Metastatic immune infiltrates correlate with those of the primary tumour in canine osteosarcoma

Our lack of understanding of the immune microenvironment in canine osteosarcoma (cOSA) has limited the identification of potential immunotherapeutic targets. In particular, our ability to utilize readily available tissue from a dog's primary tumour to predict the type and extent of immune response in their pulmonary metastatic lesions is unknown. We, therefore, collected 21 matched pairs of primary tumours and pulmonary metastatic lesions from dogs with OSA and performed immunohistochemistry to quantify T‐lymphocyte (CD3), FOXP3+ cell, B‐lymphocyte (Pax‐5), and CD204+ macrophage infiltration. We found that T‐lymphocytes and FOXP3+ infiltrates in primary tumours positively correlated with that of metastatic lesions (ρ = 0.512, P = 0.038 and ρ = 0.698, P = 0.007, respectively), while a strong trend existed for CD204+ infiltrates (ρ = 0.404, P = 0.087). We also observed T‐ and B‐lymphocytes, and CD204+ macrophages to be significantly higher in a dog's pulmonary metastasis compared to their primary tumour (P = 0.018, P = 0.018, P = 0.016, respectively), while FOXP3+ cells were only significantly higher in metastases when all primary tumour and metastasis lesions were compared without pairing (P = 0.036). Together, these findings suggest that the metastatic immune microenvironment may be influenced by that of the primary cOSA, and that primary tumour immune biomarkers could potentially be applied to predict immunotherapeutic responses in gross metastatic disease. We, therefore, provide a rationale for the treatment of cOSA pulmonary metastases with immunotherapeutics that enhance the anti‐tumour activity of these immune cells, particularly in dogs with moderate to high immune cell infiltration in their primary tumours.     

Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.     

STRAIN ELASTOGRAPHY USING DOBUTAMINE‐INDUCED CAROTID ARTERY PULSATION IN CANINE THYROID GLAND

Thyroid disease is common in dogs and conventional ultrasonography is a standard diagnostic test for diagnosis and treatment planning. Strain elastography can provide additional information about tissue stiffness noninvasively after applying external or internal compression. However, natural carotid artery pulsations in the canine thyroid gland are too weak to maintain sufficient internal compression force. The objective of the present study was to describe the feasibility of strain elastography for evaluating the canine thyroid gland and the repeatability of dobutamine‐induced carotid artery pulsation as an internal compression method. In seven healthy Beagle dogs, strain on each thyroid lobe was induced by external compression using the ultrasound probe and internal compression using carotid artery pulsation after dobutamine infusion. The thyroid appeared homogeneously green and the subcutaneous fat superficial to the thyroid lobe appeared blue. Strain values and strain ratios did not differ among dogs or between the left and right lobes. Interobserver repeatability was excellent for both compression methods. Intraobserver repeatability of the strain ratio measured using the carotid artery pulsation method (intraclass coefficient correlation = 0.933) was higher than that measured using the external compression method (0.760). Mean strain values of thyroid lobes for the external compression method (142.93 ± 6.67) differed from the internal method (147.31 ± 8.24; P < 0.05). Strain ratios between the two methods did not differ. Strain elastography was feasible for estimating thyroid stiffness in dogs. Carotid artery pulsation induced by dobutamine infusion can be used for canine thyroid strain elastography with excellent repeatability.     

Retrospective study of cecocolic intussusception (cecal inversion) in nine horses (1982–1998)

We retrospectively evaluated the medical records and obtained follow-up information for nine horses which had been treated for cecocolic intussusception (CCI) between January 1982 and April 1998. During the 16-year study period, CCI was diagnosed in nine of 748 horses in which exploratory celiotomy was undertaken for abdominal pain, representing an incidence of 1.2%. Most affected horses (78%) were less than four years of age (median age was 12 months, age range was five months to 15 years). Cecocolic intussusception affected male horses (78%) more commonly than female horses. The most common clinical presentation was abdominal pain of a severe, acute nature or milder but recurrent signs of abdominal pain persisting in spite of conservative treatment for several days. Correction of CCI by either simple reduction or reduction followed by partial typhlectomy was successful if compromise of the intestine by devitalization and adhesion formation was not found at surgery. Definitive diagnosis of CCI necessitates exploratory celiotomy, although an ultrasonographic examination of the abdomen may confirm the diagnosis in some cases. When recognized early during the course of disease, surgical correction of CCI is associated with a favorable outcome; of the eight horses which underwent surgery in our series, five horses (63%) survived surgical correction of CCI. Handling of compromised gut during reduction of CCI necessitates extreme caution because the risk of intestinal tearing is quite high.     

Needle longevity, photosynthetic rate and nitrogen concentration of eight spruce taxa planted in northern Japan

Growth characteristics of Picea glehnii Masters, P. jezoensis (Sieb. et Zucc) Carr., P. jezoensis var. hondoensis (Mayr) Rehder and P. shirasawae Hayashi from Japan, P. abies (L.) Karst. from Europe and P. glauca Voss, P. mariana Britt., Sterns and Pogg. and P. rubens Sarg. from North America were compared. The trees were grown in similar conditions at the Tomakomai Experimental Forest of Hokkaido University in northern Japan. Tree growth, needle biomass, longevity, photosynthetic rate, nitrogen concentration and specific leaf area (SLA) were measured, and photosynthetic nitrogen-use efficiency was calculated. Picea jezoensis, P. jezoensis var. hondoensis, P. abies and P. glauca had high growth rates, high photosynthetic rates in young needles, high needle nitrogen concentrations and short needle life spans. In contrast, P. glehnii, P. shirasawae, P. mariana and P. rubens had low growth and photosynthetic rates, low needle nitrogen concentrations, long needle life spans and maintained a high photosynthetic nitrogen-use efficiency in older needles. Examination of relationships between several growth parameters of the eight taxa revealed positive correlations between SLA and mass-based photosynthetic rate and between SLA and mass-based nitrogen concentration, whereas mass-based photosynthetic rate and mass-based nitrogen concentration were negatively correlated with needle longevity. The species differed greatly in growth characteristics despite being grown in similar conditions.     

EXCRETORY UROGRAPHY BY INTRAOSSEOUS INJECTION OF CONTRAST MEDIA IN A RABBIT MODEL

There are many indications for an intravenous excretory urogram. However, where intravenous access is not available, the intraosseous route to the circulation may be an alternative. We found that safe and diagnostic excretory urograms could be obtained in rabbits following the injection of different contrast media via the intraosseous route. In fact, these excretory urograms were indistinguishable from ones obtained by the conventional intravenous route. While the rabbits did not develop any abnormal clinical signs following the procedure, there were postmortem histologic lesions of osteochondrosis in 5 of 22 (22.7%) tibias receiving an intraosseous needle, but in none of the 14 tibias that did not receive an intraosseous needle. Further, the use of diatrizoate was associated with the development of osteochondrosis while the use of iopamidol was not.