Re: Winstanley AM, Mikuz G, Debruyne F, Schulman CC, Parkinson MC. Handling and reporting of biopsy and surgical specimens of testicular cancer. Eur Urol 2004;45:564-73.
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Humans↗
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Handling and pathology reporting of radical prostatectomy specimens.
Proper examination of radical prostatectomy (RP) specimens by the pathologists is critical in accurately determining the prediction of patient outcome. The pathology report should include relevant clinical information as well as provide prognostically useful data derived from the evaluation of the RP specimen.
Biopsy, Needle↗
Improved handling of structural fragile cell-biological specimens during electron microscopic preparation by the exchange method.
An exact method of preparation of soft biological specimens for electron microscopic analysis of surface fine structures is described. It allows routine preparations of fragile specimens for SEM and TEM imaging modes. With this procedure physical preparation parameters such as mechanical loads on the specimen surface or changes of temperature are controlled. The wet specimens are premounted in cheap disposable BEEM-containers or glass boats and are constantly kept under liquid in a closed system. The exchange of preparation media is performed continuously and, if necessary, over gradients. For comparative investigations with different EM-modes, at each step of the procedure parts of the specimens may be removed for individual processing. Conventionally prepared critical-point dried specimens are compared to those processed by the exchange technique and preservation of surface fine structures is demonstrated. Shadow-casted clathrin cages and stereo-replicas of virus infected cell cultures are shown in TEM preparations. For SEM, coverslip cell cultures and isolated glomerulus basement membranes are prepared and an additional flat embedding for TEM ultrathin sections is demonstrated.
Basement Membrane↗
[Handling of material, including paraffin-embedded specimens, for diagnosis by electron microscopy].
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Breast Neoplasms↗
Electrocautery: effects on steroid receptors in human breast cancer.
The determination of steroid receptors in human breast cancers has assumed increasing importance over the past several decades. Improper handling of the specimens could affect results obtained. This study details the effects excessive levels of heat that occur with the use of electrocautery can have on steroid receptor quantities and localization. Twelve resected primary and metastatic human breast cancers were analyzed for cytoplasmic and nuclear receptors by biochemical analysis. In addition, steroid binding was determined by direct fluorescent histochemical techniques. To a portion of each resected specimen a Boviec was applied to simulate electrocautery resection. Analysis of the different portions of the same tumor revealed that there was a decrease in measurable cytoplasmic receptor in all cauterized specimens and a concomitant increase in the nuclear receptor. A similar shift in steroid binding was noted in all the specimens analyzed by fluorescent histochemical techniques. The results of this study show that the application of excessive heat to human breast cancers will lead to false negative biochemical steroid receptor determination by shifting the receptors intranuclear.
Breast Neoplasms↗
An immunohistological study of factor VIII related antigen and Kaposi's sarcoma using polyclonal and monoclonal antibodies.
Immunohistochemical techniques have been used to demonstrate Factor VIII related antigen (FVIIIRA) in endothelial cells and to study a variety of tumours including Kaposi's sarcoma. Conflicting reports on the presence of this antigen within the spindle cells of Kaposi's sarcoma have been made. These differences may be due to variations in techniques, in the immunoreagents used and in the method of fixation and handling of the specimen. A major factor may also be the lack of a uniform interpretation of positive peroxidase labelling. This study was confined to paraffin-embedded tissues and compared the results and distribution of FVIIIRA labelling in Kaposi's sarcoma using two commercially available polyclonal antibodies and three monoclonal antibodies. The effects of predigesting the sections with trypsin and protease were also evaluated. Positive labelling of tumour spindle cells was accepted only when it matched that seen in labelled endothelial cells. The polyclonal antibodies provided satisfactory localization of FVIIIRA and gave positive results more frequently than the monoclonal antibodies both with and without enzyme digestion. Endothelial labelling was evident in vessels both within and around the tumour but none was seen in the tumour spindle cells. Most previous studies gave no definition of positive labelling but spindle cells were reported as labelled. However, in those few reports where a definition was given the findings are analogous to those in this report, demonstrating the importance of defining positive labelling.
Adolescent↗
Work-up and diagnosis of pediatric non-Hodgkin's lymphomas.
