Scanning electron microscopy was applied for surface analysis of unfertilized and fertilized eggs... more Scanning electron microscopy was applied for surface analysis of unfertilized and fertilized eggs of the common toad, Bufo bufo. In unfertilized eggs two types of surface protrusions were found: microvilli and microfolds. 5 min post fertilization (pf), at the place of sperm entrance, a fertilization body appears, surrounded by a group of small depressions, which gradually spread over the whole egg surface. This phenomenon is considered to reflect structural changes of the egg membrane, occurring in response to cortical granule breakdown. Another process which follows fertilization is the disappearance of microfolds and the formation of microvilli. Shortly before the first cleavage, the egg surface smooths out and a zone of spherical microvilli becomes visible at the presumptive furrow region. While the cleavage furrow penetrates into the egg, microvilli appear at the surface of the newly forming blastomeres.
In human myometrium, the formation of gap junctions at various stages of labour and in correlatio... more In human myometrium, the formation of gap junctions at various stages of labour and in correlation with the concentration of progesterone and oestradiol in maternal blood was described previously by electron microscopy and laser confocal microscopy of immunohistochemically stained myometrial sections. The present investigation focused on the effect of continuous exposure of isolated myometrial tissue to progesterone and oestradiol on the number of gap junction plaques in human myometrium cells in vitro. The presence of gap junctions was evaluated by immunocytochemistry with antibodies against gap junction protein, connexin43 (Cx43). Human myometrial cells were isolated from biopsies obtained from term pregnant women who had an elective caesarean operation in the 37th or 40th week of pregnancy. The dispersed myometrial cells that were obtained by limited enzymatic digestion of the myometrial samples were maintained in monolayer culture for 1, 3 and 6 days in the presence of medium that contained foetal bovine serum and the steroids at different concentration. In primary culture, as well as after several passages, the characteristics of these cells were morphologically and biochemically similar to those of smooth muscle cells and myometrial tissue. The obtained results showed that the cells in culture responded synchronously to the increased concentrations of oestradiol/progesterone mixtures. The number of gap junctions increased significantly on days 1, 3 and 6 in culture and showed positive correlation (p < 0.05) with the cell number when the concentration of oestradiol was raised to 1 microgram/mL in the progesterone ratio (1.0 microgram/0.5 microgram/mL). No significant correlation, however, in connexin43 gap junction number versus cell number was observed between the six experimental groups treated with progesterone only.
The expression of various connexins so far identified is metabolically and developmentally regula... more The expression of various connexins so far identified is metabolically and developmentally regulated. Examples include uterine endometrium where the expression of gap junction protein, connexin32 (Cx32) is regulated by steroid hormones. In this study we attempted to synthesise a short peptide which matches the portion of the amino acid sequence of the Cx32. Cx32 has primary structure predicted from the nucleotide sequence of cDNA clone. A fragment of Cx32 molecule was synthesised to produce anti-peptide antibody for detecting gap junction protein in mouse and rat liver and endometrium. The 12-peptide, plus Abu residue that corresponds to residues 108-119 (LEGHGDPLHLEE-Abu) of the rat Cx32 (283-mer) was synthesised by the solid phase method. Antibodies against this peptide were raised in rabbits, screened for reactivity and specificity using dot blot assays [15] and used for immunocytochemical staining at the light and at electron microscope levels. The antibodies also reacted with fish heart myocardium.
The spatial pattern of connexins26 (Cx26) and 43 (Cx43) expressions were investigated in the mous... more The spatial pattern of connexins26 (Cx26) and 43 (Cx43) expressions were investigated in the mouse digestive tract by immunocytochemistry. High levels of connexin43 in the epithelium of the oesophagus, non-glandular part of the stomach, and the circular layer of duodenal and ilea muscularis externa were detected. Cx26 was expressed in stratum granulosum of oesophagal folds and in the non-glandular part of the stomach. A low level of immunoreactivity of Cx43 was observed in the circular, and very low in the longitudinal layer of the muscularis externa in the stomach and colon. No immunoreactivity for Cx26 and Cx43 was found in the entire muscularis externa of the oesophagus or in the longitudinal muscle layers of the duodenum and ileum.
