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The article defines the fundamental questions of legal nature and functional purpose of compensation for infringement of exclusive rights. The place of compensation is assessed both in the system of civil law remedies in general and in the system of remedies for infringement of exclusive rights in particular. The authors conclude that compensation is a complex (hybrid) form of liability in which the compensatory and punitive functions of civil liability are realized with the prevalence of the punitive one. On the basis of this conclusion, the criterion for determining the amount of compensation is proposed and justified. The legislative consolidation of this criterion and its further development in doctrine and judicial practice will contribute to the correct perception of compensation as a remedy and its application in accordance with its purpose.

SummaryEnhancers are genomic DNA sequences that bind transcription factors, chromatin regulators and non-coding transcripts to modulate the expression of target genes. They have been found to act from different locations relative to a gene and to modulate the activity of promoters up to ∼1 Mb away. Here we report the first 3D genome structures of single mouse ES cells as they are induced to exit pluripotency, transition through a formative stage and undergo neuroectodermal differentiation. In order to directly study how interactions between enhancers and promoters are reconfigured genome wide we have determined 3D structures of haploid cells using single cell Hi-C, where we can unambiguously map the contacts to particular chromosomes. We find that there is a remarkable reorganisation of 3D genome structure where inter-chromosomal intermingling increases dramatically in the formative state. This intermingling is associated with the formation of a large number of multiway hubs that bring together enhancers and promoters with similar chromatin states from typically 5-8 distant chromosomal sites that are often separated by many Mb from each other. Genes important for pluripotency exit establish contacts with emerging enhancers within multiway chromatin hubs as cells enter the formative state, consistent with these structural changes playing an important role in establishing new cell identities. Furthermore, we find that different multiway chromatin hubs, and thus different enhancer-promoter interactions, are formed in different individual cells. Our results suggest that studying genome structure in single cells will be important to identify key changes in enhancer-promoter interactions that occur as cells undergo developmental transitions.

This study is carried out by analyzing the optional version of blockchain technology use to overcome vulnerabilities in the confirmation of intellectual property eligibility. It is concluded that the blockchain technology has a transnational nature of application, which can help to overcome the consequences of territoriality in the protection of IP rights. Attention is focused on the protection of IP objects existing at the moment in the digital environment as the most real way of applying the blockchain technology, aimed at strengthening IP rights.

Mice and nude rats lethally infected with T. gondii and treated with recombinant rat interferon-gamma (rIFN-gamma) or recombinant human interleukin-2 (rIL-2) were protected against death, when compared with untreated infected controls. In mice rIFN-gamma and rIL-2 played an important role in "prophylactic treatment", but not in "curative therapy". The survival rate was 42% in mice treated with 3 doses of 20,000 U of rIFN-gamma at days -2, -1, 0 before challenge and up to 66% in mice treated with 3 doses of 10,000 U of rIFN-gamma at days -2, 0, +2 before and after infection. Whereas the survival rate was 33% in mice that received 3 doses of 500 U rIL-2 at days -2, -1, 0 before infection, or -2, 0, +2 before and after infection respectively, up to 50% of the mice treated with 3 doses of 1,000 U rIL-2 at days -2, -1, 0 survived. In nude rats rIFN-gamma had a slight effect in "prophylactic treatment", whereas rIL-2 was active only in "curative treatment". The survival rate was 25% both in nude rats treated with doses of 400,000 U of rIFN-gamma at days -3, 0 before challenge, or with doses of 5,000 U of rIL-2 at days +2, +6, +9 after infection. These results lead us to hypothesise that the mechanism by which the lymphokine treatment exerts a protective effect on Toxoplasma infected mice is different from that on nu/nu rats. We conclude that these cytokines may play a notable role in modulating the host's immune defence against T. gondii infection.

The effects of placing activated monocytes in the presence of LAK cells was investigated. It was shown that the addition of monocytes to a preparation of rIL2-stimulated lymphocytes decreased LAK cell activity. This inhibition is enhanced in the presence of rIFN gamma. To analyze the mechanisms of inhibition, monocytes and lymphocytes were cultured separately, on opposite sides of a porous membrane which allowed the passage of molecules. Under such conditions, monocytes inhibited the activity of LAK cells to the same degree that a mixed culture does, suggesting a possible role of diffusible factor(s). Neither indomethacin nor PGE2 fully inhibited LAK cell activity, indicating that PGE2 is not the major monocyte-derived factor inhibiting LAK cell activity. It was also demonstrated that LAK cells can kill monocytes, but that IFN gamma can protect the monocyte from the toxic effect. This protective mechanism may be responsible for enhancing the inhibitory activity of monocytes.

