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Summary An Atlantic cod, Gadus morhua L., with unique golden pigmentation was caught during a commercial trawl on the Svalbard Bank. Stomach content analysis excluded the possibility of golden pigmentation resulting from diet. Skin morphology, mitochondrial and molecular species markers and also the concentration of melatonin (MEL) were examined. Skin samples from five areas below and above the lateral line contained correctly developed melanophores, but their distribution was different from those observed in the skin samples of standard coloured cod. Only the difference between the numbers of melanophores below the lateral line was statistically significant (8 vs 19.8 in standard coloured cod). DNA analyses allowed exclusion of the possibility of interspecies hybridisation with haddock Melanogrammus aeglefinus. MEL concentrations in the skin sample of the golden cod compared to the control sample of standard coloured cod were similar and with no statistically significant differences. Some abnormalities observed in the detailed morphology and melanophore distribution suggest a genetic or hormonal nature in the golden pigmentation of the cod. Some of these, e.g. mutations of TYR (tyrosinase) genes or of the MSH (melanophore-stimulating hormone) level can be tested in further analyses. [ABSTRACT FROM AUTHOR]

Background and Objectives: The purposes of this review are to promote better use of existing knowledge of marine pollutants, especially endocrine-disrupting compounds (EDCs), and to draw attention to the slow progression of the research on the influence of those compounds on arginine vasotocin/isotocin system (AVT/IT) in fish. EDCs are leading to the degradation of fish habitats, reducing their spawning potential and possibly their population parameters (e.g. growth, maturation), by preventing fish from breeding and rebuilding their populations. Therefore, searching for new welfare indicators such as AVT and IT and developing research procedures mimicking environmental conditions using a versatile fish model is extremely important., Discussion: Fish species such as zebrafish (Danio rerio) and round goby (Neogobius melanostomus) can be recommended as very suitable models for studying estrogenic EDCs on the AVT/IT system and other hormones involved in the neuroendocrine regulation of physiological processes in fish., Conclusion: These studies would not only improve our understanding of the effects of EDCs on vertebrates but could also help safeguard the well-being of aquatic and terrestrial organisms from the harmful effects of these compounds., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The objective of this study was to determine how the short-term exposure to a supraphysiological concentration of waterborne 17β-estradiol (E 2 ) influences on melatonin (Mel) and thyroxine (T 4 ) concentrations in plasma and E 2 and 11-ketotestosterone (11-KT) concentrations in plasma and gonads in both sexes of round goby (Neogobius melanostomus) during the pre-spawning, spawning, late spawning and non-spawning phases. The experimental protocol was based on short-term, repeated exposures of fish to a supraphysiological dose of waterborne E 2 . Mel level was unchanged on exposure to E 2 during the investigated phases, and its role in determining a time frame for spawning in both sexes of round goby seems to be stable in those conditions. T 4 and sex steroids (E 2 and 11-KT) were sensitive to the exposure of E 2 , and those changes influence gonads by accelerating oocyte development, ovulation and regression and inhibiting spermatogenesis in this species. The results demonstrate that the physiological responses of fish in all investigated phases were altered over a short window of exposure, indicating that short-term exposure to a supraphysiological dose of E 2 may impact fish in the wild. Furthermore, round goby can be recommended as a very suitable model for studying endocrine disruptors, which is sensitive to even short exposure to E 2 .

