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At least 199 records · Page 11Linked to original sources
PubMed2001

Syndecan-1 (CD138) immunoreactivity in bone marrow biopsies of multiple myeloma: shed syndecan-1 accumulates in fibrotic regions.

Syndecan-1 (CD138) mediates myeloma cell adhesion, and loss of syndecan-1 from the cell surface may contribute to myeloma proliferation and dissemination. Flow cytometry analysis of myeloma cells in bone marrow specimens shows heterogeneity in cell surface syndecan-1 expression. It is not known whether weaker expression correlates with more aggressive disease. However, recent reports suggest that variations in syndecan-1 staining intensity on myeloma cells may be an artifact of specimen handling. In this study, we evaluate syndecan-1 expression in bone marrow biopsy sections from 28 multiple myeloma patients, to elucidate the heterogeneity of syndecan-1 expression in situ. Immunoreactivity for syndecan-1, using the antibody B-B4 (CD138), was found in more than 95% of multiple myeloma cells in 27 of 28 biopsies. However, one biopsy had more than 50% CD138-negative cells and cells with weak CD138 expression were identified in the majority of cases. Loss of syndecan-1 did not appear to relate to myeloma cell differentiation. In addition, syndecan-1 was detected on intravascular and intrasinusoidal myeloma cells suggesting that loss of syndecan-1 may not be required for extramedullary dissemination. Bone marrow biopsies from nine additional patients, with variable CD138 staining intensity on myeloma cells as determined by flow cytometry, were studied by immunohistochemistry. The heterogeneous CD138 expression was confirmed in situ, with weakly positive cells concentrated in areas of reticulin fibrosis. These cells had a disrupted pattern of membrane staining in contrast to the strong linear membrane staining seen in the other multiple myeloma cells. In addition, the fibrotic stroma stained intensely for syndecan-1. Accumulation of syndecan-1 within the extracellular matrix of the marrow likely is derived by shedding of the molecule from the surface of myeloma cells. Because syndecan-1 can act to regulate the activity of heparan-binding growth factors, these reservoirs of syndecan-1 may play a critical role in promoting myeloma pathogenesis, or in regeneration of the tumor after chemotherapy.

Biopsy↗
PubMed2000

A simplified method of cleaning ixodid ticks for microscopy.

The cleaning of ixodid ticks for microscopy can be achieved quickly and efficiently using a combination of a wax solvent and an ultrasonic cleaner. The technique involves minimum handling of specimens, produces no detectable damage and is suitable for cleaning many specimens at the same time.

Acetone↗
PubMed1982

Autotransfusion of drained mediastinal blood.

The Sorenson equipment for transfusion of drained mediastinal blood has been widely used in the United States. We have tested its use postoperatively in 10 patients, assessing its ease of use from the medical and nursing views as well as analyzing the blood constituents which would be infused into the patients. The hemoglobin content of the infused drainage averaged 6 g/dl compared with 12 g/dl in the patients' blood. Platelet counts were very low, averaging 15 X 10(9)/l compared with levels of 110 X 10(9)/l for the patient. Platelet aggregation in response to ADP stimuli was assessed and was grossly deficient in the collected blood showing that the platelets that were present were not capable of aggregating. Protein and electrolyte levels were substantially the same in both series of specimens. Handling difficulties with the equipment together with these rather disappointing hematological results suggest that the equipment is only useful where excessive postoperative bleeding is encountered or where there is particular need to conserve red blood cells.

Blood Cell Count↗
PubMed1987

Asymptomatic blood methanol in emergency room patients.

Over a four-month period, methanol was found in the blood of 18 patients among 687 sequential emergency room admissions screened for alcohols by gas chromatography. In the patients with positive results, blood ethanol ranged from 6 to 570 mg/dL (1.3-123.7 mmol/L), blood methanol from 2.3 to 4.0 mg/dL (0.72-1.25 mmol/L). Methanol exposure during preparation of the sampling site or in the course of specimen handling, ingestion of denatured alcohol, as well as methanol production from the metabolism of aspartame are ruled out as causes for these findings. The authors conclude that endogenous methanol production is the probable major cause, while methanol as a fermentation congener may be a contributory minor cause.

Alcoholism↗
PubMedDate not supplied

Determination of chlorpyrifos, chlorpyrifos oxon, and 3,5,6-trichloro-2-pyridinol in rat and human blood.

