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Cell-mediated immunity (CMI) response to different antigens was examined in healthy women, in patients with cervical precancerous lesions, and in patients with cervical cancer. Cervical lesions were diagnosed by cytological (PAP) smears, from examination by colposcopy, and from "punch" biopsy material by histology. CMI response is related to specific processes in healthy and cancer cells. CMI was investigated by leukocyte adherence inhibition (LAI) assay using specific antigen (prepared from cervical carcinoma tissue) and non specific antigen (prepared from blood of mice infected by LDH--lactate dehydrogenase--virus). The CMI responses of healthy women and cancer patients to the antigens used are different: the majority of T lymphocytes display adherence and non adherence, respectively (but the CMI responses elicited by the antigens are not equal and small quantitative differences are observed). Regardless of the CIN (cervical intraepithelial neoplasia) grades, CMI responses correspond either to healthy women or to cervical carcinoma patients (at about similar ratio of cases in all the CIN groups). Effect of non specific antigen suggests that cervical carcinoma transformation may be connected with reduction of mitochondrial activity similar to processes in LDH virus infection.

We observed the immunological answer to antigens obtained from the human malignant breast tumor and from the blood of inbred mice strain C3H/H2K infected by LDH virus. We compared the modified ELISA method for humoral immunity with the leukocyte adherence inhibition (LAI) assay for cell-mediated immnunity. The modified ELISA method is suitable for early diagnosing and monitoring antibodies in a malignant breast tumor simultaneously with senological examinations which include mammography and clinical examinations, because the antibodies are determined in a high number of samples by single application.

T lymphocytes taken from healthy humans and cancer patients before and after medical treatment were exposed to the magnetic field 0.1 and 0.05 mT to study response of the cell-mediated immunity. Leukocyte adherence, which is considered to correlate with the cell-mediated immunity, was measured using an in vitro technique—leukocyte adherence inhibition (LAI) assay. Exposure to the magnetic field increases adherence of T lymphocytes especially those from cancer patients before medical treatment. The effects of exposure to the magnetic field 0.1 and 0.05 mT are similar to those of greater magnetic fields in the range of 0.5 to 10 mT. The effects of the AC and DC magnetic fields 0.05 mT do not display large differences attributable to the magnetic field.

Exposure of T lymphocytes to an external 50 Hz and 0.5 mT magnetic field was investigated in vitro using leukocyte adherence inhibition (LAI) assay which is a measure of cell-mediated immunity. Adherence of T lymphocytes taken from healthy humans and from cancer patients before and after medical treatment is enhanced after 1 h exposure to the magnetic field. The experimental findings for the magnetic field 0.5 mT are compared with published data for 1 and 10 mT. The results are consistent with suggestions of magnetic field effects on immune function in humans.

Previous work indicated that hyperstimulation of muscarinic receptors brings about profound changes not only in the density of the muscarinic receptors, but also of the beta-adrenoceptors in rat heart atria in vivo. We have now investigated whether a similar receptor cross-regulation occurs in cardiomyocytes in vitro. Cardiomyocytes from 3-4 day old rats were exposed to chemical agents on days 5-6 in culture. Densities of muscarinic and beta-adrenergic receptors were measured according to the binding of N-[3H]methylscopolamine and [ H]CGP 12177, respectively, to cell surface membranes and cell homogenates. Exposure of cells to the muscarinic agonist carbachol (1 mmol/l) brought about a profound decrease in the number of muscarinic receptors. The number of beta-adrenoceptors displayed biphasic changes, being augmented after 24 h (by 20-45% on the cell surface and by 29% in the homogenate) and diminished after 48 h and 72 h (after 48 h, decrease by 44-75% on the cell surface and by 36% in the homogenate). These effects of carbachol were not prevented by dimethylaminopropyl-bis-indolylmaleimide, the inhibitor of protein kinase C. Exposure of cells to the beta-adrenoceptor agonist isoprenaline (0.1 mmol/l) strongly diminished the number of beta-adrenoceptors on the cell surface and in the homogenate. The density of muscarinic receptors on the cell surface was diminished by 24-43% after 24 h exposure to isoprenaline and unchanged after 48 h, whereas the concentration of muscarinic receptors in the homogenate was unchanged after 24 h and increased by 20% after 48 h. The isoprenaline-induced decrease in the density of cell surface muscarinic receptors could not be simulated by forskolin and was not abolished by the protein kinase A inhibitors Rp-cAMPS and HA-1004. Dibutyryl cyclic AMP diminished the density of cell surface muscarinic receptors more than that of the beta-adrenergic receptors. Our data reveal a novel phenomenon of a biphasic change (an increase followed by a loss) in the density of beta-adrenoceptors during exposure of cardiocytes to carbachol. Activation of beta-adrenoceptors brings about less conspicuous changes in the density of muscarinic receptors. The observed phenomena of receptor cross-regulation cannot be explained by simple activations of protein kinases A and C.