Melatonin synthesis is controlled by aralkylamine N-acetyltransferase (AANAT: EC 2.3.1.87) acetylating serotonin (5-hydroxytryptamine; 5-HT) to N-acetylserotonin (NAS), and N-acetylserotonin O-methyltransferase (ASMT: EC 2.1.1.4) methylating NAS to melatonin (Mel; N-acetyl-5-methoxytryptamine). We examined the levels of expression of the aanat and asmt genes, Mel concentrations as well as AANAT isozyme activity in the eyeball (with retina) and skin of the three-spined stickleback (Gasterosteus aculeatus), at noon and midnight. We found mRNA of four genes (aanat1a, snat, asmt and asmt2) in the eyeball, and two (aanat1a and asmt2) in the skin. The presence of two transcripts of genes encoding AANAT and two of ASMT in the eyeball at noon and midnight, suggests activity of AANAT and ASMT isozymes in metabolic pathways besides "the way to melatonin", all the more so because day/night changes in Mel concentration do not follow the changes in either the expression of genes or the activity of AANAT. The high effectiveness of noon NAS synthesis in the eyeball at low substrate concentrations, which is not reflected in high Mel production, suggests the function of eye NAS beyond that of a precursor to the biosynthesis of Mel. The inhibition of AANAT isozyme activity by product observed in the eyeball may be one of the mechanisms of 5-HT husbanding in the eye (retina). The presence of transcripts of genes encoding both AANAT and ASMT and the activity of AANAT, at noon and midnight, supports a local Mel synthesis in the sticklebacks' skin., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The aim of this study was to determine changes in concentrations of melatonin (Mel) and thyroxine (T 4 ) in plasma, and 17β-estradiol (E 2 ) and 11-ketotestosterone (11-KT) in plasma and gonads of female and male round gobies (Neogobius melanostomus) from the Southern Baltic Sea in four phases of the reproductive cycle classified as pre-spawning, spawning, late spawning and non-spawning periods. The concentrations of Mel, T 4 and E 2 were determined by radioimmunoassay (RIA) whereas 11-KT was quantified using an enzyme immunoassay (EIA). The maturity stage of gonads was determined using histological analyses. The pattern of changes in Mel concentrations of females and males was similar with the greatest concentrations in the spawning and non-spawning phases. In both sexes, there was a similar tendency of change in concentrations of T 4 and E 2 with the increase being in the pre-spawning and non-spawning phases. The greatest concentrations of 11-KT were observed in the plasma and gonads of males during the spawning phase. In females, there were no changes in 11-KT concentrations either in plasma or gonads during all phases where quantifications occurred. This is the first study for determination of the pattern of changes in Mel and T 4 concentrations as well as gonadal steroids E 2 and 11-KT, supported by histological analysis of gonads, in batch spawning fish during the reproductive cycle., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The stress hormone cortisol, together with antioxidants, melatonin (Mel) and its biologically active metabolites, 5-methoxykynuramines, including AFMK, set up a local stress response system in mammalian skin. Our in vitro study of the European flounder (Platichthys flesus) was designed to examine whether Mel and AFMK would respond to cortisol while a glucocorticoid is added to the incubation medium. The concentrations of cortisol in the incubation medium mimic plasma cortisol levels seen in fish exposed to different types of stresses such as handling, confinement, high density, food-deprivation or air-exposure. We measured Mel and AFMK in skin explants and culture media using high-performance liquid chromatography (HPLC) with fluorescence detection. We also analysed melanosome response (dispersion/aggregation) in the explants subjected to the different treatments. Cortisol stimulated the release of Mel and AFMK from skin explants in a dose-dependent manner. Melanosome dispersion and a darkening of the skin explants were observed after incubation with cortisol. This study is the first to demonstrate the interrelationship between cortisol and Mel/AFMK in fish skin. Our data strongly suggest that the cutaneous stress response system (CSRS) is present in fish. The question remains whether Mel, AFMK or cortisol are synthetized locally in fish skin and/or transported by the bloodstream. The presence of the CSRS should be taken into account during elaboration of new indicators of fish welfare both in aquaculture and in the wild., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In this in vitro gradient perfusion study, we determined whether there is a functional relationship between oestradiol and the arginine vasotocin/isotocin (AVT/IT) system in the female round goby ( Neogobius melanostomus ). Brain explants were perfused in medium supplemented with 17β-oestradiol (E 2 ) at doses mimicking the plasma levels of this hormone in nature during the spawning-capable phase and regressing phase. We aimed to establish which pathway, genomic or non-genomic, is involved in this mechanism in different reproductive phases. For this purpose, brain explants were perfused in medium supplemented with Fulvestrant (ICI 182.780) or Actinomycin D (Act D) separately or in combination with E 2 The contents of AVT and IT in the perfusion media were determined using high-performance liquid chromatography (HPLC) with fluorescence and UV detection. During the spawning-capable phase, the effect of E 2 on AVT release is mediated through oestrogen receptors (ERs) via both genomic and non-genomic pathways, while IT release is mediated through ERs via a genomic pathway only. In the regressing phase, release of both nonapeptides is mediated through ERs via a genomic pathway. This is the first study to present a feasible mechanism of oestradiol action on the AVT/IT system in female fish during different phases of the reproductive cycle., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2017. Published by The Company of Biologists Ltd.)

