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PubMed1987

Montaging electron micrographs.

The preparation of overlapping electron micrographs (particularly from transmission electron microscopy) requires special forethought in planning, exceptional skills in microscopy and photographic techniques, as well as in display preparations which are unique in their handling and execution. In this report, step-by-step instructions are given on specimen preparation, micrography, darkroom printing, and mounting for montage display purposes.

Microscopy, Electron↗
PubMedDate not supplied

Brucellosis: an occupational hazard for medical laboratory personnel. Report of five cases.

Five cases of laboratory-acquired infection with Brucella melitensis are reported. This pathogen is highly contagious when handled in the laboratory. Clinicians should alert technologists when brucellosis is suspected so that specimens are handled under the most stringent safety measures. Serological surveys and instructions of laboratory workers regarding the clinical diversity of the disease are helpful for early recognition of secondary cases. On the other hand, clinicians should consider brucellosis in medical laboratory workers with unexplained signs and symptoms.

Adult↗
PubMed2004

[Histological view of ethics in medicine and handling of residual samples in clinical laboratories].

One of the important ethical issues in clinical laboratory medicine is whether organs and/or specimens should belong to the examinees. Tracing back to ancient Greece, an episode of the death of Asklepios, killed by Zeus to revive the dead, and the great contribution of Hippocrates to medicine including the vow and ethics of medicine, have been described. In the relationship between doctors and patients, the former had been superior to the latter for more than 2400 years, however, the situation has been changing from that to the same position since 1960th, along with the development of bioethics from medical ethics. For the promotion of bioethics, world medical associations have contributed declarations and continuous discussion. The declarations are based on the avoidance of actions detrimental to the life, health, privacy or dignity of examinees. On the medical use of human organs and specimens in relation to human rights, the mind and the body, discussion has continued, however, a consensus on the details has not been reached. A view on the use of residual samples for methodological study, teaching and research in the clinical laboratory was proposed by the Japanese Society of Laboratory Medicine in 2002. Briefly, it included confidentiality of the laboratory staff, responsibility of the laboratory director, the absence of a necessity to obtain consent for the use of residual samples for methodological study when they are made anonymous or pooled, and the recommendation to obtain a judgement by an ethics committee for research use. The background and discussion for the proposal and the current situation on how to obtain consent from patients in Japan are mentioned.

Bioethics↗
PubMed2000

Comparison between three quantitative assays in patients with chronic hepatitis C and their relevance in the prediction of response to therapy.

To compare three quantitative assays measuring viral load in patients with chronic hepatitis C and to determine their value in predicting response to interferon (IFN) therapy, we analysed serum from 896 patients from eight European Centres using QUANTIPLEXtrade mark bDNA, MONITOR AMPLICORtrade mark and SUPERQUANTtrade mark assays. Analyses were performed on the same sample. Viral genotype was assessed using INNO-LiPA HCV II kits. Intercentre variations were observed that were related to the handling of specimens not processed and stored within 6 h of blood sampling. Among sera with optimal handling, a stronger correlation was observed between bDNA and SUPERQUANT (0.806) than between bDNA and MONITOR (0.677) and between MONITOR and SUPERQUANT (0.632). These discrepancies were greatest with genotype 2 (bDNA/SUPERQUANT= 0.772; bDNA/MONITOR=0. 456; SUPERQUANT/MONITOR= 0.299). This correlation was influenced by viraemia level and was better at lower viral loads. The proportion of sera with undetectable viral load was 15% with bDNA, 9.7% with MONITOR and 7.7% with SUPERQUANT. For the three measurements, the best cut-offs of sustained response to IFN treatment were located at their detection threshold. Among patients with viral load below the detection level, a sustained response was observed in 35% tested with bDNA, 38% with MONITOR and 80% with SUPERQUANT. Hence a stronger correlation was observed between bDNA and SUPERQUANT than between either of these assays and MONITOR. SUPERQUANT was the most sensitive assay and this greater sensitivity was associated with a better predictive value of treatment response.

