A standard microcytotoxicity technique for quantitative analysis of lymphocyte subsets. A comparison with indirect immunofluorescence, evaluated by microscopy or flow cytometry.

A standard complement-dependent microcytotoxicity (CDC) technique was used for quantitative analysis of T-lymphocyte subsets in human peripheral blood and the results compared to those obtained by indirect immunofluorescence microscopy and flow cytometry. The monoclonal antibodies OKT3, OKT4 and OKT8 were used in the CDC method for detection of total-T cells, T-helper and T-suppressor cells respectively. The CDC technique provided reproducible results (CV, 3-7%) correlating well with both immunofluorescence techniques. This observation was valid both for healthy persons (n = 21) and for patients (n = 10) with immunological disorders. The correct antibody dilution, correction for background and the use of eosin staining are considered critical for the usefulness of this technique. The method has several advantages: it is widely used for histocompatibility testing, only simple equipment is necessary, and the amount of monoclonal antibody required per test is small.

A subroutine for rounding of numbers. Program in BASIC and for programmable calculators.

The present programs were developed to compensate for the poor precision of some microcomputer-BASICs, and to avoid printing of the non-significant figures of a numerical result. The program will round any number to a user defined number of significant digits. The subroutine is well suited for statistical programs. It enables the programmer to control the precision and format of numerical output. The program is given in two versions, one in BASIC and one for the programmable calculator TI-59.

[Gel technique, a new sensitive method for the determination of erythrocyte antibodies. Advantages and disadvantages with introduction of a new method]. / Gelteknik, en ny sensitiv metode til påvisning af erytrocytantistof. Fordele og ulemper ved indførelse af ny metode.

A gel technique for the detection of red blood cell antibodies was evaluated for antibody screening. A total of 1872 samples were screened using both gel technique and the conventional serological method, and later 18,292 samples were analysed by the gel technique and results confirmed by conventional methods. In both methods, Anti Human Globulin technique and enzyme technique were applied. The gel system gave false-negative results in 0.2% and false-positive results in 0.9% of the samples. The gel system was more sensitive than conventional technique in the detection of antibodies of clinical importance and less sensitive in detecting antibodies with low incidence of haemolytic transfusion reactions and haemolytic disease of the newborn. The gel technique seems suitable for laboratories with equipment for automatic pipetting of samples and reagents.

[Characteristics of first-time blood donors and their motives. A prospective study among voluntary and unpaid blood donors]. / Karakteristik af førstegangsbloddonorer og tilgrundliggende motiver. En prospektiv undersøgelse blandt frivillige og ubetalte bloddonorer.

We performed a questionnaire study among 413 consecutive voluntary unpaid blood donors being bled for the first time. About 50% were aged below 26 years, 93% were employed or students, and 50% of the donors came from families with other blood donors. The motives to donate blood were investigated. As motives we found: altruistic and humanitarian reasons in about 98%, to obtain a greater well-being in about 60%, tradition at the place of employment or in the family about 35% each. The importance of blood tests taken in connection with the blood letting was analysed. We could not find any support for the hypothesis that blood donors in Denmark enroll as donors to be tested for HIV antibodies.

Rhesus immunization after Rh-incompatible kidney transplantation.

Rh-immunization of a male recipient was observed following the transplantation of two kidneys from Rh-pos donors. Immunosuppression consisted of azathioprine, prednisone and antilymphocyte globulin (1st transplantation) and cyclosporin A and prednisone (2nd transplantation). The immunization was probably caused by the residual blood in the cadaver kidneys. The use of cyclosporin A might increase the risk of Rh-immunization.

Platelet cryopreservation with glycerol, dextran, and mannitol: recovery of 5-hydroxytryptamine uptake and hypotonic stress response.

Human platelets were frozen in 0.5 M glycerol, 0.5 M glycerol + 3% Dextran T40, or 0.5 M glycerol + 5% mannitol. The recovery of active transport of 5-hydroxytryptamine (5-HT) and the hypotonic stress response after freezing were dependent on the rate of cooling: the optimum range of rates was between 12 and 23 degrees C/min. The numerical recovery of cells was independent of cooling rate, but freezing altered the cell-size distribution. The combination of dextran and glycerol was no better than glycerol alone at protecting platelets against freezing damage. Mannitol, however, adversely affected platelet 5-HT uptake, and this was reflected in a low recovery of that activity after freezing platelets in glycerol supplemented with mannitol.

Red cell 2,3-DPG, ATP, and mean cell volume in highly trained athletes. Effect of long-term submaximal exercise.

