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At least 1,603 records · Page 89Linked to original sources
PubMed1978

Cell-mediated immunity to measles, myelin basic protein, and central nervous system extract in multiple sclerosis. A longitudinal study employing direct buffy coat migration inhibition assays.

Cell-mediated immunity to myelin basic protein, to an extract of central nervous system white matter, and to measles virus nuclear core, was studied nine patients with multiple sclerosis in a serial longitudinal fashion using in vitro inhibition of buffy coat migration. The mean migration index to all antigens at various times before, during, and after exacerbation of multiple scelrosis in the patients did not differ from the index in a reference group of normal subjects. The incidence of inhibition of migration induced by central nervous system white matter and basic protein was greater than in serially studied normal subjects (p less than 0.05, p less than 0.2 less than 0.1, respectively) but bore no definite relation to clinical course.

Antigens↗
PubMed2001

[Removal of PEG and salt from recombinant human tumor necrosis factor-alpha(rhTNF-alpha) with serial operation of gel filtration chromatography].

A serial gel filtration operation with the combination of G25 and G100 in one column was developed to remove salt and PEG from upper phase rich in recombinant human tumor necrosis factor alpha after simultaneous cell disruption and two-phase aqueous extraction. Buffer exchanging and primary purification were also achieved at same time. The purification factor of 4.4 was obtained with an activity recovery of 97% by one single step of serial operation.

Chromatography, Gel↗
PubMed2003

Development of real-time detection direct test for hepatitis B virus and comparison with two commercial tests using the WHO international standard.

AIMS: A highly reproducible and sensitive hepatitis B virus real-time detection direct (HBV RTD-direct) test using DNA extraction by magnetic beads coated with polyclonal anti-HBsAg, followed by the real-time detection polymerase chain reaction (PCR) method, was developed for the detection of HBV DNA. METHODS: The HBV DNA could be extracted from the HBsAg positive viral particles without resulting in viral DNA fragmentation. The HBV RTD-direct test was validated using a serial dilution panel of the WHO standard HBV DNA 97/746 I. RESULTS: The test had a dynamic range of 0.7-8.0 log10 international units (IU) per mL and the results were shown to be comparable to those obtained with two commercially available tests: the HBV DNA transcription-mediated amplification-hybridization protection assay and the Amplicor HBV Monitor test. In addition, the HBV RTD-direct test, based on magnetic extraction, successfully eliminated PCR inhibitors in clinical specimens. CONCLUSION: We conclude that the HBV RTD-direct test is an excellent alternative for monitoring patients undergoing antiviral treatment or for screening various clinical specimens.

DNA, Viral↗
PubMed1984

Glycoprotein gp118 of varicella-zoster virus: purification by serial affinity chromatography.

Glycoprotein gp118, one of the major glycosylated proteins specified by varicella-zoster virus, is biologically of great importance since it possesses an epitope which elicits a complement-independent neutralizing antibody response. To purify this glycoprotein from a Nonidet-solubilized extract of varicella-zoster virus-infected cells, we examined its affinity to a variety of ligands, including two lectins--concanavalin A and Lens culinaris, Cibacron blue and heparin, and finally an immunoadsorbent anti-gp118 monoclonal antibody. By serial affinity chromatography on three different columns consisting of, respectively (i) Cibacron blue dye-Sepharose, (ii) L. culinaris-Sepharose, and (iii) anti-gp118 murine monoclonal antibody bound to CNBr-activated Sepharose, we isolated varicella-zoster virus-specific gp118 essentially free of contamination by any other radiolabeled viral or cellular polypeptide. The fold purification was estimated at 1,025 and the percent recovery at 13.6. On the basis of its chromatographic properties, gp118 appeared to contain mainly asparagine-linked, biantennary, complex-type, and hybrid-type oligosaccharides.

Animals↗
PubMed1990

Tentorial hemorrhage associated with vacuum extraction.

The clinical and radiologic descriptions of three neonates with tentorial hemorrhage after vacuum extraction are reported. All patients were full term, with Apgar scores of 8 or more; one patient experienced fetal distress during delivery. Within 36 hours after birth, the neonates had multiple generalized seizures; computed tomography or magnetic resonance imaging outlined distinctive tentorial hemorrhages with extension over the superior surface of the cerebellum or inferior surface of the occipital lobe. One patient had diffuse hypoxic-ischemic injury, and another had bilateral temporal lobe infarcts. Treatment included medical control of seizures and intracranial hypertension; one patient had surgical evacuation of bilateral subdural hematomas. Follow-up from 1 to 5 years showed significant developmental delays in two patients. These cases demonstrate that the forces generated on the fetal cranium by vacuum extraction are similar to those produced by forceps and result in tentorial laceration, venous rupture, and subdural hemorrhage. Because these hemorrhages may be associated with significant ischemic injury, serial radiologic evaluation is recommended for the detection of persistent structural abnormalities.

