Expression of matrix metalloproteinase 2 (MMP-2), E-cadherin and Ki-67 in metastatic and non-metastatic canine mammary carcinomas

Background

The aim of the study was to demonstrate the immunohistochemical expression of proteins that affect the metastatic potential of a tumour, including matrix metalloproteinase 2 (MMP-2) and E-cadherin. Another objective was to determine their correlation with the expression of the Ki-67 antigen in metastasizing and non-metastasizing mammary carcinomas in female dogs. The study was conducted on 32 canine mammary carcinomas (12 metastatic and 20 non-metastatic), classified as simple tubular and tubulopapillary carcinomas. Immunohistochemistry was performed to evaluate the expression of MMP-2, E-cadherin and Ki-67 antigen.

Results

MMP-2 was expressed in 85 % of the non-metastatic tumours and in all the metastatic tumours, while E-cadherin was expressed in 85 % of the non-metastatic tumours and in 66 % of the metastatic tumours. The Ki-67 antigen was expressed in 65 % of the non-metastatic tumours and in 91 % of the metastatic tumours. The mean Ki-67 expression was slightly higher in tumours that had metastasized (1.5?±?0.90 vs 1.1?±?0.94; p?=?0.22). A similar relationship was found in terms of the intensity of the MMP-2 expression (2.9?±?1.9 vs 2.7?±?2.4; p?=?0.50). A decrease in the expression of E-cadherin (2.8?±?2.5) was found in metastatic tumours compared to the expression in non-metastatic tumours (3.2?±?2.3). However, these differences were not statistically significant (p?=?0.63).

Conclusion

We did not show significant differences in MMP-2, E-cadherin and Ki-67 expression between metastatic and non-metastatic tumours due to low number of cases studied, however further experiments are necessary to assess the role of these antigens in the process of canine mammary tumours metastasis.

     

“In Situ” Amendments and Revegetation Reduce Trace Element Leaching in a Contaminated Soil

Various amendments and/or a plant cover (Agrostis stolonifera L.) were assessed for their potential to reduce trace element leaching in a contaminated soil under semi-arid conditions. The experiment was carried out in field containers and lasted 30 months. Five treatments with amendments (leonardite (LEO), litter (LIT), municipal waste compost (MWC), biosolid compost (BC) and sugar beet lime (SL)) and a plant cover and two controls (control without amendment but with plant (CTRP) and control without amendment and without plant (CTR)) were established. Drainage volumes were measured after each precipitation event and aliquots were analysed for pH, electrical conductivity (EC) and trace element concentrations (As, Cd, Cu, Pb and Zn). Soil pH and trace element extractability (0.01 M CaCl₂) at three different depths (0–10, 10–20 and 20–30 cm) were measured at the end of the experiment. Incorporation of amendments reduced leaching of Cd, Cu and Zn between 40–70% in comparison to untreated soil. The most effective amendments were SL, BC and MWC. At the end of the experiment, extractable concentrations of Cd, Cu and Zn were generally lower in all amended soils and CTRP compared to CTR. Soil pH decreased and extractability of metals increased in all treatments in relation to depth. Results showed that use of these amendments combined with healthy and sustainable plant cover might be a reliable option for “in situ” stabilization of trace elements in moderately contaminated soils.     

Further studies on the etiopathology of malabsorption syndrome in broiler chickens

The paper presents two cases of malabsorption syndrome in broiler chickens. The results indicate that the etiological factor appears to be the virus agent which causes necrosis in extra-excretional parts of the pancreas. During the course of the disease disorders in the enzymatic activity of gammaglutamyltransferase, leucylaminopeptidase and alanylaminopeptidase were observed. It consequently leads to the deterioration of intestines digestion and resorption of feed components. Application of Selcewet-Polfa to the diseased birds improved the health of the birds, morphological picture of peripheral blood and mineralisation of thigh bones. No radical improvement in the compensation of the final body weight was recorded.     

