Human benign Leydig cell tumor - Biochemical evaluation.

Recently we reported expressional alterations in 219 genes and their transcripts in Leydig cell tumors but nowadays there is still a lack of full basic biochemical characteristics of these tumors. The discovery of potential biochemical markers for tumor management from early detection, treatments, and control of therapy results may markedly supplement genetic data. Leydig cell micronodules were obtained from patients with azoospermia who were qualified for testicular biopsy. The biochemistry of Leydig cell tumors was analyzed using histological staining and spectrophotometric measurements of total proteins, carbohydrates, lipids, and nucleic acids. In addition, the levels of calcium (Ca2 +), copper (Cu2 +), zinc (Zn2 +), and selenium (Se2 +) ions were measured. When compared to healthy testis we revealed, for the first time, that in the interstitial tissue with Leydig cell tumors, great amounts of proteins, carbohydrates, lipids, and acids were dislocated from the seminiferous tubules. Measurements of organic compounds showed a decrease (P < 0.05) only in the Cu2 + content in Leydig cell tumors which may be related to their altered biochemical structure. This specific result may be promising for designing further approaches to manage this tumor based on combining morphological and molecular data.

Assessment of the possibility of using biomarkers (CCL11 and TGF-beta 1) in the diagnosis of prostate gland hyperplasia in dogs.

Prostatic hyperplasia (PH) is the most common reproductive disorder in dogs and can lead to discomforting problems such as haematuria, urinary incontinence, constipation, difficulty in defecating and stiffness of the hind limbs. The diagnosis of PH is nowadays based on digital rectal examination (DRE), ultrasonography (US) and radiography (X-ray). However, markers associated with PH are barely used for diagnostic purposes. Recently, there have been reports on the use of certain biomarkers for diagnosing PH in dogs such as canine PSA (Prostate Specific Antigen), microRNA and vascular endothelial growth factor (VEGF). Nevertheless, it has been generally accepted that these biomarkers play only an auxiliary role. Accordingly, the aim of our study was to evaluate the usefulness of the CCL11 (eotaxin-1) and TGF-beta 1 markers, which are used in the diagnosis of prostate diseases in humans, in case of dogs with PH. The study was carried out on 40 dogs of different breeds divided into three groups. Group I (n = 9) comprised dogs up to 5 years of age without changes indicative of PH. Group II (n = 17) included dogs aged 5-10 that were examined and diagnosed with (PH) and Group III (n = 14) which consisted of dogs over 10 years of age who were also diagnosed with PH. The study demonstrated that CCL11 levels did not differ significantly between the study groups and the median levels were 7.27 pg/mL, 7.57 pg/mL, 6.81 pg/mL, and IQR ranges 1.55 pg/mL, 1.74 pg/mL, 2.32 pg/mL, respectively. In contrast, TGF-beta 1 levels were detectable only in 6 dogs of group III and averaged the median of 28.86 pg/mL, IQR ranges 10.07 pg/mL. The study proved that CCL11 and TGF-beta 1 markers are of a limited use when diagnosing PH in dogs as no significant correlation related to age, body weight or prostate size was found.

Transcriptome analysis of human Leydig cell tumours reveals potential mechanisms underlying its development.

Leydig cell tumours are the most common sex cord-stromal tumours. In the last years, apparent increased incidence is noted while aetiology of the tumour is still unknown. Therefore, here, we focused on the genetics of Leydig cell tumours using the next-generation sequencing. Leydig cell micronodules were revealed in patients with azoospermia who were qualified for testicular biopsy. Complete gene set of Leydig cell tumours was compared with transcriptome of healthy Leydig cells obtained from donors. Bioinformatic analysis of the obtained sequencing data revealed alterations in expression of 219 transcripts. We showed, for the first time, that a significant proportion of differentially expressed genes is directly involved in regulation of apoptotic process, which downregulation might be important to Leydig cell tumour development. Additionally, we found a significant upregulation of heat shock protein genes that might be a unique feature of Leydig cell tumours when compared to other tumour types. Our study offers fundamental transcriptomic data for future studies on human Leydig cell tumour that are crucial to determine its causes. Moreover, presented here the in-depth analysis and discussion of alterations observed in tumour transcriptome may be important for the diagnosis and therapy of this pathology.

