Comparison of supercritical fluid extraction and conventional liquid-solid extraction for the determination of benzo[a]pyrene in water-soluble smoke

In patients undergoing bone marrow transplantation cryptococcosis is rarely encountered. We report a fatal case of Cryptococcus meningitis in a 12-year-old girl with acute lymphoblastic leukemia (ALL) in second remission who had a transplant from a human leukocyte antigen (HLA)-identical unrelated bone marrow donor. The conditioning regimen was thiotepa, cyclophosphamide, and total body irradiation (TBI); graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporin A, methotrexate, and antilymphocyte globulin (ALG). The patient experienced stage III GVHD responsive to high-dose corticosteroids. On day +54 a thrombotic microangiopathy occurred. On day +64 neurological status worsened; a brain computed tomographic (CT) scan showed hyperdense lesions suggesting fungal infection. Detection of cryptococcal antigen by latex agglutination was positive but India ink stain and culture were negative. Despite treatment with amphotericin B, 5-flucytosine, and granulocyte-macrophage colony-stimulating factor, the patient died 13 days after the diagnosis.     

A comparison between solid phase extraction and supercritical fluid extraction for the determination of fluconazole from animal feed

The application of supercritical fluid extraction with carbon dioxide and modified carbon dioxide for the determination of fluconazole from an animal feed was studied. A fractional factorial design approach was used to examine the significant experimental variables for quantitative extraction of fluconazole. Gas chromatography with either flame ionisation or mass selective detection was used for quantitation of the extracts. The results indicated that modifier (methanol) had the greatest effect on the recovery of fluconazole from the animal feed.     

Packed-column supercritical fluid chromatography coupled with solid-phase extraction for the determination of organic microcontaminants in water

Fatty acids are the preferred substrate of ischemic, reperfused myocardium and may account for the decreased cardiac efficiency during aerobic recovery. Neonatal cardiac myocytes in culture respond to hypoxia/serum- and glucose-free medium by a slow decline in ATP which reverses upon oxygenation. This model was employed to examine whether carnitine palmitoyltransferase I (CPT-I) modulates high rates of beta-oxidation following oxygen deprivation. After 5 h of hypoxia, ATP levels decline to 30% control values and CPT-I activity is significantly stimulated in hypoxic myocytes with no alteration in cellular carnitine content or in the release of the mitochondrial matrix marker, citrate synthase. This stimulation was attributed to an increase in the affinity of hypoxic CPT-I for carnitine, suggesting that the liver CPT-I isoform is more dominant following hypoxia. However, there was no alteration in hypoxic CPT-I inhibition by malonyl-CoA. DNP-etomoxiryl-CoA, a specific inhibitor of the liver CPT-I isoform, uncovered identical Michaelis kinetics of the muscle isoform in control and hypoxic myocytes with activation of the liver isoform. Northern blotting did not reveal any change in the relative abundance of mRNA for the liver vs. the muscle CPT-I isoforms. The tyrosine phosphatase inhibitor, pervanadate, reversed the hypoxia-induced activation of CPT-I and returned the affinity of cardiac CPT-I for carnitine to control. Reoxygenation was also associated with a return of CPT-I activity to control levels. The data demonstrate that CPT-I is activated upon ATP depletion. Lower enzyme activities are present in control and reoxygenated cells where ATP is abundant or when phosphatases are inhibited. This is the first suggestion that phosphorylation may modulate the activity of the liver CPT-I isoform in heart.     

Straightforward solid-phase extraction method for the determination of verapamil and its metabolite in plasma in a 96-well extraction plate

Straightforward solid-phase extraction (SPE) methods were developed for the determination of verapamil and its metabolite in a plasma matrix. The spiked plasma sample was pretreated with 2% phosphoric acid followed by two different SPE methods using a Waters Oasis HLB 96-well extraction plate. Recoveries greater than 90% were obtained using both a generic and a selective SPE methods. The generic method is a good starting protocol, and it is applicable to a wide range of compounds. This generic method consists of using 5% methanol as the wash solvent, and 100% methanol for the elution. The limitation of the non-specific method is that it does not remove all plasma constituents that interfere with the quantitation of the metabolite, norverapamil. A second, more specific method was developed using the same Oasis HLB sorbent which removes more plasma interferences and provides cleaner extracts for the HPLC-UV analysis. This selective method uses both the methanol concentration and the pH advantageously to preferentially isolate the analytes of interest from a complex sample matrix. Recoveries of greater than 90% with R.S.D.s less than 3.8% were obtained with this selective method.     