Twenty-four male volunteers (mean ± SD: age 25.4±5.8 years, height 178.6±5.5 cm, body mass 72.1±7... more Twenty-four male volunteers (mean ± SD: age 25.4±5.8 years, height 178.6±5.5 cm, body mass 72.1±7.7 kg) of different training background were investigated and classified into three groups according to their physical activity and sport discipline: untrained students (group A), national and sub-national level endurance athletes (group B, 7.8±2.9 years of specialised training) and sprint-power athletes (group C, 12.8±8.7 years of specialised training). Muscle biopsies of vastus lateralis were analysed histochemically for mATPase and SDH activities, immunohistochemically for fast and slow myosin, and electrophoretically followed by Western immunoblotting for myosin heavy chain (MyHC) composition. Significant differences (P<0.05) regarding composition of muscle fibre types and myosin heavy chains were found only between groups A (41.7±1.6% of MyHCI, 40.8±4.0% of MyHCIIA and 17.5±4.0% of MyHCIIX) and B (64.3±0.8% of MyHCI, 34.0±1.4% of MyHCIIA and 1.7±1.4% of MyHCIIX) and groups A and C (59.6±1.6% of MyHCI, 37.2±1.3% of MyHCIIA and 3.2±1.3% of MyHCIIX). Unexpectedly, endurance athletes (group B) such as long-distance runners, cyclists and cross country skiers, did not differ from the athletes representing short term, high power output sports (group C) such as ice hockey, karate, ski-jumping, volleyball, soccer and modern dance. Furthermore, the relative amount of the fastest MyHCIIX isoform in vastus lateralis muscle was significantly lower in the athletes from group C than in students (group A). We conclude that the myosin profile in the athletes belonging to group C was unfavourable for their sport disciplines. This could be the reason why those athletes did not reach international level despite of several years of training.
Lipovitellin-phosvitin crystals from oocytes of the sturgeon (Acipenser boeri L.) and the tuatara... more Lipovitellin-phosvitin crystals from oocytes of the sturgeon (Acipenser boeri L.) and the tuatara (Sphenodon punctatus [Gray]) have been shown to he orthorhombic like all such known crystals from vertebrates higher than cyclostomes. Lattice parameters and projected crystal views also resembled closely the hitherto known cases. The high evolutionary conservation of this crystal structure probably reflects a cell-biological significance of the tertiary structure of crystal constituents, especially of the lipovitellines. A comparative consideration of crystallographic data from cyclostomes and higher vertebrates suggests that cyclostomes started with one species and all higher vertebrates with two species of lipovitellin molecules.
Cells of the gas gland of the perch Perca fluviatilis L ., stimulated to increased generation of ... more Cells of the gas gland of the perch Perca fluviatilis L ., stimulated to increased generation of gas by the repeated emptying of the swim-bladder, were examined in the electron microscope . Intense activity of the nuclear envelope was demonstrated . Simple vesicles originating from the external nuclear membrane and the so-called multivesicular bodies derived from the outpocketings of both membranes of the nuclear envelope were observed . The multivesicular bodies were filled with numerous fine vesiculae arising from the active proliferation of their internal membrane . The authors offer two alternative mechanisms of formation of fine vesiculae inside the multivesicular bodies and the mechanism of the tearing away of these bodies from the nuclear envelope .
The main scope of our work was to investigate the usefulness of the PROZA correction method and C... more The main scope of our work was to investigate the usefulness of the PROZA correction method and CALIBRATION CURVE construction for quantitative X-ray microanalysis of biological soft, freezedried tissues. The test samples with a known elemental concentration of NaCl, MgCl 2 , KH 2 PO 4 and Na 2 SO 4 were prepared on the basis of 20% porcine skin gelatine. A control sample containing 20% gelatine was used for sulphur subtraction. Dissolved gelatine solutions were cooled with liquid nitrogen and then cut in cryomicrotome into 16 mm thick slices. The sections were lyophilised, coated with carbon and analysed by means of a scanning electron microscope combined with an energy dispersive (ED) spectrometer. The homogeneity of the prepared samples was verified using the Fisher's test. Only homogenous samples were used for calibration. The significance of the 0(&z) (PROZA) correction method for biological sample analysis was verified by comparing the prepared standard and entering it into the Voyager computer program memory with the remaining samples of a known elemental content. The differences between the standard and the samples were noted for all elements analysed. There was no sample matching the standard after the PROZA correction procedure. A high correlation r factor (above 0.99) for all analysed elements indicates that CALIBRATION CURVES construction could be suitable for quantitative X-ray microanalysis of biological samples.