Native soil organic matter as a decisive factor to determine the arbuscular mycorrhizal fungal community structure in contaminated soils

The present study of arbuscular mycorrhizal (AM) fungi is focused on the identification of AM ecotypes associated with different plants species (Poa annua, Medicago polymorpha, and Malva sylvestris) growing in three contaminated soils with different organic matter, phosphorus, and trace element (TE; Cu, Cd, Mn, and Zn) contents. Soils were amended with biosolid and alperujo compost. Shifts in AM fungal community structure, diversity, richness, root colonization, and plant TE uptake were evaluated. Soil properties and plant species had a significant effect on AM fungal community composition as well as on root colonization. However, AM fungal diversity and richness were only affected by soil properties and especially by soil organic matter that was a major driver of AM fungal community. As soil quality increased, Glomeraceae decreased in favor of Claroideoglomeraceae in the community, AM fungal diversity and richness increased, and root colonization decreased. No effect due to amendment (exogenous organic matter) addition was found either in AM fungal parameters measured or TE plant uptake. Our results revealed that the role of TE contamination was secondary for the fungal community behavior, being the native organic matter content the most significant factor.     

Assessment of trace element phytoavailability in compost amended soils using different methodologies

Purpose

This study evaluates the effects of two soil amendments and the growth of two plant species on labile trace element (TE) fractions in two different contaminated soils.

Materials and methods

We studied the effects of two organic amendments (biosolid compost and alperujo compost) and two plant species (Medicago polymorpha and Poa annua) on pH, total organic carbon (TOC), and TE availability, by three extraction methods (CaCl₂ aqueous solution, soil pore water (SPW), and diffusive gradient in thin film (DGT)), in two contaminated soils with contrasting pH values (Aznalcázar, 6.53, and Vicario, 3.48) in a 118-day pot experiment. The effects of the composts on labile TE fractions were compared with element concentrations in plants.

Results and discussion

No relevant effects of amendments and plants were found on the physical and chemical characteristics of the Aznalcázar soil. However, the addition of amendments was essential for plant species growing in the acid Vicario soil. In this soil, amendments and plant growth increased pH and TOC and reduced substantially TE bioavailability. Although absolute values of bioavailable TE contents obtained by the three methods were very different and followed the trend CaCl₂ extraction?>?SPW?>?DGT, these values follow a similar behavior in the two studied soils and for the two species.

Conclusions

The results demonstrate that the application of organic amendments are suitable for remediating acid TE-contaminated soils, for the establishment of a vegetation cover on previously bare soils for reducing wind and water erosion and for reducing labile TE fractions to prevent leaching of pollutants into subsoil or groundwater layers. Moreover, the results obtained in this study pointed out that under microcosm conditions, the three methods tested (CaCl₂ extraction, SPW, and DGT) to predict TE bioavailability were highly correlated.

     

Factors regulating ovarian function in pigs

The hormonal interactions of the hypothalamic-pituitary-ovarian-uterine axis are accountable for a normal reproduction in female pigs. It is of importance to have knowledge of estrous symptoms and hormonal profiles around ovulation. The introduction of the transrectal ultrasonography in sows has given us the possibility to study ovarian activity in conscious animals and relate the timing of estrus to ovulation. Combining this technique with measuring of several hormones like luteinizing hormone (LH), follicle-stimulating hormone (FSH), inhibin, estradiol, progesterone, insulin-like growth hormone I (IGF-I), prostaglandin F2alpha (PGF2alpha) metabolite, oxytocin, facilitate our knowledge about the sequence of ovarian events. Evidence suggests that activation of the hypothalamic-pituitary-adrenal axis may hamper the normal gonadotropin secretion and in consequence, the ovarian function. The metabolic status during lactation, weaning of piglets and social stress might affect onset of ovarian activity and the related estrous behavior. The role of seminal plasma, artificial insemination and presence of the boar might also be included as factors regulating the temporal kinetics of ovulation, corpus luteum development, uterine function and steroid production in the ovary. Studies using a simulated stress by means of adrenocorticotrophic hormone (ACTH) administration or food deprivation are tools in understanding how the ovary is susceptible to impairment. The intention of this paper is to review current knowledge concerning the endocrine aspects of normal and stress-influenced ovarian function in pigs.     

