RESUMEN
The aim of this study was to determine the serum concentrations of acute phase proteins (APPs), including C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp) in dogs with circumanal gland tumours. Investigations were conducted on 39 male dogs of mixed breed. The animals were divided into four groups depending on the degree of tumour malignancy and type of hormones that were dominant in the bloodstream. All measurements of serum APPs were conducted by the use of commercial diagnostic kits. In dogs with benign tumours, the concentrations of each APP decreased during therapy, while in dogs with malignant tumours, despite anti-tumour therapy, concentrations of APPs in serum increased markedly. The results of this study suggest that changes in serum concentrations of CRP, SAA and Hp during anti-hormone therapy may be a reliable marker for differentiating tumour grade and degree of malignancy. Including APPs in routine diagnostics may assist with formulating a prognosis for the outcome of the disease, before implementing anti-tumour therapy. Moreover, monitoring the effectiveness of therapy may be possible based on the determination of serum APP concentrations.
Asunto(s)
Proteínas de Fase Aguda/metabolismo , Adenoma/veterinaria , Canal Anal/patología , Neoplasias de las Glándulas Anales/sangre , Carcinoma/veterinaria , Enfermedades de los Perros/sangre , Adenoma/sangre , Adenoma/patología , Neoplasias de las Glándulas Anales/patología , Animales , Biomarcadores/sangre , Carcinoma/sangre , Carcinoma/patología , Enfermedades de los Perros/patología , Perros , MasculinoRESUMEN
The purpose of this study was to determine concentrations of IL-2, IL-10, TGF-ß1 in serum and T regulatory cell (Treg) percentage in peripheral blood of dogs with perianal tumours. Investigations were conducted on 32 male dogs of mixed breed. The animals were divided into 4 experimental groups and control group. The groups were established depending on the tumour malignancy degree and the type of dominant hormones. All measurements of serum cytokine concentrations were conducted by the use of commercial diagnostic ELISA kits. Treg lymphocyte percentage was measured by flow cytometry. In both groups with benign tumours cytokine levels decreased during therapy, whilst in groups with malignant tumors, in spite of applying anti-tumour therapy, concentrations of cytokines in serum markedly increased. The mean percentage of Treg lymphocytes in dogs with benign tumours (group I and II) was significantly lower than the mean percentage of these cells in control group at all time points, but after applying of anti-hormonal therapy, the significant increase of Treg percentage was observed compared to baseline values. By contrast, in both groups with malignant tumours (group III and IV), the mean percentage of Treg lymphocytes was significantly higher at the beginning of the experiment comparing with the control group as well as both groups with benign tumours and this percentage increased during anti-tumour therapy. The results of this study suggest that monitoring changes in cytokine serum concentrations and Treg percentage in the bloodstream during anti-hormonal therapy may constitute a subsidiary marker in the monitoring of therapy effectiveness, in prognosis the outcome of a disease or in differentiating tumour degree of malignancy.
Asunto(s)
Neoplasias de las Glándulas Anales/inmunología , Citocinas/sangre , Enfermedades de los Perros/inmunología , Linfocitos T Reguladores/inmunología , Neoplasias de las Glándulas Anales/sangre , Animales , Enfermedades de los Perros/sangre , Perros , Ensayo de Inmunoadsorción Enzimática/veterinaria , Citometría de Flujo/veterinaria , Interleucina-10/sangre , Interleucina-2/sangre , Activación de Linfocitos , Masculino , Linfocitos T Reguladores/fisiología , Factor de Crecimiento Transformador beta1/sangreRESUMEN
Pyometra is a serious problem in dairy cow herds, causing large economic losses due to infertility. The development of pyometra depends mainly on the immunological status of the cow. The aim of the study was a comparative evaluation of selected indicators involving non-specific and specific immunity in cows with pyometra and in cows without inflammation of the uterus. The study was performed in 20 cows, which were divided into two groups: pyometra group and healthy group, each comprising 10 cows, based on the results of cytological and ultrasonographic tests. A flow cytometric analysis was performed for the surface molecules CD4, CD8, CD14, CD21, CD25 and CD4(+) CD25(+) on leucocytes, and the phagocytic activity was determined from granulocytes and monocytes/macrophages in the peripheral blood and uterine washings, respectively. It was demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower in cows with pyometra compared with the healthy group (p < 0.001). Significantly (p ≤ 0.001) lower percentage of CD4(+) , CD14(+) , CD25(+) and CD4(+) CD25(+) phenotype leucocytes was also observed in the peripheral blood of cows from the pyometra group, along with a significantly higher (p < 0.001) percentage of CD8(+) and CD21(+) lymphocytes as compared to the healthy group. The results of work indicate that disfunction of cell immunity coexisting with pyometra may be caused by a bacterial infection and the presence of blocking agents (IL-10), released by the increasing number of CD8(+) lymphocytes what leads to the advanced inflammation of uterus.
