The gill monogenean Sciadicleithrum variabilum induces histomorphological alterations in the gill tissues of the discus Symphysodon aequifasciatus.

High mortality is among the most serious problems and challenges in the ornamental fish trade. Examination of the discus Symphysodon aequifasciatus from ornamental fish hatchery revealed infestation with the monogenean Sciadicleithrum variabilum. Gill infestation with this monogenean induced serious damage to the gill lamellae, including clavate lamellae, vascular congestion in the peripheral blood vessels, lamellar blood sinus dilation, and other structural anomalies. Light and transmission electron microscopy showed that in all infested hosts the interlamellar cell mass (ILCM) completely filled the interlamellar space. The monogenean-associated damage combined with the ILCM led to severe impairment of respiratory efficiency of the gill. Anti-parasitic treatment was applied during breeding (hatchery), which was followed by almost complete regression of the ILCM seen in the fish. A single point of ILCM hyperplasia was observed in only one specimen at the site of parasite attachment to the gill filament. The ILCM covering the gill lamellae protected the discus against infestation with this monogenean, but considerable reduction in the gaseous exchange surface and serious damage to the gill lamellae contributed to the increased mortality of the fish in the hatchery, which reached 90%.

Quest for breathing: proliferation of alveolar type 1 cells.

There is much evidence that the vertebrate lung originated from a progenitor structure which was present in bony fish. However, critical basic elements for the evolution of breathing in tetrapods, such as the central rhythm generator sensitive to CO2/pH and the pulmonary surfactant, were present in the lungless primitive vertebrate. This suggests that the evolution of air breathing in all vertebrates may have evolved through exaptations. It appears that the capability for proliferation of alveolar type 1 (AT1) cells is the "critical factor" which rendered possible the most radical subsequent innovation-the possibility of air breathing. "Epithelial remodeling," which consists in proliferation of alveolar cells-the structural basis for gas diffusion-observed in the alimentary tract of the gut-breathing fishes (GBF) has great potential for application in biomedical research. Such a process probably led to the gradual evolutionary development of lungs in terrestrial vertebrates. Research on the cellular and molecular mechanisms controlling proliferation of squamous epithelial cells in the GBF should contribute to explaining the regeneration-associated phenomena that occur in mammal lungs, and especially to the understanding of signal pathways which govern the process.

From epithelial remodelling to carcinogenesis.

The novel cancer theory named 'the tissue organization field theory' (TOFT) suggests that carcinogenesis is a process analogous to embryonic development, whereby organs are formed through interactions among different cell types. The suggested 'morphological remodelling' of the epithelium under hypoxia in gut breathing fish (GBF) has many common features with carcinogenesis. It appears that research into the relationship among epidermal growth factor receptor (EGFR), hypoxia inducible factor (HIF) as well as hypoxia and normoxia states in GBF fishes can be crucial in learning about the steering mechanisms of squamous epithelium proliferation, leading to a better understanding of carcinogenesis.

Chemoreceptors as a key to understanding carcinogenesis process.

The tissue organization field theory (TOFT) presented completely new, different from the previous one, perspective of research on neoplasm processes. It implicates that secretory neuroepithelial-like cells (NECs), putative chemoreceptors are probably responsible for the control of squamous epithelial cells proliferation in the digestive tract during hypoxia in gut breathing fish (GBF). On the other hand, chemoreceptors dysfunction can lead to uncontrolled proliferation and risk of cancer development in mammals, including humans. The studies on NECs like cells (signal capturing and transduction) may be crucial for understanding the processes of controlling the proliferation of squamous epithelial cells in the digestive tract of GBF fish during hypoxia states. This knowledge can contribute to the explanation of cancer processes.

Differential expression of epidermal growth factor receptor (EGFR) in stomach and diverticulum of Otocinclus affinis (Steindachner, 1877) as a potential element of the epithelium remodeling mechanism.

It is suggested that due to the several stomach modifications, Otocinculus affinis (dwarf sucking catfish) possess the ability to breathe air during hypoxia, however, the exact mechanism remains unknown. The aim of this study was detailed analysis of the expression of EGFR in the stomach and diverticulum of the O. affinis at the mRNA and protein levels together with the immunohistochemical localization of EGFR in these organs. The intensity of band fluorescence corresponding to the EGFR gene expression level is significantly higher in the stomach than in the diverticulum. Further, quantitative analysis of EGFR protein abundance also revealed its higher synthesis in the stomach than in the diverticulum and the immunohistochemistry method confirmed these results. As regional localization of respiratory function in gut air-breathing fishes seems to be connected with "morphological remodeling" of the epithelium of their gut, the present research demonstrated the potential efficiency of the O. affinis stomach as a respiratory organ. Having the potential possibility to create an air-blood barrier in the gastrointestinal tract allowing gas diffusion and respiration in hypoxic states seems to be very beneficial for these fish. It seems that facultative gut air breathing fish species are a relevant vertebrate model for high throughput screening, vascular biology and evolution.

