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PubMed1989

Flow cytometric analysis of lymphomas.

Flow cytometry is rapidly developing as an important tool in the characterization of lymphomas. Analysis of cell surface antigens and DNA content in these tumors provides useful biologic information that can be applied advantageously to their diagnosis and classification. In this article we examine practical aspects of handling, preparation, and staining of lymphoid samples for flow cytometry and delineate approaches to data analysis and interpretation. We discuss specific applications, advantages, and limitations of the technique in the evaluation of neoplastic lymphoid diseases.

Evaluation Studies as Topic↗
PubMed2000

Evaluation of the von Willebrand factor antigen (vWF-Ag) assay using an immuno-turbidimetric method (STA Liatest vWF) automated on the MDA 180 coagulometer.

The standard enzyme-linked immunosorbent assay (ELISA) test for von Willebrand factor antigen (vWF-Ag), though sensitive and specific, does not deliver the flexibility to handle single sample assessments economically or provide rapid, emergency testing capability. The present study examined the performance characteristics of an immuno-turbidimetric assay kit (STA Liatest) modified for automation on the MDA(R)180 coagulation analyser using a supplied protocol. One hundred and sixteen patient samples were assessed by both the standard and the modified method. The correlation coefficient was 0.98 across the range of values 1-487 IU/dl. Above 200 IU/dl, where specimen dilution was indicated, there was greater diversity (r = 0.86) between the techniques. Plotting the difference between methods against the mean of both showed excellent agreement between methods below 100 IU/dl vWF. The intra-assay and interassay coefficients of variation were less than 3% at both low and normal range levels. The MDA(R)180 automated vWF assay merits consideration as an alternative to ELISA testing that provides random access and result availability within 30 min.

Antigens↗
PubMed1989

Effect of handling of plasma samples on histamine radio-enzyme assay.

1. Freeze-thawing of plasma samples increased the histamine level at physiological histamine concentrations as analyzed by radio-enzyme assay, but not by high performance liquid chromatography. 2. Heating of plasma samples decreased the histamine levels. 3. Enzymatic or non-enzymatic formation of histamine during handling of plasma samples was not detected. 4. Some albumin preparations contain histamine.

Adult↗
PubMed2006

Instability of calcium channel antagonists during sample preparation for LC-MS-MS analysis of serum samples.

1,4-Dihydropyridines calcium channel antagonists (1,4-DHP CCAs) are photolabile and the products of their photodecomposition have no pharmaceutical activity. In our previous work we have presented a screening procedure for eleven 1,4-DHPs in plasma by LC-MS-MS using multiple reaction motoring. The laboratory process includes preparation and storage of stock solutions, plasma storage, solid-phase extraction, reconstitution of extracts and storage time in an autosampler for LC-MS-MS analysis. Prior to validation of the analytical procedure, we have tested the stability of these compounds by exposure to light. Methanolic solutions have been exposed to laboratory and UV light and the stability of the compounds in plasma was tested by exposure of spiked plasma samples to laboratory light at room temperature. Stability during freeze-thaw cycles and stability during 2 month storage at -20 degrees C have been tested as well. Products of photodecomposition have been identified after forced degradation and the degree of degradation has been quantified using LC-UV-DAD and LC-MS-MS, respectively. A 96% degradation after only 2h has been observed when solutions of nifedipine or nisoldipine were exposed to laboratory light in clear glass vials. In plasma samples degradation was 25% in only 2h for both compounds. The main degradation product was produced by oxidation of the dihydropyridinic ring resulting in the pyridine analogue that has been described as the first metabolite in the metabolic pathway. Only minor degradation was found for the other tested compounds after 2h light exposure in methanolic solutions. Furthermore, lercanidipine and nicardipine were also degradated by esterhydrolysis. Several additional minor degradation products were found for the other tested 1,4-DHPs, however, some of them could not be identified. Preconditions for storage and handling of plasma samples prior to and during analysis for 1,4-DHP CCAs are suggested in order to avoid photodecomposition of the analytes.

Calcium Channel Blockers↗
PubMedDate not supplied

Chemical and hematological changes in stored CPD blood.

Blood was drawn from ten healthy volunteer donors into citrate-phosphate-dextrose (CPD) anticoagulant and placed on the quarantine shelf of the blood bank refrigerator. Plasma dextrose, sodium, potassium, chloride, bicarbonate, GOT, LDH, and hemoglobin as well as WBC, hematocrit, MCV, MCHC, whole blood pH, and ammonia were measured on all samples initially and at one, two, seven, 14, 21, and 28 days of storage at 4 C. Whole blood lactate also was analyzed serially on five of the units. An additional 27 units of CPD bank blood (two to 21 days of age), routinely processed, handled, and stored by the blood bank, were submitted to the same analyses on the day of administration to the patient. Five of these processed units, 21 days old, were resampled at 28 days. Results of the analyses are presented and discussed. The most pronounced changes were seen for dextrose, potassium, bicarbonate, lactate, LDH, ammonia, and hemoglobin. Plasma dextrose and bicarbonate declined in concentration while potassium, lactate, LDH, ammonia, and hemoglobin rose with storage. In general, changes in the regularly processed, singly sampled bank units were greater than those observed in the specially processed, quarantined units sampled serially. This study indicates that routine transportation, processing, and handling of bank blood may lead to increased biochemical alteration.