The main objective of our work was to investigate the possibility and usefulness of indirect meth... more The main objective of our work was to investigate the possibility and usefulness of indirect methods in X-ray microanalysis for the quanti®cation of biologically important compounds. Metallothioneinlike proteins (MT-like proteins) from kidney and liver, rich in sulfur were chosen as an indicator of heavy metal presence in cells and their environment. Tissues from gold®sh (Carassius auratus gibelio) were sampled after short and prolonged periods of exposure to Co 2 , CrO À2 4 , Pb 2 , Cu 2 and control treatment and prepared for histochemical staining for peroxidated thiolate groups. Commonly used ±S±S± bonds dye (Nitro Red) was replaced with iodine atoms and they were quanti®ed at L line by means of X-ray microanalysis combined with SEM. After ®sh treatments with heavy metal solutions changes in MT-like proteins and in I atom contents were expected. There was statistically signi®cant decrease in MT-like proteins level in kidney after lead treatment (Pb=C 0.62). In liver a statistically signi®cant increase in MT-like proteins concentration was observed after chromium, cobalt and lead ions treatment in comparison to control animals. The following ratios were noted: 3.04 for Cr=C, 2.18 for Co=C and 2.10 for Pb=C. Our ®nding indicates that the method of indirect measurement of MT-like proteins in ®sh and other animal tissues is possible. The concentration of iodine atoms is above their detection level by EDS and their changes are possible to identify. During histochemical procedures it is worth taking into account sample preparation methods which might disturb the quality and quantity of the analysed material.
The variability of Actinocyclus kuetzingii (A.S.) Sim. in the Littorina deposits of the Vistula L... more The variability of Actinocyclus kuetzingii (A.S.) Sim. in the Littorina deposits of the Vistula Lagoon and Lake Druzno were studied. During the time of the Littorina Lake Druzno was a part of the Vistula Lagoon. In the different parts of the Lagoon both the abundance of A. kuetzingii and the diameter of its valves change in a manner associated with phases of the Littorina transgression. Coscinodiscus (Actinocyclus) kuetzingii fo. subsalsa Brockmann is probably an ecotype of A. kuetzingii (A.S.) Sim.
It is generally accepted that connexin43 (Cx43) is a major constituent of heart and myometrial ga... more It is generally accepted that connexin43 (Cx43) is a major constituent of heart and myometrial gap junctions. However, the presence of Cx43 gap junctions in non-pregnant myometrium is still poorly documented. Tissue sections of porcine heart and non-pregnant uterus and myometrial smooth muscle cell cultures were immunostained with monoclonal antibody against Cx43. In the heart, intensive immunostaining was confined to the intercalated discs as previously reported. In the non-pregnant uterus, punctuate immunostaining of Cx43 was seen throughout the myometrium along cell interfaces between myocytes. The expression of Cx43 was sustained in cultured smooth muscle cells isolated from non-pregnant myometrium. Western blotting has detected single isoform of Cx43 in both, cardiac and myometrial tissues. The electrophoretic mobility of porcine heart Cx43 was similar to that of myometrial isoform but different from the pattern of mobility of Cx43 of the rat heart. Hence, porcine myometrium ma...
The synchronous contractions of the uterus in labour depend on electrical coupling of myometrial ... more The synchronous contractions of the uterus in labour depend on electrical coupling of myometrial smooth muscle cells by gap junctions. In the human myometrium, gap junctions are scarce in the non-pregnant uterus, but become abundant at term in preparation for labour. We have previously demonstrated that in the human myometrium at term, three different gap-junctional proteins are expressed, connexins 43, 45, and 40. These connexins are known to have distinctive functional capacities in in vitro expression systems but whether, in the human myometrium in vivo, they are co-assembled into the same gap junction or form different types of gap junction has previously been unclear. By applying triple immunogold labelling to sections of Lowicryl-embedded tissue for electron microscopy, together with complementary immunoconfocal microscopy, we demonstrate here that connexins 43, 45, and 40 are commonly present as mixtures within the same gap-junctional plaque. While all gap junctions contain c...