Birth Injuries↗
PubMed1982

Breakdown and reestablishment of blood-aqueous barrier with implant surgery.

Preoperative and serial postoperative anterior chamber fluorophotometry was performed after oral administration of fluorescein sodium in patients undergoing extracapsular cataract extraction combined with intraocular lens implantation. In placebo-treated eyes, the blood-aqueous barrier was reestablished at three months after surgery. The administration of topical indomethacin before and after surgery markedly decreased aqueous fluorescein concentration, and the blood-aqueous barrier was reestablished within five weeks. Because all patients were given sub-Tenon's injection of steroids immediately after surgery and intensive topical steroid therapy postoperatively, the effect of topical indomethacin was additive to or synergistic with steroids. The addition of topical indomethacin caused a small but statistically significant increase in the postoperative intraocular pressure; this effect lasted nine weeks.

Administration, Topical↗
PubMed1988

Quantitative assessment of postsurgical breakdown of the blood-aqueous barrier following administration of 0.5% ketorolac tromethamine solution. A double-masked, paired comparison with vehicle-placebo solution study.

Preoperative and serial postoperative anterior chamber fluorophotometry were performed after oral administration of fluorescein sodium in patients undergoing extracapsular cataract extraction and posterior chamber intraocular lens insertion. The administration of topical 0.5% ketorolac tromethamine solution before and after surgery markedly decreased the breakdown of the blood-aqueous barrier compared with vehicle-placebo solution administration at each time period, as measured by fluorophotometry. Corticosteroids were not given to any patients throughout the duration of the study. These fluorophotometric results correlated well with slit-lamp observations of postoperative ocular inflammation. Both ketorolac and vehicle were well tolerated by patients. No effects on intraocular pressure were seen with ketorolac administration. This study suggests that ketorolac ophthalmic solution is effective and safe as a nonsteroidal anti-inflammatory agent for topical use following cataract surgery and intraocular lens implantation.

Adult↗
PubMed1997

Model for quantitative immunohistochemical assessment of pulpal response to biomaterials.

The purpose of this study was to propose a new method for quantitative evaluation of the pulpal inflammation to restorative materials using immunohistochemistry and image analysis. Class V cavities were prepared and filled with different restorative materials in 20 healthy premolars to be extracted for orthodontic reasons. Teeth were extracted at different time intervals, fixed, demineralized, and embedded in paraffin. Six-micrometer-thick sections were cut serially and mounted on slides and every fifth section was stained using hematoxylin and eosin or Masson's trichrome, and served to localize the inflammatory reaction. The slides corresponding to the bulk of the inflammatory reaction were then used for immunohistochemical detection of the inflammatory cells using monoclonal antibodies: CD15 (granulocytes and histiocytes), CD45RO (T lymphocytes and monocytes), Pan-B cell (B lymphocytes, macrophages, and a subpopulation of T lymphocytes), CD45RA (B lymphocytes and monocytes), and EMA (plasma cells). The slides were observed and submitted to computerized image analysis using a SAMBA 2000 system for counting of the cells. The CD15, CD45RO, Pan-B-cell, and CD45RA antibodies positively stained the target cells, which could be counted with the computer. The EMA antibody did not permit staining of cells. These results indicate that polymorphonuclears, T lymphocytes, and B lymphocytes are present in inflamed human dental pulp. The immunologic detection of inflammatory cells followed by computerized image analysis allows an accurate characterization of pulpal pathology, and could be useful for the study of pulp reactions to restorative biomaterials.

Dental Materials↗
PubMed2000

Hot microscopic areas of iodine-deficient euthyroid goitres contain constitutively activating TSH receptor mutations.