Analytical methods to quantitate chlorpyrifos and two potential metabolites, chlorpyrifos oxon (oxon) and 3,5,6-trichloro-2-pyridinol (TCP), in human and rat blood are described. Chlorpyrifos and the oxon were extracted simultaneously with a methanol/hexane mixture from 0.5 mL blood that was deactivated with an acidic salt solution. The extract was then concentrated and analyzed by negative-ion chemical ionization gas chromatography-mass spectrometry (NCI-GC-MS). TCP was extracted from a separate 0.1-mL aliquot of blood, also deactivated by the addition of acid. The t-butyldimethylsilyl derivative of TCP was formed using MTBSTFA, and the analysis was performed by NCI-GC-MS. Stable isotope analogues of chlorpyrifos (-13C2-15N), oxon (-13C2-15N), and TCP (-13C2) were used as internal standards. Oxon was observed to partially degrade to TCP during the sample analysis. Accurate oxon and TCP measurements were obtained with the use of oxon-13C2-15N, TCP-13C2, and TCP-13C2-15N internal standards, which compensated for both the degradation of oxon and the formation of artifactual TCP during analysis. The limits of quantitation were 1 ng/mL blood for both chlorpyrifos and oxon and 10 ng/mL for TCP. Calibration curves were linear over the concentration range of 2.5-2500 ng/mL solvent for chlorpyrifos and oxon and between 5 and 1060 ng/mL solvent for TCP. Taking concentration factors and extraction efficiencies into account, these linear ranges represent blood concentrations of approximately 0.3-300 ng/mL blood for chlorpyrifos and the oxon and 6-1300 ng/mL blood for TCP. The lowest spike level for chlorpyrifos and the oxon was 1 ng/mL blood, and the lowest spike level for TCP was 10 ng/mL blood. Recoveries from rat blood were as follows: 106-119% for chlorpyrifos, 94-104% for oxon, and 85-102% for TCP. In addition, chlorpyrifos and oxon were incubated with rat and human blood for various time intervals before deactivation to determine precautions that needed to be taken when collecting and handling specimens. No change in chlorpyrifos concentration was observed in rat blood up to 180 min at 37 degrees C. In contrast, the oxon was rapidly hydrolyzed to TCP in both rat (t 1/2 approximately 10 s) and human (t 1/2 approximately 55 s) blood held at 37 degrees C. The hydrolysis rate for the oxon was independent of whether a rat had been administered chlorpyrifos previously, the initial oxon concentration, the presence of chlorpyrifos, and the age or gender of the human volunteers. These results suggest rapid sample preparation is critical for accurate determinations of the oxon metabolite of chlorpyrifos. These methods provide excellent tools for use in chlorpyrifos pharmacokinetic modeling studies.

Adult↗
PubMed1986

Use of monoclonal antiestrogen receptor antibody to evaluate estrogen receptor content in fine needle aspiration breast biopsies.

A monoclonal antibody prepared against estrogen receptor has been shown to be highly specific and sensitive for the detection and quantification of estrogen receptor in human breast lesions using immunohistochemical methods. A semiquantitative relationship has been shown between the intensity of staining and biochemical receptor analysis. To evaluate the usefulness of this technique in fine needle aspiration biopsies, 41 cases of breast cancer were studied. Nuclear localization of receptor antibody PAP complex was observed. Comparison of the immunocytochemical analysis of the aspiration cytologic specimens to the biochemical analysis of the excised tumor mass revealed a sensitivity and specificity of 80% and 89%, respectively. There was a significant semiquantitative correlation between the methods. These results extend the observation that antireceptor monoclonal antibody in immunohistochemical analysis is an effective tool in the evaluation of estrogen receptor content in human breast lesions and with careful specimen handling and proper controls may be extended to the evaluation of fine needle aspiration biopsies.

Antibodies, Monoclonal↗
PubMed2004

Sentinel node biopsy: ALARA and other considerations.

For a majority of solid tumors, the most powerful and predictive prognostic factor is the status of the regional lymph nodes. Sentinel lymph node sampling continues to gain in popularity as patients and their physicians seek to avoid the potential morbidity associated with standard axillary node dissection. Lymphoscintigraphy, one of the recently explored techniques of lymphatic mapping, involves pre-operative intradermal or subcutaneous administration of a radiopharmaceutical. While this approach is gaining widely spread acceptance, there is still a lack of consensus on which radiopharmaceutical agent has the most ideal properties. By far, the most commonly used agents are 99mTc labeled colloids, but other agents are also used clinically and are under investigation or development worldwide. A number of other clinical, technical, dosimetric, and logistical considerations regarding this procedure remain. They include questions such as who should be performing the procedure, what precautions to take during surgery, how to better isolate "hot" nodes and thus improve the efficacy of determining metastases to the draining lymph node, what precautions to take when handling surgical specimens, etc. There is clearly a need to review as low as reasonably achievable considerations and other issues that arise as this technique evolves and finds its role in the evaluation of various types of cancers. This paper, based on our own experiences and those of others, fills this gap.

Humans↗
PubMedDate not supplied

Development of a polymerase chain reaction assay for the detection of Haemophilus ducreyi.