Recombinant human Interleukin-1 (rhlL-1) β was found to enhance stimulus-induced granule exocytosis from human polymorphonuclear leukocytes (PMNs). PMNs were incubated with rhlL-1β and then stimulated with either heat-aggregated IgG (Hagg) or N-formyhmethionyl leucylphenylalanine (FMLP). The release of the azurophil enzyme myeloperoxidase (MPO) was measured. Low concentrations of stimuli (10 μg/ml Hagg, 2.5 x 10-9 M FMLP) did not stimulate degranulation in the absence of rhlL-1β. However, such concentrations elicited marked degranulation from PMNs preincubated with rhlL-1β (0.2-100 ng/ml). The enhancement of degranulation was dependent on the concentration of rhIL-1β employed and on the period of incubation. In other experiments, the effect of rhlL-1β on the PMN oxidative response was determined. rhlL-1β did not directly stimulate the production of superoxide anions or enhance the oxidative response to Hagg or FMLP. It is suggested that in rheumatoid joints, IL-1β may potentiate PMN degranulation, but not their oxidative response.

The stability and relative quantum yields of n -CdSe/polysulfide photoelectrochemical cells under different applied potentials, light intensities and electrolyte concentrations, and after different photoelectrode surface preparation techniques, were investigated. It was deduced that the availability at the electrode surface of polysulfide ions, which serve as hole acceptors, decreases with forward bias. On the other hand, the flow of the photogenerated holes towards the surface of the semiconductor is affected to a lesser extent by increasing (forward) bias up to the point of maximum power, beyond which the hole flux drops sharply with the bias. Consequently, the polarization of the cell (as given by the ratio between the peak and the steady-state photocurrents) is maximal near the point of maximum power and there the stability is lowest at moderate light intensities (ca. AM1). Under strong illumination, depletion of the polysulfide ions from the surface of the electrode due to photooxidation, together with increasing sulfur to sulfide ratio at the surface, leads to considerable band flattening (decrease in the flat-band potential), and consequently electrodes under forward bias show considerably smaller photocurrents and thus greater stability than electrodes under short-circuit (and reverse bias) conditions (at constant illumination). Increasing the surface area of the electrode by photoetching or increasing the concentration of the electrolyte leads to the behavior which is observed under moderate light intensities, which shows the interdependency of all these factors.

Resume La separation complete des oligonucleotides pyrimidiques correspondant a une prise de depart n'excedant ps 300 a 400 μg de DNA a pu etre realisee grâce a la chromatographie en couches micnes bidimensionnelle sur couches de silice Kodak. De cette facon seize nucleotide ont ete identifies en utilisant la spectrophotometrie et les methodes enzymatiques.

The resolution of a mixture of pyrimidine oligonucleotides obtained by acidic hydrolysis of a DNA previously labelled with [32P] in tissue culture was performed by two dimensional chromatography uncommercially available cellulose thin layers. Sixteen nucleotides were identified and their amounts determined. This method does not require the elimination of terminal phosphoryl groups. Combined with a previously described method involving chromatography on silica gel thin layers, it permits the evaluation of 20 oliganucleotides from sheep embryo kidney apurinic DNA.

The infectious units in native and alkalidenatured preparations of DNA of herpes simplex virus were characterized with respect to their sensitivity to Neurospora crassa endonuclease, their sedimentation properties in high-salt, neutral sucrose gradients, and their sensitivity to hydrodynamic shearing forces. Infectious molecules in native preparations were resistant to N. crassa endonuclease, sedimented at 56 S, and were highly sensitive to shearing forces. After alkaline denaturation, infectious molecules became sensitive to the N. crassa enzyme, sedimented at 200 S, and were relatively resistant to shear. We conclude that both intact duplex molecules ([unk]100 × 10 6 daltons) and intact single strands ([unk]50 × 10 6 daltons) are capable of initiating productive infection.

Adsorption isotherms at very high coverage of (CH3)4C and (CH3)4Si on flat gold surfaces are found to imply a potential energy of interaction between the molecule and metal which varies at the inverse square of the distance between the molecule and the metal surface.

Ornithine decarboxylase (ODC), the rate-limiting enzyme in the biosynthesis of polyamines, was measured in the brain and the liver of adrenalectomized rats after an acute S.C. treatment with glucocorticoids. The effects of corticosterone and dexamethasone were compared in three brain areas, the cerebral cortex, hippocampus, and cerebellum. These structures have similar concentrations of cytosolic glucocorticoid receptor, as measured by an in vitro exchange assay using a specific glucocorticoid ligand, [3H]RU 26988, but contain different amounts of mineralocorticoid receptor. Corticosterone and dexamethasone increased ODC activity in the liver and brain areas in a dose dependent manner, dexamethasone being more active than corticosterone in all tissues. Moreover, estradiol, progesterone, and testosterone were inactive. Aldosterone, at high doses, increased brain ODC activity. Glucocorticoids, selected for their weak binding, or lack of binding to the mineralocorticoid receptor, were tested and found to be highly active in inducing brain and liver ODC, thus showing that ODC induction by steroids is specific for glucocorticoids. These results are among the first to suggest biochemically a central action of glucocorticoids following an acute treatment and confirm that the brain is a glucocorticoid target organ.

