Cellular stress induced by photodynamic reaction with CoTPPS and MnTMPyPCl5 in combination with electroporation in human colon adenocarcinoma cell lines (LoVo and LoVoDX).

Two porphyrins, CoTPPS and MnTMPyPCl5, were tested for their photodynamic activity and potential novel use in a therapy of human cancers. We investigated an effect of photodynamic reaction (PDR), electroporation (EP) and their combination (electro-photodynamic reaction [EP-PDR]) on human colon adenocarcinoma cell lines (LoVo and resistant to doxorubicin LoVoDX), human breast adenocarcinoma (wild type MCF-7/WT and resistant to doxorubicin MCF-7/DOX), and human melanoma (Me45). The efficiency of macromolecules transport was examined with cytofluorymetry by assessing the degree of propidium iodide (PI) penetration. Additionally, cellular ultrastructure after EP was evaluated. We determined cyto- and photo-cytotoxic effect on the cells viability (MTT assay) after standard PDR and PDR combined with EP. Intracellular distribution and mitochondrial colocalization of both porphyrins was also performed. The experiments proved that both complexes exhibit desirable photodynamic properties on LoVo LoVoDX cells, and EP effectively supports photodynamic method in this type of cancer. The application of EP provided shorter time of incubation (only 10 min) and enhanced effect of applied therapy. The porphyrins did not affect the MCF-7 and Me45 cell lines.

Differentiation and growth of myotomal muscles in a non-model tropical fish Pterophyllum scalare (Teleostei: Cichlidae).

Somite differentiation, muscle fibres formation and growth were analysed in a non-model tropical fish Pterophyllum scalare. In this study, it was found that during somite differentiation, a primary myotome appears. The primary myotome is filled with multinucleated myotubes that constitute the major part of the somite. Subsequently, Pax-3 (paired-box protein)-positive cells, located externally to the myotomes, appear. In post-hatching stages, mononucleated proliferating cell nuclear antigen-positive cells are observed in the inter-myotomal spaces and within the myotomes. The mononucleated cells, situated in the myotomes, first express desmin in their cytoplasm and then Pax-7 (paired-box protein) in their nuclei. Expression of desmin indicates that they will enter myogenic pathway, whereas expression of Pax-7 suggests their role of satellite cells. We assume that mononucleated intramyotomal cells are myogenic precursors involved in muscle growth. In advanced (post-hatching) stages of myogenesis, myotomes contain both primary and new muscle fibres. Morphometric analyses show that in Pterophyllum scalare, growth of muscle fibres is mainly a result of hypertrophy.

Pax-3 and Pax-7 label muscle progenitor cells during myotomal myogenesis in Coregonus lavaretus (Teleostei: Coregonidae).

In Coregonus lavaretus, prior the mesoderm segmentation, in cells adjacent to the notochord called adaxial cells MyoD and slow myosin heavy chain (MyHC-slow) proteins were observed. After somite formation, adaxial cells migrate towards the lateral part of the myotome and form a layer of red muscles. Deeper cells differentiate into white muscle fibres. In situ hybridization using Pax-3 molecular probe revealed, that after somitogenesis, Pax-3 is expressed in a layer of cells superficial to the myotome resembling the "external cells" (found in many teleosts species) or dermomyotome described in Amniota. During later developmental stages Pax-3 gene is expressed in cells in intermyotomal space and then in myoblasts between myotubes. In these cells Pax-7 protein was also observed. Pax-3/7 positive cells which have migrated into the myotomes differentiate into satellite cells/secondary myoblasts and participate in hypertrophic and hyperplastic growth of muscles.

Mixed mesodermal and mesenchymal origin of myotomal muscles in pike (Esox lucius: Teleostei).

During the myotomal myogenesis in pike (Esox lucius) two phases of muscle differentiation can be distinguished. In the first phase, the somite cells-derived stock, the primary myoblasts (of mesodermal origin), fuse to form multinucleate myotubes. Participation of myotomal cells of mesodermal origin is insufficient for further muscle development. In the second stage mesenchymal cells migrate, via myosepts, into the myotome between myotubes. Immunocytochemical detection of proliferating cell nuclear antigen (marker of S phase of cell cycle) showed their mitotic activity. Transmission electron microscope analysis revealed that the differentiation of these cells depends on their position. Cells remaining in the myosepts develop into fibroblasts and produce collagen fibres, while those that have migrated into the myotomes transform into secondary myoblasts. Mesenchymal cells in the studied species are believed to participate in hypertrophy and hyperplasy of muscle fibres. Thus the muscle fibres in pike (E. lucius) are of mesodermal-mesenchymal origin.

