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PubMed1992

Changes in the mandibular rest position after removal of remaining teeth and insertion of complete dentures.

A 24-month study was conducted to determine changes in the mandibular rest position after removal of the remaining teeth and insertion of complete dentures. A total of 24 patients with at least 10 teeth acting as occlusal stops were included in this study. Serial lateral cephalometric radiographs were made for each patient on four occasions (1) before extraction (2) 7 to 10 days after the initial denture insertion (3) 12 months later, and (4) 24 months later. Two angular measurements and one linear measurement were made for each radiograph and compared. A statistically significant increase in the face rest height with a tendency for posterior mandibular rotation was recorded between stages 1 and 2. This initial posterior rotation was followed by a gradual forward and upward movement, which finally brought the mandibular rest position even below the preextraction levels. The findings confirm the concept of the variability of posture and suggest that the mandibular rest position is an unreliable means of reestablishing the vertical dimension of occlusion that existed before extraction of the natural teeth.

Cephalometry↗
PubMed1987

Intramural distribution of Met5-enkephalin-Arg6-Gly7-Leu8 in sphincter regions of the human gut.

The intramural distribution of Met5-enkephalin-Arg6-Gly7-Leu8 (MERGL) was studied in the oesophago-cardiac, pyloric, ileo-caecal and sigmoid-recto-anal regions of the human digestive tract. Serial samples encompassing each area were separated into mucosa, submucosa and muscularis externa and extracted for radioimmunoassay. Comparatively low levels of MERGL immunoreactivity were measured throughout the cardiac junction. Conversely, a remarkable peak of MERGL concentration was detected at the pyloric junction, in both submucosa and muscularis. A progressive decrease in tissue levels of the same peptide, most evident in the submucosa, was detected on the proximal side of the ileo-caecal region. In the distal sigmoid colon and rectum MERGL concentrations showed a rapid decline, down to very low levels in the anal canal. The results may suggest the involvement of an enkephalinergic mechanism in the control of the human pylorus.

Cardia↗
PubMed2005

Investigation of the pharmacokinetics and determination of tramadol in rabbit plasma by a high-performance liquid chromatography-diode array detector method using liquid-liquid extraction.

An HPLC system using a new, simple and rapid liquid-liquid extraction and high-performance liquid chromatography-diode array detector method (HPLC-DAD) detection was validated to determine tramadol concentration in rabbit plasma. The method described was applied to a pharmacokinetic study of intravenous tramadol injections in rabbits. The extraction with ethylacetate yielded good response. The recovery of tramadol from plasma averaged 90.40%. Serial plasma samples were obtained prior to, during and after completion of the infusion for determination of tramadol concentrations. Tramadol concentrations were measured using reverse-phase high-performance liquid chromatography and pharmacokinetic application with intravenous tramadol in rabbits revealed that tramadol followed one-compartment open model. Maximum plasma concentration (C(max)) and area under the plasma concentration-time curve (AUC) for tramadol were 14.3 microg mL(-1) and 42.2 microg h mL(-1), respectively. The method developed was successfully applied to a simple, rapid, specific, sensitive and accurate HPLC method for investigation of the pharmacokinetics of tramadol in rabbit plasma.

Analgesics, Opioid↗
PubMed2002

Dexamethasone-mediated androgen metabolism in human gingival and oral periosteal fibroblasts.

