Use of neurofilament light chain to identify structural brain diseases in dogs.

BACKGROUND: Neurofilament light chain (NfL) is released into the peripheral circulation by damaged axons. OBJECTIVES: To evaluate the diagnostic value of serum NfL concentration in dogs with intracranial diseases. ANIMALS: Study included 37 healthy dogs, 31 dogs with idiopathic epilepsy (IE), 45 dogs with meningoencephalitis of unknown etiology (MUE), 20 dogs with hydrocephalus, and 19 dogs with brain tumors. METHODS: Cohort study. Serum NfL concentrations were measured in all dogs using single-molecule array technology. RESULTS: Serum NfL concentration in dogs with each structural disease was significantly higher than in healthy dogs and dogs with IE (P = .01). The area under the receiver operating characteristic curve of NfL for differentiating between dogs with structural diseases and IE was 0.868. An optimal cutoff value of the NfL 27.10 pg/mL had a sensitivity of 86.67% and a specificity of 74.19% to differentiate the dogs with IE from those with structural brain diseases. There were significant correlations between NfL concentrations and lesion size: (1) MUE, P = .01, r = 0.429; (2) hydrocephalus, P = .01, r = 0.563. CONCLUSIONS AND CLINICAL IMPORTANCE: Serum NfL could be a useful biomarker for distinguishing IE from structural diseases in dogs and predicting the lesion sizes of MUE and hydrocephalus.

CRISPR/Cas9 targeting of passenger single nucleotide variants in haploinsufficient or essential genes expands cancer therapy prospects.

CRISPR/Cas9 technology has effectively targeted cancer-specific oncogenic hotspot mutations or insertion-deletions. However, their limited prevalence in tumors restricts their application. We propose a novel approach targeting passenger single nucleotide variants (SNVs) in haploinsufficient or essential genes to broaden therapeutic options. By disrupting haploinsufficient or essential genes through the cleavage of DNA in the SNV region using CRISPR/Cas9, we achieved the selective elimination of cancer cells without affecting normal cells. We found that, on average, 44.8% of solid cancer patients are eligible for our approach, a substantial increase compared to the 14.4% of patients with CRISPR/Cas9-applicable oncogenic hotspot mutations. Through in vitro and in vivo experiments, we validated our strategy by targeting a passenger mutation in the essential ribosomal gene RRP9 and haploinsufficient gene SMG6. This demonstrates the potential of our strategy to selectively eliminate cancer cells and expand therapeutic opportunities.

Myxomatous mitral valve disease and associated pulmonary hypertension might increase serum angiopoietin-2 in dogs.

OBJECTIVE: To evaluate the relationships between the severity of myxomatous mitral valve disease (MMVD) and pulmonary hypertension (PH) and serum angiopoietin (Ang)-1 and Ang-2 concentrations in dogs with MMVD. ANIMALS: 74 dogs (control, n = 12; MMVD, n = 62) were included. METHODS: Serum Ang-1 and Ang-2 concentrations were estimated using the canine-specific ELISA kit. The concentrations were compared between dogs with MMVD and healthy dogs, and they were analyzed according to the severity of MMVD and PH. RESULTS: The median serum Ang-1 concentration did not differ among the study groups. The median serum Ang-2 concentration was higher in dogs with stage B2 MMVD (P = .041) and acute congestive heart failure (P = .002) than in control dogs. In addition, the median serum Ang-2 concentration was higher in MMVD dogs with PH than in those without PH (P = .031). Serum Ang-2 concentration was correlated with vertebral heart score (rs = 0.36, P = .004) and vertebral left atrial score (r = 0.50, P < .001) in dogs with MMVD, and correlated with vertebral heart score (r = 0.63, P = .01), maximum E wave amplitude of the diastolic transmitral flow (rs = 0.61, P = .018), ejection fraction (rs = -0.77, P < .001) and fractional shortening (rs = -0.56, P = .032) in dogs with acute congestive heart failure. CLINICAL RELEVANCE: Circulating Ang-2 levels increase in dogs with the severity of MMVD and the presence of PH.