Constitutively activating TSH receptor mutations have been established as the most common molecular basis for the pathogenesis of toxic thyroid nodules. These mutations result in uncontrolled signalling through the TSH receptor that is likely to cause hyperfunction and proliferation. The incidence of toxic multinodular goitres has been demonstrated to be related to iodine deficiency. Moreover, scintigraphically autonomous areas are found in 40% of euthyroid goitres from iodine-deficient areas. To investigate the molecular cause of these autonomous areas, small autoradiographically hot areas were examined for somatic TSH receptor mutations using archival tissue sections from 14 patients with euthyroid goitres, which had been originally prepared nearly 20 years ago. All patients had received (125)I 17 h preoperatively for the autoradiographic investigation of their thyroid. Areas with high and low (125)I-labelling on autoradiography sections were collected separately either from serial paraffin-embedded tissue sections, or Eukitt-embedded tissue sections containing the autoradiograph. After genomic DNA extraction, the transmembrane segment of the TSH receptor was PCR-amplified and directly sequenced. Somatic TSH receptor mutations were identified in areas with high (125)I-labelling in four patients: A623I, L629P, F631L, and T632I. This is the first evidence that TSH receptor mutations occur in microscopic areas with increased (125)I-labelling in euthyroid goiters and it suggests that TSH receptor mutations in these areas confer the potential to develop into toxic thyroid nodules. It is therefore very likely that toxic thyroid nodules originate from small autonomous areas in iodine-deficient euthyroid goitres that contain a TSH receptor mutation.

Female↗
PubMed1995

Lack of presystemic metabolism of nifedipine in the rabbit.

In humans, oral bioavailability of nifedipine has been reported to be around 60%, although the organ(s) contributing to its first-pass metabolism have not been determined. The aim of this study was to determine in vivo, in anesthetized and conscious rabbits the role of the intestine, liver, and lungs in the first-pass metabolism of nifedipine. To assess the extraction of nifedipine by the intestine, liver, and lungs, nifedipine was administered before and after each organ, and serial blood samples were withdrawn from an artery. In conscious rabbits, the systemic clearance of nifedipine injected into a lateral vein of an ear was 14.6 +/- 1.6 ml/min per kg, a value that was slightly decreased by anesthesia. In anesthetized rabbits, compared to the clearance estimated when nifedipine was administered into the thoracic aorta, the administration of nifedipine into a jugular vein, into the portal vein, or into the portal vein, or into the duodenum did not increase the value of the systemic clearance. In conscious rabbits, the clearance of nifedipine estimated when the drug was administered into the duodenum, the peritoneum, the portal vein, or into the jugular vein was identical to the clearance calculated when the drug was injected into the thoracic aorta. In vitro, nifedipine was metabolized in liver and intestinal epithelial cells homogenates but not in lungs or kidneys. We concluded that in the rabbit, oral nifedipine is not subjected to a first-pass metabolism, even though the intestine and the liver may contribute to nifedipine systemic clearance.

Animals↗
PubMed2004

Isolation and biodiversity of hitherto undescribed soil bacteria related to Bacillus niacini.

The hitherto largely not described phylogenetic neighborhood of Bacillus niacini has been explored by a comprehensive cultivation experiment and genomic variety studies. Previous culture-independent studies demonstrated that approximately 15% of all Bacillus 16S rDNA directly extracted from soils worldwide was affiliated to B. niacini. Seven different media were inoculated with soil suspensions in serial dilutions and incubated at different temperatures. Then, bacterial colonies were picked and analyzed by sequencing. A mineral medium with acetate as carbon source yielded a B. niacini rate of >3% of all picked colonies. Other media were less efficient but also successful. Applying this culturing approach, we succeeded in obtaining 64 isolates from different Dutch soils. The isolates turned out to be diverse, although closely related to B. niacini as revealed by 16S rDNA sequencing. Close matches with environmental clones were also found, thus demonstrating much more diversity beyond previously known 16S rDNA sequences. The rep-PCR fingerprinting method revealed a high genomic variety, redundancy could not be observed among our isolates. Hence, the hitherto neglected B. niacini lineage, apparently among the most abundant soil Bacillus, was accessible to our cultivation approach.

Bacillus↗
PubMed1979

Improved performance of a double antibody radioimmunoassay for carcinoembryonic antigen.

A new double antibody solid-phase radioimmunoassay (RIA) for carcinoembryonic antigen (CEA) is critically analyzed. The aim of the study was 4-fold: (a) to define the level of sensitivity (a comparison of 3 different assay procedures revealed that our sequential assay was more sensitive than most previously reported RIAs, while competitive and non-equilibrium assay had wider measuring ranges); (b) to analyze recoveries of CEA in either serum, plasma or urine (the recovery, even in urine, was very close to expected values, indicating that no CEA is lost or degraded during brief storage or in the extraction procedure); (c) to evaluate inter- and intra-assay variations, since most clinical management is dependent on serial assays rather than single determinations. The coefficients of variation were low both within and between assays. A change of 3 ng CEA is required for significant change (greater than 2 S.D.) at the normal serum level which is 16 ng CEA/ml in our assay. At levels above normal, a change of 4 ng is required; (d) the assay was also developed for determination of CEA levels in a large series of perchlorid acid treated serum, plasma or urine samples. This forms the basis for an assay suitable for serial assays with high sensitivity and accuracy in various neoplastic diseases.

