[Potentialities of laboratory diagnosis in tuberculous meningitis].

The results of examination of 84 patients with tuberculosis of the central nervous system were used to make comparative clinical and laboratory studies. They revealed that lymphocytosis detected in the cerebrospinal fluid (CF), particularly the prevalence of lymphocytes (more than 50%), and decreased levels of chlorides in the cytogram were of value in the comprehensive diagnosis of tuberculous meningitis. The detection of CF Mycobacterium tuberculosis (MBT) (applying the whole currently available set of methods) is an absolute criterion for the diagnosis of tuberculous meningitis (however, with, unfortunately, few number positive results). The detection of mycobacterial DNA, antigens, and tuberculosis antibodies is an impotent component of a diagnostic complex for tuberculous meningitis. The determination of cytosis, protein, glucose, chlorides, lymphocytic subpopulations, soluble gamma-interferon mediators, mycobacteria, DNA, MBT antigens, and tuberculosis antibodies in SF is essential in treating tuberculous meningitis.

Effects of sex and hormonal status on astrocytic basic fibroblast growth factor-2 and tyrosine hydroxylase immunoreactivity after medial forebrain bundle 6-hydroxydopamine lesions of the midbrain dopamine neurons.

We examined astrocytic basic fibroblast growth factor immunoreactivity (FGF-2-IR) and tyrosine hydroxylase immunoreactivity (TH-IR) in the cell-body region of midbrain dopaminergic neurons after unilateral infusions of the neurotoxin 6-hydroxydopamine into the medial forebrain bundle in male and female rats. In addition, to determine whether neonatal exposure to gonadal hormones has consequences on the expression of astrocytic FGF-2 and cell loss in response to injury in adulthood, we studied the effects of these lesions in adult male and female rats that had been exposed or not to testosterone in the neonatal period. In both males and females there was a progressive loss of TH-expressing cells that peaked 5 weeks after the lesions. Females showed less loss of TH-expressing cells than males, but this effect was not estrogen dependent. Lesions led to an increase in expression of astrocytic FGF-2 that was greater in males than in females. Finally, it was found that, regardless of genetic sex, rats exposed to testosterone neonatally showed greater astrocytic FGF-2 expression after lesions than those not exposed, and that among those not exposed to testosterone, estrogen treatment had a modest protective effect. Analysis of behavior and striatal dopamine content showed that the percent of striatal dopamine depletion 14 days after the lesion correlated with the amount of behavioral asymmetry displayed by animals on all tests conducted after lesioning. In groups killed 2 and 5 weeks after the lesion, the amount of behavioral asymmetry correlated with the percent loss of TH-IR cells and with the percent increase in FGF-2-IR cells in the midbrain. These relationships were not evident in groups killed 3 and 7 days after the lesion, possibly because the changes in the number of FGF-2- and TH-IR cells were not fully manifested. The present findings show that hormonal events early in life can alter the response of midbrain dopamine neurons to insult and injury in adult life and suggest that the slow degeneration of these neurons may release signals triggering a sustained activation of adjacent astrocytes which, in turn, may lead to induction of astrocytic FGF-2.

Estimation of accumulated dose of radiation by the method of ESR-spectrometry of dental enamel of mammals.

ESR-spectrometry was used to investigate radiation-induced paramagnetic centers in enamel of mammals: carnivores (polar bear and fox), ungulates (reindeer, European bison, moose), and man. Values at half the microwave power saturation of the radiation signal, P1/2, evaluated at room temperature, was found to range from 16 to 26 mW for animals and man. A new approach to discrimination of the radiation induced signal from the total ESR spectrum of reindeer enamel is proposed. 'Dose-response' dependencies of enamel of different species mammals were measured within the dose range from 0.48 up to 10.08 Gy. Estimations of 'radiosensitivity' enamel of carnivores and ungulates showed good agreement with radiosensitivity enamel of man by ESR method.

[Electron spin resonance determination of copper binding site on Escherichia coli cell membrane]. / Issledovanie metodom elektronnogo paramagnitnogo rezonansa tsentrov medi v mestakh ee sil'nogo cviazyvaniia s tsytoplasmatichesko'i membrano'i bakteri'i Escherichia coli.

The electron-spin relaxation of Escherichia coli cytoplasmic membrane strong binding Cu-centers was investigated by means of the microwave power saturation of the electronic spin resonance signal. It has been established, that copper centres in the strong binding sites of the cytoplasmic membrane E.coli may be represented in the following way: 1) isolate copper complexes with small speed of the spin-lattice relaxation; 2) isolate copper complexes with increased speed of spin-lattice relaxation by means of interaction with rapidly relaxation centres; 3) dipol-binding clasters; 4) ESR-nondetectable at T = 40 K, exchange-binding clasters, which cause increasing of the spin-lattice relaxation speed for isolate copper complexes.

Energy-dependent Complex I-associated ubisemiquinones in submitochondrial particles.