BACKGROUND AND OBJECTIVES: Haemophilus ducreyi, the causative agent of chancroid is a fastidious organism difficult to culture and identify. Consequently, culture is an insensitive method for diagnosis. The polymerase chain reaction (PCR) offers a sensitive and specific nonculture method for the detection of bacterial pathogens. STUDY DESIGN: A polymerase chain reaction (PCR) assay was developed to detect the presence of Haemophilus ducreyi. A pair of primers was selected from sequences of an anonymous fragment of DNA cloned from H. ducreyi. The primers were tested in amplification reactions with both purified DNA and lysed organisms for their ability to detect H. ducreyi, and with DNA from a variety of different bacteria for their specificity. The utility of the primers for the detection of H. ducreyi in samples taken from genital ulcers was also tested and compared with culture. RESULTS: PCR was positive for 62% of the specimens that were culture-positive, however, PCR was also positive for 49% of the culture-negative specimens. Comparison of specimens that were dark field positive for T. pallidum and PCR-positive with those that were culture-positive for herpes simplex virus and PCR positive suggested that PCR was giving true and not random positive results. Additional studies demonstrated that the failure of PCR to detect H. ducreyi in all of the culture-positive specimens probably resulted from inhibitors of the Taq DNA polymerase that were present in the nucleic acids extracted from the clinical specimen. CONCLUSION: PCR should be a useful method for the detection of H. ducreyi in genital lesions, especially where culture sensitivity is poor. However, the presence of unidentified Taq polymerase inhibitors in some ulcers specimens require the development of improved methods for specimen handling.

Chancroid↗
PubMed1999

Fine needle aspiration biopsy of the liver.

The liver is a commonly targeted deep-seated organ in which fine-needle aspiration biopsy (FNAB) is performed. By far, the most frequently encountered lesions on FNAB of the liver are metastases; however, a wide variety of other lesions have been described and are likely to be encountered. Because of this wide variety, attention to architectural as well as cytologic features on smears and cell block preparations, ancillary techniques, and special studies are often required. In the setting of proper specimen handling and judicious use of ancillary procedures, FNAB of hepatic lesions can be an accurate diagnostic procedure.

Biopsy, Needle↗
PubMed2000

Recent developments in stereotactic breast biopsy methodologies: an update for the surgical pathologist.

Recently, there have been a number of new devices introduced for stereotactic biopsy of nonpalpable, mammographically detected lesions. The vacuum-assisted core biopsy (VACB) (Minimal Invasive Breast Biopsy (MIBB), U.S. Surgical, Norwalk, CT; Mammotome, Biopsys Medical, Cincinnati, OH) obtains multiple tissue cores (11-gauge) in a circumferential manner around the biopsy probe, inserted under stereotactic guidance. It provides more complete sampling of mammographic lesions than the conventional 14-gauge stereotactic core biopsy, reducing the number of unsatisfactory biopsies. The advanced breast biopsy instrumentation (ABBI) (United States Surgical Corporation, Norwalk, CT) system utilizes stereotactic technique and an oscillating blade-cutting mechanism to obtain a single large diameter (5 mm to 20 mm) tissue core, with the aim of obtaining an intact lesion in its entirety for histologic assessment. Its potential as a treatment option is still under investigation. Suggested protocols for specimen handling are presented together with a review of the recent literature. Close liaison with radiologists and surgeons performing these biopsies will allow the collection of further outcome data to evaluate the strengths and weaknesses of each technique.

Biopsy, Needle↗
PubMed2006

PCR evaluation of false-positive signals from two automated blood-culture systems.

Rapid detection of micro-organisms from blood is one of the most critical functions of a diagnostic microbiology laboratory. Automated blood-culture systems reduce the time needed to detect positive cultures, and reduce specimen handling. The false-positive rate of such systems is 1-10%. In this study, the presence of pathogens in 'false-positive' bottles obtained from BACTEC 9050 (Becton Dickinson) and BacT/Alert (Biomérieux) systems was investigated by eubacterial and fungal PCR. A total of 169 subculture-negative aerobic blood-culture bottles (104 BacT/Alert and 65 BACTEC) were evaluated. Both fungal and eubacterial PCRs were negative for all BACTEC bottles. Fungal PCR was also negative for the BacT/Alert system, but 10 bottles (9.6%) gave positive results by eubacterial PCR. Sequence analysis of the positive PCR amplicons indicated the presence of the following bacteria (number of isolates in parentheses): Pasteurella multocida (1), Staphylococcus epidermidis (2), Staphylococcus hominis (1), Micrococcus sp. (1), Streptococcus pneumoniae (1), Corynebacterium spp. (2), Brachibacterium sp. (1) and Arthrobacter/Rothia sp. (1). Antibiotic usage by the patients may be responsible for the inability of the laboratory to grow these bacteria on subcultures. For patients with more than one false-positive bottle, molecular methods can be used to evaluate the microbial DNA in these bottles. False positives from the BACTEC system may be due to elevated patient leukocyte counts or the high sensitivity of the system to background increases in CO(2) concentration.