The cluster formation of CD4Ly cells (a subpopulation of T lymphocytes) in suspension was examined. CD4Ly cells were prepared from blood taken from cancer patients and from healthy counterparts (blood donors). CD4Ly cells taken from cancer patients formed tightly bound clusters in suspension with corresponding tumour antigen or LDH (lactic dehydrogenase) virus antigen. Cluster formation was a rare event in suspensions with CD4Ly cells taken from healthy humans. Cluster formation coincides with decreased adherence of CD4Ly to solid state surfaces and suggests strong mutual interaction between CD4Ly. A tentative theoretical explanation based on the Frohlich coherent vibration hypothesis is given.

Chronic treatment with organophosphate inhibitors of cholinesterases is known to bring about down-regulation of muscarinic acetylcholine receptors in the heart while its effect on the functionally antagonistic beta-adrenergic receptors is not known. We describe experiments in which rats were exposed to daily injections of diisopropylflurophosphate (DFP) and the density of muscarinic and beta-adrenergic binding sites in their cardiac atria was measured according to the binding of (3H)quinuclidinyl benzilate ((3H)QNB) and (-)-4-(3-tert-butylamino-2-hydroxy)-propoxy-(5,7-3H) benzimidazol-2-one ((3H)CGP 12177) as subtype non-specific muscarinic and beta-adrenergic ligands, respectively. Biphasic course of changes was discovered with both ligands. With the dosage scheme applied, the density of beta-adrenoceptors was augmented 24 h after the first dose of DFP and decreased to about one half of control values after 5 days of treatment with DFP. The density of muscarinic receptors was augmented after two days of treatment with DFP and decreased to about one half of control values after 5 days of treatment. Comparatively small changes in the heart rate were observed during the treatment, they reflected changes in the muscarinic and beta-adrenergic receptor density. The finding of DFP-induced changes in the binding of (3H)CGP 12177 suggests that the mechanisms responsible for the control of the density of muscarinic and beta-adrenergic receptors in the heart cells are interconnected but potential roles of other factors involved in in vivo experiments deserve further analysis. The transient increase in the density of muscarinic receptors after two days of DFP treatment appears related to published data on transient stimulation by cholinergic agonists of the transcription of mRNAs for muscarinic receptors.

Our previous work indicated that hyperstimulation of muscarinic receptors brought about profound changes not only in the density of the muscarinic receptors, but also of the β-adrenoceptors both in vivo and in vitro. Now we try to identify the changes in receptor densities when the neuronal input from the autonomic nerves is disrupted, i.e. when neonatal rats were injected with 6-hydroxydopamine (6-OHDA; able to destroy sympathetic nerves). Although the interruption of neuronal input was successful as indicated by norepinephrine uptake, there was no change in receptor densities in the rat heart atria when 6-OHDA was applied in the first postnatal week only. When we repeated the 6-OHDA treatment consequently (on 14 th , 21 st and 28 th postnatal day) there were clear increases of both receptor types (muscarinic receptors rise to 139% of control, β-adrenoceptors to 134% of control). The atrial muscarinic receptor number was increased (to 146% of control) also when we combined the first week 6-OHDA application with another factor (when the pups abide with mother till the 56th day). The simple non-weaning of animals has no effect on the receptor densities. We can therefore conclude that the first week application of 6-OHDA was insufficient in changing the receptor number and that only repeated 6-OHDA administration was able to change them. Similarly, combination with other factor: non-weaning led to the muscarinic receptors increase. Our results can indicate the importance of the prolonged postnatal period for the heart muscarinic and β-adrenergic receptor number determination.

Motor seizures elicited by isonicotinehydrazide (INH) were studied in 188 male albino rats aged from 5 days to adulthood. INH was found to be able to induce both minimal, predominantly clonic, and major, i.e. generalized, tonic-clonic seizures during the whole course of development. Tonic hindlimb extension was observed as a part of major seizures even in the youngest group studied. Fifty-percent convulsant doses counted in 7−, 12−, 18−, 25− and 90-day-old rats did not significantly differ. The developmental changes found were a poor time resolution of minimal and major seizures during the first two postnatal weeks and significantly longer latency of major seizures in 5- and 7-day-old rat pups in comparison with all other age groups. The importance of the developmental stage of a generator of seizures as well as of the maturation of triggering mechanisms was demonstrated comparing the present results with our previous data.