Use of supercritical fluid extraction as a method of cleaning anterior cruciate ligament prostheses: in vitro and in vivo validation

The process of supercritical fluid extraction (SFE) using carbon dioxide as the mobile phase is finding increasing numbers of applications in a wide variety of industries for the extraction, separation, and cleaning of materials. This study assessed the usefulness of this approach in removing surface contaminants from a knitted polyester anterior cruciate ligament (ACL) prosthesis before packaging and sterilizing the product during manufacture. The physical, dimensional, and chemical properties of SFE treated compared with commercially scoured control samples were characterized using a number of textile test methods: electron spectroscopy for chemical analysis, Fourier transform infrared spectroscopy, differential scanning calorimetry, and solvent extraction analysis. The biocompatibility of the samples was measured in terms of their ability to generate CD18 integrin expression on activated human polymorphonuclear cells, and their inflammatory response when implanted for up to 30 days in the knee joint of rats. SFE treatment was successful in removing most of the nonpolar contaminants from the ACL prosthesis and reducing the amount of residuals to a commercially acceptable level. However, some nitrogen containing compounds and polar salts were not removed by the SFE process. The results from the biocompatibility tests demonstrated that the cleaner SFE treated prosthesis induced significantly lower CD18 expression than the scoured control fabric, and was also associated with a milder inflammatory response and a more rapid rate of healing during the 30 day animal trial. Another effect of SFE processing was to cause the polyester device to shrink and lose porosity because of yarn contraction and modification of the polymer's microcrystalline structure.     

New methods for the determination of hydrogen content of aluminum and its alloys: Part I. Improvements in the vacuum extraction method

The vacuum hot extraction method, first proposed by Ransley for the determination of hydrogen content of aluminum and its alloys, was systematically investigated to check if it could be considered as a reference procedure for the development of other methods directly applicable in routine work. For that, the use of an improved apparatus involving a mass spectrometer as gas detector and specimens intentionally charged with deuterium —a tracer whose properties are similar to those of hydrogen—permits the following conclusions: 1) The method is correct for any alloy, even for all the alloys with a high magnesium or zinc content, 2) The “getter effect” which could lead to inaccurate results does not occur, provided the pumping speed of the transfer pump exceeds the rate of outgassing from the sample, 3) The extraction from the solid phase seems to be complete, since results obtained in this condition are not significantly different from those obtained from melted samples. 4) The time required for an analytical determination (several hours) due essentially to the preliminary operations, cannot be practically reduced by extraction at higher temperature in the liquid state. The Ransley method can thus be considered as a reference method for checking other more practical methods.     

A comparison between a PCR method and a conventional culture method for detecting pathogenic Yersinia enterocolitica in food

The beige mutation is a murine autosomal recessive disorder, resulting in hypopigmentation, bleeding and immune cell dysfunction. The gene defective in beige is thought to be a homologue of the gene for the human disorder Chediak-Higashi syndrome. We have identified the murine beige gene by in vitro complementation and positional cloning, and confirmed its identification by defining mutations in two independent mutant alleles. The sequence of the beige gene message shows strong nucleotide homology to multiple human ESTs, one or more of which may be associated with the Chediak-Higashi syndrome gene. The amino acid sequence of the Beige protein revealed a novel protein with significant amino acid homology to orphan proteins identified in Saccharomyces cerevisiae, Caenorhabditis elegans and humans.     

Comparison of methods for the determination of total and free tryptophan in plasma

A comparison is made between two fluorimetric techniques for the determination of total and free plasma tryptophan. Comparison is also made between methods for the separation of the unbound and bound fraction of plasma tryptophan using ultrafiltration, Centricones and small-scale equilibrium dialysis.     

The tampon test for trichomoniasis: a comparison between conventional methods and a polymerase chain reaction for Trichomonas vaginalis in women

Vertebral artery tortuosity causing neural foraminal widening is a well described abnormality that should not be confused with other causes of neural foraminal enlargement, particularly on conventional roentgenograms. We, hereby, describe CT features of another cervical osseous change due to the vertebral artery tortuosity, the so called "tubular shaped vertebral artery canal", which is embedded in the vertebral body instead of causing neural foramen enlargement. Catheter and MR angiographic studies have also been performed to confirm the vertebral artery tortuosity causing the osseous changes.     