We analyzed the DNA content of hepatocyte and erythrocyte nuclei of the spined loach Cobitis taen... more We analyzed the DNA content of hepatocyte and erythrocyte nuclei of the spined loach Cobitis taenia (diploid) and its allopolyploid forms. Twenty triploid females and one tetraploid were used. At least 20,000 hepatocyte and erythrocyte nuclei were acquired and analyzed by flow cytometry. C. taenia erythrocyte nuclei contain 3.15 ± 0.21 pg of DNA and the hepatocyte nuclei 4.45 ± 0.46 pg of DNA. Triploid Cobitis have 5.08 ± 0.41 pg of DNA in erythrocyte nuclei and 6.11 ± 0.40 pg of DNA in hepatocyte nuclei, whereas the tetraploid erythrocyte and hepatocyte nuclei contained 6.60 and 7.40 pg of DNA, respectively. In general, the DNA contents correlate positively with the ploidy level of the fish investigated. The DNA content variation in the hepatocyte and erythrocyte nuclei may be due to differences in extent of chromatin condensation, which is more pronounced in the erythrocyte than hepatocyte nuclei, or to the several orders of ploidy that occur in the parenchymal liver cells.
The aim of this study was to compare DNA content in hepatocyte and erythrocyte nuclei of the Euro... more The aim of this study was to compare DNA content in hepatocyte and erythrocyte nuclei of the European sunbleak, Leucaspius delineatus, in relation to nuclear and cell size by means of flow cytometry and fluorescence microscopy. The DNA standards, chicken and rainbow trout erythrocytes, were prepared in parallel with both cell types, with initial separation of liver cells in pepsin solution followed by cell filtering. Standards and investigated cells were stained with a mixture of propidium iodide, citric acid, and Nonidet P40 in the presence of RNAse, and fluorescence of at least 50,000 nuclei was analyzed by flow cytometry. Average cell size was determined by flow cytometry, using fresh cell suspension in relation to latex beads of known diameter. The size of nuclei was examined on the basis of digital micrographs obtained by fluorescence microscopy after nuclei staining with DAPI. The sunbleak's erythrocyte nuclei contain 2.25 ± 0.06 pg of DNA, whereas the hepatocyte nuclei contain 2.46 ± 0.06 pg of DNA. This difference in DNA content was determined spectroscopically using isolated DNA from the two cell types. The modal diameters of the erythrocytes and hepatocytes were estimated to be 5.1 ± 0.2 and 22.3 ± 5.0 lm, respectively, and the corresponding modal dimensions of their nuclei (measured as surface area) were 15.2 and 21.4 lm 2 , respectively. The nucleoplasmic index, as calculated from diameters estimated from surface area of nuclear profiles, was 2.51 for the erythrocytes compared with 0.08 for hepatocytes.
Zeitschrift für Zellforschung und Mikroskopische Anatomie, 1970
Two types of glandular cells are present in the pars intermedia of the loach, Misgurnus fossilis.... more Two types of glandular cells are present in the pars intermedia of the loach, Misgurnus fossilis. Basophils are characterized by the presence in their cytoplasm of numerous secretory granules containing electron-dense and homogeneous material and by scarce endoplasmic reticulum. ‘Weak’ acidophils contain in their cytoplasm abundant endoplasmic reticulum and numerous granules of different electron densities.
Zeitschrift für Zellforschung und Mikroskopische Anatomie, 1969
The fine structure of the gas gland cells in Perca and Acerina was studied with the light- and el... more The fine structure of the gas gland cells in Perca and Acerina was studied with the light- and electron microscope. In some of the animals the gas glands were stimulated by the single or repeated removal of the gas from the swimbladder. In this group of fishes the changes in the organization of cytoplasm of the gas gland cells accompanying
Yolk-platelet crystals in Amia calva L., Lepisosteus osseus (L.) and Polypterus bichir have ortho... more Yolk-platelet crystals in Amia calva L., Lepisosteus osseus (L.) and Polypterus bichir have orthorhombic features with unit-cell dimensions a = 8.3 ... 8.8 nm, b = 16.4 ... 16.9 nm and c = 18.6 ... 19.8 nm as determined in electron-diffracted patterns of fixed, epoxy-resin embedded and thin-sectioned material. Electron-diffraction patterns, crystal projections and the above unit-cell data make them extremely similar to the orthorhombic yolk-platelet crystals known for amphibians and teleosts. This observation fills a gap in yolk-platelet research and supports the view that the general architecture of yolk platelets has been conserved for nearly 400 million years. It follows that the peculiar platelet architecture itself has physiological significance.
