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PubMedDate not supplied

Management pathways and the surgical diagnosis of tuberculous lymphadenitis: can they be improved? The Bradford experience.

OBJECTIVE: To study the diagnostic pathway for tuberculous lymphadenitis from initial referral to the commencement of antituberculous therapy with the object of identifying potentially avoidable delays. DESIGN: Retrospective case series study. SETTING: District General Hospital, England, UK. SUBJECTS: Thirty-four patients with head and neck tuberculous lymphadenitis diagnosed by a positive histological specimen over the 3-year period 1998-2000. RESULTS: There were no clear pathways for referral and management. There were substantial variations in the management between different specialties and even sometimes within the same specialty. There was evidence of poor communication between various teams. CONCLUSIONS: The study shows that for the management of head and neck tuberculous lymphadenitis, there is a need for a 'hospital' or 'trust' approach. It also reinforces the arguments for a dedicated 'neck lump clinic' with easy access both from the general practitioners and from other specialties within the hospital and for a protocol for handling biopsy specimens from nodes.

Adult↗
PubMed2006

Standard morphologic evaluation of the heart in the laboratory dog and monkey.

The nonrodent species most commonly utilized in preclinical safety studies are the purpose-bred beagle dog and cynomolgus macaque (Macaca fascicularis). Potential effects of a new chemical entity (NCE) on the heart pose serious concerns; consequently in vivo testing is focused on detection of functional alterations as well as morphological changes. Macroscopic and microscopic evaluation of the heart is based on a standard survey of key structures to properly assess presence of spontaneous and potential drug-induced lesions. Evaluation of historical controls to determine type and frequency of background change is valuable, as studies with non-rodent species generally have a small sample size. Archived control dog and monkey data were retrospectively reviewed, including terminal body weight (BW), heart weight (HW), and archival glass slides of heart. Control dogs had minimal background changes that included myxomatous or cartilagenous change in the cardiac skeleton and a variable degree of vacuolation in Purkinje fibers. Control monkey hearts commonly contained inflammatory cell infiltrates, myocyte anisokaryosis, and handling artifacts, while myocyte degeneration, squamous plaques, pigment, and intimal plaques were occasionally observed. These findings highlight the utility of consistently recorded and readily accessible archived control data when attempting to discern background spontaneous changes and artifacts from test-article induced changes.

Animals↗
PubMed2006

Amniotic membrane for ocular surface reconstruction: donor variations and the effect of handling on TGF-beta content.

PURPOSE: Amniotic membrane (AM) transplantation is an accepted procedure in ocular surgery. However, little is known of the interdonor and intradonor variability within the membrane. In addition, the effects of the methods of processing, storage, and preoperative preparation on the membrane are not fully elucidated. The purpose of this study was to use TGF-beta as an example to investigate interdonor and intradonor variability and to determine the effect of "handling " on TGF-beta1 within fresh, processed and stored, and transplantation-ready AM (TRAM). METHODS: Seventeen human AMs, both fresh and handled, were analyzed for TGF-beta1 by real-time polymerase chain reaction, immunohistochemistry, SDS-PAGE, and Western blotting. RESULTS: TGF-beta1 was the highest normalized expressed isoform of TGF-beta in all samples, but it varied between membranes of different donors and at different sites within the same membrane. The highest concentration was noted in the spongy layer. Removal of the spongy layer successfully removed the bulk of TGF-beta1 from TRAM. Latency-associated protein (LAP) and a latent TGF-beta-binding protein (LTBP) were also detected. CONCLUSION: TGF-beta1 is present in various regulatory forms in the AM. A degree of intermembrane and intramembrane variation is modified by handling. Unless a standardized protocol is adopted that delivers a membrane with consistent constituents, clinical outcomes may vary and comparisons may be invalid.

Amnion↗
PubMed2003

Physiologic determinants of endothelin concentrations in human saliva.

BACKGROUND: Salivary endothelin (ET) concentrations have been shown to correlate with disease severity in patients with chronic heart failure (CHF). We undertook the present study to evaluate the stability of salivary ET under different handling conditions to assess its suitability as a biochemical marker in screening, diagnosis, and management of CHF. METHODS: Saliva samples were collected from healthy individuals and/or CHF patients, subjected to different handling conditions, and then stored at -80 degrees C until assayed by an ELISA for ET. RESULTS: Salivary ET concentrations showed a time-dependent increase during storage at room temperature. After 72 h of incubation at room temperature, ET increased approximately 2.8-fold (P = 0.03). Simultaneously, salivary big ET showed a time-dependent 11.2-fold decrease (P <0.0001). This activity was blocked by an ET-converting enzyme (ECE) inhibitor, suggesting that these changes were attributable to ECE-dependent cleavage of endogenous big ET in saliva. Ex vivo conversion was also observed when samples were stored at 4 degrees C, but the magnitude of these changes was markedly smaller (P <0.0001). Posture also affected salivary ET concentrations in CHF patients. With a change from supine to seated rest, salivary ET concentrations increased 1.5- and 1.8-fold after 20 and 40 min, respectively (P = 0.01). With a return to supine rest, salivary ET concentrations returned to baseline concentrations (P = 0.008). CONCLUSIONS: These data suggest that saliva sampling and handling conditions could markedly affect measurement of salivary ET. In particular, care should be taken to minimize ECE-dependent enzymatic conversion of endogenous big ET in saliva.