The presence of clusters of morphologically complete sperm in vaginal smears taken in the morning from a stimulus female seems to be sufficient for determination of the ejaculation of a male accompanying the female the previous night. Ten sexually experienced male rats were exposed to three different experimental conditions. Each experimental condition lasted for 4 consecutive days. There was a 9-day interval between conditions. In Experiment 1, males were allowed to copulate with four females. The presence of sperm was determined in vaginal smears taken from all stimulus females. Males were able to ejaculate daily, but the number of inseminated females was lower on the 2nd and 3rd day. In Experiment 2, the number of stimulus females was increased from one to four over 4 days. Again, the males ejaculated daily. The “Coolidge effect” did not take place under these conditions. On the contrary, a “conditioned preference” for the first female developed. In Experiment 3, males were accompanied by a single female: first, the most “preferred” female from Experiment 1 and, 1 week later, the “nonpreferred” female, the least successful one in Experiment 1. In both situations, males were able to ejaculate continuously. There was a slight tendency for the “preferred” females to be inseminated on more days than the “nonpreferred” ones. The difference was most pronounced during the third day. In comparison to the 1st and 4th day, the number of inseminated females during the 2nd and 3rd day decreased (Experiments 1–3). It is concluded that animals are not so exhaustible as was indicated on the basis of one-session exhaustion tests.

Motor and electrocorticographic (ECoG) phenomena induced by pentamethylenetetrazol (PTZ) were directly compared in adult as well as in young rats (age groups of 7, 12, and 18 days). The technique for implantation of cortical electrodes was elaborated in young rats. Myoclonic jerks were elicited by PTZ in all age groups studied. In adult animals they were accompanied by isolated spikes and/or spike-and-wave complexes, whereas in 7-day-old rats there was a poor correlation between ECoG abnormalities and myoclonic jerks. With increasing age an improved correlation between ECoG abnormalities and behavioral changes were noted. After further injections of PTZ, rats had generalized tonic-clonic motor seizures. Ictal ECoG activity usually started in one cortical region in 7-day-old rats and became progressively generalized whereas in older rats generalized electrical discharges accompanied the motor seizures from onset. The clonic phase of motor seizures was not always accompanied by specific ECoG changes in young rats. At all ages the correlation between ECoG activity and individual phases of the generalized tonic-clonic seizures was poor.

Three rhythmic cortical phenomena (rhythmic after-discharges, incremental responses and self-sustained after-discharges formed by spike-and-wave rhythm) were elicited by electrical stimulation of thalamic nuclei during ontogenesis in rats. All three phenomena could be recorded for the first time at the age of 12 days and they matured until the 18th postnatal day. Possible explanation for the delayed development of these rhythmic thalamo-cortical phenomena in comparison with single thalamo-cortical responses are discussed.

Posthypoxic fluctuations in the levels of two excitatory amino acids, glutamate and aspartate, may be related to changes in mechanisms(s) which are responsible for their reuptake. As gamma-glutamyl transpeptidase (GGT) plays a role in mediating the uptake of glutamate and aspartate into various compartments of the brain, we studied changes in the activity of this enzyme in main regions of the brain in young and adult rats. We found a posthypoxic increase in bound GGT activity in some brain regions of 18-day-old animals after acute exposure, but no changes were observed after prolonged altitude hypoxia, with the exception of a decrease in cortical GGT activity. In contrast, acute hypoxia decreased GGT activity in the cortical capillaries to 59%, but prolonged hypoxic exposure was ineffective. However, the activity of soluble GGT in the cerebrospinal fluid of both groups of rats was several-times elevated in comparison with controls. At the same time, bound GGT activity was increased in the liver after acute or prolonged altitude hypoxia. The soluble GGT activity in plasma was only increased after prolonged exposure. Ninety days after prolonged hypoxic exposure the bound GGT activity was reduced in all brain regions to about 60–70% of controls (significantly higher in females than in males) as long-term developmental sequel from early postnatal hypoxia.

Response of leukocytes to exposure to an external magnetic field with frequency 50 Hz and sinusoidal waveform was investigated in vitro using the leukocyte adherence inhibition (LAI) assay developed as a measure of cell-mediated immunity. Leukocytes taken from healthy humans adhere, but their adherence decreases after 1 hr of exposure to the magnetic field with magnetic induction of 1 and 10 mT. The majority of leukocytes taken from cancer patients before any medical treatment do not adhere, and exposure to the magnetic field increases adherence. Correlation between the LAI assay results and the cell-mediated immunity suggests an effect of magnetic fields on leukocyte immune function in humans.