The determination of delta-5-androstenediol and its sulphate in serum and urine by gas chromatography-mass spectrometry

OBJECTIVES We developed an assay for delta-5-andro-stenediol (adiol) and delta-5-androstenediol-3-sulphate (adiol-3S) in serum and adiol and total adiol sulphate (adiol-S) in urine. DESIGN An analytical procedure using HPLC and gas chromatography-mass spectrometry was devised and tested for its reliability. MEASUREMENTS After addition of deuterated androstenediol as internal standard, serum and urine samples were extracted. Steroid sulphates were hydrolysed. The extracts and hydrolysates were purified on HPLC, adiol was derivatized using heptafluorobutyric anhydride and finally quantified by gas chromatography-mass spectrometry. RESULTS The assay is accurate and reproducible. The coefficient of variation (CV) for the determination of adiol in serum samples is 4% (intra-assay) and 9% (interassay) and for urine samples 3 and 8% respectively. The intra-assay CV for the adiol-3S analyses is 5% for serum and 2% for urine samples while the interassay CV values for adiol-3S are 10% for serum and 7% for urine samples. The recovery of adiol and adiol-3S from serum and urine samples is 97%. CONCLUSIONS The developed assay meets the analytical demands needed for clinical applications.     

Correlation between urinary levels of histamine metabolites in 24-hour urine and morning urine samples of man: influence of histamine-rich food

In this study, we have determined and correlated the excretion of the 2 most important histamine metabolites, N tau-methylhistamine and N tau-methylimidazoleacetic acid, in 24-hour urine and morning urine samples of 14 normal healthy persons. We found no significant difference between morning urine and 24-hour urine samples, provided that the subjects were on a histamine-poor diet for at least 24 hours. We also studied the influence of the consumption of histamine-rich foodstuffs on the reliability of these parameters. The morning urinary N tau-methylhistamine excretion is less affected by histamine-rich food-stuffs and therefore proposed to be the most reliable parameter for endogeneous histamine release.     

CD40 ligand triggered interleukin-6 secretion in multiple myeloma

Previous studies have suggested that interleukin-6 (IL-6) may mediate growth of multiple myeloma (MM) in either an autocrine or paracrine growth mechanism. However, those molecules which can trigger IL-6 secretion either by tumor cells or non-MM marrow cells are not well characterized. In the present study, we have examined the expression and functional significance of CD40 on MM and plasma cell leukemia (PCL) cells and derived cell lines, as well as long-term bone marrow stromal cells (BMSCs) and derived cell lines. CD40 was expressed on the majority of MM cells (> 90%) and BMSCs (> 70%). Triggering via CD40 using NIH3T3 CD40 ligand transfectant (CD40LT) cells increased (> 30%) cell surface CD80, CD18, CD11a, CD11b, and CD11c expression on MM cell lines. Culture with either fresh or paraformaldehyde fixed NIH3T3 CD40LT cells upregulates IL-6 secretion in MM cells and MM-derived cell lines, as well as normal and MM bone marrow mononuclear cells (BMMCs), BMSCs, and BMSC lines; this effect can be specifically blocked by anti- CD40 monoclonal antibody (MoAb). BMMCs and BMSCs from patients with MM secreted significantly more IL-6 than those from healthy donors (n = 3, P < .001); moreover, after stimulation using CD40L, IL-6 secretion was fourfold greater (n = 3, P < .001) from MM BMMCs and BMSCs than from normal BMMCs and BMSCs. Myeloma (CD38+CD45RA-) cells and non-MM (CD38+CD45RA+, CD38-CD45RA+, and CD38-CD45RA-) BMMCs were separated by dual fluorescence cell sorting. The latter secreted fourfold more IL-6 than the former (n = 2, P < .001). Increased IL-6 secretion (up to 28- fold) and proliferation (Stimulation index 10) by CD38+CD45RA-MM cells was triggered by culture with NIH3T3 CD40LT cells. Finally, anti- CD40MoAb partially (30%) blocked tumor cell to BMSC adhesion-induced IL- 6 secretion. These studies support the view that CD40L may trigger IL-6 secretion by both MM cells and BMSCs and that IL-6-mediated autocrine and paracrine growth mechanisms may be possible in MM.     