Scanning electron microscopy was applied for surface analysis of unfertilized and fertilized eggs... more Scanning electron microscopy was applied for surface analysis of unfertilized and fertilized eggs of the common toad, Bufo bufo. In unfertilized eggs two types of surface protrusions were found: microvilli and microfolds. 5 min post fertilization (pf), at the place of sperm entrance, a fertilization body appears, surrounded by a group of small depressions, which gradually spread over the whole egg surface. This phenomenon is considered to reflect structural changes of the egg membrane, occurring in response to cortical granule breakdown. Another process which follows fertilization is the disappearance of microfolds and the formation of microvilli. Shortly before the first cleavage, the egg surface smooths out and a zone of spherical microvilli becomes visible at the presumptive furrow region. While the cleavage furrow penetrates into the egg, microvilli appear at the surface of the newly forming blastomeres.
In human myometrium, the formation of gap junctions at various stages of labour and in correlatio... more In human myometrium, the formation of gap junctions at various stages of labour and in correlation with the concentration of progesterone and oestradiol in maternal blood was described previously by electron microscopy and laser confocal microscopy of immunohistochemically stained myometrial sections. The present investigation focused on the effect of continuous exposure of isolated myometrial tissue to progesterone and oestradiol on the number of gap junction plaques in human myometrium cells in vitro. The presence of gap junctions was evaluated by immunocytochemistry with antibodies against gap junction protein, connexin43 (Cx43). Human myometrial cells were isolated from biopsies obtained from term pregnant women who had an elective caesarean operation in the 37th or 40th week of pregnancy. The dispersed myometrial cells that were obtained by limited enzymatic digestion of the myometrial samples were maintained in monolayer culture for 1, 3 and 6 days in the presence of medium that contained foetal bovine serum and the steroids at different concentration. In primary culture, as well as after several passages, the characteristics of these cells were morphologically and biochemically similar to those of smooth muscle cells and myometrial tissue. The obtained results showed that the cells in culture responded synchronously to the increased concentrations of oestradiol/progesterone mixtures. The number of gap junctions increased significantly on days 1, 3 and 6 in culture and showed positive correlation (p &lt; 0.05) with the cell number when the concentration of oestradiol was raised to 1 microgram/mL in the progesterone ratio (1.0 microgram/0.5 microgram/mL). No significant correlation, however, in connexin43 gap junction number versus cell number was observed between the six experimental groups treated with progesterone only.
The expression of various connexins so far identified is metabolically and developmentally regula... more The expression of various connexins so far identified is metabolically and developmentally regulated. Examples include uterine endometrium where the expression of gap junction protein, connexin32 (Cx32) is regulated by steroid hormones. In this study we attempted to synthesise a short peptide which matches the portion of the amino acid sequence of the Cx32. Cx32 has primary structure predicted from the nucleotide sequence of cDNA clone. A fragment of Cx32 molecule was synthesised to produce anti-peptide antibody for detecting gap junction protein in mouse and rat liver and endometrium. The 12-peptide, plus Abu residue that corresponds to residues 108-119 (LEGHGDPLHLEE-Abu) of the rat Cx32 (283-mer) was synthesised by the solid phase method. Antibodies against this peptide were raised in rabbits, screened for reactivity and specificity using dot blot assays [15] and used for immunocytochemical staining at the light and at electron microscope levels. The antibodies also reacted with fish heart myocardium.
The spatial pattern of connexins26 (Cx26) and 43 (Cx43) expressions were investigated in the mous... more The spatial pattern of connexins26 (Cx26) and 43 (Cx43) expressions were investigated in the mouse digestive tract by immunocytochemistry. High levels of connexin43 in the epithelium of the oesophagus, non-glandular part of the stomach, and the circular layer of duodenal and ilea muscularis externa were detected. Cx26 was expressed in stratum granulosum of oesophagal folds and in the non-glandular part of the stomach. A low level of immunoreactivity of Cx43 was observed in the circular, and very low in the longitudinal layer of the muscularis externa in the stomach and colon. No immunoreactivity for Cx26 and Cx43 was found in the entire muscularis externa of the oesophagus or in the longitudinal muscle layers of the duodenum and ileum.