Adult↗
PubMed2003

Multipurpose robot for automated cycle sequencing.

Paramagnetic beads have the superior advantages of easy separation and resuspension by controlling the magnetic filed. Previously, we have developed Magtration technology to automate paramagnetic bead handling and have built several automated instruments that handle 1-12 samples simultaneously. To achieve more high-throughput sample processing, two types of a 96-arrayed Integrated Magtration Unit (IMU) were developed, one installed with electromagnets and the other with thin rod-shaped magnets made of neodymium. A multipurpose robot (SX-96GC) equipped with the IMU was also developed for fully automatic processing of 96 samples in parallel. The cleanup of dye-terminator sequencing products was performed using the robot installed with the permanent magnet version of IMU. The results had quality comparable to those by the same protocol in manual handling or to those by the conventional protocols. The robot processed 96 samples in a microplate within 30 min. The protocol that can purify 384 samples within 1 h by processing two microplates concurrently was successfully designed.

Equipment Design↗
PubMed1996

Percutaneous needle biopsy and synovial histology.

Percutaneous needle biopsies of synovium are successfully used for diagnosis and investigation of joint disease by an increasing number of groups around the world. This procedure can be done in the office with little morbidity; a large number of samples can minimize the potential limitation of sampling error. Clinical indications for 'imaging the joint' by looking at morphological and other features of the actual tissue include undiagnosed acute or chronic mono- or oligoarthritis, haemarthrosis, suspected deposition diseases, new developments in previous stable disease and less often unexplained polyarthritis. Research into any joint disease can be helped by study of synovium especially using newer immunohistochemical, EM and molecular techniques. This report has reviewed other methods used for obtaining synovium, described the different percutaneous biopsy needles, detailed the methods used for biopsy with the Parker-Pearson needle and described how our group handles tissue so as to obtain maximal impact. The very few side effects of needle biopsy include haemarthrosis and, rarely, needle breakage. Finally, we have provided a brief overview of normal synovium and some aspects of synovium in a variety of joint diseases.

Arthritis↗
PubMed1995

Tissue handling in suspected Creutzfeldt-Jakob disease (CJD) and other human spongiform encephalopathies (prion diseases)

Despite many sensational and intimidating reports in the mass media, transmissible spongiform encephalopathies (prion disease) are not contagious in the usual sense. Successful transmission requires both specific material (an affected individual's tissue, from or adjacent to CNS) and specific modes (mainly penetrating contact with the recipient). Nevertheless, specific safety precautions are mandatory to avoid accidental transmission and to decontaminate any infectivity. Autopsy is essential for definite diagnosis of these disorders. Recommendations are given here for performance of the autopsy, for neuropathology service and appropriate decontamination; they are based on the current literature and on precautions taken in most laboratories with experience in handling tissue from transmissible spongiform encephalopathies. In particular, special care must be taken to avoid penetrating wounds, possible contamination should be kept to a minimum, and potential infectious material must be adequately decontaminated by specific means.

Autopsy↗
PubMed2002

Effects of handling on heat shock protein expression in rainbow trout (Oncorhynchus mykiss).

As part of an effort to validate the use of heat shock proteins (HSPs) as biomarkers of exposure to and effects of contaminants, we evaluated the effect of two handling regimens on the induction of HSP 60 and 70 in rainbow trout (Oncorhynchus mykiss). Fish were acclimated to laboratory conditions for several weeks before the beginning of the experiment. Fish were then captured by net, placed in a cooler for 1 h while being transported in a truck, returned to their original tanks, then sacrificed 6 to 8 h later. Tricaine methane sulfonate (MS-222) was used during different phases of handling to reduce handling stress. Heat-stressed fish were included in the experiment as a positive control. Muscle, liver, gills, and heart were analyzed for HSP 60 and 70 by immunoblotting. We found no effect of any handling regimen on the induction of HSPs. These findings suggest that the capture and transport of fish for environmental monitoring purposes should not interfere with the use of stress proteins as biomarkers.

