The Density of CD8+ Tumor-infiltrating Lymphocytes Correlated With Akt Activation and Ki-67 Index in Canine Soft Tissue Sarcoma.

BACKGROUND/AIM: The activation of phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway has been implicated in canine soft tissue sarcoma (STS) and may serve as a prognostic marker. This study investigated the correlation between PI3K/Akt activation in tumor cells and tumor-infiltrating lymphocytes (TILs). MATERIALS AND METHODS: A total of 59 STS samples were labeled via immunohistochemistry to calculate the density of TILs, including CD3+ T cells, CD8+ T cells, CD20+ B cells, and FOXP3+ regulatory T cells. RESULTS: Forty-eight samples (81.3%) had intra-tumoral TILs with a high density of CD3+ T cells (mean: 283.3 cells/mm2) and CD8+ T cells (mean: 134.8 cells/mm2). Conversely, CD20+ B cells (mean: 73.6 cells/mm2) and FOXP3+ regulatory T cells (mean: 9.2 cells/mm2) were scarce. The abundance of CD3+/CD8+, CD3+/CD20+, and CD8+/CD20+ TILs were highly correlated in multivariate analyses (r=0.895, 0.946, and 0.856, respectively). Nonetheless, TIL density was unrelated to clinicopathological parameters (sex, age, tumor location, breed) and tumor grade. The abundance of CD8+ T cells was positively correlated with the activation of PI3K/Akt, indicating that samples with high levels of phospho-Akt and phospho-S6 tend to have a higher CD8+ T cell density (p=0.0032 and 0.0218, respectively). Furthermore, TIL density was correlated with the Ki-67 index, a tumor proliferation and growth marker. Samples with a high Ki-67 index had a significantly higher abundance of CD3+ T cells, CD8+ T cells, and CD20+ B cells (p=0.0392, 0.0254, 0.0380, respectively). CONCLUSION: PI3K/Akt pathway activation may influence the infiltration of CD8+ T cells within the tumor microenvironment in canine STS. Prospective studies involving a higher number of cases are warranted to confirm these findings.

Activation of the Akt signalling pathway as a prognostic indicator in canine soft tissue sarcoma.

Canine soft tissue sarcoma (STS) is relatively common in dogs and is the generic term for tumours that originate from mesenchymal cells. While histopathological grade and immunolabelling with Ki-67 have been used for estimating prognosis, additional indicators are needed for predicting prognosis. Aberrant cell signalling pathways may contribute to disease activity and, therefore, prognostic markers. However, their role in canine STS remains poorly understood. The aim of this study was to investigate expression of phosphorylated Akt (phospho-Akt) and phosphorylated S6 (phospho-S6) as potential prognostic indicators. Immunohistochemical labelling was conducted on clinical samples of canine STS (n = 67). We found that phospho-Akt expression was positively correlated with histopathological grade (P = 0.001) and Ki-67 index (P <0.01). There was no apparent relationship between the type of STS and the expression of phospho-Akt. The number of cases that expressed phospho-S6, which is the downstream molecule of the Akt signalling pathway, was higher in immunopositive phospho-Akt cases than in immunonegative phospho-Akt cases (P <0.0001). Furthermore, phospho-Akt expression was significantly higher in recurrent and metastatic cases. We also confirmed that phosphorylation of Akt occurred in conjunction with S6 phosphorylation in three canine STS cell lines. These results suggest that immunolabelling for phospho-Akt, phospho-S6 and Ki-67 could potentially be used as a prognostic indicator and therapeutic target in canine STS.

Investigation of chlamydophilosis from naturally infected cats.

BACKGROUND: Chlamydophila felis, formerly known as Chlamydia psittaci var. felis, is frequently associated with ocular, respiratory, and occasionally reproduction tract infections. Even though the infection is sometimes asymptomatic, it potentially results in a latent immunosuppressive infection. OBJECTIVE: This study aimed to identify occurrences of feline chlamydophilosis, rarely reported in cats in Indonesia. METHODS: The observation was conducted in three cats with clinical signs of Cp. felis infection, particularly relapsing conjunctivitis. The cats' histories were recorded based on owners' information. Conjunctival swabs were sampled for cytology examination and molecular assay detection. A phylogenetic tree was generated using MEGA-X software to reveal group clustering. A post-mortem examination was performed on the cat that died during an examination. RESULTS: Cp. felis was detected in both cytological examination and polymerase chain reaction assay. The phylogenetic tree demonstrated that the Cp. felis isolated in this study clustered with several other isolates from the other countries. Cp. felis can be isolated from cats with different clinical manifestations and levels of severity. The chronic fatal infection demonstrated interstitial broncho-pneumonia under histopathological examination. CONCLUSIONS: Molecular assay of Cp. felis is always recommended to obtain a definitive diagnosis of feline chlamydophilosis since the disease can have various clinical manifestations. Even though it may be subclinical and is often not fatal, an infected cat may be a carrier that could spread the pathogen in the surrounding environment. Serious disease management is suggested to avoid high costs associated with regularly relapsing disease.

Evaluation of lower urinary tract disease in the Yogyakarta cat population, Indonesia.