Two distinct species of Complex I-associated ubisemiquinones (SQNf and SQNs) were detected by cryogenic EPR analysis of tightly coupled submitochondrial particles oxidizing NADH or succinate under steady-state conditions. The g = 2.00 signals from both fast-relaxing SQNf (P1/2 = 170 mW at 40 K) and slow-relaxing SQNs (P1/2 = 0.7 mW) are sensitive to uncouplers, rotenone and thermally induced deactivation of Complex I. At higher temperatures the SQNf signal is broadened and only the SQNs signal is seen (P1/2 = 7 mW at 105 K). The spin-spin interaction between SQNf and the iron-sulfur cluster N2 was detected as split peaks of the g parallel 2.5 signal with a coupling constant of 1.65 mT, revealing their mutual distance of 8-11 A. The data obtained are consistent with a model in which N2 and two interacting bound ubisemiquinone species are spatially arranged within the hydrophobic domain of Complex I, participating in the vectorial proton translocation.

[New prospects in assessing the absorbed radiation dose by electron paramagnetic resonance]. / Novye vozmozhnosti v otsenke pogloshcheniia radiatsionnoi dozy metodom élektronnogo paramagnitnogo rezonansa.

Spin-lattice relaxation rates of radiation induced and background centers in dental enamel have been investigated by electron spin resonance techniques. It has been found that these centers differ in the value of the saturation half-power and the saturation process. Based on these experimental results a new approach to discrimination of the radiation induced signal from the total ESR spectrum of dental enamel is proposed. Comparison of the current methods of radiation dose estimation by the ESR analysis has been performed using the new approach.

The source of non-heme iron that binds nitric oxide in cultivated macrophages.

In cultured macrophages (J 774 line) a decrease in iron-sulfur centers (ISC) was not observed after 5 min treatment with nitric oxide (NO) (10(-7) M NO/10(7) cells). The content of these centers was measured by electron spin resonance (ESR) spectroscopy at 16-60 K. However, the appearance of a characteristic ESR signal at g(av) = 2.03 indicated the formation of dinitrosyl iron complex (DNIC) in these cells. These findings suggest that loosely bound non-heme iron (free iron) but not iron from ISC is mainly involved in DNIC formation. ISC might release iron for DNIC formation after their destruction induced by the products of NO oxidation (NO2, N2O3, etc).

[The source of the iron binding with nitric oxide in activated macrophages]. / Ob istochnike zheleza, sviazyvaiushchegosia s oksidom azota v aktivirovannykh makrofagakh.

No decrease in iron-sulphur centers was found in cultured macrophage cells (J774) after the treatment with nitric oxide (10(-7) M NO/10(7) cells) during 5 min. The center content was controlled by the electron spin resonance (ESR) method. The macrophages pretreated with dithionite + methyl viologen showed the formation of dinitrosyl iron complexes (DNIC) with a characteristic ESR signal at g approximately 2.03. The data suggest that loosely bound nonheme iron (free iron) mostly contributes to the formation of these complexes. Iron from iron-containing proteins does not release from these centers under the direct action of nitric oxide. The iron-sulphur centers can be destroyed by the products of nitric oxide oxidation (NO2, N2O3, etc.) as oxidizing and acid agents.

Several forms of chromaffin granule cytochrome b-561 revealed by EPR spectroscopy.

Low-temperature EPR spectra of chromaffin granule membranes from bovine adrenal medulla reveal 3 different signals of the ferric cytochrome b-561. A typical gZ signal of a low-spin cytochrome observed at g approximately 3 is comprised of a high-potential component with gZ = 3.14 and a low-potential one with gZ = 3.11, the low-potential signal showing significantly faster relaxation. In addition, a highly temperature-sensitive heme signal at g = 3.7 is observed which is fully retained in the preparation of granule membranes with b-561 reduced by 50% but disappears upon full reduction of the cytochrome by ascorbate. The signal is strikingly similar to that of the mitochondrial low-potential cytochrome b heme (bL or b-566). The presence of several forms of b-561 in chromaffin granule membranes may provide a structural basis for the transmembrane electron transfer believe to be catalyzed by this hemoprotein.

[Preparation of tumor cells from human lung cancer tissue for the purpose of cloning]. / Poluchenie opukholevykh kletok iz tkanei raka legkogo cheloveka s tsel'iu klonirovaniia.

Various approaches to isolation of tumor cells are analyzed on the basis of examination of 46 human pulmonary tumors of different histologic types. Mechanical treatment alone resulted in the death of the majority of tumor cells. Short trypsin treatment (for 10-15 min in several stages) proved to be the most suitable for disaggregation of small-cell carcinoma, adenocarcinoma, and carcinoid samples. A specific approach has been developed for isolation of cells from squamous-cell carcinoma, consisting in tumor treatment with 0.25 percent trypsin at 4 degrees C for 16-18 h, followed by DNAase, collagenase, and trypsin treatment, and then again trypsin treatment at 37 degrees C for 10-15 min. The suggested approaches permit a harvest of at least 6 x 10(6) tumor cells from 1 g of tissue, with more than 70 percent of these cells viable.

[A comparative evaluation of 2 methods for cloning human lung tumors]. / Sravnitel'naia otsenka dvukh sposobov klonirovaniia opukholei legkogo cheloveka.