Dexamethasone modulates the effects of other hormones and mediates cell function; the periodontium is a target tissue for androgens. It was therefore relevant to investigate the modulation of androgen metabolism by dexamethasone in cultured human gingival (HGF) and oral periosteal fibroblasts (HPF). Each cell line was incubated in Eagle minimum essential medium with [(14)C]testosterone/[(14)C]4-androstenedione as substrates and serial concentrations of dexamethasone (0.5-50 microg/ml), for 24h; the medium was solvent-extracted, analyzed and quantified for steroid metabolites. In response to dexamethasone, both HGF (n=6) and HPF (n=4) showed up to two-fold increases in the formation of 5alpha-dihydrotestosterone and 4-androstenedione (P<0.01, one-way ANOVA), and 3.6- to 5-fold increases in the formation of testosterone (P<0.001), from [(14)C]4-androstenedione, with some inhibition at higher concentrations. Dexamethasone stimulated the formation of physiologically active androgen metabolites in a dose-dependent manner. These metabolites might therefore contribute to dichotomous effects in connective tissues of the periodontium, dependent on effective concentrations of dexamethasone.

Adult↗
PubMed1978

Isolation of an infectious agent from bone-marrows of patients with multiple sclerosis.

Bone-marrow aspirates from 5 patients with recent episodes of multiple sclerosis (M.S.) contained increased numbers of plasma cells, and aspirates from 3 of these patients contained atypical reticulum cells. When bone-marrow from 4 of the patients was inoculated into cell cultures a cytopathic effect (C.P.E.) was observed, and this effect could be serially passaged to further cultures. The C.P.E. was not observed when ether-treated extracts of cells showing C.P.E. were used for passage or when the passage material was filtered through a 100 nm filter. Appearance of C.P.E. was delayed in cultures inoculated with 220 nm filtrates of inoculum was passed through filters of greater pore size. No C.P.E. was observed after culture of bone-marrow aspirated from each of 7 patients under clinical investigation for disorders other than M.S.

Adult↗
PubMed1999

Human pulp response after an adhesive system application in deep cavities.

OBJECTIVES: To evaluate the pulpo-dentin complex response to a dentin adhesive application in deep cavities performed in human teeth. METHODS: Deep class V cavities were prepared on the buccal surface of 46 premolars. The remaining dentin of the axial wall received 10% phosphoric acid and dentin adhesive (group DA), or was protected before the acid and dentin adhesive application with calcium hydroxide cement (group CH). Half of the teeth, which received the acid application directly over the axial wall, were contaminated prior to the procedures with dental plaque collected from the patient's own teeth (group DAC). The plaque was placed on the dentin for 5 min and then the cavity was washed. All teeth were restored with a light-cured composite resin. The teeth were extracted after 7, 30 or 60 days and prepared according to normal histologic techniques. Serial sections were stained with H/E, Masson's trichrome and Brown & Brenn technique for demonstration of bacteria. RESULTS: The histopathologic evaluation showed that in groups DA and DAC, the inflammatory response was more evident than in group CH. Also, the intensity of the pulp reaction increased as the remaining dentin thickness decreased. There was no statistical difference in the inflammatory response between the groups DA and DAC. CONCLUSION: Based on the experimental conditions, we concluded that the All Bond 2 adhesive system, when applied on dentin in deep cavities, showed an acceptable biocompatibility. However, the intensity of the pulpo-dentin complex response depends on the remaining dentin thickness.

Acid Etching, Dental↗
PubMed2003

Morphological analysis and chemical content of natural dentin carious lesion zones.