Non-Hodgkin's lymphomas (NHL) in children and adolescents represent about 10% of childhood cancers. Although the types of NHL commonly seen in this population are relatively limited to lymphoblastic lymphomas, Burkitt's and Burkitt-like lymphomas, and large cell lymphomas, correct diagnosis and classification are essential for optimal therapy. Careful handling of pathologic specimens, along with collection of proper materials for ancillary studies such as immunophenotyping, cytogenetics, or molecular studies, will aid the pathologist in reaching a correct diagnosis. Specific morphologic, immunophenotypic, and genetic features of the commonly seen types of pediatric NHL are described.
Adolescent↗
Simplified fixative medium for direct immunofluorescence in skin biopsies.
The main limitation for the use of direct immunofluorescence in skin biopsies was the necessity for frozen tissue. In 1973 Michel et al reported use of a liquid fixative, consisting of ammonium sulfate, N-ethyl-maleimide, magnesium sulfate, and potassium citrate, for handling skin biopsy specimens. The present study introduces a simplified liquid fixative medium consisting only of ammonium sulfate and saline. Comparisons of direct immunofluorescence findings in biopsy specimens transported in these two transport media and in fresh-frozen specimens revealed only minimal differences between the three different procedures.
Ammonium Sulfate↗
Lassa fever: response to an imported case.
In February, 1976, a Peace Corps worker returned to the United States from Sierra Leone with an undiagnosed illness later recognized as Lassa fever. To assess the risk of transmission and to contain a potential outbreak, we identified 552 contacts as having had exposure to the patient before the start of strict isolation procedures, and maintained intensive surveillance on these contacts for 21 days. At the end of the surveillance period, no illness had developed in contacts. One month later, a serologic survey among 29 of the contacts judged to be at high risk gave no evidence of infection. In response to the importation of this communicable and highly fatal disease, procedures for the isolation of the patient, the identification, surveillance and management of contacts and the handling of laboratory specimens were developed and implemented. These procedures could be adapted to future introductions of highly contagious diseases.
Adult↗
Effects of crystalloid solutions on circulating lactate concentrations: Part 1. Implications for the proper handling of blood specimens obtained from critically ill patients.
OBJECTIVES: a) To test the hypothesis that circulating lactate concentrations are the same in simultaneously collected arterial and central venous blood specimens; b) to test the hypothesis that even small amounts of crystalloid solutions, which are inadequately "cleared" from these indwelling arterial and venous catheters, can lead to clinically important and misleading changes in the measured lactate values. DESIGN: A prospective, multiexperiment study. SETTING: A critical care research laboratory and a 20-bed intensive care unit (ICU). PATIENTS: Three hundred fifty-five patients. INTERVENTIONS: Blood samples were collected. MEASUREMENTS AND MAIN RESULTS: Experiment 1: Simultaneously collected arterial and central venous blood specimens were obtained on 148 occasions from 48 medical ICU patients receiving no lactated Ringer's solution (RL). Arterial and central venous lactate values were nearly identical in these patients. The correlation between the arterial and central venous lactate concentrations was excellent (r2 = .85; p < .0001) and the agreement between the arterial and central venous lactate concentrations was also excellent (bias and precision = 0.04 mmol/L and +/- 0.38 mmol/L, respectively). Experiment 2: Arterial and mixed venous blood samples were obtained from 100 percutaneous transluminal coronary angioplasty (PTCA) and 75 cardiac surgical patients immediately before the performance of these cardiac procedures. We found the central venous lactate concentrations to be higher than arterial lactate values in the cardiac surgical group, and there was a very poor correlation (r2 = .07) between arterial and central venous lactate values in the cardiac surgical group. The correlation between central venous and arterial lactate concentrations in the PTCA patients was excellent (r2 = .84) and similar to the findings of experiment 1. Since the cardiac surgical patients received RL and the PTCA patients received no RL, we speculated that the intravenous infusion of RL in the cardiac surgical group accounted for these discordant findings. To test this speculation, we performed experiments 3 and 4. Experiment 3: In a large bench study, blood specimens were divided into multiple 1-mL aliquot portions, to which 0.01, 0.05, 0.10, 0.50, or 1.0 mL of various crystalloid solutions, containing or not containing RL, were added. In a volume-dependent and linear manner, solutions containing RL increased the circulating lactate concentration from 10% to > 400% of the baseline lactate value. In a volume-dependent and linear fashion, the non-RL crystalloid solutions decreased the lactate concentration by 0 to 66% of the baseline nondiluted lactate concentration. Experiment 4: In 30 different cardiac surgical patients, we simultaneously obtained central venous and arterial blood specimens. Patients this time received no RL, and catheter lines were adequately cleared (removal > 5 mL) of crystalloid solutions. We found a correlation (r2 = .82; p < .0001) that was virtually identical to the findings of experiment 1 and to the findings in the PTCA group of experiment 2. CONCLUSIONS: a) Arterial and central venous lactate concentrations are similar in hemodynamically stable critically ill patients, b) Even small amounts of RL-containing solutions in catheters used for blood sampling may cause false increases in the circulating lactate concentration. c) Even small amounts of non-RL crystalloid solutions in catheters used for blood sampling may falsely decrease circulating lactate values. d) When blood specimens are drawn from indwelling catheters, all crystalloid solutions must be cleared from the line.