Case report: Fundic gland polyps caused by long-term omeprazole use in a Maltese dog.

Long-term use of proton-pump inhibitors can induce fundic gland polyps in the human stomach. However, this phenomenon has not been described in the veterinary literature. A 5-year-old intact female Maltese dog was referred to our hospital with chronic intermittent vomiting. The dog had been administered omeprazole (0.7-1.0 mg/kg PO q24 h) for the management of hydrocephalus for over 4 years; the omeprazole dose was increased to 10 mg/kg PO q24 h 8 months prior to presentation at referring hospital. Abdominal ultrasonography revealed marked thickening of the gastric wall with multi-lobulated, thickened folds. Subsequent endoscopy revealed marked polypoid lesions, and histological examination of the biopsy samples was consistent with the fundic gland polyps associated with proton-pump inhibitor use in humans. The lesions resolved after cessation of omeprazole, as assessed by ultrasonography. This report describes a case of fundic gland polyps following the long-term administration of omeprazole in a dog.

Case report: Central-pituitary hypothyroidism concurrent with hyperadrenocorticism without pituitary macroadenoma in a Miniature Schnauzer dog.

Multiple endocrine disorders are uncommon in veterinary medicine, and the disease combination is usually related to hypercortisolism or autoimmunity. Central-pituitary hypothyroidism, also refer to secondary hypothyroidism, can be caused by hypercortisolemic conditions and is well-recognized in human medicine. However, central hypothyroidism, including pituitary hypothyroidism, concurrent with hyperadrenocorticism, is rarely reported in veterinary medicine. A 7-year-old, intact female Miniature Schnauzer presented with generalized alopecia, scale, and pruritus and was diagnosed with superficial pyoderma and Malassezia dermatitis. Hormonal tests were performed, and the results indicated multiple endocrinopathies with a combination of non-adrenal dependent hyperadrenocorticism and central-pituitary hypothyroidism. Magnetic resonance imaging (7 T) and high-resolution research tomography positron emission tomography were performed to differentiate neuroendocrine tumors; however, no lesion was found in the hypothalamic to pituitary region. Hyperadrenocorticism was managed first to control endocrinopathy. After controlling hypercortisolism, a weak elevation of free thyroxine (T4) was revealed, whereas total T4 and thyroid-stimulating hormone (TSH) were still undetectable, and hypothyroidism management was added. About 9 months after the management, both endocrine diseases were well controlled, and clinical signs improved; however, serum TSH was unmeasured consistently. This case study describes a case of multiple endocrinopathies in a Miniature Schnauzer dog diagnosed with central-pituitary hypothyroidism concurrent with non-adrenal dependent hyperadrenocorticism without pituitary macroadenoma.

Case report: Evaluation of hindlimb ischemia using 18F-fluorodeoxyglucose positron emission tomography in a cat with cardiogenic arterial thromboembolism.

A 12-year-old castrated male domestic shorthair cat weighing 6.7 kg presented with acute hindlimb paralysis and tachypnea. The femoral pulse was absent bilaterally. Thoracic radiography showed finding compatible with cardiogenic pulmonary edema. Echocardiography revealed hypertrophic cardiomyopathy phenotype and a spontaneous echocardiographic contrast in the left atrium, suggesting cardiogenic arterial thromboembolism. Oxygen supplementation, diuretics, and antithrombotic and thrombolytic agents were also administered. However, hindlimb motor function was not restored. Severely increased aspartate aminotransferase and creatinine phosphokinase, as well as neutropenia with a degenerative left shift were identified, and amputation was considered to prevent sepsis caused by necrosis of the ischemic tissues. 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET)/computed tomography was performed to evaluate the metabolic activity of the muscle tissues and determine the level of amputation. There was no 18F-FDG uptake in the extremities of either the hind limbs or the caudal parts of the bilateral femoral muscle mass, suggesting a loss of metabolic activity in the area. Considering the wide affected area, a decreased quality of life was predicted postoperatively, and the cat was euthanized at the owner's request. Postmortem muscle biopsy confirmed weak atrophy of the left femoral muscle and prominent atrophy of the right calf. This case report describes the use of 18F-FDG PET in a cat with ischemia caused by cardiogenic arterial thromboembolism.