Dentin is one of the earliest bio-mineralization products to appear in the evolution of vertebrates. Dentin reactions to infection mimic earlier phylogenetic patterns, and carious lesions are divided into different zones which reflect the natural patho-morphological reaction of dentin to the carious attack. It was the aim of this study to investigate deep dentin carious lesions of human molars with combined polarization light microscopy, scanning electron microscopy and energy dispersive X-ray element analysis (EDX) for the determination of different zones of the carious lesions, their extent and the chemical content. Sixteen extracted teeth with deep dentin carious lesions were embedded in Technovit 9100 (Kulzer) and serial sections of 80 microm thickness were made. These sections were then examined with polarized light microscopy to identify the different zones of the lesions. The outlines of the zones were traced consecutively and 3D-reconstructions were made for the determination of the extent and calculation of the volumes of the different zones. From the volumes of the demineralizing dentin and the translucent zone a Dentin Demineralization Index (DDI) was calculated. Three sections of each lesion were then coated with carbon and studied with a scanning electron microscope. 3D-reconstruction of the teeth showed the rather stable translucent zone, interrupted by remnants of dead tracts, and very different volumes of demineralizing dentin. Therefore, with increasing size of the demineralizing dentin, the DDI increased. The chemical content was measured using energy dispersive X-ray analysis (EDX) in areas of intertubular dentin. The content of Ca, P, and C was significantly different in all zones. The Ca/P ratio was significantly different between sound dentin and demineralizing dentin. From the results we conclude that the mineral content of intertubular dentin of the translucent zone and demineralizing dentin is different from that of sound dentin, and the unique mineralization pattern of the translucent zone is a biological reaction to the carious attack. Because active dentin lesions exhibit many non-occluded open dentin tubules, further bacterial invasion or, in case of dentin treatment, the penetration of bonding agents towards the pulp is morphologically not prevented and therefore of clinical importance.

Dental Caries↗
PubMed1999

Purification and characterization of a novel cyclohexylamine oxidase from the cyclohexylamine-degrading Brevibacterium oxydans IH-35A.

Cyclohexylamine oxidase (CHAO) from a cell extract of Brevibacterium grown on cyclohexylamine was purified 50.2-fold, to electrophoretic homogeneity, by serial chromatographies. The molecular mass of the native enzyme was estimated to be approximately 50 kDa by gel filtration and SDS-PAGE. The optimum pH was 7.4 and the stable pH range was 6.0 to 7.0. The enzyme was thermostable up to 30 degrees C. The enzyme was found to be highly specific for the deamination of alicyclic monoamines such as cyclopentylamine, cycloheptylamine, and N-methylcyclohexylamine and aliphatic monoamines, such as sec-butylamine. The apparent K(m) value for cyclohexylamine was 1.23 mM. The enzyme was inhibited by flavin enzyme inhibitors such as quinine and quinacrine. The N-terminal 27 amino acid residues were determined as Gly-Ser-Val-Thr-Pro-Asp-Pro-Asp-Val-Asp-Val-Ile-Ile-His-Gly-Ala-Gly-Ile-Ser-Gly-Ser-Ala-Ala-Ala-Lys-Ala-Leu-, revealing homology to conventional flavin-containing amine oxidases (EC 1.4.3.4).

Journal Article↗
PubMed2003

Recombinant phospholipase Czeta has high Ca2+ sensitivity and induces Ca2+ oscillations in mouse eggs.

Sperm-specific phospholipase Czeta (PLCzeta) is known to induce intracellular Ca(2+) oscillations and subsequent early embryonic development when expressed in mouse eggs by injection of RNA encoding PLCzeta (Saunders, C. M., Larman, M. G., Parrington, J., Cox, L. J., Royse, J., Blayney, L. M., Swann, K., and Lai, F. A. (2002) Development 129, 3533-3544). The present study addressed characteristics of purified mouse PLCzeta protein that was synthesized using the baculovirus/Sf9 cell expression system. Microinjection of recombinant PLCzeta protein into mouse eggs induced serial Ca(2+) spikes quite similar to those produced by the injection of sperm extract, probably because of repetitive Ca(2+) release from the endoplasmic reticulum caused by continuously produced inositol 1,4,5-trisphosphate. Recombinant PLCdelta1 also induced Ca(2+) oscillations, but a 20-fold higher concentration was required compared with PLCzeta. In the enzymatic assay of phosphatidylinositol 4,5-bisphosphate hydrolyzing activity in vitro at various calcium ion concentrations ([Ca(2+)]), PLCzeta exhibited a significant activity at [Ca(2+)] as low as 10 nm and had 70% maximal activity at 100 nm [Ca(2+)] that is usually the basal intracellular calcium ion concentration level of cells. On the other hand, the activity of PLCdelta1 increased at a [Ca(2+)] between 1 and 30 microm. EC(50) was 52 nm for PLCzeta and 5.7 microm for PLCdelta1. Thus, PLCzeta has an approximately 100-fold higher Ca(2+) sensitivity than PLCdelta1. The ability of purified PLCzeta protein to induce Ca(2+) oscillations qualifies PLCzeta as a proper candidate of the mammalian egg-activating sperm factor. Furthermore, such a high Ca(2+) sensitivity of PLC activity as PLCzeta that can be active in cells at the resting state is thought to be an appropriate characteristic of the sperm factor, which is introduced into the ooplasm upon sperm-egg fusion, triggers Ca(2+) release first, and maintains Ca(2+) oscillations.