RU 24722, a compound selected for its protective effect against cerebral anoxia and ischemia in rats, induced a dose-dependent increase in brain ornithine decarboxylase (ODC), a rate limiting enzyme in the biosynthesis of polyamines. This action already 2 hr after injection, increased at 4 and 6 hr and ODC activity returned to pretreatment levels at 16 hr. Since it has been shown previously that glucocorticoids stimulate brain ODC, it was considered necessary to know if corticosterone could play a role in the action of RU 24722. This compound increased serum corticosterone levels 1 hr after injection, its effect being nil at 4 hr. Nevertheless, the effect of RU 24722 on brain ODC does not appear to be entirely dependent on the increase of serum corticosterone. the delays needed to obtain a stimulation of the enzyme by the steroid are longer (6 hr) than those necessary to observe the action of the drug (2 hr)on brain ODC. Furthermore, RU 24722 increased brain ODC even in adrenalectomized animals. In order to get an insight on the interaction of RU 24722 with putative transmitters, we have studied the effect of the compound on brain ODC in the presence of different pharmacological agents. Experiments performed using noradrenergic agonists and antagonits suggest that the action of RU 24722 on brain ODC is due to the blockade of inhibitory post-synaptic alpha-2 adrenoceptors. We have studied the action of other compounds, used for the treatment of senile cerebral insufficiency: codergocrine, dihydroergocristine, dihydroergocryptine, dihydroergocornine, dihydroergocristine and piribedil increased rat brain ODC; vincamine, piracetam and nicergoline were devoid of any action.

Five Monoclonal Antibodies (MAbs) coded respectively #111, 122, 206, 209 and 609 were produced against human recombinant Interleukin-1 beta (rIL-1 beta, amino acids 117-269). Four of these MAbs (#111, 122, 206 and 609) have been identified able to inhibit the biological activity of recombinant and natural Interleukin-1 beta. Competition studies suggested that three non-overlapping epitopes of the molecule were recognized by the MAbs. MAbs #609 and 206 have been selected on the basis of high affinity and used together in a two-site ELISA to detect IL-1 beta. The sensitivity of this ELISA was 1 pg/well (10-20 pg/ml). The assay did not detect rHuIL-1 alpha, rHuIL-2, rHuTNF-alpha, rHuIFN-gamma, rHuIL-6, and natural a- and b-FGF. The immunoassay was then compared to current assay such as co-mitogenic effect on murine thymocytes for detection of IL-1 beta in the intra- and extracellular compartments of IFN-gamma and/or LPS-stimulated human blood monocytes. We confirm that IFN-gamma potentiates IL-1 beta secretion in response to LPS (even with high dose 1 microgram/ml) but not the intracellular precursor of IL-1 beta. This immunoassay could be used also to detect IL-1 beta in plasma, sera and synovial fluids.

Expression of the neuronal marker 14-3-2 or NSE (neuron-specific enolase) has been studied during in vitro differentiation of cells in culture. The 14-3-2 protein of neuroblastoma cells is immunologically identical with that found in mouse brain extract. The lack of detectable 14-3-2 in cultures of non-neuronal lines shows that this protein, as has been already shown in vivo, is also a specific marker of neurons in vitro. The presence of 14-3-2 in a differentiated hypothalamic clone—but not in its presumptive precursor—indicates selective initial derepression of 14-3-2. Moreover, modulation of the amount of 14-3-2 already present in dividing neuroblastoma cells is related to the confluent phase of growth or morphological differentiation of neuroblasts. Both mechanisms may be related to the mechanisms underlying initial differentiation and subsequent maturation of neurons in vivo. In dividing neuroblastoma cells modulation of the basal level of 14-3-2 is not necessarily associated with expression of the morphological differentiation, but seems generally concomitant with an arrest of cell division.

We demonstrate the presence of high affinity receptors specific for interferon-gamma (IFN-gamma) in human lymphoblastoid Namalva cells. The presence of these receptors, whose binding affinity and cross-linking characteristics were not distinguishable from those of the corresponding receptors in sensitive cells, was not consistent with the lack of responsiveness of Namalva cells to IFN-gamma as regards growth inhibition, induction of 2'-5' oligoadenylate synthetase activity and inhibition of virus multiplication. Nevertheless, IFN-gamma enhanced the expression of two genes, HLA class II and c-myc. Although the mechanism of these IFN-gamma-mediated modifications is not understood, these results provide evidence that the IFN-gamma receptors present in Namalva cells are functional.