Twenty-four male volunteers (mean ± SD: age 25.4±5.8 years, height 178.6±5.5 cm, body mass 72.1±7... more Twenty-four male volunteers (mean ± SD: age 25.4±5.8 years, height 178.6±5.5 cm, body mass 72.1±7.7 kg) of different training background were investigated and classified into three groups according to their physical activity and sport discipline: untrained students (group A), national and sub-national level endurance athletes (group B, 7.8±2.9 years of specialised training) and sprint-power athletes (group C, 12.8±8.7 years of specialised training). Muscle biopsies of vastus lateralis were analysed histochemically for mATPase and SDH activities, immunohistochemically for fast and slow myosin, and electrophoretically followed by Western immunoblotting for myosin heavy chain (MyHC) composition. Significant differences (P<0.05) regarding composition of muscle fibre types and myosin heavy chains were found only between groups A (41.7±1.6% of MyHCI, 40.8±4.0% of MyHCIIA and 17.5±4.0% of MyHCIIX) and B (64.3±0.8% of MyHCI, 34.0±1.4% of MyHCIIA and 1.7±1.4% of MyHCIIX) and groups A and C (59.6±1.6% of MyHCI, 37.2±1.3% of MyHCIIA and 3.2±1.3% of MyHCIIX). Unexpectedly, endurance athletes (group B) such as long-distance runners, cyclists and cross country skiers, did not differ from the athletes representing short term, high power output sports (group C) such as ice hockey, karate, ski-jumping, volleyball, soccer and modern dance. Furthermore, the relative amount of the fastest MyHCIIX isoform in vastus lateralis muscle was significantly lower in the athletes from group C than in students (group A). We conclude that the myosin profile in the athletes belonging to group C was unfavourable for their sport disciplines. This could be the reason why those athletes did not reach international level despite of several years of training.
Lipovitellin-phosvitin crystals from oocytes of the sturgeon (Acipenser boeri L.) and the tuatara... more Lipovitellin-phosvitin crystals from oocytes of the sturgeon (Acipenser boeri L.) and the tuatara (Sphenodon punctatus [Gray]) have been shown to he orthorhombic like all such known crystals from vertebrates higher than cyclostomes. Lattice parameters and projected crystal views also resembled closely the hitherto known cases. The high evolutionary conservation of this crystal structure probably reflects a cell-biological significance of the tertiary structure of crystal constituents, especially of the lipovitellines. A comparative consideration of crystallographic data from cyclostomes and higher vertebrates suggests that cyclostomes started with one species and all higher vertebrates with two species of lipovitellin molecules.
Cells of the gas gland of the perch Perca fluviatilis L ., stimulated to increased generation of ... more Cells of the gas gland of the perch Perca fluviatilis L ., stimulated to increased generation of gas by the repeated emptying of the swim-bladder, were examined in the electron microscope . Intense activity of the nuclear envelope was demonstrated . Simple vesicles originating from the external nuclear membrane and the so-called multivesicular bodies derived from the outpocketings of both membranes of the nuclear envelope were observed . The multivesicular bodies were filled with numerous fine vesiculae arising from the active proliferation of their internal membrane . The authors offer two alternative mechanisms of formation of fine vesiculae inside the multivesicular bodies and the mechanism of the tearing away of these bodies from the nuclear envelope .
The main scope of our work was to investigate the usefulness of the PROZA correction method and C... more The main scope of our work was to investigate the usefulness of the PROZA correction method and CALIBRATION CURVE construction for quantitative X-ray microanalysis of biological soft, freezedried tissues. The test samples with a known elemental concentration of NaCl, MgCl 2 , KH 2 PO 4 and Na 2 SO 4 were prepared on the basis of 20% porcine skin gelatine. A control sample containing 20% gelatine was used for sulphur subtraction. Dissolved gelatine solutions were cooled with liquid nitrogen and then cut in cryomicrotome into 16 mm thick slices. The sections were lyophilised, coated with carbon and analysed by means of a scanning electron microscope combined with an energy dispersive (ED) spectrometer. The homogeneity of the prepared samples was verified using the Fisher's test. Only homogenous samples were used for calibration. The significance of the 0(&z) (PROZA) correction method for biological sample analysis was verified by comparing the prepared standard and entering it into the Voyager computer program memory with the remaining samples of a known elemental content. The differences between the standard and the samples were noted for all elements analysed. There was no sample matching the standard after the PROZA correction procedure. A high correlation r factor (above 0.99) for all analysed elements indicates that CALIBRATION CURVES construction could be suitable for quantitative X-ray microanalysis of biological samples.