The presence and expression of the HIF-1α in the respiratory intestine of the bronze Corydoras Corydoras aeneus (Callichthyidae Teleostei).

Bronze corydoras (Corydoras aeneus) is a small diurnal activity fish from South America. Under hypoxia conditions, it uses the posterior part of the intestine as an accessory respiratory organ. The present PCR studies demonstrated higher expression of HIF-1α (hypoxia-inducible factor) gene in the respiratory than that in digestive part of bronze corydoras intestine. Further, immunolocalization studies using antibodies specific to HIF-1α and transmission electron microscopy (TEM) revealed the presence of HIF-1α epitopes in the intestine of Corydoras aeneus. In the respiratory intestine, the numerous clusters of gold particles visualizing HIF-1α antibody were observed within fibroblasts, whereas in the digestive tract of this species, single gold grains in the epithelial cells were noted. On the other hand, the presence of HIF-1α and the cytoplasmic domain of the epidermal growth factor receptor (EGFR) in the respiratory intestine of bronze corydoras assumes their interactions in the system where these factors appeared for the first time. The non-obligatory air-breathing fishes using their digestive tract as an accessory respiratory organ during hypoxia conditions are very interesting for the studies of the processes that control HIF-1α expression and squamous cell proliferation.

Confirmation of the immunoreactivity of monoclonal anti-human C-terminal EGFR antibodies in bronze Corydoras Corydoras aeneus (Callichthyidae Teleostei) by Western Blot method.

Bronze corydoras (Corydoras aeneus) uses the distal part of the intestine as accessory respiratory organ. Our previous study showed the presence of epidermal growth factor receptor (EGFR) cytoplasmic domain in the digestive tract of the bronze corydoras. In this study, using Western Blot method, we validated the results presented in the previous research. In detail, results of Western Blot analysis on digestive and respiratory part of bronze corydoras intestine homogenates confirmed the immunoreactivity of anti-cytoplasmic domain (C-terminal) human EGFR antibodies with protein band of approximately 180kDa (EGFR molecular weight). This indicates a high homology of EGFR domain between these species and the possibility of such antibody use in bronze corydoras. Statistically significantly higher EGFR expression was observed in the respiratory part of intestine when compared to the digestive part. This implies higher proliferation activity and angiogenesis of epithelium in this part of intestine, creating conditions for air respiration. Therefore, Corydoras aeneus may be considered as a model organism for the molecular studies of the mechanisms of epithelial proliferation initiation and inhibition depending on hypoxia and normoxia.

Squamous epithelium formation in the respiratory intestine of the bronze Corydoras Corydoras aeneus (Callichthyidae Teleostei).

Accessory respiratory organs in fish exhibit great diversity but share the presence of numerous capillaries covered by a simple squamous epithelium. The adoption of the intestine for respiratory function needs certain special modifications. In this study, we explored immunohistochemical and metabolic fingerprint features that could underlay this adaptation in bronze corydoras Corydoras aeneus. Immunohistochemical localization of the cytoplasmic domain of epidermal growth factor receptor (EGFR) in the respiratory part of intestine demonstrated a strong positive immunoreaction in epithelial cells and connective tissue. Fourier Transfer Infrared (FTIR) spectroscopy coupled with chemometrics discriminated between anterior and posterior region of intestine in terms of secondary structure of proteins and the abundance of p-cresol and other phenolics. The latter were reduced in the posterior part of intestine, indicating the cessation of digestive function in this region. It has been suggested that aquatic hypoxia via endocrine cells (hypoxia-sensitive) activate EGFR, which induce proliferation of squamous epithelial cells, thereby enabling gas diffusion in the posterior part of intestine. It seems that hypoxia and normoxia are opposed conditions adjusting the production of squamous epithelial cells in this intestine. The physiological role of EGFR in the respiratory intestine of bronze corydoras is of interest not only from an evolutionary aspect but also in terms of a potential model for observations process proliferation squamous epithelial cells. Future investigations on the molecular responses to different water oxygen levels in air-breathing bronze corydoras fish are required to clarify the mechanism responsible for squamous cell proliferation.

Expression patterns and quantitative assessment of neurochemical markers in the lung of the gray bichir, Polypterus senegalus (Cuvier, 1829).