Blood Specimen Collection↗
Use of different anticoagulants in test tubes for analysis of blood lactate concentrations: Part 2. Implications for the proper handling of blood specimens obtained from critically ill patients.
OBJECTIVES: a) To test the hypothesis that the measurement of the circulating lactate concentration is influenced by the anticoagulant in the test tube that contains the blood sample; b) to test the hypothesis that the measurement of the circulating lactate concentration is influenced by the tissue used for analysis. DESIGN: A prospective, controlled study. SETTING: A critical care research laboratory, a 20-bed intensive care unit (ICU), and the general wards. SUBJECTS: Twenty-three ICU and ward patients with hyperlactatemia and 19 healthy volunteers. INTERVENTIONS: Blood samples were collected for determination of blood lactate concentration. MEASUREMENTS AND MAIN RESULTS: Venous blood samples (12 mL) were obtained from each of the 19 normal subjects and each 12-mL specimen was evenly divided into six aliquot portions (six test tubes). Experiment 1: Of the six tubes, two tubes were set aside for experiment 2. The other four tubes were used to test four anticoagulants (one anticoagulant per tube). The anticoagulants tested were: sodium heparin; EDTA; lithium heparin; and sodium citrate. Lactate concentrations were analyzed using an ion-selective, amperometric electrode that we have previously validated. There were no statistically significant differences between the lactate concentrations derived from blood samples stored in sodium heparin, EDTA, or lithium heparin (p > .05; n = 19; Student-Newman-Keuls' multiple comparisons test). The lactate concentration of blood stored in sodium citrate, however, was lower than all other anticoagulants (p < .001; n = 19; Student-Newman-Keuls' multiple comparisons). Experiment 2: Of the remaining two test tube samples from each subject, one tube contained sodium heparin and the other tube did not contain an anticoagulant. Each of these two tubes was centrifuged at 50 degrees F (10 degrees C) for 15 mins to obtain plasma and serum samples. Lactate concentrations were measured in the serum and plasma and compared with those concentrations found in whole blood samples from the tube containing sodium heparin from experiment 1. The plasma and serum lactate concentrations were consistently higher than the whole blood lactate values from the same specimen (p < .05; n = 42; Student-Newman-Keuls' multiple comparisons test). Since experiment 1 involved the collection of blood from healthy volunteers with normal lactate concentrations, we chose to investigate whether this discordance between plasma or serum and whole blood was dependent on the lactate concentration. To answer this question, we studied 23 patients with known hyperlactatemia and found that in subjects with a lactate concentration of < 2.2 mmol/L, there was a difference of 0.11 mmol/L in the mean values between plasma and whole blood concentrations (p < .0004; n = 19; paired t-test). In subjects with a lactate concentration of > 2.2 mmol/L, there was a difference of 0.14 mmol/L (p < .0001; n = 23; paired t-test) in the mean values between plasma and whole blood. In all samples at all concentrations, there was no significant difference between serum vs. plasma samples (p > .05; Student-Newman-Keuls' test). CONCLUSIONS: a) Sodium citrate, as an anticoagulant, caused lower lactate concentrations to be measured as compared with heparin or EDTA; b) the measurement of lactate concentrations in plasma or serum samples yields a higher value than the concentration found in the original whole blood specimen.