Studies on nucleolar organiser regions (AgNors) in selected spontaneous and transplantable animal tumors

Behaviour of argyrophilic nucleolus organising regions (AgNOR) was estimated in various types of spontaneous and transplantable tumors in animals. The studies were performed on spontaneous epithelial and mesenchymal tumors, malignant and non-malignant, as well as transplantable tumors: Morris hepatoma, mammary gland carcinoma and Yoshid sarcoma. The examinations were made on paraffin sections, using silver-staining method according to Ploton et al. Quantitative assessment was made with computer-aided microscopic image analysis system Multi-Scan Base V.8 for Windows, coupled with Carl Zeiss microscope. It was demonstrated that AgNOR index reflects malignancy of the tumor, since it increases clearly in cancers and sarcomas, both spontaneous and transplantable. The highest AgNOR index--0.13--was noted in the group of spontaneous tumors in epithelial malignant tumors, and in the group of transplantable tumors in mesenchymal tumors (Yoshid sarcoma) it was 0.15. Classification of the studied spontaneous and transplantable tumors into groups of the same histogenesis, though phenotypically different, was aimed at demonstration of the increasing tendency of AgNOR index.     

Impact of Exogenous ACTH During Pro-Oestrus on Endocrine Profile and Oestrous Cycle Characteristics in Sows

Sows housed in freely moving groups have elevated cortisol levels until the rank order is established, which takes place within approximately 48 h. The aim of this investigation was to study the effect of repeated administration of synthetic adrenocorticotropic hormone (ACTH; Synacthen Depot), during the follicular phase (pro-oestrus) on oestrus, ovulation and endocrine parameters. Four multiparous sows were used. Follicular growth and ovulation were recorded by ultrasonography. The first oestrous cycle after weaning was used as control cycle. Onset of oestrus in the sow occurs 3-4 days after the time when plasma progesterone reaches a concentration of 8 nmol/l. The progesterone profile in the control cycle of the individual sow was used for estimation when the ACTH injections should start. In the third pro-oestrus ACTH (2.5 microg/kg) was given via an indwelling catheter every 2 h for 48 h. The sows were euthanased 4-6 days after onset of the third oestrus and the ovaries were examined. Cortisol levels were elevated during the treatment period (p < 0.05). The second cycle, in which the sows were injected with ACTH, was prolonged with 2.5 days compared with the control cycle (p < 0.05). The oestradiol pattern during oestrus was similar in the control and the treatment cycle in ovulating sows. Three sows had ovulated (fresh corpora lutea), but the ovaries contained additionally one or several luteinized follicles/cysts. In conclusion, ACTH administration during pro-oestrus caused a prolongation of the oestrous cycle and a disturbed follicular development.     

Effects of Exogenous ACTH during Oestrus on Early Embryo Development and Oviductal Transport in the Sow

This study was conducted to assess the effects of ACTH injections on the early development of embryos and their transportation to the uterus. Fifteen sows were monitored for ovulation using transrectal ultrasonography during the first two oestrous periods after weaning. The sows were randomly divided into a control group (C group, n = 8) and an ACTH-treated group (ACTH group, n = 7), and were all surgically fitted with intra-jugular catheters. From the onset of the second standing oestrus after weaning, the sows were injected (NaCl/synthetic ACTH) every 4 h. Blood samples were collected immediately before and 45 min after each injection. All sows were inseminated once 10-33 h before ovulation in their second oestrus after weaning. At 48 (n = 4) or 60 (n = 11) h after ovulation during their second oestrus, the sows were killed and the embryos retrieved from the oviduct and uterus. The embryos were counted and compared with the number of corpora lutea, cleavage rate was noted and, finally, the embryos were prepared for confocal laser scanning microscopy and transmission electron microscopy. There was no difference between the groups regarding cleavage rate, the cytoskeleton, or the number of active nucleoli. However, the ACTH group had significantly (p < 0.05) fewer ova/embryos retrieved (51%) than the C group (81%), and there was a tendency towards faster transportation to the uterus in the ACTH group, possibly because of high progesterone concentrations during treatment. To conclude, administration of ACTH every 4 h from onset of oestrus to 48 h caused significant loss of oocytes or embryos, and possibly faster transportation through the oviduct.