BACKGROUND AND AIM: This paper reports a retrospective study performed in 185 cats diagnosed with feline lower urinary tract disease (FLUTD). The analyzed population involved feline patients at the Veterinary Clinic of Universitas Gadjah Mada, Indonesia. This research aimed to evaluate the clinical indications and causes of FLUTD in the Yogyakarta cat population. MATERIALS AND METHODS: The medical data of all feline patients were obtained to conduct this study. FLUTD diagnoses were based on physical examinations, urinalyses, ultrasound examinations, and bacterial cultures. Only cats with a complete examination were used in the study. The clinical signs were evaluated and accompanied by the results of laboratory tests in cats that showed symptoms of FLUTD. The medical history of all feline patients was analyzed thoroughly. Most of the feline's urine samples were collected by catheterization. RESULTS: The most commonly diagnosed of FLUTD in the Yogyakarta cat population were: feline idiopathic cystitis (FIC) (56%), urinary tract infection (UTI; 25%), urolithiasis (13%), urethral plugs (UP) (4.9%), and neoplasia (0.4%), respectively. The prevalence of UTI is higher than that reported in Europe or the US. Older cats more often show symptoms of UTI and neoplasia, whereas young cats more often show symptoms of FIC and UP. The prevalence of male cats experiencing FLUTD in Yogyakarta is much higher than female cats. CONCLUSION: The incidence rate of FLUTD cases in Yogyakarta is related to age and sex. The results of this study are similar to those of the previous research studies conducted in other countries.

Clinical and pathological features of aspergillosis due to Aspergillus fumigatus in broilers.

BACKGROUND AND AIM: Aspergillus fumigatus is a ubiquitous pathogen causing aspergillosis in poultry. This research aimed to evaluate the clinical and pathological features of aspergillosis infection in broilers. MATERIALS AND METHODS: A. fumigatus infection was induced experimentally by intra-air sac inoculation of a 1.7×108 spore suspension into broilers. Infected and non-infected birds were closely observed for the development of clinical signs of infection twice daily. Pathological samples were collected 5, 14, and 30 days post-infection (dpi) and examined by hematoxylin-eosin staining. RESULTS: A total of 160 birds were included in this study. Clinical signs emerged at 3 dpi and became consistent at 5 dpi. A considerable decrease in severity and number of birds showing infection symptoms followed. The clinical signs of aspergillosis included anorexia (n=40; 50%), lethargy (n=32; 40%), dyspnea (n=38; 48%), and gasping (n=29; 36%). Macroscopic changes in the air sacs at 3 dpi included the development of minor lesions showing cloudiness, slight membrane thickening, and local exudates. Histopathological examination of the air sacs collected at 3 dpi indicated local inflammation surrounded by hyphae and spores. At 5 dpi, infected birds developed nodules, necrosis, and parenchymal consolidation of the lungs. Pulmonary changes, such as bronchopneumonia, spores, septate hyphae, and mild granulomatous inflammation, were also observed. At 14 dpi, multiple caseous nodules and plaques were found in the air sacs; plaque and necrosis in large areas of the lungs and severe multifocal granulomatous inflammation were noted. CONCLUSION: The clinical symptoms of aspergillosis emerged at 3 dpi and gradually decreased beginning at 7 dpi. Similar pathological changes were observed in the air sacs and lungs. The results of this work provide additional information on the pathogenesis of aspergillosis.

Effect of shell as natural testosterone boosters in Sprague Dawley rats.

AIM: This study aimed to evaluate the effect of shell supplementation on the regulation of male reproduction in rats. MATERIALS AND METHODS: The zinc (Zn) level of shell from blood clam (Anadara granosa), green mussel (Perna viridis), and conch shell (Telescopium telescopium) was analyzed. The highest Zn content shell was fed to male Sprague Dawley rats for 0, 9, 30, and 50 days at the dose of either 0.09 mg/200 g BW or 0.18 mg/200 g BW. To determine the testosterone levels, blood was collected through the infraorbitalis sinus just before the rat was sacrificed. Testicular and brain were also collected for Cyp19 aromatase receptor analysis. RESULTS: The Zn level in the shell of blood clam, green mussel, and conch shell 61.55 mg/kg, 2.78 mg/kg, and 3.93 mg/kg, respectively. The testosterone level of T1 group receiving 0.18 mg/200 g BW for 0, 9, 30, and 50 days was 1.42±0.59, 2.15±1.58, 2.98±2.53, and 8.11±2.03 ng/mL, respectively. The testosterone level of T2 group receiving 0.09 mg/200 g BW for 0, 9, 30, and 50 days was 2.50±0.32, 1.25±0.60, 3.87±3.27, and 3.54±0.23 ng/mL, respectively. The T3 group receiving Na-CMC showed the level of testosterone at days 0, 9, 30, and 50 days was 0.77±0.22, 1.99±1.65, 4.12±0.07, and 2.19±1.30 ng/mL, respectively. Finally, the T4 group receiving Zn showed testosterone levels at days 0, 9, 30, and 50 days was 0.51±0.58, 2.24±3.16, 4.58±1.97, and 2.89±0.20 ng/mL, respectively. There was a significant difference (p<0.05) between the T1 group compared to the other groups. However, the absence of expression of Cyp19 aromatase both in Leydig cells and the brain indicated no conversion of testosterone to estradiol. To add, this finding showed the potential use of the shell to boost the testosterone level in male rats. CONCLUSION: Shell acted as an aromatase blocker to boost the testosterone level in male rats. This also indicates its promising application in birds to manipulate the quality of song and feather.