The main objective of our work was to investigate the possibility and usefulness of indirect meth... more The main objective of our work was to investigate the possibility and usefulness of indirect methods in X-ray microanalysis for the quanti®cation of biologically important compounds. Metallothioneinlike proteins (MT-like proteins) from kidney and liver, rich in sulfur were chosen as an indicator of heavy metal presence in cells and their environment. Tissues from gold®sh (Carassius auratus gibelio) were sampled after short and prolonged periods of exposure to Co 2 , CrO À2 4 , Pb 2 , Cu 2 and control treatment and prepared for histochemical staining for peroxidated thiolate groups. Commonly used ±S±S± bonds dye (Nitro Red) was replaced with iodine atoms and they were quanti®ed at L line by means of X-ray microanalysis combined with SEM. After ®sh treatments with heavy metal solutions changes in MT-like proteins and in I atom contents were expected. There was statistically signi®cant decrease in MT-like proteins level in kidney after lead treatment (Pb=C 0.62). In liver a statistically signi®cant increase in MT-like proteins concentration was observed after chromium, cobalt and lead ions treatment in comparison to control animals. The following ratios were noted: 3.04 for Cr=C, 2.18 for Co=C and 2.10 for Pb=C. Our ®nding indicates that the method of indirect measurement of MT-like proteins in ®sh and other animal tissues is possible. The concentration of iodine atoms is above their detection level by EDS and their changes are possible to identify. During histochemical procedures it is worth taking into account sample preparation methods which might disturb the quality and quantity of the analysed material.
The variability of Actinocyclus kuetzingii (A.S.) Sim. in the Littorina deposits of the Vistula L... more The variability of Actinocyclus kuetzingii (A.S.) Sim. in the Littorina deposits of the Vistula Lagoon and Lake Druzno were studied. During the time of the Littorina Lake Druzno was a part of the Vistula Lagoon. In the different parts of the Lagoon both the abundance of A. kuetzingii and the diameter of its valves change in a manner associated with phases of the Littorina transgression. Coscinodiscus (Actinocyclus) kuetzingii fo. subsalsa Brockmann is probably an ecotype of A. kuetzingii (A.S.) Sim.
It is generally accepted that connexin43 (Cx43) is a major constituent of heart and myometrial ga... more It is generally accepted that connexin43 (Cx43) is a major constituent of heart and myometrial gap junctions. However, the presence of Cx43 gap junctions in non-pregnant myometrium is still poorly documented. Tissue sections of porcine heart and non-pregnant uterus and myometrial smooth muscle cell cultures were immunostained with monoclonal antibody against Cx43. In the heart, intensive immunostaining was confined to the intercalated discs as previously reported. In the non-pregnant uterus, punctuate immunostaining of Cx43 was seen throughout the myometrium along cell interfaces between myocytes. The expression of Cx43 was sustained in cultured smooth muscle cells isolated from non-pregnant myometrium. Western blotting has detected single isoform of Cx43 in both, cardiac and myometrial tissues. The electrophoretic mobility of porcine heart Cx43 was similar to that of myometrial isoform but different from the pattern of mobility of Cx43 of the rat heart. Hence, porcine myometrium ma...
The synchronous contractions of the uterus in labour depend on electrical coupling of myometrial ... more The synchronous contractions of the uterus in labour depend on electrical coupling of myometrial smooth muscle cells by gap junctions. In the human myometrium, gap junctions are scarce in the non-pregnant uterus, but become abundant at term in preparation for labour. We have previously demonstrated that in the human myometrium at term, three different gap-junctional proteins are expressed, connexins 43, 45, and 40. These connexins are known to have distinctive functional capacities in in vitro expression systems but whether, in the human myometrium in vivo, they are co-assembled into the same gap junction or form different types of gap junction has previously been unclear. By applying triple immunogold labelling to sections of Lowicryl-embedded tissue for electron microscopy, together with complementary immunoconfocal microscopy, we demonstrate here that connexins 43, 45, and 40 are commonly present as mixtures within the same gap-junctional plaque. While all gap junctions contain c...