Social relationships are crucially dependent on individual ability to learn and remember ecologically relevant cues. However, the way animals recognize cues before engaging in any social interaction and how their response is regulated by brain neuromodulators remains unclear. We examined the putative involvement of arginine vasotocin (AVT) and isotocin (IT), acting at different brain regions, during fish decision-making in the context of cooperation, by trying to identify how fish distinguish and recognize the value of other social partners or species. We hypothesized that the behavioural responses of cleaner fish clients to different social contexts would be underlain by changes in brain AVT and IT levels. We have found that changes in AVT at the level of forebrain and optic tectum are linked with a response to allopatric cleaners (novel or unfamiliar stimuli) while those at cerebellum are associated with the willingness to be cleaned (in response to sympatric cleaners). On the other hand, higher brain IT levels that were solely found in the diencephalon, also in response to allopatric cleaners. Our results are the first to implicate these nonapeptides, AVT in particular, in the assessment of social cues which enable fish to engage in mutualistic activities.

The 2015 Tissue Engineering Congress, held in London, UK on 8-10 September 2015, brought together the principles of engineering and life sciences in tissue development and regenerative medicine to discuss scientific research and developments of clinical applications from leading experts in the field. The newest research and developing technology were presented in the field of stem cell biology, tissue regeneration, 3D culture and scaffolds and biomaterials. The focus was on interdisciplinary approaches based on the combination of new-generation biomaterials and cell-based therapies, which can lead to breakthroughs in regenerating tissues in the future.

The teleost fish nonapeptides, arginine vasotocin (AVT) and isotocin (IT), have been implicated in the regulation of social behavior. These peptides are expected to be involved in acute and transient changes in social context, in order to be efficient in modulating the expression of social behavior according to changes in the social environment. Here we tested the hypothesis that short-term social interactions are related to changes in the level of both nonapeptides across different brain regions. For this purpose we exposed male zebrafish to two types of social interactions: (1) real opponent interactions, from which a Winner and a Loser emerged; and (2) mirror-elicited interactions, that produced individuals that did not experience a change in social status despite expressing similar levels of aggressive behavior to those of participants in real-opponent fights. Non-interacting individuals were used as a reference group. Each social phenotype (i.e. Winners, Losers, Mirror-fighters) presented a specific brain profile of nonapeptides when compared to the reference group. Moreover, the comparison between the different social phenotypes allowed to address the specific aspects of the interaction (e.g. assessment of opponent aggressive behavior vs. self-assessment of expressed aggressive behavior) that are linked with neuropeptide responses. Overall, agonistic interactions seem to be more associated with the changes in brain AVT than IT, which highlights the preferential role of AVT in the regulation of aggressive behavior already described for other species., (Copyright © 2016 Elsevier Inc. All rights reserved.)

There were two aims of this in vitro perfusion study. Firstly, to determine which class of receptors, glucocorticoid (GRs) or mineralocorticoid (MRs), are involved in cortisol regulation of arginine vasotocin (AVT) and isotocin (IT) release from the hypothalamo-pituitary (H-P) complex of round goby (Neogobius melanostomus). Secondly, to determine which pathways, genomic or non-genomic, are involved in the aformentioned process.The H-P explants were perfused with cortisol (1.4 × 10(-) (7)  M, 2.8 × 10(-7) M, 0.4 × 10(-6) M); only the highest dose significantly increased a release of both nonapeptides. In the perfusion of H-P explants, we used cortisol (0.4 × 10(-6) M) in combination with GRs antagonist RU486 (0.3 × 10(-6)  M) or MRs antagonist C03DA01 (0.36 × 10(-6)  M) or transcription inhibitor Actinomycin D (1 × 10(-7)  M). All inhibitors were also tested seperately. The contents of AVT and IT in the perfusion media was determined by high-performance liquid chromatography (HPLC) with UV detection. This study suggested that different mechanisms were involved in the regulation of AVT and IT release from H-P complex in round goby. Apparently it was GRs but not MRs that were involved in cortisol regulation of AVT and IT release. In the case of AVT, our data points to both genomic and non-genomic pathways mediating the effect of cortisol; in the case of IT, it is only the non-genomic pathway. This study presents the first feasible mechanisms of cortisol action on AVT and IT release from the H-P complex in round goby., (© 2015 Wiley Periodicals, Inc.)

