptide removal from sOmpA-IL1 beta. Moreover, these E. coli secretion vectors allowed us to produce, in high levels, IL1 beta fragments which otherwise could not be stably accumulated within the cytoplasmic compartment.

Amino Acid Sequence↗
PubMed1985

Structure of a murine alpha interferon pseudogene with a repetitive R-type sequence in the 3' flanking region.

A murine alpha interferon pseudogene was identified in a mouse genomic library. The nucleotide sequence revealed several in-phase termination codons within the gene and repetitive oligonucleotides in the flanking regions. The nucleotide sequences and the amino acids of the peptide signal sequences were compared with known human alpha interferon genes and the pseudogene.

Amino Acid Sequence↗