Histological and Molecular Biological Changes in Canine Skin Following Acute Radiation Therapy-Induced Skin Injury.

Radiation therapy is a crucial cancer treatment, but it can damage healthy tissues, leading to side effects like skin injuries and molecular alterations. This study aimed to elucidate histological and molecular changes in canine skin post-radiation therapy (post-RT) over nine weeks, focusing on inflammation, stem cell activity, angiogenesis, keratinocyte regeneration, and apoptosis. Four male beagles received a cumulative radiation dose of 48 Gy, followed by clinical observations, histological examinations, and an RT-qPCR analysis of skin biopsies. Histological changes correlated with clinical recovery from inflammation. A post-RT analysis revealed a notable decrease in the mRNA levels of Oct4, Sox2, and Nanog from weeks 1 to 9. VEGF 188 levels initially saw a slight increase at week 1, but they had significantly declined by week 9. Both mRNA and protein levels of COX-2 and Keratin 10 significantly decreased over the 9 weeks following RT, although COX-2 expression surged in the first 2 weeks, and Keratin 10 levels increased at weeks 4 to 5 compared to normal skin. Apoptosis peaked at 2 weeks and diminished, nearing normal by 9 weeks. These findings offer insights into the mechanisms of radiation-induced skin injury and provide guidance for managing side effects in canine radiation therapy.

Comparison of bone subtraction CT angiography with standard CT angiography for evaluating circle of Willis in normal dogs.

BACKGROUND: Bone subtraction computed tomography angiography (BSCTA) is a useful alternative technique for improving visualization of vessels surrounded by skull bone. However, no studies have compared computed tomography angiography (CTA) and BSCTA for improving the visibility of canine cerebral blood vessels. OBJECTIVES: To evaluate the potential benefit of BSCTA for better delineation of brain arteries of the circle of Willis (CoW) in dogs by comparing BSCTA with non-subtraction computed tomography angiography (NSCTA). METHODS: Brain CTA was performed for nine healthy beagle dogs using a bolus tracking method with saline flushing. A total dose of 600 mgI/kg of contrast agent with an iodine content of 370 mgI/mL was injected at a rate of 4 ml/s. Bone removal was achieved automatically by subtracting non-enhanced computed tomography (CT) data from contrast CT data. Five main intracranial arteries of the CoW were analyzed and graded on a scale of five for qualitative evaluation. RESULTS: Scores of basilar artery, middle cerebral artery, and rostral cerebral artery in the BSCTA group were significantly higher than those in the NSCTA group (p = 0.001, p = 0.020, and p < 0.0001, respectively). Scores of rostral cerebellar artery (RcA) and caudal cerebral artery (CCA) did not differ significantly between the two groups. However, scores of RcA and CCA in the BSCTA group were higher than those in the NSCTA group. CONCLUSIONS: BSCTA improved visualization of intracranial arteries of the CoW with close contact to bone. Thus, it should be recommended as a routine scan method in dogs suspected of having brain vessel disease.

Optimizing contrast protocol for bone-subtraction CT angiography of intracranial arteries in normal dogs using 160-slice CT.

BACKGROUND: Bone-subtraction computed tomography angiography (CTA) (BSCTA) is a new technique designed to overcome the limitation of three-dimensional CTA, where the vessels surrounded by bone and calcification can be obscured. An optimal contrast CT protocol for intracranial artery visualization with BSCTA has yet to be established in dogs. OBJECTIVES: The purpose of this study was to determine the optimal contrast protocol of CTA for visualizing intracranial artery using an automatic bone-subtraction technique in dogs. METHODS: Brain CTA was performed four times for each of nine healthy beagle dogs to cover all the contrast protocols: two different contrast iodine concentrations (300 and 370 mgI/mL) and two different contrast media injection rates (2 and 4 mL/s). Bone removal post-processing was performed automatically by subtracting the non-enhanced CT data from the contrast CT data using a dedicated workstation. The bone-subtracted intracranial vessels were analysed for quantitative and qualitative evaluation. RESULTS: Quantitative evaluation showed significantly higher CT attenuation values for the group with a 370 mgI/mL iodine content at a rate of 4 mL/s than the two groups with a 300 mgI/mL iodine content at the rates of 2 and 4 mL/s (p < 0.001). Qualitative assessment revealed significantly higher mean scores for the 370 mgI/mL groups than the 300 mgI/mL groups and significantly higher mean scores for the 4 mL/s groups than the 2 mL/s groups (p < 0.05). CONCLUSIONS: The optimal contrast protocol for BSCTA suggests that high iodine material concentration and high injection rate should be used for strong arterial attenuation and great visualization of the intracranial arterial structure in dogs.

