Non-alcoholic duct destructive chronic pancreatitis: a histological, immunohistochemical and in-situ apoptosis study of 18 cases.

AIMS: To assess retrospectively pancreatic changes in non-alcoholic duct-destructive chronic pancreatitis and to investigate the role of apoptosis in duct destruction. METHODS AND RESULTS: Eighteen patients (mean age 46 years, nine women and nine men) underwent pancreatic resection for suspected pancreatic tumour and were diagnosed as having non-alcoholic duct-destructive chronic pancreatitis. We performed a morphological study either semiquantitatively (fibrosis and inflammation) or quantitatively (CD3+ intraepithelial lymphocytes, M30 and TUNEL+ apoptotic cells). The results were compared with those obtained in 10 cases of chronic alcoholic pancreatitis and nine cases of chronic obstructive pancreatitis. Pancreatic changes were diffuse and heterogeneous in 14 cases, but segmental in four cases. The main pancreatic lesions were ductal epithelial alteration, periductal inflammation and fibrosis. There were no cysts or calcifications. We found a marked increase in mast cells in the infiltrate, a slight increase in TiA1+ lymphocytes and in intraepithelial lymphocytes compared with other types of chronic pancreatitis. No significant increase in ductal apoptosis was observed. CONCLUSION: Non-alcoholic duct-destructive chronic pancreatitis is a well-defined pathological entity, distinct from alcoholic and obstructive chronic pancreatitis. Our results indicate that apoptosis probably does not play a major role in ductal alterations.

Does deep endometriosis infiltrating the uterosacral ligaments present an asymmetric lateral distribution?

OBJECTIVE: To investigate whether deeply infiltrating endometriosis occurs with equal frequency between left and right uterosacral ligaments. DESIGN: Retrospective analysis of consecutive cases. SETTING: Department of gynaecological surgery in a tertiary care university hospital in Paris, France. POPULATION: One hundred and thirty consecutive women with laparoscopic resection of histologically proven deep endometriosis infiltrating the uterosacral ligaments. METHODS: Laterality, intraoperative aspect of the uterosacral ligaments, and associated endometriosis were recorded during laparoscopy. Deep endometriosis infiltrating the uterosacral ligaments was considered as histologically proven in the presence of endometrial glands and stroma. MAIN OUTCOME MEASURE: Frequency of left- and right-sided deep endometriosis infiltrating the uterosacral ligaments. RESULTS: The left uterosacral ligament alone was involved in 69 cases; the right uterosacral ligament alone was involved in 38 cases; both were involved in 23 cases. For patients with unilateral deep endometriosis infiltrating the uterosacral ligaments the observed proportion of endometriosis involving the left uterosacral ligament (69/107, 64.5%) was significantly different from the expected proportion of 50% (chi2 = 8.98; P < 0.01). CONCLUSION: Anatomical differences between left and right hemipelvis and differences in the frequency of ovulation between right and left ovary could explain these results.

The use of free cortisol index for laboratory assessment of pituitary-adrenal function.

We developed a time-resolved-fluoro-immunoassay to measure cortisol binding globulin (CBG) in serum. It is a microtitre plate, solid phase, reagent excess, sandwich assay in which the same polyclonal antiserum is used as a source of capture and labeled antibodies. The results of this assay were shown to be reliable and were fully comparable with those obtained by a commercially available kit. As a reflection of the free cortisol concentration we measured cortisol and CBG concentrations in serum and calculated the Free Cortisol Index (FCI) = [cortisol]serum/[CBG]serum.100. The clinical use of this parameter, as a screening test for disturbances of the pituitary-adrenal axis, was investigated in different groups of subjects: healthy men and women, women using oral contraceptives, pregnant women at term, patients with thyroidal illnesses, patients using anti-epileptic drugs and patients suffering from adrenal diseases. In a number of groups we compared the FCI results with measurements of cortisol in saliva, another parameter used as an estimate of the concentration of free cortisol in blood. Our conclusion is that the FCI, in contrast to a total cortisol measurement alone, can prevent unnecessary further testing.

Temperature effects on free-thyroxine measurements: analytical and clinical consequences.

We compared the analytical and clinical performance of two free-thyroxine (FT4) assays--a solid-phase radioimmunoassay, Spectria, and a time-resolved fluoroimmunoassay, Delfia, both of them two-step methods--with the performance of a direct radioimmunoassay, Nichols, to measure FT4 concentration in equilibrium dialysate of undiluted serum. The three assays showed comparable analytical performance. We tested clinical utility in sera from 135 healthy subjects with and without thyroxine-binding abnormalities and in 61 patients with and without thyroidal illnesses. We found significant differences for FT4 measured by different assays in sera from the same euthyroid patients. To explain the differences, we studied the influence of temperature on performance and calibration. Most important was the neglected fact that the association constant for the binding of thyroxine to thyroxine-binding globulin decreases when the temperature rises from 20 to 37 degrees C, causing a doubling of FT4. The two-step assays, if performed at room temperature without a well-defined calibration, can give misleading FT4 concentrations. This is the case when sera from patients with thyroxine-binding abnormalities are measured against kit standards, made up in normal human sera. If an assay is to reflect the in vivo FT4 concentration at body temperature in all types of samples, it should be performed at body temperature. For practical reasons 37 degrees C is recommended, and reference values should be defined at 37 degrees C. The same might be valid for other free-hormone assays.