Asunto(s)
Enfermedades de los Bovinos/patología , Granulocitos/fisiología , Macrófagos/fisiología , Monocitos/fisiología , Fagocitosis/fisiología , Piómetra/veterinaria , Animales , Bovinos , Femenino , Citometría de Flujo/veterinaria , Piómetra/patologíaRESUMEN
Endometritis in dairy cow herds is a serious economic problem all over the world due to the large economic losses. The aim of the study was a comparative evaluation of selected indicators of non-specific and specific immunity in cows with subclinical endometritis and in cows without inflammation of the uterus. The study was performed on 40 cows on day 65 after delivery. Based on the results of cytological tests, the cows were divided into two groups: experimental (subclinical endometritis) and control (20 cows in each group). A flow cytometric analysis was performed for the leukocyte surface molecules CD4, CD8, CD14, CD21, CD25. Moreover the phagocytic activity of granulocytes and monocytes/macrophages in peripheral blood and uterine washings was determined. It has been demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower for cows with subclinical endometritis when compared to cows undergoing a normal puerperal period (p<0.001). A significant (p≤0.001) decrease in the percentage of CD4+, CD14+, CD25+ and CD4+CD25+ leukocytes was also observed in peripheral blood of the cows from the experimental group. In uterine washings a significant decrease (p<0.001) in CD21+ and increase in CD8+ lymphocytes was detected. The results indicate that dysfunction of cell immunity coexisting with subclinical endometritis may be the main factor causing advanced inflammation of the uterus. Knowledge of immunological mechanisms observed in cows with subclinical endometritis could aid in choosing the right adjuvant therapy using immunomodulating agents.
Asunto(s)
Biomarcadores/metabolismo , Enfermedades de los Bovinos/inmunología , Endometritis/inmunología , Inmunidad/fisiología , Animales , Enfermedades Asintomáticas , Biomarcadores/análisis , Biomarcadores/sangre , Sangre/metabolismo , Líquidos Corporales/metabolismo , Estudios de Casos y Controles , Bovinos/inmunología , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/metabolismo , Endometritis/diagnóstico , Endometritis/metabolismo , Femenino , Técnicas Microbiológicas , Pronóstico , Útero/inmunología , Útero/metabolismo , Útero/microbiologíaRESUMEN
This study was a comparative evaluation of selected immunological parameters in peripheral blood and uterine wash samples from cows with a normal postpartum period compared with cows with endometritis. We aimed to determine the usefulness of these parameters in monitoring the puerperium. In total, 40 cows were included in the study: 20 had endometritis (experimental group), and 20 did not have uterine inflammation (control group). Animals were chosen on the basis of cytological and bacteriological test results. The tests were conducted 5, 22, and 40 days postpartum. In both groups, flow cytometric analysis of the surface molecules CD4, CD8, CD21, CD25, and CD14 in the peripheral blood and uterine washings was performed. Granulocyte and monocyte phagocytic activity was determined using a commercial Phagotest kit that was adapted for flow cytometry. The percentage of phagocytic granulocytes and monocytes in both the peripheral blood and the uterine washings was significantly lower for cows in the experimental group compared with the control group (P < 0.01). A significant decrease (P < 0.01) in the percentage of CD4+, CD25+, CD14+, and CD4 + CD25(high) leukocyte subpopulations was also observed in the peripheral blood of cows with endometritis. A significant decrease (P < 0.01) in CD21+ lymphocytes and an increase in CD8+ lymphocytes was detected in uterine washings. The results of this work indicate that cell immunity dysfunction may be the main factor causing advanced inflammation of the uterus in endometritis. Knowledge of the immunological mechanisms observed in cows with endometritis might aid in choosing the correct immunomodulating agent-based adjuvant therapy.
Asunto(s)
Enfermedades de los Bovinos/patología , Endometritis/veterinaria , Granulocitos/citología , Leucocitos/citología , Monocitos/citología , Fagocitosis , Animales , Bovinos , Enfermedades de los Bovinos/sangre , Endometritis/sangre , Endometritis/patología , Femenino , Inmunidad Celular , Inmunidad Humoral , Fenotipo , Periodo Posparto , Útero/patologíaRESUMEN
The objective of the study was to evaluate cytometrically the percentage of apoptotic and necrotic spermatozoa in fresh semen of silver foxes in the breeding season. In males F3 and F4 with high percentages of early apoptotic (A+Pi-), late apoptotic (A+Pi+) and necrotic (A-Pi+) spermatozoa as well as 56-65% of living spermatozoa (A-Pi-) with progressive motility, the semen was characterised by reduced fertility. In males F1 and F2 with spermatozoa showing the motility and viability of 89-90% and high percentages of living cells that do not bind Annexin V and propidium iodide, the semen was assessed as valuable and useful for artificial insemination. Amongst 16 females of group I and II inseminated with semen from F1 and F2 males, 15 (93.75%) had multi-cub litters - on average 6.1 and 4.8, respectively. In contrast, amongst 16 females of group III and IV inseminated with semen from F3 and F4 males, only 10 (62.5%) had litters with few cubs (on average 2.6 in group III and 2.1 in group IV). Our findings explicitly indicate that semen of farm male foxes should be evaluated before the breeding season, as one of the causes of reproduction failures is likely to be a high percentage of apoptotic and necrotic spermatozoa. Thanks to flow cytometry, fresh ejaculates can be speedily evaluated and their usefulness for artificial insemination determined.