Optic disc oval ness,refractive error and axial length of Hong Kong Chinese

Myopic crescent, refractive error and axial length were previously investigated in Hong Kong Chinese subjects. The myopic crescent was found to correlate with axial length and myopic refraction. In this study, three groups of Hong Kong Chinese with different degrees of myopia were assessed for optic disc ovalness, refractive error and axial length. The axial length was significantly correlated with the degree of myopia, indicating that the myopia was axial in nature. The regression line shows that 0.44 mm of axial elongation would give about one dioptre of increase in myopia. The elliptical ratio of the optic disc was defined as the maximal disc diameter divided by the minimal disc diameter. All three groups showed an oval disc with vertical axis greater and an increased ovalness for the high myopic group with an elliptical ratio from 1.11 in low myopia to 1.29 in high myopia. There is a small amount (about four degrees) of temporal rotation of this vertical oval orientation, which is independent of the amount of myopia. This result shows an association between axial elongation of the globe and optic disc ovalness, in addition to the previously described temporal myopic crescent. Therefore, in myopic subjects, a vertically oval disc may be associated with a myopic refraction rather than glaucoma.     

乳酸中毒和pH对狗肺血管张力和气体交换功能影响的实验研究

急性代谢性酸中毒对狗血管张力和气体交换功能的影响进行了研究。当滴入乳酸或盐酸造成急性代谢性酸中毒（ｐＨ７．００）时，可引起肺血管收缩，肺动脉压力明显升高，血氧饱和度下降，并且乳酸和盐酸引起的酸中毒对肺动脉平均压影响程度相似，提示肺血管收缩和肺动脉压升高的程度与血中ｐＨ值下降关系密切，而与此时血中乳酸水平的高低无关。     

Short-term growth in children with allergic rhinitis treated with oral antihistamine, depot and intranasal glucocorticosteroids

Short-term growth was studied during the grass pollen season with weekly knemometry in 44 schoolchildren with allergic rhinitis. The design was a randomized, parallel group study. After a four-weeks run-in period, the children were allocated to six weeks' treatment with either a single im injection of methylprednisolone acetate 60 mg at the beginning of the period, intranasal budesonide 200 μg bid (aerosol spray) or terfenadine tablets 60 mg daily. Treatment with methylprednisolone acetate was open, whereas treatment with budesonide and terfenadine was double-blinded. Twelve children in the methylprednisolone acetate group, 11 in the budesonide group and 12 in the terfenadine group completed the study. Compared with the run-in period, treatment with methylprednisolone acetate and budesonide (run-in growth velocities 0.46 and 0.59 mm/week, respectively) was associated with a reduction in mean lower leg growth velocity of 0.28 and 0.54 mm/week ( p <0.01, t = 3.3, 95% confidence interval 0.09–0.47 mm/week; and p < 0.001, t = 6.1, 95% confidence interval 0.34–0.72 mm/ week, respectively). Terfenadine (run-in and treatment mean growth velocity 0.35 and 0.51 mm/week) did not influence lower leg growth significantly. Short-term lower leg growth is suppressed in children with allergic rhinitis treated with intranasal and depot steroids in the doses investigated. No conclusions can be drawn with respect to long-term statural growth. *     

MECHANISMS OF HYPOPHOSPHATAEMIA IN ALCOHOLIC PATIENTS

The aim of our study was to determine the possible pathophysiological mechanisms of hypophosphataemia in a group of 127 patients admitted to hospital for alcohol-related causes. Blood and fresh urine specimens were taken to determine acid-base and electrolyte parameters. Thirty-seven patients (29.1%) had hypophosphataemia (serum phosphorus <0.77 mmol/l) with a range of serum phosphorus of 0.32-0.74 mmol/l. In 17 hypophosphataemic patients inappropriate phosphaturia (FEP04 >20%, TmPO₄/GFR<0.80 mmol/l) was evident, possibly due to hypomagnesaemia, metabolic acidosis, metabolic alkalosis, or a proximal tubular defect in phosphate transport. The causes of hypophosphataemia in the remaining 20 patients were alcohol withdrawal syndrome, respiratory alkalosis and diarrhoea. Patients with hypophosphataemia were more often found to have hypomagnesaemia and respiratory alkalosis than normophosphataemic patients. In conclusion, hypophosphataemia is frequently observed in alcoholic patients due to various pathophysiological mechanisms, such as inappropriate phosphaturia, increased phosphorus entry into cells and increased gastrointestinal loss of phosphate.