Evaluation of progression of chronic kidney disease in dogs with myxomatous mitral valve disease.

Introduction: Cardiovascular and renal diseases are known to affect each other in the cardiovascular renal axis disorder (CvRD). Although CvRD, which includes myxomatous mitral valve disease (MMVD) and chronic kidney disease (CKD), has been described in dogs, there are only a few reports on the progression of CKD in accordance with the severity of MMVD. The aim of this study was to evaluate whether the presence of MMVD is associated with the rate of progression of CKD in dogs. The time from the initial diagnosis to the worsening of the International Renal Interest Society (IRIS) stage and the time for the occurrence of hyperphosphatemia and isosthenuria were evaluated. Materials and methods: In this retrospective study, CKD progression was determined as an increase in the IRIS stage by at least one level and the development of hyperphosphatemia or isosthenuria. The CKD progression was compared in dogs with and without comorbid MMVD. Results: Dogs with CKD were divided into two groups: dogs with and without MMVD (n = 63, concurrent group; n = 52, CKD group, respectively). The concurrent group was further divided into two subgroups based on the American College of Veterinary Internal Medicine guidelines (B1 group, n = 24; B2 group, n = 39). The time for progression of CKD from IRIS stage 1 to IRIS stage 2 was significantly shorter in the concurrent group than in the CKD group (log-rank test, p < 0.001). MMVD was associated with an increased risk of progression from stage 1 to stage 2 (hazard ratio, 6.442; 95% confidence interval (CI), 2.354 to 18.850; p < 0.001). The timing of the onset of hyperphosphatemia or isosthenuria in the concurrent group and the CKD group was not significantly different. Conclusion: The results of this study suggest that MMVD could be a risk factor for the progression of CKD. Our findings may help predict the prognosis of dogs with both CKD and MMVD compared to CKD only.

T-cell lymphoma infiltrating the uterus and ovaries of a Golden Retriever: a case report.

BACKGROUND: To the best of our knowledge, this is the first report of female genital system infiltration of T-cell lymphoma in veterinary literature. CASE PRESENTATION: A 1.5-year-old, intact female Golden Retriever was referred due to melena and hyporexia that lasted for three weeks. Fever (40.5℃), tachycardia, tachypnoea, pale mucous membranes, and purulent vaginal discharge were identified on physical examination. Blood analyses revealed leucocytosis, anaemia, hypoalbuminemia, and increased lactate and C-reactive protein levels. On abdominal radiography, the small intestine was moderately deviated because of an oval-shaped mass (13 cm × 8.7 cm) located in the mid-abdomen. An enlarged tubular-shaped structure that had the opacity of soft tissue located in dorsal to the bladder to the middle of the abdomen, and an oval-shaped mass (5.28 cm × 3.26 cm), which was suspected to be a medial iliac lymph node located at the sixth to seventh lumbar level. Abdominal ultrasonography revealed gas and fluid in the lumen of the uterine horn with a severely thickened wall, round enlarged lymph nodes around the genitourinary system, and free fluid in the abdominal cavity. Based on these results, pyometra was suspected, and an exploratory laparotomy was performed for ovariohysterectomy. The resected ovary and uterus were macroscopically hypertrophied. Histopathological examination of the ovary and uterus revealed neoplastic proliferation of large round cells with strong immunoreactivity for CD3, indicating T-cell lymphoma. Therefore, the young dog was diagnosed with genital lymphoma. CONCLUSIONS: The present report describes T-cell lymphoma infiltrating the uterus and ovaries in a young dog, which is rarely diagnosed and could aid in the differential diagnosis of genital diseases in young dogs.

