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Background The relationships among gastric lymphoid follicular hyperplasia (GLFH), Helicobacter‐like organisms (HLOs), and clinical signs have not been established in dogs. Objectives To evaluate the epidemiologic, clinical, endoscopic, and histopathologic findings associated with GLFH in dogs, and determine the association of GLFH with HLOs and the French Bulldog (FB) breed. Animals Two hundred eighty‐eight dogs that underwent gastroscopy between 2013 and 2016. Methods Retrospective, cross‐sectional study. Gastric biopsy samples were reviewed and scored for inflammation and HLOs. Dogs were divided into 3 groups: group 1 (63 FBs), group 2 (45 non‐FB brachycephalic dogs), and group 3 (180 nonbrachycephalic dogs). Variables were evaluated for their association with GLFH. Results Univariate analysis determined that intact males, young age, vomiting, gastroscopic findings (discoloration, hemorrhage, and ulcers), and histopathologic findings (gastric lamina propria lymphocytic infiltration and HLO score) were associated with GLFH (P ≤ .03). In the multivariate analysis, GLFH was associated with the HLO score (odds ratio [OR] > 5 for HLO scores 1‐2 and >15 for HLO score of 3; P < .001), with vomiting (OR > 4; P = .01) but not with FB breed (P = .76) and age (P = .1). The HLO score was associated with younger age (P < .001). Conclusion and Clinical Importance The HLO score was associated with a high GLFH score. Vomiting was associated with GLFH. Helicobacter‐like organisms are highly prevalent in young dogs and GLFH is indirectly associated with this factor. Clinical relevance of the identification of GLFH and HLO remains to be determined.

Background Despite its low incidence, the aggressive nature and high metastatic potential of canine prostate cancer (PC) make it a serious problem in veterinary medicine. However, knowledge of the molecular mechanisms and regulatory networks underlying the development and progression of canine PC is still limited. Using high-throughput methods, many PC-related genes have been identified by comparing gene expression profiles in malignant and non-malignant prostate tissues. In these studies, the reference group containing both normal and hyperplastic prostate tissues specimens was compared to the group composed of cancerous tissues specimens. However, this approach does not allow the exclusion of inflammation-related genes from those directly involved in PC development and progression. Furthermore, the RNA-Seq and microarray data were not validated by additional analyses, including immunohistochemistry (IHC). Therefore, to identify PC-specific genes, a comparative analysis of gene expression profiles in PC and benign prostate hyperplasia (BPH) tissue samples was performed using RNA-Seq, and the results were validated by Nano String technology, Western blotting (WB) and IHC. Results Using the NanoString assay, significant differences were identified for 12 genes, of which ASPM , CLU , TOP2A , TSPAN8/TM4SF3 and HELLS were overexpressed in PC compared to BPH, while ATP6V0A4 , CACNA2D1 , CLDN10 , GOLGA4 , KLK2 , NKX3.1 and SERPINB6 were downregulated in PC. Validation of the differentially expressed genes (DEGs) at the protein level using WB and IHC revealed a highly decreased expression of the voltage-gated calcium channel subunit alpha-2 delta-1 (α2δ1), encoded by the CACNA2D1 gene, in PC samples compared to BPH. Conclusions Our findings make α2δ1 protein a novel and promising diagnostic biomarker to differentiate PC from BPH.

Pseudoplacentational endometrial hyperplasia (PEH) is a common uterine lesion in dogs. A high frequency of pyometra has been associated with PEH in dogs, suggesting that PEH might be related to the pathogenesis of pyometra. This study aimed to assess transcription levels and expression of Toll like receptors (TLR) 1, 2 and 4; alpha estrogen receptors (ESR1), progesterone receptors (PR) and prolactin receptors (PRLR) in uteri with PEH. Furthermore, the inflammatory response of dog endometrium with PEH against ex vivo bacterial stimulus was also investigated. Uteri were classified as controls or with PEH. Uterine explants were cultured for 6 and 12 hours after in vitro stimulus with inactivated Escherichia coli . Transcription of receptors and proinflammatory cytokines, namely interleukin-6 ( IL-6 ) and CXCL8 were evaluated. Expression of receptors was also evaluated in uterine explants and uteri from biopsy archives. CXCL8 concentration was measured in supernatants from all cultured explants. Transcription levels and expression of both PR and ESR1 were lower in uteri explants with PEH not stimulated and cultured for 6 hours. Expression of PRLR was higher in uteri with PEH from biopsy archives. Proinflammatory response by transcription levels of interleukin 6 demonstrated downregulation in uteri with PEH at 6 hours of stimulation followed by upregulation at 12 hours. However, no differences between groups were observed. Both control and uteri with PEH secreted similar concentrations of CXCL8 at 6 hours of bacterial stimulation. At 12 hours, no response to stimulation was observed in the PEH group and supernatant concentrations of CXCL8 were higher in the control group. The inflammatory response to bacterial stimulus in uteri with PEH had different pattern than the control group, with an inversion in IL-6 transcription levels between 6–12 hours of culture. Additionally, CXCL8 production ceased earlier in explants with PEH than in control.