Anatomical and functional studies of the autonomic innervation and the putative oxygen receptors-the neuroepithelial (NEC)-like cells of the bichirs are lacking. The present paper describes the distribution of both NEC-like cells and the polymorphous granular cells (PGCs) that populate the mucociliated epithelium of the lung in the air breathing fish Polypterus senegalus. By using confocal immunohistochemistry we determined the coexpression of specific neurochemical markers. Colocalization studies showed that 5HT is coexpressed with calbindin and nNOS in the NEC-like cells and PGCs, and choline acetyltransferase (ChAT) is coexpressed with nNOS in both the two types of cells. Distribution of neurotransmitters (5HT, NO) and neurochemical marker ChAT is also investigated in the lung muscle. The role of these transmitters may be the autonomic control of circulation and respiration. However, the importance of these signals for the respiratory responses in the species studied is still not known. The present study also shows for the first time the simultaneous occurrence of piscidin 1 and 5HT in the PGCs. The function of these cells being equivalent to ones found in fish gill subepithelial parenchyma, is still not known. Due to the importance of piscidin 1 in local immune defense, more research is useful to understand a possible interaction of PGCs with immune response in the bichir lung.

Morphology and innervation of the teleost physostome swim bladders and their functional evolution in non-teleostean lineages.

Swim bladders and lungs are homologous structures. Phylogenetically ancient actinopterygian fish such as Cladistians (Polypteriformes), Ginglymods (Lepisosteids) and lungfish have primitive lungs that have evolved in the Paleozoic freshwater earliest gnathostomes as an adaptation to hypoxic stress. Here we investigated the structure and the role of autonomic nerves in the physostome swim bladder of the cyprinid goldfish (Carassius auratus) and the respiratory bladder of lepisosteids: the longnose gar and the spotted gar (Lepisosteus osseus and L. oculatus) to demonstrate that these organs have different innervation patterns that are responsible for controlling different functional aspects. The goldfish swim bladder is a richly innervated organ mainly controlled by cholinergic and adrenergic innervation also involving the presence of non-adrenergic non-cholinergic (NANC) neurotransmitters (nNOS, VIP, 5-HT and SP), suggesting a simple model for the regulation of the swim bladder system. The pattern of the autonomic innervation of the trabecular muscle of the Lepisosteus respiratory bladder is basically similar to that of the tetrapod lung with overlapping of both muscle architecture and control nerve patterns. These autonomic control elements do not exist in the bladders of the two species studied since they have very different physiological roles. The ontogenetic origin of the pulmonoid swim bladder (PSB) of garfishes may help understand how the expression of these autonomic control substances in the trabecular muscle is regulated including their interaction with the corpuscular cells in the respiratory epithelium of this bimodal air-breathing fish.

The simultaneous presence of neuroepithelial cells and neuroepithelial bodies in the respiratory gas bladder of the longnose gar, Lepisosteus osseus, and the spotted gar, L. oculatus.

Anatomical and functional studies on the autonomic innervation as well as the location of airway receptors in the air-bladder of lepisosteids are very fragmentary. These water-breathing fishes share in common with the bichirs the presence of a glottis (not a ductus pneumaticus) opening into the esophagus. In contrast to a high concentration of neuroepithelial cells (NECs) contained in the furrowed epithelium in the lung of Polypterus, these cells are scattered as solitary cells in the glottal epithelium, and grouped to form neuroepithelial bodies (NEBs) in the mucociliated epithelium investing the main trabeculae in the air-bladder of Lepisosteus osseus and L. oculatus. The present immunohistochemical studies also demonstrated the presence of nerve fibers in the trabecular striated musculature and a possible relation to NEBs in these species, and identified immunoreactive elements of this innervation. Tyrosine hydroxylase (TH), choline acetyltransferase (ChAT), 5-HT and neuropeptide immunoreactivities were detected in the intramural nerve fibers. 5-HT and VIP immunopositive nerve fibers are apparently associated with NEBs. TH, VIP and SP immunoreactivities are also present in nerve fibers coursing in the radially arranged striated muscle surrounding the glottis and its submucosa. 5-HT positive neurons are also found in submucosal and the muscle layers of the glottis. The physiological function of the adrenergic and inhibitory innervation of the striated muscle as well as the neurochemical coding and morphology of the innervation of the NEBs are not known. Future studies are needed to provide evidence for these receptors with the capacity of chemoreceptors and/or mechanoreceptors.

[Over-the-counter Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) in the material of Poison Information Centre]. / Niesteroidowe leki przeciwzapalne (NLPZ) dostepne bez recepty w materiale Osrodka Informacji Toksykologicznej.

