Jan Danko

Bendiocarbamate induced structural alterations in rabbit thymus after experimental peroral administration

In this study histological structure of rabbit thymus after bendiocarbamate (2,3–isopropyledene–dioxyphenyl methylcarbamate) administration was studied. Bendiocarbamate was perorally administered for 90 days. At Day 3, 10, 20, 30, 60, 90 morphometric analysis was realized. Quantitative evaluation showed that in the control group thymus cortex forms 57.94 ± 7.10% and medulla 35.94± 7.38%. In almost all experimental groups significantly higher relative volume of cortex and lower relative volume of medulla was detected. Detail morphometric analysis found that the number of thymocytes per constant area and the diameter of tymocytes was decresed after bendiocarbamate administration. The number and diameter of reticular cells was not affected. Results of this study suggest negative effect of bendiocarbamate on the formation of thymus structures.

In vitro effect of nickel on bovine spermatozoa motility and annexin V-labeled membrane changes

In this study the effect of in vitro culture of bovine spermatozoa with nickel (NiCl2) on spermatozoa motility and membrane changes was analyzed. The spermatozoa motility significantly decreased after 120 min of culture at the concentration of 1000 μm Ni ml−1 (P < 0.05) and after 240 min of culture at the concentration of 500 and 1000 μm Ni ml−1 (P < 0.001) as compared with control. The progressive motility was the highest in the control group and in the groups with the lowest nickel concentrations (7.8 and 125 μm Ni ml−1). The progressive spermatozoa motility was significantly altered even after 30 min of culture in the group with the highest nickel concentration (1000 μm Ni ml−1). A significant decrease in progressive motility from the concentration of 250 μm Ni ml−1 was detected after 240 min of culture. Concentrations from 125 μm Ni ml−1 in various time periods of culture stimulated spermatozoa motility after 30 min (P < 0.001), but later an inhibitory effect was noted. After 240 min of in vitro spermatozoa culture with 125 μm Ni ml−1 a typical Annexin V fluorescence reaction was detected. Fluorescence was detected in mitochondrial segment of bovine spermatozoa. In spermatozoa exposed to higher nickel concentrations the Annexin V‐positive reaction was detected also on the spermatozoa head membrane. In the group with the highest concentration and the longest time of exposure (1000 μm Ni ml−1; 240 min) the apoptotic Annexin‐positive regions were detected not only in the mitochondrial part, but also in the spermatozoa head (acrosomal and postacrosomal part), showing significant alteration of spermatozoa membrane integrity. Copyright

In vitro effects of radiofrequency electromagnetic waves on bovine spermatozoa motility

In this study the effects of 1800 MHz GSM–like radiofrequency electromagnetic waves (RF–EMW) exposure on bovine semen was monitored. The experimental samples were analyzed in vitro in four time periods (0, 30, 120 and 420 min) and compared with unexposed samples (control). Spermatozoa motility was determined by computer assisted semen analyzer (CASA). Evaluation of the percentage of motile spermatozoa showed significant (P < 0.001) decrease in experimental groups after 120 and 420 min of culture when exposed to microwaves, in comparison to control. Similar spermatozoa motility inhibition was detected for the percentage of progressively motile spermatozoa, too. Average path distance decreased significantly (p < 0.001) in experimental groups after 30 and 420 min of culture. Path velocity increased in the experimental groups exposed to RF–EMW after 30 minutes of culture, but subsequently decreased after 420 min of culture, in comparison to control. This indicates a possible initial stimulation and subsequent velocity inhibition of bovine spermatozoa under RF–EMW exposure. Changes in spermatozoa motility were also detected for some fine parameters, too. A significant decrease (P < 0.001) was noted for amplitude of lateral head displacement in the experimental group after 420 minutes of culture. Detailed in vitro motility analysis of bovine spermatozoa exposed to microwave radiation suggested that the parameters of path and velocity at the beginning of the culture significantly increase, but after longer culture (420 minutes) a significant decrease occur in the experimental group as compared to control. In general, results of this experiment indicate a negative time–dependent effect of 1800 MHz RF–EMW radiation on bovine spermatozoa motility.

In vivo and in vitro effect of bendiocarb on rabbit testicular structure and spermatozoa motility

In this study the effect of bendiocarb on the rabbit testicular structure and spermatozoa motility was investigated. For testicular structure evaluation the animals were fed with bendiocarb tablets daily at a dose of 5 mg/kg of body weight for 13 days. The relative volume of the germinal epithelium, interstitium and lumen was measured. The testicular structure evaluation showed decreased relative volume of germinal epithelium in both experimental groups in comparison with the control group. The relative volume of the interstitium was increased in both experimental groups. An increase of the relative volume of the lumen was registered also in both experimental groups. Qualitative analysis detected a dilatation of blood vessels in the interstitium, undulation of the basal membrane and some empty spaces in the germinal epithelium after bendiocarb administration. The spermatozoa motility was evaluated by the computer assisted semen analyzer (CASA) method in various time intervals (0–180 minutes) and the bendiocarb concentration in the culture medium added to experimental groups varied from 0.054 to 0.268 mg/mL. Spermatozoa motility and progressive motility significantly decreased with increased bendiocarb administration and with extending the period of incubation. For other fine motility parameters, a decrease dependent on the time of incubation and on the bendiocarb concentration almost in all experimental groups in comparison to the control was detected. These results clearly suggest that in vitro also in vivo bendiocarb administration decrease male fertility.