Animals↗
PubMed2005

Epithelium and bacteria in periapical lesions.

OBJECTIVE: The purpose of this study was to evaluate 50 human periapical lesions for bacteria and epithelium in a case study in dental practice. STUDY DESIGN: Specimens were obtained from the extraction of 50 untreated teeth that had lesions attached to their apices. The specimens were histologically evaluated using serial sections. RESULTS: Bacteria were found in all teeth, colonizing necrotic tissue in the main canal, dentinal tubules, or apical ramifications, and in the body of the periapical lesion in 18 abscesses or cysts. Twenty-one lesions were epithelialized; 14 abscesses, 20 granulomas, and 16 cysts were distinguished. In 18 root canals inflamed tissue was found in the apical part of the canal. A single foramen was present in 13 cases while apical ramifications were found in 37 cases. CONCLUSIONS: Granulomas were most common, and most epithelialized lesions were cysts. Bacteria were only detected periapically in abscesses or cysts. Inflamed tissue was present in the apical root canal in one third of cases.

Adolescent↗
PubMed1994

Bacterial retention in canal walls in vitro: effect of smear layer.

When dentin is planed by endodontic instruments, a smear layer forms. Whether this layer should be removed is unknown and controversial. This study was conducted to assess the effect of the smear layer on retention of bacteria using an in vitro root canal bacterial colonization model. Canals of 26 extracted human canines were step-back prepared using 2.5% NaOCl. Teeth were then randomly divided into two groups based on the type of high volume final flush: 1-20 ml of sterile saline (0.85% wt/vol) or (2-10 ml of 17% EDTA followed by 10 ml of 2.5% NaOCl which removes smear layer. Streptococcus anginosus (milleri) was cultured in trypticase soy broth supplemented with 0.5% yeast extract at 37 degrees C in 5% CO2. Cells were harvested by centrifugation and resuspended in fresh media. Serial dilutions were performed to achieve inocula of 10(6) colony-forming units in a 30-microliters volume. Teeth were inoculated and incubated for 2 h in 5% CO2 at 37 degrees C. Following incubation, teeth were split and processed for microbiological analysis. Numbers of colonizing bacteria were determined by a spiral-plating system. Enumeration of the numbers of bacteria revealed a reproducible, order of magnitude difference (p = 0.0002) between teeth with smear layer (10(4) colony-forming units) versus teeth without smear layer (10(5) colony-forming units). This suggests that smear layer produced during root canal therapy may inhibit bacterial colonization of root canals. One suggested mechanism is that smear layer may block bacterial entry into dentinal tubules.

Bacterial Adhesion↗
PubMed1985

The type-specific polysaccharide and the R protein antigens of the L-phase from a group B, type III Streptococcus.

The type-specific polysaccharide and the R protein antigens from filtered culture supernatants of the bacterial phase and L-phase of the group B, type III streptococcal strain 76-043 were studied by several immunological methods. In the L-phase of growth, the two antigens were separate and distinct molecules which were found principally in the culture supernatant even on the 254th serial subculture in the cell-wall-defective state. Only trace amounts of these antigens were detected in extracts of L-phase cells. The type III polysaccharide antigens in the supernatant of cultures of the parent bacterium and the L-phase gave reactions of identity in immunodiffusion. Precipitin bands obtained by immunoelectrophoresis (IEP) revealed that the type-specific antigen of the bacterial phase of growth migrated toward the anode, whereas that of the L-phase remained near the antigen well. The R protein antigen in the L-phase supernatant was immunologically identical to the R protein of the supernatant and 1% trypsin-extracted antigens from whole cells of the parent bacterial strain, and other groups A, B and C streptococcal strains sharing a common R antigen. Immunologically, the R antigen appeared to be the species R4. The R protein of the L-phase and bacterial phase cultures was resistant to 5% trypsin but sensitive to 0.5% pepsin at 37 degrees C/2hr. Antiserum prepared in rabbits against L-phase cells contained an antibody reactive with the R protein antigens of the bacterial and L-phase cultures. The soluble, naturally released type III and R protein streptococcal antigens of the L-phase of growth permitted immunological confirmation of its bacterial origin.

Antigens, Bacterial↗
PubMed1985

Transcutaneous oxygen tension measurements during graded hemorrhage and reinfusion.