There were 10 636 toxicological information given in 2004-2008 by the Poison Information Center Jagiellonian University Medical College. 538 of them concerned NonSteroidal Anti-Inflammatory Drugs (NSAIDs); 336 cases of NSAIDs ingested alone and 202 co-ingested with other drugs and/or with ethanol. 58,9% of them were intentional and 29,6% were accidental. Children (1-6 years) and adolescents (15-19 years) were mostly involved.

[System of biocide products control in Poland]. / System kontroli zatruc biocydami w Polsce.

In this article we presented actual legislation concerning biocide products in Poland. Rules of reporting and archiving of biocide products exposure were discussed. We presented the first results of monitoring of biocide poisoning cases in Poland from July 2007 to June 2008.

[Stinging catfish spine envenamation]. / Narazenie na jad suma indyjskiego.

The stinging catfish Heteropneustes fossilis (Bloch, 1794) has become a popular aquarium fish in Poland and is available in almost every pet shop. The toxicological centers in Cracow, Gdansk and Warsaw have recorded twelve cases of envenomations caused by stinging catfish in the years 1994-2008. In most cases the injury was accompanied by oedema, erythema, intense pain, numbness of the site and dizziness. Schema of medical management was prepared using analysis of cases.

Lionfish envenomations in Poland.

14 injuries caused by lionfish Pterois volitans (L) kept in home aquariums were noted over a time period of 3 years. All the punctures occurred in the victims' hands. Intense pain and systemic findings observed in 11 patients. Enveno-mations had a mild course, the patients were treated for their symptoms. Toxicology information centers have also recorded 3 cases of exposure to tank-bred lionfish in which typical envenomation symptoms did not appear. Those cases were probably what is referred to as "empty stings", in which no toxin is injected into the victim's body. Empty stings are known to be made by snakes, which can consciously inject no venom, while for lionfish, the lack of exposure to venom was probably caused by the structure of the venom organ in which there was no channel to deliver the venom.

[Medical treatment during fish envenomation]. / Postepowanie w zatruciu jadami ryb.

Expositions to fish venoms should be treated as a separate group of intoxications because of their different diagnostic procedure. Until now, there are over 220 venomous fish species described, but skin excretions are potentially toxic for humans. Cases of fish envenomations (37), consulted by Poison Information Centres in Poland, as well as described in literature and contained in Micromedex database were analyzed. The course of envenomation, medical management during exposition to venomous of Chondrichthyes and Osteichthyes, freshwater and marine fishes were resolved. Injuries caused by venoms fishes were similarly treated, usually symptomatic. Specific antivenoms are available only for two fish species. Each patient exposed to sting or bite should be examined and observed. If characteristic sings and symptoms of envenomation are present, proper medical management should be proceed.

[Clinical practice guidelines in acetaminophen poisoning--pre-hospital management]. / Wytyczne postepowania w przypadku zatrucia acetaminofenem--postepowanie przedszpitalne.

We described guidelines for personel working in poison information service in case of acute paracetamol overdose. The guidelines were created with respect to EBM (Evidence Based Medicine) by the American Association of Control Centers.

Green tobacco sickness in Poland.

Green tobacco sickness (GTS) is an illness associated with exposure to nicotine originating directly from the growing tobacco plant. The exposure takes place in the course of activities during tobacco farming. Nausea, vomiting, headache and dizziness are the symptoms typical of GTS. The GTS cases have been most commonly reported in the USA, Japan, India and Italy. The first case of GTS in Poland has been diagnosed in a young man working on a Virginia bright tobacco plantation. The patient had symptoms typical of GTS. The toxicological analysis of urine demonstrated the presence of cotinine at the level of 869 ng/ml. Intravenous fluids, anticholinergic agents and potassium supplementation were used in the treatment. Interestingly, diplopia at the initial stage of tobacco poisoning and horizontal nystagmus, which resolved on the 2nd day of hospital stay were observed. We believe that cases of GTS occur in Poland; however patients do not seek medical care. Given that the majority of activities during tobacco farming in Poland are performed manually, it seems necessary to launch a public awareness campaign on GTS and decontamination methods.

Diplopia in Green Tobacco Sickness.

This paper examines the first reported case of Green Tobacco Sickness (GTS) in Poland. A 25-year-old man who worked in a tobacco field for 14 hours with no protective measures experienced symptoms of GTS, and additionally, diplopia, after leaving the field. Upon hospital admission, diplopia was no longer observed. Diplopia was most probably caused by disturbances to the cholinergic neuromuscular transmission, secondary to nicotine. These neurological disturbances, the first of their kind observed in the course of GTS, deserve special attention. The case shows a potential adverse health effect related to tobacco harvest, as most Polish tobacco plantations are not mechanized. Polish farmers should be obligated to protect their workers with protective clothing, shoes, gloves and masks. Recommendations for tobacco harvester health are put forth in the paper.