We analyzed the DNA content of hepatocyte and erythrocyte nuclei of the spined loach Cobitis taen... more We analyzed the DNA content of hepatocyte and erythrocyte nuclei of the spined loach Cobitis taenia (diploid) and its allopolyploid forms. Twenty triploid females and one tetraploid were used. At least 20,000 hepatocyte and erythrocyte nuclei were acquired and analyzed by flow cytometry. C. taenia erythrocyte nuclei contain 3.15 ± 0.21 pg of DNA and the hepatocyte nuclei 4.45 ± 0.46 pg of DNA. Triploid Cobitis have 5.08 ± 0.41 pg of DNA in erythrocyte nuclei and 6.11 ± 0.40 pg of DNA in hepatocyte nuclei, whereas the tetraploid erythrocyte and hepatocyte nuclei contained 6.60 and 7.40 pg of DNA, respectively. In general, the DNA contents correlate positively with the ploidy level of the fish investigated. The DNA content variation in the hepatocyte and erythrocyte nuclei may be due to differences in extent of chromatin condensation, which is more pronounced in the erythrocyte than hepatocyte nuclei, or to the several orders of ploidy that occur in the parenchymal liver cells.
The aim of this study was to compare DNA content in hepatocyte and erythrocyte nuclei of the Euro... more The aim of this study was to compare DNA content in hepatocyte and erythrocyte nuclei of the European sunbleak, Leucaspius delineatus, in relation to nuclear and cell size by means of flow cytometry and fluorescence microscopy. The DNA standards, chicken and rainbow trout erythrocytes, were prepared in parallel with both cell types, with initial separation of liver cells in pepsin solution followed by cell filtering. Standards and investigated cells were stained with a mixture of propidium iodide, citric acid, and Nonidet P40 in the presence of RNAse, and fluorescence of at least 50,000 nuclei was analyzed by flow cytometry. Average cell size was determined by flow cytometry, using fresh cell suspension in relation to latex beads of known diameter. The size of nuclei was examined on the basis of digital micrographs obtained by fluorescence microscopy after nuclei staining with DAPI. The sunbleak's erythrocyte nuclei contain 2.25 ± 0.06 pg of DNA, whereas the hepatocyte nuclei contain 2.46 ± 0.06 pg of DNA. This difference in DNA content was determined spectroscopically using isolated DNA from the two cell types. The modal diameters of the erythrocytes and hepatocytes were estimated to be 5.1 ± 0.2 and 22.3 ± 5.0 lm, respectively, and the corresponding modal dimensions of their nuclei (measured as surface area) were 15.2 and 21.4 lm 2 , respectively. The nucleoplasmic index, as calculated from diameters estimated from surface area of nuclear profiles, was 2.51 for the erythrocytes compared with 0.08 for hepatocytes.
Zeitschrift für Zellforschung und Mikroskopische Anatomie, 1970
Two types of glandular cells are present in the pars intermedia of the loach, Misgurnus fossilis.... more Two types of glandular cells are present in the pars intermedia of the loach, Misgurnus fossilis. Basophils are characterized by the presence in their cytoplasm of numerous secretory granules containing electron-dense and homogeneous material and by scarce endoplasmic reticulum. ‘Weak’ acidophils contain in their cytoplasm abundant endoplasmic reticulum and numerous granules of different electron densities.
Zeitschrift für Zellforschung und Mikroskopische Anatomie, 1969
The fine structure of the gas gland cells in Perca and Acerina was studied with the light- and el... more The fine structure of the gas gland cells in Perca and Acerina was studied with the light- and electron microscope. In some of the animals the gas glands were stimulated by the single or repeated removal of the gas from the swimbladder. In this group of fishes the changes in the organization of cytoplasm of the gas gland cells accompanying
Yolk-platelet crystals in Amia calva L., Lepisosteus osseus (L.) and Polypterus bichir have ortho... more Yolk-platelet crystals in Amia calva L., Lepisosteus osseus (L.) and Polypterus bichir have orthorhombic features with unit-cell dimensions a = 8.3 ... 8.8 nm, b = 16.4 ... 16.9 nm and c = 18.6 ... 19.8 nm as determined in electron-diffracted patterns of fixed, epoxy-resin embedded and thin-sectioned material. Electron-diffraction patterns, crystal projections and the above unit-cell data make them extremely similar to the orthorhombic yolk-platelet crystals known for amphibians and teleosts. This observation fills a gap in yolk-platelet research and supports the view that the general architecture of yolk platelets has been conserved for nearly 400 million years. It follows that the peculiar platelet architecture itself has physiological significance.
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Papers by Wincenty Kilarski