In animals, estrogens and progestins modulate food intake, running wheel activity and sexual behavior, androgens stimulate sexual behavior and aggression, and corticoids influence cerebral electrolyte regulation, sleep and memory. In this and other studies, we have established that soluble receptors binding steroid hormones are present, often in high concentration, in various central structures such as hypothalamus, cortex, hippocampus and amygdala and have shown that some of the effects of steroids on behavior can be related to their interaction with these receptors and the resultant biochemical events. Thus, in the rat, the induction of sexual behavior by estrogens and progesterone can be related to the increase in progestin receptor induced in the hypothalamus by estrogens. The action of glucocorticoids has been implicated in memory processes involving protein synthesis and this study shows that glucocorticoids stimulate protein synthesis in the cortex of hypophysectomized rats, a structure rich in glucocorticoid receptors. Since the hormonal specificity of receptor binding is similar in peripheral and central target tissues, it is suggested that competition experiments on a peripheral target tissue can be used as a simple test to select compounds (agonists or antagonists) able to replace endogenous hormones at the receptor site in the treatment of psychiatric disorders due to endocrine dysfunction (hormone deficit or excess).

We report the characterization of two monoclonal antibodies (mab) directed against unglycosylated human IL2. Mice were immunized with recombinant IL2 (rIL2) and two of the hybridomas obtained (referred to as 15.2 and 17.2) were selected for further analysis. Both 15.2 and 17.2 antibodies bound to recombinant IL2 and weakly to natural IL2 in an enzyme linked immuno assay (ELISA). Both antibodies were very efficient in retaining rIL2 on affinity columns with a yield of 95% purified material after acid elution. In high concentrations both mabs strongly inhibited the growth of CTLL-2 cells induced by the immunizing-rIL2, while the growth of CTLL-2 cells in the presence of natural-IL2 was only slightly inhibited. Neither mab blocked the effect of rat IL2 in this test. Both mabs were used in an immuno-radiometric assay (IRMA) to quantify recombinant IL2. Cross inhibition experiments performed in this last assay indicate that the two mabs recognize two different epitopes on the recombinant IL2 molecule.

In 1979, we first reported that in order to inhibit prostaglandin (PG) synthesis, anti-inflammatory steroids required glucocorticoid receptor occupancy, ribonucleic acid (RNA) and new protein synthesis (Russo-Marie et al., 1979). As it had been previously demonstrated that steroids were inhibiting PG synthesis by inhibiting the release of arachidonic acid from their membrane phospholipid stores, probably by inhibiting the enzyme responsible for the phospholipid deacylation (Gryglewski et al., 1975; Hong & Levine, 1976), we advanced the hypothesis that this induced protein was able to inhibit phospholipase A2. In the meantime, Flower & Black-well (1979) reported also that anti-inflammatory steroids induced the biosynthesis of a phospholipase A2 inhibitor which prevents prostaglandin generation. In the model they used, they were able to demonstrate that the protein was not only induced but was also released by the cell which had been synthesizing it.

The capacity of free polyribosomes from rat cerebral cortex to incorporate labeled amino acids into protein (polyribosome activity) was compared in normal, hypophysectomized, and treated hypophysectomized rats. Polyribosome activity was measured in a cell-free system using a pH 5 supernatant from sham-operated rats. The polyribosome activity of hypophysectomized rats was 10-20% less than that of sham-operated rats. Subcutaneous treatment of hypophysectomized rats daily for 10 days with ACTH, corticotropic ACTH fragments [10-2- microgram of ACTH-(1-24), ACTH-(1-23), or Hoe 433 (ACTH-(1-17))], or glucocorticoids (1 mg corticosterone or 10 microgram dexamethasone) stimulated activity to a level 25-50% higher than that in sham-operated rats. Polyribosome aggregation, as measured by sucrose density gradient analysis, was also greater in treated hypophysectomized rats than in sham-operated rats. On the other hand, a daily sc dose of 100 microgram of the noncorticotropic fragment, ACTH-(1-10), did not stimulate brain polyribosome activity in hypophysectomized rats but merely restored it to the level observed in sham-operated rats. The present study suggests that stimulation of cerebral protein synthesis by ACTH and corticotropic ACTH analogs may be partly due to their ability to promote adrenal steroid secretion. Their stimulatory effect and that of glucocorticoids might explain their physiological roles during stress and learning.