Blood volume determination with sodium fluorescein and radioactive chromium--a clinical comparison of methods

OBJECTIVE: There exists no method so far for the determination of circulating blood volume as an important parameter of circulatory function widely usable under clinical conditions. Therefore, the present study was designed to investigate whether identical distribution spaces could be measured by two methods for blood volume determination using sodium fluorescein (SoF) and radioactively labelled red blood cells (51Cr*). DESIGN: Comparative study. SETTING: Operating theatre, recovery room, or intensive care unit of a university hospital. PATIENTS: 35 patients undergoing abdominal, urological or vascular surgery. INTERVENTIONS: Simultaneous determinations of blood volume using SoF and 51Cr* in the intra- and postoperative period. RESULTS: There were no significant differences between the calculated means of blood volume (4,445 vs. 4,407 ml), red cell volume (1,554 vs. 1,540 ml), and plasma volume (2,891 vs. 2,807 ml) for 51Cr*-vs. SoF-stained red blood cells. The coefficient of correlation between the two methods was r = 0.95. The mean percentage error was -0.6% between the two methods, the precision 5.6%. CONCLUSIONS: SoF-stained erythrocytes allow a determination of the same distribution space as the well-established radioactive method using 51Cr*. Therefore, SoF-staining may replace 51Cr* labelling of red blood cells for the determination of blood volume in patients.     

Direct comparison of three methods for the determination of radon in well water

Radon concentrations obtained using a bubbler device developed to collect and bubble water samples in the laboratory and field were compared with results from conventional liquid scintillation counting. Measurements from standard solutions with a wide concentration range showed excellent agreement between liquid scintillation and results obtained using the bubbler device in conjunction with alpha-scintillation cells. Measurements of waterborne radon concentrations in 110 community and private wells in New York State ranged from 1 to 4,100 Bq L-1, with arithmetic and geometric means of 200 and 30 Bq L-1, respectively. Excellent agreement between the analytical techniques was obtained for each field site.     

Automated fluorometric method for determination of total vitamin C in food products

A specific microfluorometric method for the determination of ascorbic acid, dehydroascorbic acid, and total vitamin C in food products has been automated. The procedure developed is an adaptation of the official AOAC method (secs. 43.056-43.062), except that N-bromosuccinimide is used instead of Norit to oxidize vitamin C. Ascorbic acid is selectively oxidized by N-bromosuccinimide before other interfering substances that may be present, so this method is a highly sensitive and specific technique with extensive applicability. The proposed automated method is simple, rapid, reliable, and sufficiently sensitive to analyze as little as 2 x 10-3 to 0.1 mg ascorbic acid/ml. Analytical results obtained for ascorbic acid, dehydroascorbic acid, and total vitamin C in a wide variety of food products are reported. The analytical system developed has the capability of analyzing 50 samples/hr.     

The sensitivity of hyaluronan analysis of pleural fluid from patients with malignant mesothelioma and a comparison of different methods

This retrospective chart review describes the efficacy and safety of long-term administration of intravenous alpha1-antitrypsin (AAT) in 14 patients with hereditary AAT deficiency and COPD. During the 12- to 48-month observation period, 12 to 14 patients had stabilization of functional status; 4 patients had reductions in hospitalizations. Thirteen of 14 patients had no decline in pulmonary function. Three patients had self-limited adverse reactions to the AAT with one patient requiring a brief hospitalization.     

A simple radioimmunoassay for plasma cortisol: comparison with the fluorimetric method of determination

A quick and simple method for the radioimmunoassay of plasma cortisol is described. The mean morning plasma cortisol concentration in 43 normal subjects was 9.8 +/- 3.1 (S.D.) microgram/100 ml with a range of 5.0-19.5 microgram/100 ml. Mean midnight concentration in 24 normal subjects was 4.3 +/- 2.3 (S.D.) microgram/100 ml with a range of 1.4-9.6 microgram/100 ml. When compared with the fluorimetric method the mean results by radioimmunoassay of 154 routine specimens were 23% lower. In samples from unstimulated patients, regression analysis of results obtained by the two methods gave a correlation coefficient (r) of 0.93, regression line slope of 1.1, and intercept of 1.4 microgram. Mean radioimmunoassay results were 15% lower. When plasma cortisol concentration was above the normal range (greater than 30 microgram/100 ml) the regression line slope was 0.87, the intercept 17.9 microgram, r = 0.87 and mean radio immunoassay results were 37% lower. Plasma cortisol concentration in patients after insulin or Synacthen stimulation exhibited similar responses when measured by either method. Plasma cortisol concentration in normal subjects given metyrapone was lower when measured by radioimmunoassay (mean +/- S.D. = 8.7 +/- 2.7 microgram/100 ml) than when measured by fluorimetry (18.5 +/- 10.8 microgram/100 ml). The diagnostic usefulness of the two methods, ease of assay, and costs are compared.     