Adult↗
PubMed1996

United Kingdom national audit of breast fine needle aspiration cytology in 1990-91--organization and level of activity.

A survey amongst members of the British Society for Clinical Cytology (BSCC) was performed in 1990 and 1991 into their experience of fine needle aspiration (FNA) cytology of the breast. Here we present the findings of that part of the survey covering organizational aspects of the service and methods of specimen handling. The replies demonstrate differences in approach to many of the topics covered. There is a preference for the examination of direct smears (rather than cytospins) with air dried Giemsa staining and wet fixation Papanicolaou staining finding nearly equal favour. Some perceived problems were highlighted, including availability and funding of training and quality assurance as well as facilities for liaison between pathologists, clinicians and radiologists.

Biopsy, Needle↗
PubMed1977

A simple low-cost critical point dryer with continuous flow dehydration attachment.

A simple low-cost critical point dryer with continuous flow dehydration attachment is described. By using the critical point pressure chamber for all fluid exchanges, loss of material and air-drying artefacts due to specimen handling are reduced. An operating schedule that allows the optimum path around the critical point process to be followed is proposed.

Cells, Cultured↗
PubMed1981

Preparation and assessment of frozen-hydrated sections of mammalian tissue for electron microscopy and X-ray microprobe analysis.

A system is described for preparing and examining frozen-hydrated sections of mammalian tissue, using commercially available equipment. An essential part of the system is a liquid nitrogen cooled vacuum transfer system. Minor modifications were required to interface the various manufacturers' equipment, and also to facilitate handling of specimens in the cryoultramicrotome. A variety of criteria indicates that our sections are truly frozen-hydrated, but negligible image contrast in the sections is a serious problem both for morphological work and X-ray microanalysis.

Animals↗
PubMed1982

A simple method for the handling and orientation of small specimens for electron microscopy.

Cyanoacrylic glue (Eastman 910) was used to affix small pieces of nasal scrapings to lens paper immediately before fixation in the glutaraldehyde. The lens paper not only served to hold specimens together so that they were not lost during tissue processing, but also functioned as a 'landmark' for the specimens, so that specimens could be oriented in a specific manner during embedding and subsequent sectioning.

Adhesives↗
PubMed1994

A simple method for processing individual oocytes and embryos for electron microscopy.

A simple method for handling individual specimens that must be processed either for scanning or transmission electron microscopy studies is described. For scanning microscope processing, dehydration is carried out with samples enclosed in small cages made from TAAB capsules in which top and bottom are substituted by plankton nets, and for transmission electron microscopy, samples are preembedded in agarose. This procedure significantly reduces mouth pipetting, dissecting microscope observations, is less labour intensive and, most importantly, reduces sample loss.

Animals↗
PubMed1997

Effects of delayed serum separation and long-term storage on the measurement of thyroid hormones in equine blood samples.

Studies were conducted to determine the effects of delaying the separation of serum from the clot and of long-term storage of serum samples on the measurement of thyroid hormones in blood from horses using a fluorescence polarization immunoassay. The measured concentrations of T3 and T4 were not affected by leaving serum on the clot for as long as 24 hours at room temperatures. Storage of serum for 19 to 22 months at -20 degrees C resulted in significant increases of measured T4, but not T3. These studies support previous work demonstrating that thyroid hormones are resistant to degradation, immunologically stable, and reasonably insensitive to potential problems of routine specimen handling when measured with an immunoassay.

Journal Article↗
PubMed1984

Antibodies to the core protein of lymphadenopathy-associated virus (LAV) in patients with AIDS.

Lymphadenopathy-associated virus ( LAV ), a human T- lymphotrophic retrovirus isolated from a homosexual man with lymphadenopathy, has been causally associated with acquired immunodeficiency syndrome (AIDS). A sensitive and specific radioimmunoprecipitation test was developed for the detection of antibodies to the major core protein of LAV , p25 (molecular weight 25,000). Antibody to LAV p25 was found in the serum of 51 of 125 AIDS patients, 81 of 113 patients with lymphadenopathy syndrome, 0 of 70 workers at the Centers for Disease Control (some of whom had handled specimens from AIDS patients), and 0 of 189 random blood donors. Of a group of 100 homosexual men from San Francisco whose serum was obtained in 1978, only one had antibody to LAV p25; in contrast, of a group of 50 homosexual men in the same community whose serum was obtained in 1984, 12 had antibodies to LAV p25.

Acquired Immunodeficiency Syndrome↗