Animals↗
PubMed2000

A miniature integrated device for automated multistep genetic assays.

A highly integrated monolithic device was developed that automatically carries out a complex series of molecular processes on multiple samples. The device is capable of extracting and concentrating nucleic acids from milliliter aqueous samples and performing microliter chemical amplification, serial enzymatic reactions, metering, mixing and nucleic acid hybridization. The device, which is smaller than a credit card, can manipulate over 10 reagents in more than 60 sequential operations and was tested for the detection of mutations in a 1.6 kb region of the HIV genome from serum samples containing as few as 500 copies of the RNA. The elements in this device are readily linked into complex, flexible and highly parallel analysis networks for high throughput sample preparation or, conversely, for low cost portable DNA analysis instruments in point-of-care medical diagnostics, environmental testing and defensive biological agent detection.

DNA↗
PubMed1975

The distribution of placental lactogen in the human term placenta.

Serial frozen sections of human term placentae were examined by quantitative morphology and, after extraction, by a radioimmunoassay for human placental lactogen (HPL). The total quantity of HPL in the placenta was estimated as 720 mg. The amount of HPL was related to the amount of trophoblast in any area, but otherwise did not vary in different parts of the placenta. A semi-logarithmic relationship was observed between the amount of HPL and the percentage trophoblast in a section. It is suggested that the amount of HPL in a given area of the placenta is related to the blood-flow in that area.

Cell Count↗
PubMedDate not supplied

A new system for gastric emptying analysis using impedance measurement.

A tetra-polar electrical impedance device has been constructed to permit study of gastric emptying using a microcomputer. An alternative current of 100 kHz, 4 mM peak-to-peak, is injected through a pair of surface electrodes on the upper abdomen. The voltage variations picked up across the second pair of electrodes correspond to the variations of epigastric impedance in response to the applied current. The low frequency voltage corresponding to gastric emptying and recording noise is then conditioned and digitised at a sampling frequency of 1 Hz. A parallel 8-bit signal is finally converted to standard serial form and sent, in real time, to a microcomputer via a serial port. In a process of off-line analysis, the emptying trace was extracted from measured traces by taking successive Fast Fourier Transform (FFT) of length 64 points and was then fit by three mathematical models: linear, exponential and Weibull. The rate of emptying was calculated in terms of the time needed to achieve 50% emptying (T 1/2) from the best fit model. A clinical experiment was done in 20 healthy volunteers to investigate the reproducibility of the method and compared to a scintigraphic method. Successive measurements on the same subject gave statistically similar results and were statistically independent. No correlation has been observed between impedance and scintigraphic methods.

Adult↗
PubMed1991

[The study of chemical mediators in the patients with allergic rhinitis 2). Histamine, leukotriene and kinins in the nasal secretion during dual phase response].