Background There is a remodeling of the central airways in horses with severe asthma but whether a similar process occurs in horses with the mild or moderate asthma (MMA) is unknown. Objectives To evaluate lesions affecting the central airways of horses with MMA. Animals Twelve horses with MMA and 8 control horses. Methods Case‐control retrospective study of horses classified as MMA affected or controls based on history and bronchoalveolar lavage fluid cytology. Endobronchial biopsies were analyzed using histomorphometry and a semiquantitative histologic scoring system. Results Histomorphometry identified epithelial hyperplasia (47 μm2/μm [34‐57 μm2/μm]; P = .02), a thickened lamina propria (166 μm [73‐336 μm]; P = .04), and smooth muscle fibrosis (42% [33%‐78%]; P = .04) in horses with MMA when compared to controls horses (24 μm2/μm [21‐80 μm2/μm]; 76 μm [36‐176 μm]; and 33% [26%‐52%], respectively). The semiquantitative score results indicated, in horses with MMA, the presence of epithelial hyperplasia (7 of the 12 horses with MMA and only 1 of the 8 control horses had a score of 1/1), and submucosal inflammatory leucocytes in the central airway (11 of the 12 horses with MMA and only 4 of the 8 control horses had a score ≥ 1/2). Conclusions and Clinical Relevance Tissue remodeling of the bronchial lamina propria, epithelium, and smooth muscle was present in horses with MMA.

Here we report the case of a cow with two ovaries that each exhibited hyperplasia but that otherwise had normal gross morphology. Both ovaries had a large number of tertiary follicles on the ovarian surface. Oocytes from one ovary were studied in more detail. The transcriptome was largely similar to other oocytes. Oocytes could undergo cleavage at a rate consistent with other oocytes and result in blastocyst-stage embryo formation after in vitro maturation and fertilization. Review of the literature from cattle and other species did not reveal reports of a similar type of spontaneous ovarian abnormality. Whole genome sequencing revealed many single nucleotide polymorphisms with predicted large effects on protein structure that could potentially be causative for the phenotype. The variant considered most likely to cause the observed alteration in ovarian function was a mutation in the glycoprotein-modifying enzyme MAN1A2.

To explore the shape characteristics of ablation lesions created via laser ablation (LA), radiofrequency ablation (RFA) and microwave ablation (MWA) in canine prostates and the clinical significance of these characteristics, six adult male beagles were randomly assigned to the LA, RFA, and MWA groups. These ablations were performed with common parameters applied in clinical practice (LA, 3 W/1200 J; RFA and MWA, 30 W/120 s). One ablation lesion was created in each lobe of the prostate via the ablation technique, resulting in a total of twelve ablation lesions. Transrectal ultrasound (TRUS) was used as guidance during puncture and to monitor changes in the ablation lesions. Finally, the ablation efficacy was assessed using transrectal contrast-enhanced ultrasonography (CEUS), and the transverse diameter (TRD), anteroposterior diameter (APD) and longitudinal diameter (LD) of each ablation lesion were measured. The volume (V) and the ratio (R) value were calculated. R reflects the shape characteristic of the ablation lesion (the R value close to 1.0 indicates a more spherical shape). The R values of the ablation lesions were 0.89 ± 0.02, 0.72 ± 0.01, and 0.65 ± 0.03 for RFA, MWA and LA, respectively, and they were significantly different (P = 0.027). The volumes of the ablation lesions were 2.17 ± 0.10 ml, 1.51 ± 0.20 ml, and 0.79 ± 0.07 ml for MWA, LA and RFA, respectively, and they were also significantly different (P = 0.001). The three abovementioned thermal ablation techniques with common parameters in clinical practice can be used for ablation in the prostate. The shapes and volumes of the ablation lesions of the three techniques were varied: The RFA-created lesions had the lowest volumes and were more spherical in shape, demonstrating that RFA could be used for the treatment of relatively small lesions or tumours adjacent to vital organs. The MWA lesions had the largest size with a spherical shape, which could be advantageous for the ablation of tumours with relatively large sizes. The sizes of the ablation lesions created via LA were between those of RFA and MWA but presented more oval in shape, suggesting that this method is highly appropriate for the ablation of benign prostatic hyperplasia (BPH).