Anticoagulants↗
Spongiform encephalopathies: the physician's responsibility.
The spongiform encephalopathies encompass several diseases affecting humans and animals. In the United States, the most common of these disorders in humans is Creutzfeldt-Jakob disease. The most frequent manifestations include dementia, pyramidal tract signs, and extrapyramidal movement disorder. Several clinically distinct syndromes can be identified. Often the diagnosis is confused with other forms of dementia, and the only definitive method for establishing the diagnosis is autopsy evaluation of brain tissue. Unfortunately, since the recognition of the infectious etiology of Creutzfeldt-Jakob disease, fear has often unreasonably interfered with clinical care and autopsy evaluation of affected patients. In actuality, because of the low and restricted infectivity of the responsible agent, affected individuals present minimal risks to clinical caretakers, and handling of patient specimens is not dangerous if appropriate precautions are taken. These precautions are well established, and physicians and other health care workers should not refuse care of appropriate evaluation (including autopsy) to individuals with suspected Creutzfeldt-Jakob disease.
Aged↗
Wooden sticks as the source of a pseudoepidemic of infection with Rhizopus microsporus var. rhizopodiformis among immunocompromised patients.
Wooden sticks used to suspend feces obtained for surveillance cultures were found to be the source of Rhizopus microsporus var. rhizopodiformis causing a pseudo-outbreak among 17 immunocompromised patients cared for in three different wards. Nonsterile wooden products should therefore not be used for collecting, handling, and processing specimens for microbiological examination.
Cross Infection↗
ACP Best Practice No 157. Guidelines for the laboratory handling of laryngectomy specimens.
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Dissection↗
[Recommendations for anatamo-pathologic management of soft tissue sarcomas in the adult. Pathologists of the FNCLCC Sarcoma Group (Fédération Nationale des Centres de Lutte Contre le Cancer)].
Adult soft tissue sarcomas comprise a heterogeneous group of rare tumors having a wide range of clinical, gross and histological presentations. Outcome is variable. The pathologist's task is difficult but must be based on a rigorous method. The aim of these recommendations is to describe the main steps of management: collection of clinical information, performing biopsy, handling the fresh specimen, gross examination, histological examination, use of special techniques, definition of the main prognostic factors and key features of the pathology report.
Adult↗
[Management of ductal carcinoma in situ by the pathologist: current questions].
As a consequence of increased screening mammography, ductal carcinoma in situ represents a growing percentage of breast cancer diagnoses. The management of per-cutaneous biopsies as well as wire-localized surgical biopsies is a difficult task for pathologists. In this Article, we describe the diagnostic criteria of these lesions as well as the histopathological characteristics of their two principal differential diagnoses: atypical ductal hyperplasia and micro-infiltrating carcinoma. We also detail the utility of immunohistochemistry in the diagnosis of these lesions. This paper reviews the accepted prognostic factors for ductal carcinoma in situ and describes the important techniques needed to characterize them (grade and extent, status of margins and exclusion of microinvasion). Correct handling of pathological specimens, as detailed here, is one of the main keys for therapeutic success with ductal carcinomas in situ. Indeed, if dealt with adequately, such lesions should almost never affect the patient's long-term survival.
Adult↗
[Handling of surgical specimens containing a non-palpable minute mammary carcinoma (author's transl)].
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Adult↗
[Levels of manganese in biological media: value, samplings, physiopathological variations. Bibliographic review].
The great diversity of methods for manganese analysis in biological materials (serum, plasma, whole blood, urine, spinal fluid and hair) reveals the difficulty in measuring extremely small quantities of this element. An examination of these methods demonstrate that the most used one is flameless atomic absorption spectrometry. This review offers a comparison of the different instrument settings. Authors' recommendations for the collecting of samples are reported in detail. A description of the preliminary steps, the handling of the specimen samples, the preparation of the standards, procedures used, analytical criteria and perturbations is also included. Normal values and their physiological and pathological variations are also described.
Body Fluids↗