Ammonia↗
PubMed1999

Absorbable mesh envelope facilitates handling of pediatric en bloc kidneys during transplantation.

There is increasing evidence to favor the use of pediatric en bloc kidneys in renal transplantation. However, from a technical standpoint, pediatric en bloc renal transplantation is more complex because of difficulty in optimal intraoperative handling and positioning, as well as a higher potential for vascular complications. We describe our experience with a technique that has helped us to minimize the technical difficulties associated with the surgical procedure. Use of a tailored absorbable mesh envelope improves handling of the en bloc pediatric kidneys during performance of the vascular anastomoses and reduces the risk of torsion of renal pedicles.

Absorbable Implants↗
PubMedDate not supplied

Pakistan's experience of a bioterrorism-related anthrax scare.

From November 2001 to March 2002, the National Institute of Health, Islamabad, Pakistan, received 230 samples from 194 different sources for analysis for anthrax spores. These samples were taken from letters/packages suspected of containing anthrax and from individuals exposed to them. When cultured on sheep blood agar, 141 samples yielded growth suggestive of Bacillus species. On the basis of growth characteristics, absence of beta-haemolysis, absent or doubtful motility and morphological characters of the isolates on Gram stain, 62 isolates were considered suspicious and were inoculated into guinea-pigs. Inoculated animals remained healthy well beyond the required observation period of 5 days. All the samples were therefore reported as negative for B. anthracis. Systems for handling and analysing suspected anthrax-contaminated materials are discussed.

Anthrax↗
PubMed1997

From delivery suite to laboratory: optimizing returns from placental examination in medico-legal defence.

While examinations of the placenta may reveal significant lesions that are helpful in a medico-legal context, its value maybe diminished by a lack of attention to detail in the period between delivery of the baby and receipt of the placenta in the pathology department. These fall generally into 2 groups: initial description and handling of the placenta and lack of relevant clinical information. This communication describes some of these pitfalls. By observing these details, the onus then falls on the pathologist to provide a meaningful assessment of the placenta.

Australia↗
PubMed2006

Novel technique for use of cyanoacrylate in Mohs surgery.

BACKGROUND: Many Mohs procedures involve the handling and manipulating of thin, fragile skin tissue, particularly eyelid skin. After excision of such tissue, proper handling of the tissue becomes critical for tissue orientation and histologic processing. OBJECTIVE: The authors present a novel way to handle thin, friable tissue after excision in Mohs surgery with the direct application of cyanoacrylate onto freshly excised tissue. METHODS: Nonsterile generic cyanoacrylate can be applied along the epidermal surface edge to excised thin tissue sections before inking the margins, sectioning, and processing. CONCLUSIONS: The superglue increases tissue rigidity and provides for easier handling while staining and orienting the tissues. The added glue does not alter tissue integrity and shows minimal artifact upon histologic examination. This technique furthermore is inexpensive and adds little time to the Mohs procedure. Clinical photographs and histologic pictures are presented.

Cyanoacrylates↗
PubMed1997

Brain natriuretic peptide is stable in whole blood and can be measured using a simple rapid assay: implications for clinical practice.

OBJECTIVES: To compare the stability of brain natriuretic peptide (BNP) to that of N-terminal atrial natriuretic peptide (NT-ANP) in whole blood and plasma stored under different conditions. To compare a rapid, simple, direct (unextracted) BNP assay to a conventional assay using plasma extraction. DESIGN: Blinded, prospective, comparative study. SETTING: Tertiary referral cardiology department. SUBJECTS: Forty two subjects (24 men, 18 women) comprising 28 patients with left ventricular systolic dysfunction (LVSD) ranging from mild to severe and 14 healthy volunteers. MAIN OUTCOME MEASURES: Stability of NT-ANP and BNP when stored as whole blood or plasma at room temperature over three days. Reproducibility of measurements. RESULTS: BNP was stable in whole blood stored at room temperature for three days; mean change in concentration -7.4% (95% CI 0.6 to -14.8), (direct), -6.3% (5.0 to -16.4), (extracted); whereas a significant decline in BNP concentration was noted in plasma stored at room temperature; -23.2% (-13.7 to -31.6), (direct); -14.4% (-3.2 to -24.3), (extracted). By contrast a small nonsignificant rise in NT-ANP concentration was noted both in whole blood and plasma stored at room temperature for three days; whole blood +8.6% (+22.3 to -3.5), plasma +6.3%, (23.2 to -8.4). The reproducibility of the BNP measurements, and particularly the rapid, direct, measurement, was superior to that for NT-ANP. CONCLUSIONS: BNP is shown to be stable in whole blood for three days and can be measured using a rapid, simple assay. Routine assay of BNP is feasible in ordinary clinical practice and may be of value to general practitioners and hospital based physicians in the diagnosis and management of patients with LVSD. Samples can be sent to a central laboratory without special handling requirements.