Severity of myxomatous mitral valve disease in dogs may be predicted using neutrophil-to-lymphocyte and monocyte-to-lymphocyte ratio.

OBJECTIVE: To investigate the neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), and platelet-to-lymphocyte ratio (PLR) in dogs with myxomatous mitral valve disease (MMVD). ANIMALS: 106 dogs with MMVD and 22 healthy dogs were included in the study. PROCEDURES: CBC data were obtained retrospectively, and NLR, MLR, and PLR were compared between dogs with MMVD and healthy dogs. The ratios were also analyzed according to MMVD severity. RESULTS: NLR and MLR were significantly higher in dogs with MMVD C and D (NLR of 4.99 [3.69-7.27]; MLR of 0.56 [0.36-0.74]) than in healthy dogs (NLR: 3.05 [1.82-3.37], P < .001; MLR: 0.21 [0.14-0.32], P < .001), MMVD stage B1 (NLR: 3.15 [2.15-3.86], P < .001; MLR: 0.26 [0.20-0.36], P < .001), and MMVD stage B2 dogs (NLR: 3.22 [2.45-3.85], P < .001; MLR: 0.30 [0.19-0.37], P < .001). The area under the receiver operating characteristic curves of the NLR and MLR to distinguish dogs with MMVD C and D from those with MMVD B were 0.84 and 0.89, respectively. The optimal cutoff value for NLR was 4.296 (sensitivity, 68%; specificity, 83.95%), and the MLR value was 0.322 (sensitivity, 96%; specificity, 66.67%). NLR and MLR were significantly decreased after treatment in dogs with congestive heart failure (CHF). CLINICAL RELEVANCE: NLR and MLR can be used as adjunctive indicators of CHF in dogs.

Use of 18 F-2-deoxy-2-fluoro-D-glucose positron emission tomography/computed tomography for staging thyroid carcinoma in a cat.

Thyroid nodules are common in older cats and are mostly benign; however, carcinomas may occur infrequently. In cats, thyroid carcinomas tend to be highly metastatic. The role of 18 F-2-deoxy-2-fluoro-D-glucose (FDG) positron emission tomography (PET)/computed tomography (CT) in human thyroid carcinoma has been well established. However, guidelines have not yet been established for veterinary medicine. Metastasis assessment has typically been performed using CT in veterinary medicine; however, it is poorly sensitive in detecting regional lymph nodes or distant metastases if these lesions are not abnormally contrast-enhanced, enlarged or cause overt mass effects. This case suggested that FDG PET/CT may be used for staging feline thyroid carcinoma, and the results contributed to treatment recommendations.

18F-fluorodeoxyglucose positron emission tomography findings of peripheral nerve sheath tumour of the nasal cavity in a dog.

An 8-year-old Miniature Poodle presented with chronic sneezing and unilateral epistaxis. A left-sided intranasal mass was identified on computed tomography. 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) was performed to evaluate the metabolic activity of the mass. The intranasal mass showed mildly increased 18F-FDG uptake. The maximal and mean standardized uptake values (SUVs) of the mass were 3.4 and 2.6, respectively. The maximal SUV of the mass/mean SUV of the normal liver was 2.5. The 7-cm soft, pink mass was easily removed through rhinoscopy, with subsequent dramatic improvement in clinical signs. Histopathological and immunohistochemical analyses determined that the mass was an intermediate-grade malignant peripheral nerve sheath tumour (PNST). This is the first report of 18F-FDG PET findings in a PNST in dogs.

Successful Treatment With Prednisolone and Mycophenolate Mofetil in a Dog With Recurrent Erythema Multiforme Minor.