Antiviral Agents↗
PubMed1989

Application of a new, simple method for quantitative collection of 24-hour urines in small laboratory animals: determination of basal excretion of proteins, creatinine, urea, electrolytes, and of free steroids.

The application of a simple method for collection of 24-hour urines in laboratory animals is described and basal excretion values of urinary proteins, creatinine, urea, electrolytes and of free steroids (glucocorticosteroids, DHEA, progesterone, testosterone) in small laboratory animals like the Mongolian gerbil, rat, guinea pig and tree shrew were measured. The major advantage of this method is that cages are identical in size to animal's home cages, are easily constructed and cheap, and that dried and liquid portions of each 24-hour urine are quantitatively removed from aluminium sheets by a simple washing step. This facilitates the monitoring of changes of urinary excretion rates over longer time periods without stressing the animals by handling.

Animals↗
PubMed1984

Sex steroid receptor concentration in breast carcinoma tissue: effect of devascularization during mastectomy.

The accurate determination of sex steroid receptors at the time of mastectomy (MX) for breast carcinoma is important for the determination of subsequent therapy of patients who develop metastases in inaccessible sites. The estrogen (E) and progesterone (P) receptor (R) proteins are heat labile, and measured levels may be vulnerable to alterations once the tumor is devascularized. To evaluate potential differences in ER and PR determinations in tumor tissue acquired at biopsy as compared with tumor from the MX specimen, quantitative analyses of ER (21 patients) and PR (17 patients) were performed on dual samples acquired from the initial biopsy (BX) and the subsequent MX specimen. Receptor concentrations were determined both by sucrose density gradient analysis and titration analysis, and results were expressed as fmol/mg cytosol protein. ER values were classified as receptor-rich (greater than 10 fmol/mg), intermediate (3 to 10 fmol/mg), or receptor-poor (less than 3 fmol/mg); PR values greater than 3 fmol/mg were considered positive. ER BX values were found to be rich or intermediate in 18 patients. When compared with BX values, MX ER values were quantitatively unchanged in 11 patients, lower (MX less than BX) in four patients, and higher in three patients (MX greater than BX). In no patient was the BX ER rich or intermediate and the concomitant MX ER poor. In two patients the PR value was "positive" at BX but "negative" at MX. Accordingly, malignant tissue from a pre-MX biopsy specimen is preferred for receptor analysis although it is apparent that tumor tissue from a properly handled MX specimen is satisfactory for the determination of ER status for clinical purposes.

Adult↗
PubMed2004

Gene expression analysis by real-time reverse transcription polymerase chain reaction: influence of tissue handling.

Factors such as warm ischemia and time at room temperature before tissue treatment may influence the results of mRNA expression analyses on tissue specimens obtained during surgery. We evaluated the effect of these factors on RNA integrity and mRNA expression levels by incubating freshly obtained mouse liver tissue at 25 or 37 degrees C for periods of 0-4 h. Changes in the mRNA expression levels of seven genes, Tbp, Eef1a, Fos, Junb, Myc, Vegf, and Glut2, were determined by real-time reverse transcription-polymerase chain reaction. Incubation at 25 degrees C for up to 4 h only slightly altered (by a factor of less than 2) levels of mRNA for Tbp, Eef1a, Junb, Myc, Vegf, and Glut2. This result is consistent with limited RNA degradation at this temperature. Incubation at 37 degrees C strongly affected the levels of these mRNAs. Four hours of incubation at this temperature resulted in extensive RNA degradation, with mRNA levels falling to 1/10th those before incubation. When relative quantification was performed, i.e., quantification of the target gene transcripts in comparison to an endogenous housekeeping transcript (Tbp or Eef1a), the changes in mRNA levels were reduced to less than 2.5-fold. Fos behaved very differently from the other genes tested on incubation, with Fos mRNA levels increasing considerably following incubation at either 25 or 37 degrees C. Our data suggest that, with the exception of certain genes induced by tissue injury, relative quantification of mRNA, even on degraded RNA samples, can provide a reliable estimate of in vivo mRNA levels.

Animals↗