Fish may respond to different social situations with changes in both physiology and behaviour. A unique feature of fish is that social interactions between males and females strongly affect the sexual characteristics of individuals. Here we provide the first insight into the endocrine background of two phenomena that occur in mono-sex groups of the black molly (Poecilia sphenops): masculinization in females and same-sex sexual behaviour, manifested by gonopodial displays towards same-sex tank mates and copulation attempts in males. In socially controlled situations, brain neurohormones impact phenotypic sex determination and sexual behaviour. Among these hormones are the nonapeptides arginine vasotocin (AVT) and isotocin (IT), counterparts of the well-known mammalian arginine vasopressin and oxytocin, respectively. To reveal potential hormone interactions, we measured the concentrations of bioactive AVT and IT in the brain, along with those of the sex steroids 17β-estradiol and 11-ketotestosterone in the gonads, of females, masculinized females, males displaying same-sex sexual behaviour and those who did not. These data were supplemented by morphological and histological analyses of the gonads. Correlations between brain nonapeptides and gonadal steroids strongly suggest a cross talk between hormonal systems. In the black molly, the masculinization process was associated with the production of brain AVT and gonadal steroids, whereas same-sex sexual behaviour involves both brain nonapeptides, but neither of the sex steroids. This study extends current knowledge of endocrine control of phenotypic sex and sexual behaviour in fish and for the first time links brain nonapeptides with the occurrence of male-male sexual behaviour in lower vertebrates., (© 2015. Published by The Company of Biologists Ltd.)

The aim of this in vitro study was to determine whether arginine vasotocin (AVT) and isotocin (IT) release from nerve endings is affected by urotensin II (UII) in gilthead sea bream pituitary. Primary cultures of pituitary cells were exposed to 10(-12), 10(-10), and 10(-8) M UII for 6, 24, and 48 hr. AVT and IT contents were determined in the culture media by high performance liquid chromatography (HPLC). UII at all doses decreased AVT release after 6, 24, and 48 hr of incubation. IT release was increased by UII only after 24 hr of incubation. This study, for the first time, indicates that UII affects AVT and IT release from nerve endings in the pituitary of Sparus aurata. It is presumed that UII together with AVT and IT may control response to different salinities in fishes., (© 2014 Wiley Periodicals, Inc.)

In natural spawning grounds, breeding round goby, Neogobius melanostomus, males are exposed to various social stimuli, including high density of same-sex competitors and separation from females. We hypothesize that breeding males subjected to overcrowding in the wild experience high stress that affects their socio-sexual behavior and their relationships among conspecifics. We designed an experiment to mimic natural stimulation when highly aggregated breeding males are subjected to same-sex opponents. Males were sampled sequentially from experimental tank stocked at decreasing fish densities of 15 fish/m(2), 9 fish/m(2) and 4 fish/m(2). We studied the effects of overcrowding on male behavior and selected hormones, brain arginine vasotocin (AVT) and isotocin (IT) and plasma 11-ketotestosterone (11-KT) and cortisol as these are known to play roles in reproduction and related social interactions. The highest brain AVT and plasma cortisol levels were measured in non-aggressive males kept in the overcrowded group of 15 fish/m(2). IT level was elevated in fish kept at the lower density of 9 fish/m(2), and at which the males began to display territoriality and aggression. The plasma level of 11-KT was similar in all the males. Brain AVT and IT and plasma cortisol along with behavioral observations can be applied as species-specific indicators of the well-being of round goby males., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Objective: Dendritic cells (DC) have been reported among inflammatory infiltrating cells in muscle tissue in idiopathic inflammatory myopathies (IIM), but to our knowledge no studies concerning the expression of langerin (CD207) or fascin (markers of immature and mature DC, respectively) in IIM have been published., Methods: Immunohistochemical analyses of langerin and fascin expression were performed on specimens from normal muscles, as well as those affected by polymyositis (PM) and dermatomyositis (DM). The results were analyzed by Mann-Whitney U test., Results: In PM and DM, fascin-positive cells were numerous in the majority of the studied samples in perimysial, endomysial, and perivascular cellular infiltrates. Sporadic langerin-positive cells were detected., Conclusion: Fascin-positive DC predominance in inflammatory infiltrates in myositic muscles confirms the prevalence of mature forms and indicates that there are conditions stimulating DC maturation in both PM and DM. The induction of immunological tolerance by inhibiting DC maturation may be a promising direction for studies of myositis treatment.