In order to clarify the details of a late phase response (LPR) in allergic rhinitis, serial changes of nasal symptoms and concentration of chemical mediators (histamine, leukotriene (LT) C4 and kinins) in nasal lavage fluid were investigated after the nasal antigen challenge in ten patients with perennial allergic rhinitis with a positive intradermal skin reaction to mite extract. Samples were collected periodically for 8 hours following the challenge. Nasal airway resistance (NAR) and clinical symptoms were also recorded serially. Three out of ten patients showed only early phase response (EPR) and the other seven patients showed both early and late phase response after the challenge. The chief nasal symptom in LPR was nasal obstruction. In EPR, concentrations of chemical mediators in the nasal lavage fluid always increased. In dual (early and late) phase responders, increases of chemical mediators in nasal lavage fluids were notable during both early and late phase responses. In EPR, there was a significant correlation between severity of sneezing attack and concentration of histamine. In LPR, increase of LTC4 level was significantly correlated with the increase of NAR. RAST score and daily nasal symptoms tended to be higher and more sever in dual phase responders compared with early phase responders. We suggest that the amounts of released chemical mediators during allergic reaction denoted the LPR in allergic rhinitis patients.

Adult↗
PubMedDate not supplied

Relationship between V3 genotype, biologic phenotype, tropism, and coreceptor use for primary isolates of human immunodeficiency virus type 1.

OBJECTIVES: The predictive value of positively charged amino acids at positions 11 and 25 within the V3 loop region of the human immunodeficiency virus type 1 (HIV-1) envelope gene for the syncytium-inducing (SI) phenotype was assessed. STUDY DESIGN/METHODS: Sequencing was performed on DNA extracted from primary peripheral blood mononuclear cells (PBMCs) and complementary DNA (cDNA) prepared from serial viral isolates from 10 HIV-1-seropositive subjects. Proviral DNA sequencing was also performed on biologic clones from most of these subjects. RESULTS: Positive charge at position 11 and/or 25 in 257 isolate cDNA, PBMC DNA, and biologic clone PBMC DNA sequences was compared with 69 phenotypic determinations, of which 62.3% were SI. V3 genotype was 51.2% sensitive and 85.8% specific for the SI phenotype, with positive and negative predictive values of 62.8% and 79.0%, respectively. Cellular tropism failed to correlate with V3 genotype, coreceptor use, or biologic phenotype. Exclusive use of CCR5 was associated with the nonsyncytium-inducing (NSI) phenotype. Overall, V3 loop charge was higher in SI than in NSI isolates (5.01 and 3.78, respectively; p = 0.0211). CONCLUSIONS: The predictive power of SI phenotype from V3 genotype is relatively weak, especially in a low SI prevalence population. The direct measurement of viral phenotype, cellular tropism, and coreceptor use in HIV-1 isolates is essential for accurate biologic characterization.

Amino Acid Sequence↗
PubMed2005

Evaluation of Western blotting methods using samples with or without sodium phosphotungstic acid precipitation for diagnosis of scrapie and chronic wasting disease.

The purpose of this study was to enhance the sensitivity of the Western blot (WB) test for use as an alternative and confirmatory method for the diagnosis of scrapie and chronic wasting disease (CWD) in Canada by comparing 2 sample preparation procedures: an abnormal prion protein (PrPSc) concentration procedure using sodium phosphotungstic acid (PTA) precipitation and a procedure using crude sample without precipitation. A total of 100 cerebrum samples (52 sheep and 48 elk), including 66 negative (31 sheep, 35 elk) and 34 positive (21 scrapie and 13 CWD positive) samples diagnosed by using immunohistochemistry (IHC) on retropharyngeal lymph node (RPLN) and medulla oblongata at obex, were tested by using WB with the 2 sample preparation procedures. The WB using non-PTA enriched sample (crude extract) detected, on average, only 71.7% (9 of 15, 60.0% for scrapie, 5 of 6, 83.3% for CWD) of the samples that tested positive by using WB with PTA enriched samples. No case was positive by WB using crude extract but negative by WB using PTA enriched sample. No false positive was found. Serial dilution of PTA precipitated samples demonstrated that the technique increases the detection limit approximately 100 fold. Additionally, the comparison of the WB and IHC on cerebrum from all the positive cases demonstrated that WB following PTA precipitation and IHC had 100% agreement by detecting 6 positive for CWD on cerebrum; while IHC detected scrapie in only 14 out of 15 positive cerebrum samples by using WB following PTA precipitation. Phosphotungstic acid precipitation is therefore a useful adjunct to WB analysis of scrapie and CWD and tissues.