Asunto(s)
Apoptosis/fisiología , Cruzamiento , Citometría de Flujo , Zorros/fisiología , Semen/citología , Espermatozoides/fisiología , Animales , Femenino , Inseminación Artificial/veterinaria , Masculino , Espermatozoides/citologíaRESUMEN
Since numerous studies indicate that 2-methoxyestradiol (2-ME) as a metabolite of 17beta estradiol (17ß-E(2)) may exert antitumor activity by unclear mechanism, we undertake the study to elucidate the effect of 2-ME on oval cells (OC) activated by a carcinogenic choline deficient ethionine supplemented diet (CDE diet). Isolated OC were treated with different concentrations of 2-ME for 24, 48 and 72 hours. In these periods of time phenotypic studies, apoptosis detection and proliferative activity of cells were performed. A marked inhibition of OC proliferation was observed at the presence of 1.0 µM of 2-ME, with the lowest value obtained after 48 h. However, at the end of the cells' incubation, maximally reduced proliferative response of OC was attributed to 2.0 µM of 2-ME. Simultaneously with the time of incubation the amount of Thy-1-positive cells decreased slightly from 50.5±1.4% to 31.5±3.6%. Contrary to 1.0 and 2.0 µM of 2-ME, its lowest value (0.5 µM) reduced Thy-1 positive cells after 48 hours. The same 2-ME concentration resulted in the elevation of the cell number expressing CK-19. In turn, the marked increase of albumine-positive cells was observed under 1.0 µM of 2-ME and reaching 21.5±6.2 % and 23.9±5.7% after 48 and 72 hours, respectively. Although the presence of 1.0 µM of 2-ME dramatically intensified apoptosis within 24 h of cell culture, the percentage of apoptotic cells remained unchanged under 2.0 µM of 2-ME. When subjected to the carcinogenic effect of CDE, 2-ME exerts anti-proliferative, proapoptotic, and differentiation effects in OC.
Asunto(s)
Apoptosis/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Deficiencia de Colina/patología , Dieta , Estradiol/análogos & derivados , Etionina/administración & dosificación , Hígado/efectos de los fármacos , 2-Metoxiestradiol , Animales , Biomarcadores/metabolismo , Células Cultivadas , Deficiencia de Colina/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Estradiol/farmacología , Femenino , Hígado/metabolismo , Hígado/patología , Ratas , Ratas Wistar , Factores de TiempoRESUMEN
The aim of the study was to assess the effect of selected isolation methods on the viability and metabolism of bovine leukocytes. The cells were isolated using a Ficoll 1077, Histopaque 1083 gradient and osmotic shock method, and Ficoll or Histopaque with osmotic shock. Evaluation were made of the total number of cells, viability after isolation and in 24h culture on RPMI 1640 medium and metabolism with NBT reduction assay. Microscopic and cytometric evaluation of the leukocytes revealed that the isolation methods applied had an influence on their number and viability. Based on the results it can be concluded that isolation methods of cells in a Histopaque or Ficoll yield highly pure cell fractions with high viability.
Asunto(s)
Leucocitos/fisiología , Animales , Bovinos , Separación Celular/métodos , Separación Celular/veterinaria , Supervivencia Celular , Centrifugación por Gradiente de Densidad , Diatrizoato , Ficoll , Presión OsmóticaRESUMEN
The aim of this study was to assess changes in serum concentrations of C-reactive protein (CRP), serum amyloid A component (SAA), and haptoglobin (Hp) in bitches with pyometra undergoing ovariohysterectomy that developed postoperative wound infection-related complications. The study revealed that difficulties in postoperative wound healing were induced by infections with Escherichia coli and Staphylococcus spp. leading to re-increased levels of CRP and SAA immediately after surgery and persistently high Hp levels throughout the experiment. Our results indicate that acute-phase proteins in bitches undergoing surgery because of pyometra are useful markers for monitoring the postoperative period. Moreover, they enable prompt therapeutic management once complications develop.