The study was designed to develop a new procedure for perfusion of brain and pituitary explants collected from three-spined stickleback (Gasterosteus aculeatus) and round goby (Neogobius melanostomus). The procedure was elaborated for studies of arginine vasotocin (AVT) and isotocin (IT) release from explants of both species. AVT and IT, analogs of mammalian vasopressin and oxytocin, are neurohormones produced in hypothalamus and released in neurohypophysis of Teleostei. Both nonapeptides are used as biomarkers of fish well being. Three perfusion sets were applied to test the method of medium transport into gradient container, without or with aeration. Medium supply to the gradient container from the top, without aeration is recommended only for short-term studies. Aeration of the medium with a mixture of 95% O(2) and 5% CO(2) at a pressure of 127.51 mm Hg is necessary for a long-term research. Transport of one or two media in the gradient container from the top and the bottom, simultaneously, requires aeration with a mixture of 95% O(2) and 5% CO(2) at a pressure of 315.03 mm Hg. Although the presented procedure has been elaborated for studies of AVT and IT in fish explants, after only minor modification, if any, it can serve many other purposes., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The following disease entities are being included into the idiopathic inflammatory myopathy group (IIM): dermatomyositis (DM), polymyositis (PM), inclusion body myositis (IBM). These are primary inflammatory muscle diseases with substantial muscle weakening in their course. Everyone of the entities belonging to IIM is possessed of a particular clinical picture, as well as of histological and immunological features. In this paper, there have been briefly described classification, clinical pictures, supplementary diagnostic tests, treatment and up-to date views on IIM etiopathogenesis.

In the normal striated muscle, tissue transglutaminase (TG2) content is vestigial. However, this protein's presence has been reported to occur in myoblasts and myotubes during the fetal period. Its increased expression has been also found in the muscle tissue in the course of sporadic inclusion body myositis, as well as in polymyositis (PM) and dermatomyositis (DM), which are considered to be diseases of immunological origin. Based on in vitro studies, a substantial TG2 role in the infiltration of some T cell subsets into inflamed tissues has been suggested lately. In this study, the immunohistochemical reactions in the guinea pig experimental myositis specimens and in the ones from PM/DM patients were compared. The guinea pig tissue specimens were taken from muscles affected by experimental myositis induced by intramuscular injections of: 1/sera from 30 neoplasm patients with no metastases; 2/sera from 10 healthy people; 3/sera from 2 DM patients; 4/neuropeptides (SP, NPY or VIP) and from 5/the muscles affected by the reversed passive Arthus reaction (RPAR). The immunostaining for TG2 revealed substantial presence of this protein in single, damaged muscle fibers and a weak reaction in regenerating fibers appearing in PM/DM patients' specimens. From among experimental myositis specimens, a very intensive reaction appeared only in the damaged and regenerating muscle fibers present in the slides from guinea pig muscles injected with DM patients' sera. Such results suggest some presence of a specific factor(s) (the one(s) responsible for TG2 expression in the damaged muscle fibers) in DM patients' sera. The results suggest that transglutaminase can be a marker of inflammatory myopathies. A probable correlation between TG2 expression in muscles and organismal immunological factors, including the complement activation status, requires additional studies.

Effects of histamine (HA) on cyclic AMP production and its action upon the effects evoked by vasoactive intestinal peptide (VIP) were studied in the chick hypothalamus. HA (0.1-1000 microM) potently stimulated cyclic AMP formation in the hypothalamic slices, reaching maximal effect (2.5-3.5-fold increase) at a 100 microM concentration, and displaying an EC50 value of approximately 6.5 microM/ The stimulatory action of HA was mimicked by agonists of HA receptors, with the following rank order of potency: HA>4-methylHA (H2)>or=Nalpha,Nalpha-dimethylHA (H3>>H1=H2)>or=2-methylHA (H1)>>amthamine (H2)>>dimaprit (H2) approximately tele-methylHA. The HA (100 microM)-evoked increase in cyclic AMP production was concentration-dependently antagonized by selective H2-HA receptor blockers (aminopotentidine>>cimetidine>or=ranitidine>>zolantadine) and was not affected by mepyramine and thioperamide, a selective H1- and H3-HA receptor antagonist, respectively. The pharmacological profile of HA receptors linked to the cyclic AMP-generating system in the chick hypothalamus indicates that they represent either an avian-specific H2-like HA receptor or a novel subtype of HA receptors. Chicken VIP (cVIP; 0.1-3 microM) potently stimulated cyclic AMP synthesis in the chick hypothalamus in a concentration-dependent manner. A combination of cVIP with HA produced cyclic AMP response more than additive, and such a synergistic interaction was antagonized by ranitidine. It is suggested that in the avian brain HA and VIP may play in concert to regulate neuroendocrine processes.