A 3-year-old neutered male miniature poodle dog was referred with a 19-month history of unresolved dermatological signs despite long-term treatment. On physical examination, the dog had severe multifocal erythematous non-blanching patches and scales in the ventral trunk. Dermatological examination revealed Malassezia infection. Considering the history, clinical signs, and degree of infection, the possibility of a drug eruption appeared higher than that of Malassezia dermatitis. Therefore, bathing in lukewarm water was performed for 4 weeks without any other treatment, but there was no improvement. Subsequently, treatment for Malassezia dermatitis and differentiation from allergic dermatitis were performed, but there was still no improvement. A biopsy was performed, with the histopathology revealing lymphocytic interface dermatitis with keratinocyte apoptosis. Based on the histopathologic evaluation and clinical signs, the dog was diagnosed with erythema multiforme (EM) minor. Immunosuppressive therapy with prednisolone (1 mg/kg PO, twice daily) was initiated and had a good therapeutic effect. However, the lesion recurred after tapering the prednisolone dose (0.5 mg/kg PO, every other day). Therefore, steroid-sparing agents were added to the prednisolone regimen. Ciclosporin, azathioprine, and human intravenous immunoglobulin were administered in combination with prednisolone. Yet again, the lesion recurred when the dose of prednisolone was tapered to 0.5 mg/kg once daily. Mycophenolate mofetil (20 mg/kg PO, twice daily) was then added to the immunosuppressive regimen as a steroid-sparing agent, and complete remission was achieved and maintained even when the dose of prednisolone was tapered to 0.5 mg/kg every other day. This is the first reported case of recurrent EM successfully treated with a combination of prednisolone and mycophenolate mofetil, and this treatment option should be considered for recurrent EM.

Case report: Absent fluorodeoxyglucose uptake in a dog with unexpected brain death.

A 5-year-old male Maltese dog was presented with generalized tonic seizures and hypermetria. Multiple nodular subcortical cerebellar enhancements and meningeal enhancement were observed on magnetic resonance imaging. Fluorodeoxyglucose-positron emission tomography/computed tomography was performed due to suspicion of neoplastic disease, and no fluorodeoxyglucose uptake was observed in the intracranial structures. In PET images of this dog, absent fluorodeoxyglucose uptake was identified in the brain indicating no cerebral metabolism, strongly suggested brain death. The dog had no spontaneous breathing and no brainstem reflexes for more than 24 h after the termination of anesthesia. Through these results, this dog was diagnosed with unexpected brain death, and it is presumed that the cause was anesthesia. We report herein a case of brain death in a dog diagnosed using fluorodeoxyglucose-positron emission tomography/computed tomography.

Selective cytotoxicity of a novel mitochondrial complex I inhibitor, YK-135, against EMT-subtype gastric cancer cell lines due to impaired glycolytic capacity.

Epithelial-to-mesenchymal transition (EMT)-subtype gastric cancers have the worst prognosis due to their higher recurrence rate, higher probability of developing metastases and higher chemoresistance compared to those of other molecular subtypes. Pharmacologically actionable somatic mutations are rarely found in EMT-subtype gastric cancers, limiting the utility of targeted therapies. Here, we conducted a high-throughput chemical screen using 37 gastric cancer cell lines and 48,467 synthetic smallmolecule compounds. We identified YK-135, a small-molecule compound that showed higher cytotoxicity toward EMT-subtype gastric cancer cell lines than toward non-EMT-subtype gastric cancer cell lines. YK-135 exerts its cytotoxic effects by inhibiting mitochondrial complex I activity and inducing AMP-activated protein kinase (AMPK)-mediated apoptosis. We found that the lower glycolytic capacity of the EMT-subtype gastric cancer cells confers synthetic lethality to the inhibition of mitochondrial complex I, possibly by failing to maintain energy homeostasis. Other well-known mitochondrial complex I inhibitors (e.g., rotenone and phenformin) mimic the efficacy of YK-135, supporting our results. These findings highlight mitochondrial complex I inhibitors as promising therapeutic agents for EMT-subtype gastric cancers and YK-135 as a novel chemical scaffold for further drug development. [BMB Reports 2022; 55(12): 645-650].

WNK3 inhibition elicits antitumor immunity by suppressing PD-L1 expression on tumor cells and activating T-cell function.