不锈钢中镍含量的三种测定方法比较

本文比较了不锈钢中镍含量的三种测定方法：丁二酮肟光度法、丁二酮肟重量法和火焰原子吸收光度法。结果显示,丁二酮肟光度法的回收率约为98．7％,RSD为4．36％,丁二酮肟重量法的回收率约为97．0％,RSD为11．74％,火焰原子吸收方法的回收率约为99．7％,RSD为1．32％,由此可见,火焰原子吸收方法测量不锈钢中的镍准确度、精密度最高。     

Improved high-performance liquid chromatographic method for the separation and quantification of lipid classes: application to fish lipids

The demyelination process that occurs in the central nervous system (CNS) of patients with multiple sclerosis (MS) is in part due to an inflammatory response in which CD4+ and CD8+ T cells and macrophages infiltrate white matter. In this study, we have identified a peptide sequence derived from the CNS-specific myelin protein proteolipid protein (PLP) which could bind to HLA-A3 and induce a HLA-A3-restricted CD8+ CTL response from HLA-A3+ donors. These PLP peptide-specific CTL could lyse HLA-A3+ target cells pulsed with a homologous peptide derived from the CRM1 protein of Saccharomyces cerevisae. These findings demonstrate the immunogenic potential of a PLP-derived peptide for generation of autoreactive HLA-A3-restricted CD8+ CTL, and further show that these CTL can be activated by a peptide derived from a common environmental microorganism.     

Integration of inulin determination in the AOAC method for measurement of total dietary fibre

A suitable modification of the standard AOAC method for the measurement of dietary fibre is proposed to quantitatively include beta-fructans in the determination of the soluble dietary fibre fraction and as a consequence in the related total dietary fibre fraction. The standard AOAC method is modified by including a preheated commercial inulinase, Novozym SP 230, to the amyloglucosidase incubation step. It was previously outlined that this commercial inulinase contains some pectolytic activity. It is now demonstrated that a heat pretreatment of this enzyme preparation at 60 degrees C for 2 h eliminates this pectolytic activity while keeping sufficient activity to hydrolyse all the inulin from the soluble fibre fraction.     

Rapid shell vial culture technique for detection of enteroviruses and adenoviruses in fecal specimens: comparison with conventional virus isolation method

Detection of enteroviruses and adenoviruses mainly in fecal specimens by rapid culture with inoculation onto cell monolayers in flat-bottom tubes by centrifugation and immunofluorescence staining with genus-specific monoclonal antibodies was compared with that by the conventional virus isolation procedure. For both conventional culture and shell vial culture human lung fibroblast cells and tertiary monkey kidney cells were used. For enterovirus detection, 979 clinical specimens (916 stool specimens, 56 cerebrospinal fluid specimens, and 7 nasopharyngeal swabs) were used. Conventional culture detected 74 enterovirus isolates. A cytopathic effect compatible with the presence of an enterovirus after 3 days of incubation occurred in 25 of the 74 (34%) specimens that eventually became positive. The detection rate for enteroviruses by rapid cell culture after 2 to 3 days of incubation was 42 of 74 (57%). The genus-specific enterovirus monoclonal antibody did not react with strains of echovirus types 22 and 23 or enterovirus type 71. Rapid cell culture for the detection of adenoviruses was performed with 567 clinical specimens (536 stool specimens, 25 cerebrospinal fluid specimens, and 6 miscellaneous specimens), in which 42 adenoviruses were found by conventional culture. Nine of the 42 (21%) adenovirus isolates were detected by conventional culture within 3 days after inoculation, whereas 21 (50%) were found by rapid cell culture within 2 to 3 days. Only two of the nine specimens found to be positive for the enteric adenovirus type 41 by conventional culture as well by a type-specific enzyme-linked immunosorbent assay (ELISA) tested positive by rapid cell culture. In conclusion, the rapid shell vial assay allows the early detection and identification of enteroviruses and adenoviruses in clinical specimens but is markedly less sensitive than the conventional isolation procedure according to the eventual results of the conventional isolation procedure. Conventional cell culture remains a prerequisite for serotyping of enteroviral isolates. On the basis of the results for adenovirus type 41, the rapid detection of adenoviruses was not considered to be useful for the detection of clinically relevant adenoviruses in fecal samples.