Adult↗
PubMed2004

A simple method for bone-graft handling during spine surgery.

Morsellized bone-graft handling during spine surgery to get vertebral fusion can be a slow, laborious and time-consuming procedure. It is not absolutely exempt from complication risk. An easy, quick and inexpensive alternative technique is described.

Bone Transplantation↗
PubMedDate not supplied

Safe handling of tissue containing radioactive substances.

Patients recently treated with radioactive isotopes may present problems or even hazards during physical examination, operation or autopsy, especially following the use of iodine isotopes and 198Au. 32P is rarely a significant hazard. Contamination of victims of radiation accidents may be a problem initially, but they are usually promptly decontaminated. Guidance of the hospital's radiation safety officer is helpful, particularly with regard to handling of contaminated persons or materials. Long-lived isotopes, such as radium or thorotrast, are usually present in too low concentration to be dangerous. Projected use of artificial hearts powered by plutonium should not be hazardous because of adequate shielding of the source.

Autopsy↗
PubMed1976

Laboratory diagnosis of fungal infections.

Most hospitals do not perform mycologic examinations because laboratory personnel still believe in the "old wives' tale" that mycology is too difficult to do and that the fungi are too infectious to handle. This discussion presents methods suitable for use in laboratories of all sizes. It is hoped that laboratories will take the initiative in offering diagnostic mycology services routinely. Medical mycology can be both a challenge and a rewarding experience to those who choose to become involved.

Culture Media↗
PubMed2003

Powder dispensing robot for sample preparation.

An automated powder dispensing station capable of transferring milligram quantities (1-100 mg) of powder for sample preparation was developed and integrated into a commercial robotic workstation (Zymark Prelude). The system's performance was optimized with respect to vacuum flow rate and powder transfer tube cross sectional area, and shown to possess excellent powder dispensing accuracy (RSD = < or = 0.1% for target weights < or = 15 mg) and precision (RSD = 3.43%) for a vanillin sample. Using the commercial features of the Zymark Prelude workstation (liquid handling, weighing, and vortexing/mixing) and the custom powder dispensing station, multiple sets of analytical calibration standards were prepared and subsequently analyzed by FIA in order to assess the system's robustness for sample preparation.

Drug Industry↗
PubMed1973

The effect of ethylene on (1-14C)glycerol incorporation into phospholipids of etiolated pea stems.

The effect of ethylene (10 p.p.m.) on the rate of incorporation of [1-(14)C]glycerol into phospholipids of etiolated pea stems was studied. After 2-3h treatment with ethylene, incorporation was decreased by 50%. It remained at this value for as long as ethylene was supplied (8h). Handling the plants also caused a temporary decrease in incorporation, which we attribute to the production of endogenous ;wound' ethylene. The percentage decrease in incorporation was the same in four major phospholipid fractions, i.e. phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol.

Carbon Radioisotopes↗
PubMed1999

Multicenter evaluation of the Abbott LCx Mycobacterium tuberculosis ligase chain reaction assay.

Four Australian hospital laboratories evaluated the performance of the Abbott LCx Mycobacterium tuberculosis assay with 2,347 specimens (2,083 respiratory and 264 nonrespiratory specimens) obtained from 1, 411 patients. A total of 152 specimens (6.5%) were culture positive for Mycobacterium tuberculosis complex (MTBC); of these, 79 (52%) were smear positive. After resolution of discrepant data, the overall sensitivity, specificity, and positive and negative predictive values for the LCx assay were 69.7, 99.9, 99.1, and 97.7% respectively. For smear-positive respiratory specimens that were culture positive for MTBC, the values were 98.5, 100, 100, and 98.4%, respectively, while the values for smear-negative respiratory specimens were 41.5, 99.9, 96.4, and 98%, respectively. Relative operating characteristic curves were constructed to demonstrate the relationship between sensitivity and specificity for a range of possible cutoff values in the LCx assay. These graphs suggested that the assay sensitivity for respiratory samples could be increased from 70.2 to 78.6%, while the specificity would be reduced from 99.9 to 99.4% by inclusion of a grey zone (i.e., LCx assay values of between 0.2 and 0.99). An algorithm is presented for the handling of specimens with LCx assay values within this grey zone.

DNA Ligases↗
PubMed1979

An improved method for embedding brain tissue in albumin-gelatin.

A new modification of the Snodgrass-Dorsey (1963) albumin embedding method is described. Formalin fixed brains of various ages of rhesus monkey (Macaca mulatta) were sunk in 10% phosphate buffered formalin which contained 30% sucrose, and then embedded in a 3% gelatin, 30% egg albumin solution which had been centrifuged to ensure uniformity. The albumin-gelatin was hardened in formaldehyde fumes and blocks cut frozen at 10-40 micron. Sections thus prepared can be handled easily and mounted without damage to the tissue. Modifications of conventional cell and fiber stains produce high quality finished slides in which the stained brain tissue is surrounded by a colorless albumin-gelatin matrix.

Animals↗