MicroRNA and vascular endothelial growth factor (VEGF) as new useful markers in the diagnosis of benign prostatic hyperplasia in dogs.

Numerous specific biomarkers with a prognostic and diagnostic value comparable to histopathological findings are now used for non-invasive diagnosis of prostate diseases in humans. Meanwhile, as far as dogs are concerned, the diagnosis of prostate disorders is based solely on clinical examination and ultrasound (USG). Therefore, the aim of the study was to assess the usefulness of two biomarkers, i.e. miRNA-129 and VEGF for the diagnosis of BPH in dogs. The study involved 40 dogs divided into three groups. Group I (n = 9) comprised healthy dogs up to the age of 5 years, Group II (n = 17) comprised dogs between the ages of 5-10 suffering from BPH as confirmed by the examination and Group III (n = 14) comprising dogs over 10 years of age, which also had BPH confirmed. The results demonstrated that dogs in group II and III exhibited a significant decrease in miRNA expression (P < 0.0001) and a significant increase in serum VEGF levels (P = 0.025) when compared to the dogs in group I. There was also a positive correlation between the prostate size and VEGF level. The findings led to the conclusion that the determination of miRNA-129 and VEGF can significantly contribute to the diagnosis of prostate disorders in dogs.

The Level of Prolactin, Serum Amyloid A, and Selected Biochemical Markers in Mares Before and After Parturition and Foal Heat.

The aim of the study was to determine the level of prolactin (PRL), serum amyloid A (SAA), and selected biochemical markers (T-Chol, AST, TP, Mg2+, P+, and Ca2+) in the blood of mares during the perinatal period. The study involved 14 mares of the Polish Coldblood Horse breed, which were in the third trimester of pregnancy. Blood was collected for testing 2 weeks before parturition and then 24 hours after delivery and in the foal heat (9 days) and 9 days after ovulation and breeding. The research revealed significant differences in the level of PRL and SAA before and after delivery. The highest PRL level was found 24 hours after delivery, lowest in foal heat and 9 days after ovulation. Serum amyloid A concentration was within the accepted norms; however, on day 9 after foaling, a significant increase of this protein was observed. All biochemical markers were within physiological limits. However, significant increases in T-Chol, AST, and TP levels was observed 24 hours after the delivery, whereas in foal heat and after ovulation levels of T-Chol and TP significantly decreased and the AST level remained at a similar level. There were no significant changes in electrolyte levels such as Mg2+, P+, and Ca2+. The pregnancy rate in the foal heat was at 43%. Collectively, the results of this study in conjunction with clinical observations demonstrated that when the perinatal period was normal, no disturbances in health related to pregnancy, parturition, lactation, and reproductive status during the postpartum period were found.

Changes in Blood Lymphocyte Subpopulations and Expression of MHC-II Molecules in Wild Mares Before and After Parturition.

INTRODUCTION: Pregnancy is a physiological state in which the immune system undergoes certain changes. On the one hand, by depleting cell defence mechanisms, it favours development and maintenance of the pregnancy. At the same time cells of the immune system ensure resistance to many risk factors, including infectious agents. MATERIAL AND METHODS: The study was carried out on 24 Polish Konik breed mares which were divided into two equal groups. The first group (group I) included mares living in the reserve. The second group (group II) comprised mares maintained under conventional conditions in the stables. The blood samples were collected for the first time in the perinatal period, i.e. 2 weeks before parturition (trial 0), then within the first 24 h after delivery, and then on 7th and 21st day after foaling. Flow cytometric analysis of lymphocyte expressing TCD4+, TCD8+, CD2+, and MHC class II antigens was performed. RESULTS: Before the delivery, in group I there was a significantly higher CD4:CD8 ratio compared to group II (P ≤ 0.05). Similarly, significantly increased CD4:CD8 ratio in group I was noted within 24 h after parturition (P ≤ 0.001) and it was also observed on 7th day (P ≤ 0.03) and 21st day after foaling (P ≤ 0.02). In the first 24 h after parturition, a significant decline of lymphocytes CD8+ (P ≤ 0.02) was noted. No significant differences in terms of lymphocytes CD2+ and CD3+ were observed. Expression of MHC-II molecules before and after the parturition was higher in group I compared to group II; however, the difference between the groups was not significant. CONCLUSION: The results obtained indicate that mares living in the reserve display higher activity of cell defence mechanisms.