Animals↗
PubMed2000

Lymph node revealing solution and traditional 10% buffered formaldehyde for detecting lymph nodes in colorectal carcinoma.

BACKGROUND: Cancer stage has been the most important factor in evaluating the prognosis and treatment of colorectal carcinoma, and the presence of lymph node metastasis in the resected specimen plays a key role in serial staging. We evaluated a newly proposed "lymph node revealing solution" (LNRS) that proponents claim is inexpensive, easy, rapid, and innocuous, and that could help to establish the stage of disease more accurately than buffered formaldehyde (formalin). Our study compared traditional 10% formalin and LNRS as second search solutions to determine whether LNRS is a more useful and practical solution than formalin. METHODS: Thirty randomly selected colorectal carcinomas from low anterior resection (LAR) were studied. The specimens were handled routinely on the first day. Then, the entire fat of odd-numbered cases was immersed in three times its volume of LNRS, and the entire fat of even-numbered cases was immersed in 10 times its volume of formalin overnight. The chi-squared test was used to evaluate the results. RESULTS: Of 145 cassettes in the second search by LNRS, seven had no lymph nodes. Of 107 cassettes in the second search by formalin, 18 had no lymph nodes. Totally, 792 lymph nodes were found with an average number of 26.4 per case and 1.53/cm (per centimeter of specimen) in all 30 cases. By the LNRS method, 138 lymph nodes were found in the 15 odd-numbered cases, and by the formalin method, 89 lymph nodes were found in the 15 even-numbered cases. We were able to adjust the stage upward in two cases by LNRS-fixation method. There were no changes in staging in the formalin-fixation method cases. CONCLUSIONS: LNRS showed no statistically significant difference from formalin in the numbers of total lymph nodes, positive lymph nodes, or minute (less than 1 mm) lymph nodes found. Nonetheless, we do believe that LNRS is a potential replacement solution for formalin due to its fast fixation, ease of searching, high identification rate and time-saving procedure.

Colorectal Neoplasms↗
PubMed1998

Handling and evaluation of pediatric renal tumors.

Optimal pathologic evaluation of pediatric renal tumors, which share many features in common with their adult counterparts, also includes some relatively unique considerations, in large part related to specialized classification, grading or staging systems, and ongoing collaborative clinical and biologic studies. Important factors in the gross evaluation, procurement of tissue for special studies, and selection of tissue blocks for light microscopy are briefly reviewed.

Adolescent↗
PubMed2006

Direct MALDI-MS analysis of cardiolipin from rat organs sections.

Cardiolipins (CL) are mitochondria specific lipids. They play a critical role in ATP synthesis mediated by oxidative phosphorylation. Abnormal CL distribution is associated with several disease states. MALDI-MS and MALDI-MS/MS were used to demonstrate in situ analysis and characterization of CL from tissue sections of organs containing high concentrations of mitochondria. Once the experimental parameters were established, a survey of CL distribution in heart, liver, kidney, leg muscle, and testis was undertaken. The major CL specie in the heart muscle, leg muscle, liver, and kidney is the (18:2)(4) CL, while liver and kidney also contain a minor specie, (18:2)(3)/(18:1) CL. The major CL specie in testis is the (16:0)(4) CL. The CL species distribution in various organs appeared to be in agreement with prior reports. Overall, proper matrix selection, tissue section handling, instrument tuning, and the inclusion of cesium ion in matrix ensured successful in situ MALDI-MS and MALDI-MS/MS analysis of CL. Upon modification and standardization, this method could be streamlined for rapid pathological diagnosis with short turnaround time in clinical settings.

Animals↗
PubMed1977

Contained indirect viable-cell membrane immunofluorescence microassay for surface antigen analysis of cells infected with hazardous viruses.

A microtechnique for an indirect viable-cell membrane immunofluorescence titration assay was developed using Friend murine leukemia virus (F-MuLV)-producing cells and monospecific rabbit antisera to F-MuLV structural antigens. The assay was sensitive and displayed little variation within or between assays. Since moderate-risk tumor viruses, such as recently discovered primate oncornaviruses or feline leukemia virus (FeLV), may be hazardous to laboratory personnel, the assay was adapted for containment of cells infected with such viruses. Cells producing gibbon ape lymphoma virus or FeLV were grown in class III containment cabinets and transferred in sealed flasks to a class II laminar-flow cabinet, where the assay was performed. This micromethod not only conserved reagents but also minimized the numbers of moderate-risk tumor virus-infected cells handled at one time. Centrifugation was contained using custom-made devices shown to form a gas-tight seal over microtiter plates. Interspecies reactivity of monospecific rabbit antisera against F-MuLV structural antigen gp71, but not against p12, was demonstrated for surface antigens on FeLV-producing cells.