Measurement of transcutaneous oxygen tension (PtCO2) has been suggested as a useful monitoring tool in the hypovolemic patient. Our study was undertaken to evaluate changes in PtCO2 that occur during graded hemorrhage and reinfusion, and to compare PtCO2 values to standard cardiorespiratory and biochemical parameters during hypovolemia. Seven mongrel dogs were bled 50% of their estimated blood volume (44 mL/kg) over one hour. This was followed by a one-hour monitoring period, a 30-minute reinfusion period, and an additional one-hour monitoring period. Pulmonary capillary wedge pressure (PCWP), central venous pressure (CVP), cardiac output (CO), mean arterial pressure (MAP), mixed venous oxygen tension (MvO2), arterial blood gases, and PtCO2 were measured serially throughout the study period. Cardiac index (CI), peripheral vascular resistance (PVR), O2 consumption, delivery, and percentage of extraction were calculated for each sampling period. A statistically significant fall in CI, MvO2 and PCWP occurred following the first 10% of blood loss; PtCO2 and MAP fell significantly after 20% hemorrhage; CVP fell after 30% hemorrhage. PtCO2 rose significantly after the first 10% of reinfusion, and it continued to rise during the entire reinfusion period, as did MvO2, CO, MAP, CVP, and PCWP. In contrast to the other measured variables, the elevations in PtCO2, and MvO2 were more pronounced early in the reinfusion period. During postreinfusion monitoring, PtCO2, MvO2, CO, and PCWP fell significantly despite maintenance of prehemorrhage MAP and CVP. Overall PtCO2 correlated well with MvO2 and the O2 extraction ratio, and to a lesser extent with CI, MAP, and O2 delivery.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2003

Ketone and pyruvate Ringer's solutions decrease pulmonary apoptosis in a rat model of severe hemorrhagic shock and resuscitation.

BACKGROUND: Resuscitation fluids containing beta-hydroxybutyrate (BHB) have been shown to decrease cellular injury after hemorrhagic shock and resuscitation through an unknown mechanism. We tested whether this effect was related to BHB-induced metabolic modulations. METHODS: Male Sprague Dawley rats (n=30) were subjected to volume-controlled hemorrhage (27 mL/kg during 10 minutes followed by 75 minutes of shock during which another 8 mL/kg of blood was withdrawn). Experimental groups included the following: (1) sham, (2) no resuscitation (NR), (3) racemic lactated Ringer's (DL-LR) solution, (4) LR containing L-isomer only (L-LR), (5) ketone Ringer's solution with lactate substituted by BHB (KR), and (6) pyruvate Ringer's solution with lactate substituted by pyruvate (PR). The resuscitation fluids were infused during 45 minutes simultaneously with additional hemorrhage of 8 mL/kg. Hemodynamic and physiologic parameters and the plasma levels of BHB were serially measured. The animals were killed 2 hours after resuscitation, and tissues were frozen instantaneously for cellular adenylate extraction and adenosine triphosphate (ATP) and adenosine diphosphate analysis. Pulmonary apoptosis was studied using Western blotting, immunohistochemistry, and reverse transcriptase-polymerase chain reaction. Expression of enzymes involved in ketogenesis and ketolysis was analyzed by reverse transcriptase-polymerase chain reaction. RESULTS: NR and resuscitation with DL-LR increased the expression of apoptotic markers, whereas resuscitation with KR and PR significantly decreased the expression of apoptotic markers in rat lungs. Resuscitation with KR was followed by a profound increase in plasma BHB levels; however, the expression levels of ketolytic enzymes were essentially unaffected. KR infusion did not induce significant improvements in tissue ATP levels. CONCLUSIONS: Resuscitation with KR and PR protects against pulmonary apoptosis without improving tissue ATP content. Therefore, metabolic modulation is unlikely to be the major mechanism by which BHB exerts its protective effects during reperfusion.

3-Hydroxybutyric Acid↗
PubMed1986

Bovine parathyroid cells: cultures maintained for more than 140 population doublings.

Primary cultures of bovine parathyroid cells were developed using Coon's modified Ham's F-12 medium containing low (0.3 mM) concentrations of calcium and supplements of bovine hypothalamic extract, bovine pituitary extract, epidermal growth factor, insulin, transferrin, selenous acid, hydrocortisone, triiodothyronine, retinoic acid, and galactose. These cells were cultured serially on serum-coated dishes for 140 population doublings before signs of senescence were detected. The cells were epithelioid and maintained a high degree of differentiation as evidenced by calcium regulation of both growth and secretion and by prostaglandin E1 stimulation of cAMP formation and hormone release.

Alprostadil↗