Cystic endometrial hyperplasia-pyometra complex (CEH-P) is a common disease in sexually mature bitches. Disease progression leads to oxidative stress, resulting in the depletion of uterine antioxidants and lipid peroxidation of associated cells, which further aggravates the condition. The concentration of antioxidant enzymes, the level of lipid peroxidation within the uterine tissue, and its reflection in the serum and urine need to be elucidated. The aim of this study was to analyze the concentration of antioxidants such as superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), glutathione peroxidase (GPx), and the lipid peroxidation marker malonaldehyde (MDA) in three types of samples, i.e., serum, urine, and uterine tissue. For this purpose, 58 pyometra-affected and 44 healthy bitches were included in the present study. All animals underwent ovariohysterectomy (OVH). Our data indicated highly significant difference (p<0.01) in the antioxidant concentrations of uterine, serum and urine samples. Furthermore, there was a highly significant (p<0.01) difference in the serum levels of ferric reducing antioxidant power (FRAP) and free radical scavenging activity (FRSA) indicated poor capacity to overcome oxidative stress in the CEH-Pyometra condition. We showed that CEH-P induces oxidative stress, which further depletes the antioxidant enzyme reserves in the uterus. Thus, the weak antioxidant defence predisposes to uterine damage and disease progression. The simultaneous depletion of antioxidants and an increase in lipid peroxidation in the serum and urine may also act as early indicators of uterine pathology.

Background Nesfatin-1 is a neuropeptide that regulates the hypothalamic-pituitary-gonadal axis and may play a role in uterus function. It is co-expressed with other peptides, such as phoenixin, which can influence sex hormone secretion. Our previous research has confirmed that phoenixin-14 is involved in the development of cystic endometrial hyperplasia (CEH) and pyometra in dogs. Therefore, based on the similarities and interactions between these neuropeptides, we hypothesized that nesfatin-1 might also regulate the reproductive system in dogs. This study aimed to determine the expression of nesfatin-1 and its interaction with phoenixin-14 in dogs with CEH or pyometra compared to healthy females, and concerning animals’ body condition score (BCS 4–5/9 vs. BCS > 5/9). Results The analysis of nesfatin-1 in the uterus of bitches consisted of qPCR, western blot and immunofluorescence assays, and ELISAs. The results showed significantly higher nesfatin-1 encoding gene, nucleobindin-2 mRNA ( Nucb2 ) and nesfatin-1 protein expression in overweight females and those suffering from CEH or pyometra compared to healthy animals. The immunoreactivity of nesfatin-1 was elevated in the uteri of bitches with higher BCS > 5/9. Moreover, nesfatin-1 blood concentrations increased in all examined overweight bitches. In the case of phoenixin signals, we found opposite results, regardless of the female body condition score. Conclusion The etiology of CEH and pyometra are not fully known, although we have expanded the level of knowledge with respect to the possible interaction of nesfatin-1 and phoenixin in female dogs’ uteri. They interact oppositely. With increasing female body weight, the expression of nesfatin-1 in the uterus and its peripheral blood concentration increased. However, for female dogs affected by CEH and pyometra, a decreased level of phoenixin-14, irrespective of their body condition score is characteristic. This knowledge could be crucial in the development of biomarkers for these conditions, which may lead to earlier recognition.

A 13‐year‐old Labrador retriever was diagnosed with Cushing's syndrome (CS) caused by primary bilateral nodular adrenocortical hyperplasia with adrenocorticotropic hormone (ACTH) expression. The pituitary origin of CS was ruled out by suppression of plasma ACTH concentration and absence of a proliferative lesion on histological evaluation of the pituitary gland using periodic acid‐Schiff (PAS) staining, reticulin staining, and immunostaining for ACTH. A pheochromocytoma also was found at necropsy examination. On histological evaluation of both adrenal glands, at the junction of the fascicular and glomerular zones, multiple cell clusters distributed in both hyperplastic adrenal cortices expressed ACTH, whereas the pheochromocytoma cells did not. These results indicate that a disease similar to primary bilateral macronodular adrenocortical hyperplasia in humans also occurs in dogs, with intra‐adrenocortical expression of ACTH, glucocorticoids excess, and clinical signs of CS. Therefore, the term ACTH‐independent could be inappropriate in some cases of bilateral adrenocortical hyperplasia and suppressed plasma ACTH concentration in dogs.

The role of M1 macrophages (M1M)-derived exosomes in the progression of neointimal hyperplasia remains unclear now. Using a transwell co-culture system, we demonstrated that M1M contributed to functional change of vascular smooth muscle cell (VSMC). We further stimulated VSMCs with exosomes isolated from M1M. Our results demonstrated that these exosomes could be taken up by VSMCs through macropinocytosis. Using a microRNA array assay, we identified that miR-222 originated from M1M-derived exosomes triggered the functional changes of VSMCs. In addition, we confirmed that miR-222 played a key role in promoting VSMCs proliferation and migration by targeting Cyclin Dependent Kinase Inhibitor 1B (CDKN1B) and Cyclin Dependent Kinase Inhibitor 1C (CDKN1C) in vitro. In vivo, M1M-derived exosomes significantly aggravated neointima formation following carotid artery ligation injury and wire injury and these effects were partly abolished by miR-222 inhibitor 2′OMe-miR-222. Our findings thus suggest that exosomes derived from M1M could aggravate neointimal hyperplasia through delivering miR-222 into VSMCs. Future studies are warranted to validate if the post-injury vascular neointimal hyperplasia and restenosis could be attenuated by inhibiting miR-222.