Quantitative Analysis of Brain CT Perfusion in Healthy Beagle Dogs: A Pilot Study.

Brain computed tomography (CT) perfusion is a technique that allows for the fast evaluation of cerebral hemodynamics. However, quantitative studies of brain CT perfusion in veterinary medicine are lacking. The purpose of this study was to investigate the normal range of perfusion determined via CT in brains of healthy dogs and to compare values between white matter and gray matter, differences in aging, and each hemisphere. Nine intact male beagle dogs were prospectively examined using dynamic CT scanning and post-processing for brain perfusion. Regional cerebral blood volume (rCBV), regional cerebral blood flow (rCBF), mean transit time, and time to peak were calculated. Tissue ROIs were drawn in the gray matter and white matter of the frontal, temporal, parietal, and occipital lobes; caudate nucleus; thalamus; piriform lobe; hippocampus; and cerebellum. Significant differences were observed between the white matter regions and gray matter regions for rCBV and rCBF (p < 0.05). However, no significant differences were identified between hemispheres and between young and old groups in brain regions. The findings obtained in this study involving healthy beagle dogs might serve as a reference for regional CT perfusion values in specific brain regions. These results may aid in the characterization of various brain diseases in dogs.

HPRT1 Most Suitable Reference Gene for Accurate Normalization of mRNA Expression in Canine Dermal Tissues with Radiation Therapy.

Reference genes are crucial in molecular biological studies as an internal control for gene re-search as they exhibit consistent expression patterns across many tissue types. In canines, radiation therapy is the most important therapeutic tool to cure various diseases like cancer. However, when using radiation for therapeutic strategy, radiation exposure to healthy tissues leads to some possible side effects such as acute radiation-induced skin injury and alters gene expression. Therefore, the analysis of a change in reference gene expression during the skin recovery process after radiation therapy is essential in healthy canine tissue. In the present study, we analyzed eight reference genes (ACTB, GAPDH, YWHAZ, GUSB, HPRT1, RPL4, RPS5, and TBP) in canine dermal tissues at 0, 1, 2, 3, 4, 5, 7, and 9 weeks of radiation exposure that affected the skin condition of canines. The stability of reference genes is determined by evaluating radiation therapy's effect on healthy canine dermal tissue. Epidermal marker, Keratin 10 expression varies each week after irradiation, and HPRT1 is found to be the most suitable for normalization of mRNA expression in radiation-exposed canine dermal tissues. Changes in the gene expression level were evaluated by using a reliable tool such as quantitative real-time polymerase chain reaction (qRT-PCR). In order to achieve a valid qRT-PCR result, the most stable reference genes used for normalization after the radiation exposure process are important. Therefore, the current study was designed to evaluate the most stable reference gene for the post-irradiation canine tissues. After radiation exposure, the alternation of reference gene expression was estimated by three algorithms (geNorm, Normfinder, and Bestkeeper). The RG validation programs (GeNorm and NormFinder) suggested that HPRT1, RPL4, and TBP were suitable for normalization in qRT-PCR. Furthermore, three algorithms suggested that HPRT1 was the most stable reference gene for normalization with qRT-PCR results, regardless of before and after radiation exposure. Whereas GAPDH was found to be the most unstable reference gene. In addition, the use of stable or unstable reference genes for the normalization of Keratin 10 expression showed statistical differences. Therefore, we observed that, to obtain accurate and suitable PCR results of the canine tissues with and without radiation exposure, the HPRT1 reference gene is recommended for normalization with its high stability. Additionally, the use of RGs such as HPRT1, RPL4, and TBP for normalization in qRT-PCR experiments is recommended for post-radiation canine tissues to generate more accurate and reliable data. These results will provide fundamental information regarding internal controls for gene expression studies and can be used for the analysis of gene patterns in regenerative medicine.