Laboratory test form design influences test ordering by general practitioners in The Netherlands.

The effect of listing fewer laboratory tests on the test request form on test-ordering behavior of a group of 47 Dutch general practitioners was studied. The number of laboratory tests ordered by this experimental group during a 12-month period was recorded. The usual, old standard form was used by a control group of 28 general practitioners in a different region. After having used the old, standard form with 178 tests for 5 months, the experimental group received forms listing only 15 hematological and chemical tests plus several urine and feces tests, and space was allowed for "others." A comparison of the experimental and control periods showed that the number of tests ordered monthly in the experimental group was reduced by 18%. When the usual standard form was re-introduced, the general practitioners quickly returned to their old pattern. Results revealed that fixed and often unsound routine influences the use of additional diagnostic procedures. In addition, limiting and restructuring the test-ordering forms may break the routine but does not essentially modify the rationale of test-ordering behavior.

Improved procedure for the second-derivative spectrophotometric analysis of carboxyhaemoglobin.

We describe a modification of the second-derivative spectrophotometric assay for carboxyhaemoglobin in blood. Using the original procedure we have often observed a time-dependent change in the signal leading to unreliable results. By using phosphate-buffered-saline (pH 7.4) instead of ammonia to dissolve the deoxygenating agent (sodium dithionite), we obtained stable and reproducible second-derivative signals from which the percentage carboxyhaemoglobin in the patients' samples can be calculated.

Interpretations of five monoclonal immunoassays of lutropin and follitropin: effects of normalization with WHO standard.

Five mono(oligo)clonal immunometric assays for lutropin (LH) and follitropin (FSH)--bioMérieux, IRE-Medgenix, Serono Diagnostics, Diagnostics Products Corp. (DPC), and LKB--were evaluated in comparison with two polyclonal RIAs (DPC and Amersham). Detection limits varied from 0.04 to 0.32 int. unit/L and 0.06 to 0.86 int. unit/L for LH and FSH, respectively. Intra- and interassay precision (CV) at three concentrations varied from 2.0% to 29.8%, showing that not all kits tested gave acceptable results, especially for LH. Linearity and parallelism were acceptable, except for the DPC FSH kit and the bioMérieux LH kit. High-dose "hook" effects were seen in some kits at LH concentrations of 250 int. units/L, but not in the FSH kits up to concentrations of 350 int. units/L. Reagents in some kits cross-reacted with choriogonadotropin. The clinical validity of the assays was tested in 25 pre- and 25 postmenopausal healthy women and in 66 patients with polycystic ovary disease. In contrast to FSH, LH values varied significantly not only between polyclonal and monoclonal assays but also between the various monoclonal assays, despite the fact that all manufacturers state that their kits are calibrated on the same standards: WHO International Reference Preparation (IRP) 68/40 for LH and 78/549 for FSH. We normalized the results by using new WHO standards: IRP 80/552 for LH and IRP 83/575 for FSH. This decreased significantly the between-kit differences in LH results for individuals. The much-used LH/FSH ratio greater than 3 for diagnosing patients with polycystic ovary disease is not valid when monoclonal assays are used, and is kit-dependent. However, using the normalized results yields a "new" LH/FSH ratio, which is kit-independent and differs significantly between patients and healthy subjects.

Analytical and clinical evaluation of three sensitive thyrotropin assays.

Three sensitive assays (Behring, Organon and LKB-Pharmacia) for measurement of serum thyrotropin (TSH) were evaluated. All three assays showed good precision, sensitivity, linearity and for practical purposes negligible high dose hook effects. The correlations between the assays were excellent, but due to standardisation and/or matrix effects there were incomplete recoveries in two kits and the kits showed systematic differences of up to 20% in the TSH values obtained. The clinical utility of the three assays was investigated in patients with thyroidal and non-thyroidal illnesses and in healthy subjects with thyroxine binding abnormalities. According to the results of the analytical and clinical evaluation, all three kits are acceptable and reliable as first-line thyroid function tests.

Evaluation and modification of a radioimmunoassay for pregnancy-specific beta 1-glycoprotein.

The radioimmunoassay available from Behringwerke for pregnancy-specific beta 1-glycoprotein (SP 1) was tested for its ability to detect pregnancy prior to the first missed menstrual period. It was found that the equine serum, used as solvent for the standards, did not react like human serum. The standard solvent was replaced by bovine serum albumin 50 g/1 and pooled human serum respectively. Equilibrium and sequential incubation procedures were compared. The latter appeared to be more sensitive in the low value range and was therefore more suitable for the early detection of pregnancy. Also, with standards in albumin, the sequential assay was more specific. SP1 could be detected in sera of men and non-pregnant women, using albumin as standard solvent. This could be due to different cross reacting material of the protein matrix, or to the presence of SP1- like material in human sera. The choice of human male serum seemed to be the most practical. It has also been adopted by Behring, and a commercial kit has been prepared.