Chicken, guinea pig and mammalian (human/porcine/rat) vasoactive intestinal peptides (VIP; 0.001-3 microM) were compared with respect to their ability to stimulate adenosine 3', 5'-cyclic monophosphate (cAMP) formation in the cerebral cortical slices of rat and guinea pig. Of the tested peptides, i.e. chicken VIP (cVIP), guinea pig VIP (gpVIP) and human/rat/porcine (mammalian) VIP (mVIP), the strongest stimulator of cAMP synthesis was cVIP, and the weakest one - the gpVIP. Pituitary adenylate cyclase-activating polypeptide (PACAP) used as a reference drug at 0.1 microM concentration strongly stimulated cAMP formation in the cerebrum of both species, being, however, significantly more potent in the guinea pig model. The obtained data demonstrate significant differences in biological activity between cVIP and two distinct mammalian VIPs in the cerebral cortex of rat and guinea pig.

By means of immunoperoxidase and immune-alkaline phosphatase methods the immunoreactivities to neuropeptides: neuropeptide Y (NPY), calcitonin gene related peptide (CGRP) and a pan-neuronal marker, the protein gene product 9.5 (PGP 9.5), were evaluated in guinea pig deep dorsal skin specimens. CGRP immunoreactive (CGRP-IR) and NPY-immunoreactive (NPY-IR) nerve fibres were dispersed in the papillary dermis and sometimes inside the hair roots and among the sebaceous gland cells. Such localized nerve fibres have not so far been described. In the subcutaneous layer nerve trunks were found composed of CGRP-IR and NPY-IR nerve fibres. Some of these indicated vestigial or negative immunoreactivity to PGP 9.5.

The commercially available antibodies for immunohistochemical purposes, though numerous and diversified, are mostly designed to detect human antigens. This study was undertaken to evaluate the usefulness of anti-human antigen antibodies in guinea pig tissues. The subjects of our interest were mostly skin and striated muscles and it was important for us to stain vascular and neural elements and to visualize separate cell kinds, including also skin Langerhans cells and the cells, contributing to inflammatory infiltrations.

This study was undertaken to evaluate tissue transglutaminase (TG2) expression in guinea pig tissues, using an antibody against the guinea pig liver TG2. The tests were performed by means of a few immunohistochemical methods in specimens from: gut, skin, the lungs, lymph nodes, the heart, the thymus, the spleen, skeletal muscles of control guinea pigs and from inflamed skeletal muscles and skin. TG2 expression was found in artery, vein and lymphatic vessel endothelia, in mesothelia of pleura, pericardium and peritoneum, and in smooth muscle cells. Spleen deserves a special attention, since this organ, especially rich in TG2, may be much more involved in celiac disease and in liver diseases than it has been accepted so far. The subject calls for further elucidation.

The aim of this study was to examine a morphological picture of guinea pig skin that had been injected with neuropeptides (NPS)(2)- substance P (SP) and guinea pig vasoactive intestinal peptide (VIP) - to elucidate their local influence. Routine histological stainings were performed, together with immunohistochemical reactions for T cells and for macrophages. In the deeper layers of the skin, T cell and macrophagic infiltrations were observed. The intensity of these changes was greater 24 hours after injections than that observed at the third hour of the experiment.

We tested whether intramuscular injections of dermatomyositis (DM) patients' sera into guinea pig muscles can be used to transfer myositic alterations to these animals. Additionally, similar tests were performed using neoplastic patients' sera and sera from non-neoplastic, non-myositic patients. The DM patients' sera induced idiopathic inflammatory myopathy (IIM) type histological changes in muscle fibres in guinea pig quadriceps muscles, which were especially evident 72 h after sera injections. Immunohistochemical stainings of myositic guinea pig muscles were done for guinea pig pan-T-cells, monocytes/macrophages, the neuronal marker-protein gene product 9.5 (PGP 9.5) and protein S-100. Our studies proved that the factor(s) responsible for the appearance of characteristic alterations in diseased muscles during the course of DM is/are present in patient sera.