Animals↗
PubMed2000

Integrated sample collection and handling for drug discovery bioanalysis.

An integrated sample handling process for drug discovery bioanalysis is described. The streamlining of study design, sample collection and automatic bioanalytical sample processing is demonstrated. Specific details for the entire procedure regarding the time saved, ease of automation and integration are defined. Details of sample handling involved a sample collection map, sample collection formatting and volume, dilution schemes for high concentration samples, choice of biological fluid and evaluating the capabilities of two liquid-handling workstations. Numerous comparisons were conducted between the new approaches and the conventional sample handling approaches. The precision and accuracy obtained from the new integrated sample handling process were comparable to those obtained from a conventional approach, as were pharmacokinetic profiles and parameters. This new sampling process greatly improved the efficiency of drug discovery bioanalysis. The integration of pre-clinical protocol design, sample collection and bioanalysis processes was also achieved.

Animals↗
PubMed1995

Anti-HLA antibody screening with extracted platelet HLA antigens by the mixed passive hemagglutination method.

For the detection of anti-HLA antibody in the serum of patients receiving frequent platelet transfusions, we developed a new method in which platelet antigens are extracted into physiological saline containing 3% sucrose and coated as a layer on a U-type Terasaki plate. In the present study, screening of anti-HLA antibodies was conducted with this plate by the mixed passive hemagglutination test although the plate contained both HLA and HPA. The reactivity determined by this method correlated well with antihuman immunoglobulin-lymphocyte cytotoxicity test (AHG-LCT) results (r = 0.963). The plate can be preserved for at least 2 years at -80 degrees C and is easier to handle than the frozen lymphocyte panels used in the LCT test in which lymphocytes must be kept alive. This new method is an alternative way to screen HLA antibodies. In addition, it is expected that this method will be used to screen anti-HPA antibodies.

Blood Platelets↗
PubMed1978

Management of the rape victim.

If a rape crisis center and specialized personnel are not available, the family physician can adequately examine and treat the rape victim as well as collect and properly handle all the required medical evidence. The detailed history has several facets. Laboratory confirmation can be obtained for prior consensual intercourse, sodomy and other special aspects. Treatment is indicated for the prevention of venereal disease and pregnancy. Psychologic support is essential. The physician's examination and testimony can play an important legal role.

Female↗
PubMed2003

Direct tissue analysis using matrix-assisted laser desorption/ionization mass spectrometry: practical aspects of sample preparation.

Practical guidelines for the preparation of tissue sections for direct analysis by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry are presented. Techniques for proper sample handling including tissue storage, sectioning and mounting are described. Emphasis is placed on optimizing matrix parameters such as the type of matrix molecule used, matrix concentration, and solvent composition. Several different techniques for matrix application are illustrated. Optimal instrument parameters and the necessity for advanced data analysis approaches with regards to direct tissue analysis are also discussed.

Animals↗
PubMed2001

Determination of folates in human plasma using hydrophilic interaction chromatography-tandem mass spectrometry.

Folic acid is an essential nutrient, and folate deficiency is associated with a variety of disorders including neural tube defects (during pregnancy) and heart disease. A fast, sensitive, and robust HPLC-tandem mass spectrometry (LC-MS-MS) method was developed for the quantification of free folic acid, tetrahydrofolate, 5'-methyltetrahydrofolate, and 5'-formyltetrahydrofolate in human plasma. Sample preparation required only acetonitrile precipitation of proteins followed by filtration instead of solid-phase extraction or solvent-solvent extraction as in other methods. The rapid and streamlined sample handling procedure minimized degradation of the highly unstable folate species. Hydrophilic interaction chromatography was used for additional sample cleanup on-line, and baseline separation and detection of all four folate species was achieved in less than 30 min. The folate species were detected using negative ion electrospray-tandem mass spectrometry with multiple reaction monitoring of the diagnostic fragment ions of each deprotonated molecule. The predominately organic (hydrophobic) solvent system combined with the microbore flow rate (50 microL/min) used for the chromatography resulted in enhanced electrospray signal response compared to reversed-phase HPLC using a wider bore column. The recovery of all folate species (from spiked plasma) was >97% over a concentration range from 300 pg/L to 12 mg/L with intraday precision (RSD, n = 5) of 3.7-6.5%. Stability studies were carried out for spiked samples in order to define storage and handling conditions. The folic acid limit of quantification (LOQ) in human plasma was 80 pmol/L +/- 10%, and the limit of detection (LOD) was 37.5 pmol/L. The LOQ and LOD for tetrahydrofolate, 5'-methyltetrahydrofolate, and 5'-formyltetrahydrofolate were 1250, 400, and 360 pmol/L of plasma and 425, 165, and 140 pmol/L of plasma, respectively.

Chromatography↗