Position, structure and function of pulsatile vesicles in parasitic leeches Codonobdella truncata Grube, 1873 and Caspiobdella fadejewi Epshtein, 1961 (Hirudinea, Piscicolidae, Piscicolinae).

Histological analysis of frontal section through the body of parasitic leeches (Codonobdella truncata, Caspiobdella fadejewi) of the subfamily Piscicolinae revealed the presence of 11 pairs of subepidermal pulsatile vesicles. The vesicles of C. truncata are poorly visible on the body surface, contrary to those of C. fadejewi, which results from the fact that in the former species the vesicles are located in a layer of subepidermal connective tissue thicker (82. 82 microm) than in C. fadejewi (20.6 microm). In the studied species the pulsatile vesicles have a club-like shape, are surronded by a muscular tunicle and connected with a short canal that penetrates the body wall. Some of the sections show that the vesicles are divided in two chambers (lower and upper) with an incomplete membrane.

Myotomal myogenesis in Triturus vulgaris L. (Urodela) with special reference to the role of mesenchymal cells.

Two stages can be distinguished in the differentiation of myotomal muscle fibres in Triturus vulgaris. In the first stage only synchronously differentiating myotomal cells are engaged; in the second stage mesenchymal cells also take part in the process. Myotomal cells (primary myoblasts) fuse to form 2-3 nucleate myotubes. Only in the caudal part of the embryo mononucleate myotubes persist. The mononucleate myotubes, like polynucleate ones, occupy the whole length of the myotome. The differentiation of myotubes is accompanied by vitellolysis. At further development stages mesenchymal cells enter the intermyotomal fissure, after which they migrate to the myotomes, between the myotubes. The cells that remain in the intermyotomal fissures retain their fibroblastic potential (they synthesise collagen). Their daughter cells adjoining the myotubes acquire myogenic abilities. Their myoblastic potential is evidenced by their ability to fuse with the myotube. Fusion of secondary myoblasts (of mesenchymal origin) with the myotube results in further growth of the myotubes. In T. vulgaris myotomal myotubes and muscle fibres developing from them are of myotomal-mesenchymal origin.

Variability of the reproductive system of Piscicola fasciata (Kollar, 1842 (Hirudinea, Piscicolidae).

The studies on the reproductive system of P. fasciata were based on 43 specimens. Two main types of the reproductive system can be distinguished in the studied population. Two types of reproductive system were the most frequent (20.93% each): with symmetrical ejaculatory ducts without a loop and symmetrical seminal vesicles reaching the end of the first pair of testes, situated next to the testes (adult individuals) (fig. 23) and with symmetrical ejaculatory ducts without a loop (fig. 21) and symmetrical poorly developed seminal vesicles situated asymmetrically anterior to the first pair of testes (juvenile individuals). The structure of the reproductive system corresponding with the description of the type specimen (BRUMPT, 1900) was not observed in the studied population.

Variability of the reproductive system of Piscicola respirans (Troschel, 1850) (Hirudinea, Piscicolidae).

The studies on the reproductive system of P. respirans, based on 115 specimens, have revealed its considerable variability. Two main types of the system can be distinguished: 1) ovaries reaching the first pair of testes, ejaculatory ducts symmetrical, no seminal vesicles (juvenile individuals), 2) ovaries reaching the first pair of testes, ejaculatory ducts symmetrical, seminal vesicles reaching the ends of ovaries (adults individuals). The structure of the reproductive system is correlated with the individual size and age. Because the reproductive system of P. respirans as described by BRUMPT (1900) and SKET (1968) based on type specimens designated by them, is found only in a small fraction (c. 4-8%) of specimens, a redescription of the species is necessary, best based on a neotype series.

[Parasites leech Batracobdelloides moogi (Nesemann & Csanyi, 1995) (Hirudinea, Glossiphoniidae vaillant, 1850) of Polish fauna?]. / Pasozytnicza pijawka Batracobdelloides moogi (Nesemann & Csanyi, 1995) (Hirudinea, Glossiphoniidae Vaillant, 1850) W faunie Polski?

In the light of recent studies (NESEMANN and CSANYI 1995) on the genus Batracobdella (VIRGUIER 1879) Batracobdella paludosa (CARENA 1824)--a component of the Polish fauna--is actually Batracobdelloides moogi (NESEMANN and CSANYI 1995).