Arterial Peculiarities of the Thoracolumbar Spinal Cord in Rabbit

The aim of this study was to investigate the arterial blood supply of the thoracolumbar spinal cord in rabbit. The study was carried out on twenty adult New Zealand white rabbits. Ten rabbits were used in the corrosion technique and ten rabbits in the dissection technique. After the killing, the vascular network was perfused with saline. Batsons corrosion casting kit no. 17

Changes of the immunological and haematological parameters in rabbits after bendiocarbamate application

The effect of bendiocarbamate application (5 mg/kg b.w.) on the haematological and immunological parameters in rabbits was evaluated. Total leukocyte cell count, erythrocyte cell count, differential cell count were determined during the period of three months of bendiocarbamate application and compared with those in healthy animals. The immunotoxic effect was evaluated by the test of ingestion ability of phagocytes (phagocytic activity and index of phagocytic activity) and proliferation activity of lymphocytes after mitogen stimulation. The significant decrease of total leukocytes, lymphocytosis and neutropaenia were found after bendiocarbamate application. The functional activities of phagocytes (expressed as phagocytic activity) and lymphocytes (proliferative activity) were significantly suppressed in rabbits treated with bendiocarbamate compared with those in control groups and values before the experiment.

Anatomical description of the renal arteries and veins in the European rabbit

The aim of this study was to describe origin, localisation and variations of renal arteries and veins in the rabbit. The study was carried out on 40 adult European rabbits. We prepared corrosion casts of the rabbit arterial and venous system. Spofacryl was used as the casting medium. In 75% of cases the origin of arteriae renales was located at the level of the third lumbar vertebra and in remaining 25% of cases arteria renalis dextra branched off at the level of the second lumbar vertebra. In 10% of cases we observed that the number of arteria renalis sinistra was doubled. We recorded also in one case the presence of arteria renalis accessoria for ren dexter. In 10% of cases we observed that the number of vena renalis sinistra was doubled. In 5% of cases two venae renales sinistrae arose from the kidney and subsequently, about 1 cm from opening to vena cava caudalis, they united to form a single vein. In 5% of cases two venae renales sinistrae arose from the kidney and subsequently, approximately 1 cm away from hilus renalis, they united. The obtained variations of the number of renal arteries were partially homologous to the human, but variations of renal veins were localized on the other side as in human.

The Effect of Transgenesis on Rabbit Thyroid Tissue Structure

With 1 figure and 3 tables

Dynamics of Evans blue clearance from cerebrospinal fluid into meningeal lymphatic vessels and deep cervical lymph nodes

Abstract Objectives Recently, it has been confirmed, that excess fluid and waste products from the brain are drained into the cerebrospinal fluid (CSF) and afterwards cleared via the olfactory route and/or lymphatic vessels in the brain dura and corresponding extracranial lymphatic structures. Therefore, the aim of present study was to monitor time-dependent uptake of Evans blue (EB) tracer from subarachnoid space into the meningeal lymphatic vessels and extracranial lymph nodes in rats during 3 hours–12 days. Methods EB was injected into the cisterna magna of anesthetized rats and after required survival, plasma, brain dura matter and corresponding lymph nodes (cervical, thoracic and lumbar) were dissected and processed for lymphatic vessels analyses using immunofluorescence and immunohistochemistry. Furthermore, we have used sensitive ultra-high-performance liquid chromatography (UHPLC) method for the determination of EB concentrations in selected samples. Results Using a combination of imaging methods, we have detected two different types of the vascular structures in the brain dura and in deep cervical lymph nodes. The blood vessels, which were RECA-1 + positive and the lymphatic-like vessels, expressing bright intense red fluorescence of EB tracer. Subsequently, using UHPLC with UV detection, we have quantified the EB concentration in positive structures by 3 hours up to 12 days after tracer delivery. A significant increase of EB concentration was detected in deep cervical lymph nodes already at 3 hours with a peak at 1 day that decreased to about one-tenth of its peak value by 12 days. Similar pattern was detected in brain dura. On the contrary, the brain tissue and plasma were almost negative for EB tracer during all tested time periods. Conclusion Our results demonstrate the dynamic changes of EB in meningeal lymphatic vessels and in deep cervical lymph nodes, thus recapitulating the downstream outflow of intracisternally injected tracer during 3 hours–12 days via dura mater lymphatic vessels towards corresponding extracranial draining system, particularly the deep cervical lymph nodes.

The Anatomical Correlation between the Internal Venous Vertebral System and the Cranial Venae Cavae in Rabbit.

The aim of this study was to describe the possible variations in the connection between the internal venous vertebral system and the cranial vena cava in rabbit using corrosion technique. The study was carried out on 40 adult New Zealand white rabbits. The venous system was injected by using Batsons corrosion casting kit number 17. We found the connection between the internal venous vertebral system and the cranial vena cava by means of the vertebral veins and the right azygos vein. The vertebral vein was present as independent tributary in 36 cases (90%). In the rest of the cases, it was found as being double, being triple, or forming a common trunk with other veins. The azygos vein was present as independent tributary of the cranial vena cava in 39 cases (97.5%). We found also a common trunk formed by the junction of the deep cervical vein, the right vertebral vein, and the azygos vein in one case (2.5%). The azygos vein received 6, 7, 8, or 9 pairs of dorsal intercostal veins. Documenting the anatomical variations in the rabbit will aid in the planning of future experimental studies and determining the clinical relevance on such studies.