Immune checkpoint therapies, such as programmed cell death ligand 1 (PD-L1) blockade, have shown remarkable clinical benefit in many cancers by restoring the function of exhausted T cells. Hence, the identification of novel PD-L1 regulators and the development of their inhibition strategies have significant therapeutic advantages. Here, we conducted pooled shRNA screening to identify regulators of membrane PD-L1 levels in lung cancer cells targeting druggable genes and cancer drivers. We identified WNK lysine deficient protein kinase 3 (WNK3) as a novel positive regulator of PD-L1 expression. The kinase-dead WNK3 mutant failed to elevate PD-L1 levels, indicating the involvement of its kinase domain in this function. WNK3 perturbation increased cancer cell death in cancer cell-immune cell coculture conditions and boosted the secretion of cytokines and cytolytic enzymes, promoting antitumor activities in CD4+ and CD8+ T cells. WNK463, a pan-WNK inhibitor, enhanced CD8+ T-cell-mediated antitumor activity and suppressed tumor growth as a monotherapy as well as in combination with a low-dose anti-PD-1 antibody in the MC38 syngeneic mouse model. Furthermore, we demonstrated that the c-JUN N-terminal kinase (JNK)/c-JUN pathway underlies WNK3-mediated transcriptional regulation of PD-L1. Our findings highlight that WNK3 inhibition might serve as a potential therapeutic strategy for cancer immunotherapy through its concurrent impact on cancer cells and immune cells.

Case Report: 18F-Fluoro-L-Phenylalanine Positron Emission Tomography Findings and Immunoreactivity for L-Type Amino Acid Transporter 1 in a Dog With Meningioma.

A 12-year-old intact female Miniature Pinscher dog weighing 5.4 kg presented with a history of seizures. On neurological examination, postural reactions were decreased in the left-sided limbs, and menace responses were bilaterally absent. Magnetic resonance imaging (MRI) of the brain was performed, and a solitary amorphous mass (2.7 × 1.9 × 2.2 cm) was observed on the right side of the frontal lobe. Based on the signalment, clinical signs, and MRI findings, a brain tumor was tentatively diagnosed, and meningioma was suspected. The dog was treated with hydroxyurea, prednisolone, and other antiepileptic drugs. One week after the treatment began, postural reactions returned to normal, and the menace response improved. At 119 days after treatment, 18F-fluoro-L-phenylalanine (18F-FDOPA) positron emission tomography (PET) was performed. Marked 18F-FDOPA uptake was observed in the lesion. The mean and maximal standardized uptake values of the lesion were 2.61 and 3.72, respectively, and the tumor-to-normal tissue ratio was 1.95. At 355 days after the initial treatment, a second MRI scan was performed and the tumor size had increased to 3.5 × 2.8 × 2.9 cm. The dog died 443 days after the initial treatment and was definitively diagnosed with grade 1 meningioma by histopathological examination. Immunohistochemical staining for Ki67 and L-type amino acid transporter 1 was positive and negative for p53, respectively. The labeling index of Ki67 was 2.4%. This is the first case to demonstrate 18F-FDOPA PET findings in a clinical case of a dog histologically diagnosed with a meningioma.

Evaluation of the blood neutrophil-to-lymphocyte ratio as a biomarker for meningoencephalitis of unknown etiology in dogs.

BACKGROUND: The neutrophil-to-lymphocyte ratio (NLR) has been identified as a biomarker in several inflammatory and autoimmune diseases. Multiple sclerosis (MS) has been found to be associated with changes in the NLR in humans. OBJECTIVES: To examine the diagnostic value of the NLR in meningoencephalitis of unknown etiology (MUE) in dogs. ANIMALS: Thirty-eight MUE dogs, 20 hydrocephalic dogs, 10 brain tumor (BT) dogs, 32 idiopathic epilepsy (IE) dogs, and 41 healthy dogs. METHODS: Retrospective study. Medical records were reviewed to identify dogs with a diagnosis of neurologic disease. The NLR was determined in all dogs. RESULTS: The median NLR was significantly higher in MUE dogs (6.08) than in healthy (1.78, P < .001), IE (2.50, P < .05), and hydrocephalic dogs (1.79, P < .05). The area under the receiver operating characteristic curve of the NLR for differentiation between MUE and healthy dogs was 0.96, and between the MUE dogs and dogs with other forebrain diseases was 0.86. An optimal cutoff of 4.16 for the NLR had a sensitivity of 71.1% and specificity of 83.9% to differentiate the MUE dogs from the dogs with other forebrain diseases. CONCLUSIONS AND CLINICAL IMPORTANCE: The NLR could be a biomarker for diagnosing MUE and distinguishing it from other intracranial diseases in dogs.