Animals↗
PubMed1990

Methodology of ECG interpretation in the Lyon program.

To provide an in-depth interpretation of the spatial QRS-T contour, we established a set of vectorcardiographic parameters computed octant by octant. For each statement the criteria constitute a model which is closely related to pathophysiological patterns. The diagnostic strategy is of the heuristic type. For each diagnosis a table lists several criteria providing both specific and differential diagnostic information. Inter-table competition is handled by specific logic. The program classifies diagnoses according to the number of non-satisfied criteria. Three versions are available including 124, 51 and 7 diagnoses, respectively. The first, intended for interpretation of complex situations, has given satisfactory results in such situations. Rhythm analysis is performed from 8-second recordings. All functions--acquisition, quality control, choice of a dominant complex, wave recognition, parameter extraction, interpretation, editing, storage--are integrated into a mobile cart. A complementary program performs serial analysis of successive tracings. It highlights with great precision morphological changes in the spatial loops and interval variations.

Diagnosis, Computer-Assisted↗
PubMed1988

Changes in expression of oncogenes and endogenous retroviral-like sequences during colon carcinogenesis.

The possible roles in experimental colon carcinogenesis of two protooncogenes (c-myc and c-H-ras), two endogenous retrovirus-related DNA sequences [rat leukemia virus (RaLV) and the 30S sequence], and two cell cycle related genes (beta-actin and ornithine decarboxylase) were studied by analyzing the levels of their corresponding RNAs during the course of azoxymethane induced and high fat promoted colon carcinogenesis. F-344 male rats received three s.c. injections of azoxymethane (15 mg/kg) or normal saline and were then subdivided into high or low fat diet groups. During subsequent serial sacrifices normal colon mucosa, adenomas, and carcinomas were harvested for histology and RNA extraction. Seventy-one RNA samples were analyzed by the Northern blot hybridization procedure using the appropriate 32P-labeled DNA probes. A marked increase in the abundance of c-myc, RaLV, and 30S RNAs were seen in all of the colon tumors, including adenomas and invasive carcinomas. No or a very low level of expression of RaLV and c-myc RNA was found in the flat grossly normal mucosa adjacent to the tumors and in the mucosa of the control rats. Some of the colon tumors also displayed increased levels of c-H-ras, ornithine decarboxylase and beta-actin RNAs but these findings were less striking and more variable than those seen with c-myc, RaLV, and 30S RNAs. These results suggest that increased expression of the c-myc protooncogene and of the endogenous retrovirus-like sequences (RaLV) and 30S are hallmarks of colon carcinogenesis in this model system.

Animals↗
PubMedDate not supplied

Sensitivity of cultured human and mouse fibroblasts to trichothecenes.

Cultured human diploid fibroblasts and a cloned permanent line of Swiss mouse fibroblasts (3T3) have been adapted to a biological assay of trichothecenes and other toxic fungal metabolites both in purified form and as crude extracts. The assays are carried out in multi-well tissue culture trays with serial dilutions of toxins in 200 microL volumes of medium. Cells are cultured with the toxins for 3-4 days, fixed, and stained, providing a permanent record of results. The sensitivity of the assay ranges from 0.1 ng/mL (20 pg/assay) for T-2 toxin on mouse fibroblasts to no detectable toxicity at 5 micrograms/mL for cis-zearalenone and verrucarol. Human skin fibroblasts exhibit patterns of relative sensitivities similar to that observed with mouse fibroblasts, but the absolute sensitivities are much greater with mouse fibroblasts because they can be cultured at much lower cell densities.

Animals↗