Histochemical studies on the activity of some neuropeptides (PGP, CT, NPY, SP and betaE) and enzymes (AP, AlP, SDH and MAO) were performed on guinea pig parathyroid glands after injections of histamine, histamine receptor blockers and muscarinic receptor blocker. Under conditions of histamine shock, the immunoreactivity of CT and the reactivity of SDH and MAO were found to decrease together with an increase in the activity of betaE and AP. The reactions for SP, NPY, PGP and AlP did not change in any of the groups.

The aim of the study was to examine a morphological picture of guinea pig skeletal muscles injected with neuropeptide Y (NPY), substance P (SP) or vasoactive intestinal peptide (VIP), to evaluate the influence of a single injection of the mentioned neuropeptides (NPS) on muscle morphology and mast cell, T lymphocyte and macrophage chemotaxis. There were different degrees of muscle fibre injuries as well as different intensities and compositions of infiltrations inside the muscle after the introduction of the NPS. The observed changes did not disappear, but increased after 24 hours, comparing to the 3-hour post-injection changes, suggesting that NPS are proinflammatory rather than antiinflammatory factors in skeletal muscles. The local, particularly delayed action of NPS in vivo requires further studies.

The aim of the study was to analyze serum tumor necrosis factor alpha (TNF-alpha) levels in patients with different neoplasm types qualified for surgical treatment and to evaluate their possible correlations with circulating immune complexes (CIC), IgG, IgM, and the complement (C) compounds: C1 inhibitor (C1i), C3c and C4 levels. Studies were performed in sera from 30 neoplasm patients before surgical treatment and in 10 persons from a control group (CG) with no malignancy. Serum TNF-alpha levels were measured with the Cytogen ELISA kit. Average TNF-alpha levels measured in neoplastic patient groups qualified for surgical treatment were not significantly different from the average TNF-alpha level in the CG group.

In the sera of 30 neoplasm patients without metastases, the average IgG level was higher than in the control group (CG) (18.16+/-5.10 vs. 12.62+/-2.14 g/l or 12.22+/-2.14 after excluding an outier). Average concentrations of circulating immune complexes (CIC), IgM, complement 1 inhibitor (Cli), C3c and C4 did not statistically differ between the groups. Dividing the patients' group into: breast or ovary cancer (BC), melanoma (M), digestive tract cancer (DT) and other neoplasms (ON) subgroups revealed that the IgG increase did not apply to the BC group. Relatively decreased CIC concentrations in the BC and DT group and an increased Cli in the DT group were found. Several diversities detected in the humoral immunity indices' distributions and correlations suggest activation of different mechanisms depending on the neoplasm types.

Very suggestive evidence for possible role of beta-endorphin (BE) in immune activity has been continuously accumulated in the literature in spite of contradictory results about the involvement of the opiate system in immunological response. The reversed passive Arthus reaction (RPAR) was performed in the dorsal area of the guinea pig skin and immunohistochemical PAP method was applied for visualisation of BE. The positive BE immunoreactivity was observed in sebaceous gland cells (SGC) , mainly localized in the middle layer of the glands, both in the normal and experimental skin. The maximal intensity of immunoreactivity to BE in SGC during RPAR was stronger than normal. In control skin, 50% of SGC revealed positive immunoreactivity comparing to 60-66% in the late phase of RPAR. Sebaceous glands (SGs) probably take part in local homeostasis in the skin, also during immune inflammation.

The reversed passive Arthus reaction (RPAR) was performed in the dorsal area of the skin and skeletal muscles of the pelvic girdle of male guinea pigs. Biopsies were taken 3, 24 and 48 hours after antibody administration. In the dermal type of RPAR attention was paid to the structure of striated muscles of the subcutaneous panniculus carnosus. Histological and electron microscopic studies were performed. Changes in the dermal RPAR were more rapid and more intensive than those directly in the muscles. In both types of RPAR the affected muscle fibres demonstrated vacuolar and hyaline degeneration, segmental necrosis, internal nuclei, cellular infiltration, phagocytosis and regeneration in the late phase of reaction. Neutrophils predominated in the infiltrations after 3 hrs but after 24 and 48 hrs the main cells were macrophages and lymphocytes. Ultrastructural studies showed various stages of damage to the mitochondria, dispersion and disruption of the myofibrils and formation of tubular structures. A similar morphological appearance is characteristic of polymyositis (pm) and dermatomyositis (dm) suggesting a similar i.e. immune complex mechanism of muscle fibre damage in RPAR and in pm and dm.