Snail acetylation by autophagy-derived acetyl-coenzyme A promotes invasion and metastasis of KRAS-LKB1 co-mutated lung cancer cells.

BACKGROUND: Autophagy is elevated in metastatic tumors and is often associated with active epithelial-to-mesenchymal transition (EMT). However, the extent to which EMT is dependent on autophagy is largely unknown. This study aimed to identify the mechanisms by which autophagy facilitates EMT. METHODS: We employed a liquid chromatography-based metabolomic approach with kirsten rat sarcoma viral oncogene (KRAS) and liver kinase B1 (LKB1) gene co-mutated (KL) cells that represent an autophagy/EMT-coactivated invasive lung cancer subtype for the identification of metabolites linked to autophagy-driven EMT activation. Molecular mechanisms of autophagy-driven EMT activation were further investigated by quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting analysis, immunoprecipitation, immunofluorescence staining, and metabolite assays. The effects of chemical and genetic perturbations on autophagic flux were assessed by two orthogonal approaches: microtubule-associated protein 1A/1B-light chain 3 (LC3) turnover analysis by Western blotting and monomeric red fluorescent protein-green fluorescent protein (mRFP-GFP)-LC3 tandem fluorescent protein quenching assay. Transcription factor EB (TFEB) activity was measured by coordinated lysosomal expression and regulation (CLEAR) motif-driven luciferase reporter assay. Experimental metastasis (tail vein injection) mouse models were used to evaluate the impact of calcium/calmodulin-dependent protein kinase kinase 2 (CAMKK2) or ATP citrate lyase (ACLY) inhibitors on lung metastasis using IVIS luciferase imaging system. RESULTS: We found that autophagy in KL cancer cells increased acetyl-coenzyme A (acetyl-CoA), which facilitated the acetylation and stabilization of the EMT-inducing transcription factor Snail. The autophagy/acetyl-CoA/acetyl-Snail axis was further validated in tumor tissues and in autophagy-activated pancreatic cancer cells. TFEB acetylation in KL cancer cells sustained pro-metastatic autophagy in a mammalian target of rapamycin complex 1 (mTORC1)-independent manner. Pharmacological inhibition of this axis via CAMKK2 inhibitors or ACLY inhibitors consistently reduced the metastatic capacity of KL cancer cells in vivo. CONCLUSIONS: This study demonstrates that autophagy-derived acetyl-CoA promotes Snail acetylation and thereby facilitates invasion and metastasis of KRAS-LKB1 co-mutated lung cancer cells and that inhibition of the autophagy/acetyl-CoA/acetyl-Snail axis using CAMKK2 or ACLY inhibitors could be a potential therapeutic strategy to suppress metastasis of KL lung cancer.

Serum concentrations of leptin and adiponectin in dogs with chronic kidney disease.

