[Urinary excretion of neopterin and plasma levels of interferon gamma in patients with multiple myeloma]. / Mocová exkrece neopterinu a plazmatické hladiny interferonu gama u pacientu s mnohocetným myelomem.

Neopterin, a pteridine derivate, is produced and released by lymphocytes and monocytes/macrophages after stimulation by T-cell derived interferon (IFN) gamma. The urinary excretion of neopterin and plasma IFN-gamma levels were measured in 35 patients with multiple myeloma (MM) and 21 patients with other monoclonal gammapathies (MG). Significantly higher excretion of neopterin was found in patients with MM in the time of diagnosis and/or progressive phase of disease (median 963,7 mumol/mol creatinine, range 183,9-2 376,3) comparing the patients in stable phase of MM (median 407,4, range 184,3-826,4), monoclonal gammapathy of undetermined significance (406,2, 226,9-1 689,3) and other MG (452,5, 240, 4-913,6) (p < 0,05). Elevation of urinary neopterin levels preceded changes of other parameters of disease activity by several months, persistent high urinary excretion was associated with refractory disease respectively. The differences of neopterin levels between patients of various clinical stages of MM was observed in newly diagnosed and progressive disease (III. vs. I and II. clinical stage, p < 0.05), but not in stable phase of MM. The plasma concentration of IFN-gama were insignificantly higher in MM than in health controls, however, the difference between all groups of patients was not found. We conclude, that neopterin is a good marker of disease activity in patients with MM. The plasma levels of its regulatory cytokine, IFN-gama, did not correlate with urine neopterin levels in our study.

[Potential side-effects of oral contraception: hirsutism and hypertension]. / Potenciální vedlejsí úcinky perorální antikoncepce: hirzutismus a hypertenze.

Since we observed the development of hirsutism and hypertension in a series of middle-aged women using peroral contraceptive drugs, we tried to ascertain whether there is any correlation between body mass index, blood pressure and urinary elimination of a series of steroid hormones. Although most correlations tested were statistically insignificant, we found a statistically significant correlation between the systolic blood pressure and the sum of 11 beta-hydroxyandrosterone + oxoetiocholanolone + 11 beta-hydroxyetiocholanolone urinary excretion, as assessed by thin-layer chromatography of urinary 17-oxosteroids, using dansylhydrazine as a prelabelling reagent.

[Correlation between urinary excretion of neopterin and levels of thyroid gland hormones in patients with goiter]. / Korelace mezi mocovou exkrecí neopterinu a hladinou hormonu stítné zlázy u nemocných se strumou.

The authors assessed in 18 patients with goitre the urinary excretion of neopterin and the blood level of thyroid hormones. They revealed a significant correlation between urinary neopterin excretion and the thyroxine (T4) and triiodothyronine (T3) blood level. This correlation did not apply in female patients treated with large doses of Carbimazol: these patients had a high neopterin excretion, while the thyroid hormone levels were only slightly elevated. This suggests again that Carbimazol has in addition to an antithyroid effect also an immunomodulating action.

Thin-layer chromatography of urinary 17-oxosteroids using dansylhydrazine as a prelabelling reagent.

This paper describes a modification of a high-performance liquid chromatographic method for measurement of 17-oxosteroids in biological fluids for use with thin-layer chromatography and fluorometric scanning detection. After extraction from urine samples with Separon-C18 microcolumns, free oxosteroids were labelled with dansylhydrazine in acetonitrile-acetic acid and chromatographed on silica gel F-254 plates with the solvent system chloroform-methanol (97:3). Linearity of fluorescence detection (Shimadzu CS-9000 densitometer) was obtained between 30 and 1000 ng.

[Group determination of 17-oxosteroids on thin-layer chromotography]. / Skupinové stanovení 17-oxosteroidu chromatografií na tenké vrstve.

17-oxosteroids from human urine are measured by thin layer chromatography (Merck Kieselgel F-254 plates) with fluorometric detection at 310 nm (densitometer SHIMADZU CS-9000): 11-OH-etiocholanolone, 11-OH-androsterone + oxoetiocholanolone, oxoandrosterone, dehydroepiandrosterone and androsterone. Fluorescence of hydrazone derivatives after the reaction of steroids with dansylhydrazine represents the principle of the method. The recoveries of steroids added to human urine samples are 97-101.3% and the coefficients of variation are 0.78-11.77.

Stroma-free hemoglobin solutions purified by chloroform and pasteurization.

Several approaches to the processing of native stroma-free hemoglobin solutions (SFH) were reconsidered regarding present requirements for SFH production and quality. Treatment of outdated red blood cells (RBCs) with chloroform and/or by pasteurization were evaluated for technical ease, speed and efficacy in removing stromata, phospholipids and non-heme proteins from RBC hemolysates. The influence of both procedures upon spontaneous hemiglobin formation in stored, preferably freeze-dried SFH was compared. Among other analytical methods, sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS PAGE) and isoelectrofocusing were used for mutual comparison of the purification procedures. Pasteurized samples were significantly better purified, more homogeneous but also more susceptible to spontaneous oxidation, probably due to heat inactivation of enzymic scavengers of oxygen radicals. On the other hand, the chloroform-treated, unheated SFH samples were less purified from non-heme proteins, but were more stable. Fructose and sucrose were equally active in protecting SFH from oxidation during freeze-drying. At present, the easy chloroform treatment and freeze-drying of thus purified SFH with fructose of sucrose seems to offer a plausible technological compromise which merits further investigation.