Clonogenic capacity of tumor cells from 59 human lung cancer specimens was investigated in Petri dishes (24 specimens) and in capillaries (40 specimens), and parallel cloning was done in 5 cases. The results were as follows: (a) bacterial contamination in capillaries was noted less frequently than in Petri dishes (12.5 vs. 25%); (b) cells did not form colonies in capillaries in 42.5% and in Petri dishes in 25%; (c) colony-forming effectiveness in capillaries was 3.46-fold higher than that in Petri dishes; (d) the amount of tumor cells for analysis in capillaries is 10-fold less than that for analysis in Petri dishes. However, the absolute number of colonies, indispensable for assessment of tumor sensitivity to antitumor action (not less than 15 colonies per capillary or Petri dish) was obtained in 67% in Petri dishes and in 33% only in capillaries.

Coupling site I and the rotenone-sensitive ubisemiquinone in tightly coupled submitochondrial particles.

The rotenone-sensitive g = 2.00 low temperature EPR signal attributed to ubisemiquinone is observed in submitochondrial particles during coupled electron transfer from NADH to oxygen and from succinate to NAD+. The signal is seen only in the presence of oligomycin added to induce the respiratory control (7-9 with NADH and 3-4 with succinate) and it disappears in the presence of uncouplers (CCCP or gramicidin D). No reduction of the iron-sulfur center N-2 in the presence of 20 mM succinate and cyanide is observed, thus suggesting that N-2 is not in equilibrium with the ubiquinone pool. A hypothesis is proposed on delta mu H+ generation coupled with electron transfer between iron-sulfur center N-2 and the ubiquinone pool.

[Several parameters of growth of heterografts of human lung tumor in the subrenal capsule of CBA mice]. / Nekotorye parametry rosta geterotransplantatov opukholei legkogo cheloveka pod kapsuloi pochki myshei linii CBA.

28 species of human lung cancer of the III-IV stages were investigated. The maximal growth of heterotransplants was found for them with the initial area of 1.2-1.5 mm2. The analysis of the data obtained has shown that the samples of lung adenocarcinomas grow more slowly than those of squamous cell and small-cell carcinomas and also of carcinoid.

[Assessment of individual sensitivity to chemotherapy of human pulmonary tumors transplanted to subrenal capsule of CBA mice]. / Otsenka individual'noi chuvstvitel'nosti k khimiopreparatam opukholei legkogo cheloveka pri ikh transplantatsii pod kapsulu pochki myshei linii CBA.

The individual response to chemotherapy was investigated in 17 heterotransplants of human lung tumours of different histogenesis by the subcapsule test. It is established that the subcapsule test is of high informativity only when testing the individual resistance of lung tumours, especially adenocarcinomas, to cytostatics.

[The clonogenic capacity of human lung cancer cells]. / Klonogennaia sposobnost' kletok raka legkogo cheloveka.

Comparative studies of clonogenic ability of twenty four lung cancer samples have been performed in semisolid agar in vitro and in diffusion chambers in vivo. It is concluded that plating efficiency of lung cancer cells varies from 0.00002% to 0.03% in vitro assays and from 0.01% to 0.11% in vivo. A small number of colonies (less than 20) in 15 specimens from 16 does not permit using this method for preclinical individual sensitivity test. The cloning of lung cancer in vitro gives the efficiency more than twenty colonies per a dish in 5 cases of 11. There is some evidence that these models can be used for prediction of treatment response of individual tumors to chemotherapy.

[Evaluation of the proliferative activity of malignant human tumors by an indirect immunofluorescent method using antithymidine antibodies]. / Otsenka proliferativnoi aktivnosti nekotorykh zlokachestvennykh opukholei cheloveka nepriamym immunofluorestsentnym metodom s pomoshch'iu antitimidinovykh antitel.

155 samples of human malignant tumours were studied by the immunofluorescent method and the antithymidine antibodies index-labelling (IL--% of tumour cells in the S-phase). Wide individual variability (0.1-73%) of IL of the studied tumours was shown to be present and to be preserved inside the tumours which were homogeneous by the histological type and developmental stage. A correlation between the labelling index and tumour differentiation in the gastric adenocarcinoma is observed.

[A specific rosette formation method in assessing the immunological status of acute leukemia patients]. / Metod spetsificheskogo rozetkoobrazovaniia v otsenke immunologicheskogo statusa bol'nykh ostrym leikozom.

The paper deals with the evaluation of immunologic vigor in 88 patients suffering acute lymphoblastic leukemia. It involved a modified test of specific rosette formation of blood-circulating lymphocytes and those of the bone marrow with erythrocytes bearing leukemia cell extracts. The highest levels of rosette-forming lymphocytes were registered in the acute period prior to treatment and in recurrence. The said levels decreased gradually following chemoimmunotherapy, falling to nil in complete remission. Patients in whom the treatment had failed revealed no changes in lymphocyte response. The control group (healthy subjects and cases of nonmalignant hematologic pathology) showed levels of rosette-forming lymphocytes similar to those in acute lymphoblastic leukemia patients in remission.