BACKGROUND: An imbalance in adipokines is associated with the progression of chronic kidney disease (CKD) in humans. However, alterations in adipokines in dogs with CKD remain unclear. OBJECTIVES: To examine whether adipokine concentrations in serum differ between healthy dogs and dogs with CKD and to determine the correlation between serum adipokine concentrations and CKD severity in dogs. ANIMALS: Twenty dogs with CKD and 10 healthy dogs. METHODS: In this cross-sectional study, serum concentrations of leptin, adiponectin, interleukin (IL)-6, IL-10, IL-18, and tumor necrosis factor (TNF)-α were measured in healthy dogs and dogs with CKD, which were classified according to the International Renal Interest Society guidelines. RESULTS: Serum leptin concentrations were positively correlated with systolic arterial blood pressure (r = .41), creatinine concentrations (r = .39), and symmetric dimethylarginine concentrations (r = .73). Serum adiponectin concentrations (median [range]) in CKD dogs with borderline or non-proteinuric (20.25 [14.9-45.8] ng/mL) were significantly higher than those in proteinuric CKD dogs (13.95 [6.4-22.1] ng/mL; P = .01). Serum IL-6 (median [range]; 43.27 [24.30-537.30] vs 25.63 [6.83-61.03] pg/mL; P = .02), IL-18 (median [range]; 25.98 [11.52-280.55] vs 10.77 [3.53-38.45] pg/mL; P = .01), and TNF-α (median [range]) concentrations (11.44 [8.54-38.45] vs 6.105 [3.97-30.68] pg/mL; P = .02) were significantly different between proteinuric and borderline or non-proteinuric CKD dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: leptin and adiponectin concentrations in serum might be associated with severity of CKD and proteinuria in dogs with CKD, respectively.

Evaluation of 18F-fluorodeoxyglucose uptake of beagle dogs for different durations of isoflurane anesthesia.

18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) is used for tumor evaluation. In veterinary medicine, anesthesia is an essential tool during the PET scanning process. However, the changes in FDG uptake in dogs that have undergone anesthesia for a longer duration have not been studied. This study aimed to analyze the influence of isoflurane anesthesia on FDG uptake in dogs undergoing PET. A crossover design was implemented by exposing 3 groups of 6 dogs to different durations of anesthesia (60, 90, and 150 minutes). Inhalation anesthesia was maintained throughout the scanning process (30 minutes) and FDG was injected 1 hour before the start of the PET scan. The standard uptake value of FDG was obtained for the 7 gross structures (whole brain, lung, salivary gland, liver, spleen, mediastinal blood pool, and kidney cortex) as well as for the 7 intracranial structures (frontal, parietal, temporal and occipital lobes, cerebellum, brain stem, and caudal colliculus). The whole brain and intracranial structures showed significantly lower FDG uptake in dogs with a longer duration of anesthesia, whereas other gross structures did not. Our results suggest that the duration of anesthesia should be considered when evaluating the uptake of FDG by the brain.

La tomographie par émission de positrons (PET) au 18F-fluorodésoxyglucose (FDG) est utilisée pour l'évaluation des tumeurs. En médecine vétérinaire, l'anesthésie est un outil essentiel lors du processus de PET. Cependant, les modifications de l'absorption du FDG chez les chiens ayant subi une anesthésie de plus longue durée n'ont pas été étudiées. Cette étude visait à analyser l'influence de l'anesthésie à l'isoflurane sur l'absorption du FDG chez les chiens subissant une PET. Un modèle croisé a été mis en oeuvre en exposant trois groupes de six chiens à différentes durées d'anesthésie (60, 90 et 150 minutes). L'anesthésie par inhalation a été maintenue tout au long du processus de numérisation (30 minutes) et le FDG a été injecté 1 heure avant le début de la PET. La valeur d'absorption standard du FDG a été obtenue pour les sept structures macroscopiques (cerveau entier, poumon, glande salivaire, foie, rate, pool sanguin médiastinal et cortex rénal) ainsi que pour les sept structures intracrâniennes (frontale, pariétale, temporale et lobes occipitaux, cervelet, tronc cérébral et colliculus caudal). L'ensemble du cerveau et les structures intracrâniennes ont montré une absorption de FDG significativement plus faible chez les chiens avec une durée d'anesthésie plus longue, contrairement aux autres structures. Nos résultats suggèrent que la durée de l'anesthésie doit être prise en compte lors de l'évaluation de la captation du FDG par le cerveau.(Traduit par Docteur Serge Messier).