[Is it possible to predict the development of Nelson's syndrome in Cushing's disease after bilateral adrenalectomy?]. / Je mozná predikce vývoje Nelsonova syndromu u Cushingovy choroby po bilaterální adrenalektomii?

A detailed analysis of clinical and laboratory findings in a group of 66 patients with Cushing's disease treated with bilateral adrenalectomy, out of whom 15 developed Nelson's syndrome, gave evidence that Nelson's syndrome mostly affects children and young patients compared to older persons. Bilateral adrenalectomy performed in patients aged over 40 years was never accompanied by Nelson's syndrome. Basal plasma ACTH values prior to adrenalectomy (30.9 +/- 4.53 pmol/l in the absence of Nelson's syndrome and 31.3 +/- 5.41 pmol/l in patients with Nelson's syndrome) as well as the degree of their suppressibility with 8 mg dexamethane (to 18.8 +/- 3.43 pmol/l in Nelson's syndrome and 19.0 +/- 3.44 pmol/l in patients without it) did not provide sufficient evidence for the prediction of later development of Nelson's syndrome. The same is true of the plasma cortisol concentrations. A tendency to a significant plasma ACTH increase after adrenalectomy within a 6-month period, and especially its lesser suppressibility raises a strong suspicion of an incipient development of Nelson's syndrome. These findings will have to be taken into account when deciding on the surgical treatment of Cushing's disease.

[Analysis of estrogen levels in the urine of pregnant women using liquid chromatography]. / Stanovení estrogenu v moci tehotných zen plynovou chromatografií.

The gas chromatography estimation using CHROM IV device enables a simultaneous investigation of oestrone, oestradiol and oestriol in urine. It is possible to use sorption on a solid sorbent PRESEP C 18 TESSEK for steroid extraction from biological material which shortens and simplifies the investigation method. The oestriol results gained by the described technique are compared with the values of total oestrogens estimated by the classical fluorometric method after Ittrich. The linear regression analysis gives the following result: Y = 0.7637x-2.7531, r = 0.9431, p less than 0.001 and confirms the correlation of both methods.

[Increased urinary excretion of neopterin in patients with Graves-Basedow disease]. / Zvýsená exkrece neopterinu mocí u nemocných s Graves-Basedowovou nemocí.

In 12 patients with Graves'-Basedows' disease treated with carbimazole, the mean urinary neopterin, excretion (indicator of stimulation of cellular immunity) was 757.17 +/- 184.17 (95% confidence limit) mumol/mol creatinine with a range of 367-1420. In 12 healthy subjects of matching age and sex who served as controls the mean excretion was 422.0 +/- 96.38 (range 115 to 711, p less than 0.01). It cannot be ruled out that this increase is due to the immunomodulating action of carbimazole.

[Simple chromatographic determination of the neopterin marker in immunopathies]. / Jednoduché chromatografické stanovení neopterinu-markeru imunopatií.

Estimation of neopterin in urine has become part of examination in phenylketonuria, malignities, immunodeficiency incl. AIDS and HIV infection. The authors describe a simple chromatographic method for estimation of neopterin which does not call for highly efficient liquid chromatography nor imported kits. The di- and tetrahydro- forms of neopterin are oxidized with MnO2 and stable neopterin is obtained. The specimen is purified on a Dowex 50WX4 (Fluka) column in a H+ cycle and on PRESEPC18 (TESSEK) columns. Then thin-layer chromatography on Merck Kieselgel 60F254 plates is used in a system of ethyl acetate--isopropanol--1 N NH4OH (35:45:25). Fluorescence is assessed on a densitometer (Shimadzu CS-920 at 367.5 nm). The RF of neopterin is 0.214. The fluorescence is linear within the range of 5-200 ng. The method is sufficiently sensitive also for the estimation of normal neopterin excretion. The authors submit the results of estimations in controls, immunopathies and AIDS.

Separation of urinary steroids by gas-liquid chromatography on packed and mixed-bed columns.

Gas-liquid chromatography (GLC) is one of the most widely used and effective techniques for group analysis of hormonal steroids. Packed columns with liquid phases OV-1, OV-17 and SE-30 are most often used and are most thermally stable. OV-1 methylsiloxane polymer is a non-selective phase employed for separating steroids according to their molecular weights and shapes. The selective phase OV-225 is a methylphenylpolysiloxane containing 75% of phenyl groups: it is used for further differentiation of functional groups of steroid molecules. We mixed nine parts of OV-1 (1%) and one part of OV-225 (3%) both on Gas-Chrom Q (100-120 mesh) and thus obtained a mixed-bed column, which enabled a higher degree of separation of some clinically important steroid metabolites.