20 male elite long distance runners were compared to a control group of blood donors to determine the effect of training on red blood cells. The acute effects of exercise on red cells were investigated in 11 of the runners following a race of 15-30 km. The runners had elevated resting values of red cell 2,3-DPG (P less than 0.05) and mean cell volume (P less than 0.01); blood Hb and ATP were not different from concentrations in the control group. The red cell status of the athletes may be explained by an increased proportion of young erythrocytes in runners. No statistically significant changes in red cell 2,3-DPG, ATP, mean cell volume or blood Hb were found post exercise.

Oral testosterone load related to liver function in men with alcoholic liver cirrhosis.

The relation between liver function and an oral testosterone load was examined in 42 consecutive patients with alcoholic liver cirrhosis. Administration of an oral load of 400 mg micronized free testosterone increased the serum concentration of testosterone (range, 31.9-694.4 nmol/l; median, 140.8 nmol/l) in male patients with alcoholic liver cirrhosis to significantly (P less than 0.01) higher levels than in male subjects without liver disease (range, 25.4-106.6 nmol/l; median, 61.5 nmol/l). The increase of testosterone after the load (log delta testosterone) in patients correlated inversely with galactose elimination capacity (r = 0.54; P less than 0.001), serum albumin (r = -0.53; P less than 0.001), plasma factor II + VII + X (r = 0.62; P less than 0.001), indocyanine green clearance (r = -0.71; P less than 0.001), and hepatic blood flow (r = -0.61; P less than 0.01) and correlated directly with wedged-to-free hepatic vein pressure (r = +0.54; P less than 0.01). The increase of testosterone after the load did not correlate significantly with sex hormone-binding globulin (r = +0.35; P greater than 0.05). It is concluded that the hepatic extraction of testosterone is significantly decreased in patients with alcoholic cirrhosis. This decrease seems to be due to decreased liver function, decreasing hepatic blood flow, and increased portosystemic shunting. Oral testosterone loading may therefore be of prognostic significance in patients with alcoholic liver cirrhosis.

Pituitary-testicular function in patients with alcoholic cirrhosis of the liver.

In forty-two patients with alcoholic liver cirrhosis and without recent alcohol ingestion the pituitary-testicular function was studied in an effort to relate the endocrine abnormalities with the degree of liver cell dysfunction, evaluated on a quantitative basis. Compared with values in twenty-one healthy controls, we found significantly elevated serum oestrone, oestradiol, follicle-stimulating hormone, luteinizing hormone and prolactin (P less than 0.01). Serum dehydro-epiandrosterone and dehydroepiandrosterone sulphate were significantly reduced in the cirrhotics (P less than 0.01), whereas serum testosterone was not significantly different from that in the controls. Raised levels of sex-hormone binding globulin were found in 71% (22/31) of the patients (median 8 x 10(-18) mol/l, range 3-17 x 10(-8) mol/l). The incidence of gynaecomastia (38%), cutaneous spiders (67%), testicular atrophy (24%) and reduced axillary hair (71%) was without significant relation to raised levels of sex-hormone binding globulin or progressively reduced liver function. In the presence of clinical or hormonal hypo-gonadism we found evidence of a state of primary hypogonadism together with an inadequate secretion of gonadotropins. The state of hyperoestrogenaemia and the concentration of gonadotropins were significantly correlated to the hepatic synthesis of coagulation factors.

Hypothalamic-pituitary-gonadal function in relation to liver function in men with alcoholic cirrhosis.

Serum concentrations of oestrone, oestradiol, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and sex hormone-binding globulin (SHBG) were significantly (P less than 0.01) raised in men with alcoholic liver cirrhosis (no. = 42) compared with age-matched controls (no. = 20). No significant difference was observed when comparing serum testosterone concentrations. Patients were divided into three groups in accordance with the severity of liver cirrhosis, using biochemical and clinical criteria. Patients with the best-preserved liver function (no. = 11) and patients with moderately affected liver function (no. = 18) had significantly (P less than 0.05) raised serum concentrations of testosterone, FSH, and LH when compared with both controls and patients with severely affected liver function (no. = 13). Serum concentrations of testosterone, FSH, and LH in the latter group showed no significant differences from the controls. Serum concentrations of oestrone and oestradiol were significantly (P less than 0.05) increased in all patient groups, and serum oestrone increased with decreasing liver function. No significant differences were observed concerning SHBG concentrations in the three groups of patients. Dexamethasone suppression did not change the concentration of testosterone significantly, but oestrone and oestradiol concentrations decreased significantly (P less than 0.01) in controls and patients. In patients, but not in controls, a significant (P less than 0.01) increase in FSH and LH concentrations was observed after dexamethasone suppression. The mean percentage increase of FSH and LH was higher the greater the severity of liver cirrhosis.