Cystatin C and Neutrophil Gelatinase-Associated Lipocalin as Early Biomarkers for Chronic Kidney Disease in Dogs.

Early diagnosis of chronic kidney disease (CKD) could facilitate timely and appropriate monitoring and therapy. Traditional biomarkers have limitations. Thus, new biomarkers are needed. The objective of the present study was to compare renal biomarkers (including symmetric dimethylarginine [SDMA], cystatin C [CysC], and urine neutrophil gelatinase-associated lipocalin [NGAL]-creatinine ration [UNCR]) and creatinine (CREA) for early detection of CKD in dogs. Nine healthy dogs and 32 dogs with CKD were included in this study. All dogs underwent physical examination, blood analysis (included CREA and SDMA), urinalysis, and imaging examinations. CysC and NGAL levels were measured in serum and urine, respectively. SDMA, CysC, and UNCR were significantly elevated in dogs with CKD and IRIS stage Ι (P < .0001) than in controls. CysC demonstrated a strong correlation with CREA (r2 = 0.6556, P < .0001). CysC (sensitivity 93.55%, specificity 100%) had the highest sensitivity for detecting CKD, followed by UNCR (sensitivity 90%, specificity 100%), SDMA (sensitivity 84.37%, specificity 100%), and CREA (sensitivity 43.75%, specificity 100%). Additionally, CysC and UNCR (sensitivity 88.89%, specificity 100%) exhibited higher sensitivity and specificity than CREA (sensitivity 88.89%, specificity 66.67%) and SDMA (sensitivity 88.89%, specificity 88.89%) in dogs with CKD International Renal Interest Society (IRIS) stage Ι. CysC as a marker of glomerular filtration rate (GFR) and urinary NGAL as a marker of tubular damage could be used to detect CKD early in dogs better than CREA and SDMA.

Cutaneous adverse drug reaction in a dog following firocoxib treatment.

A 9-year-old intact female toy poodle was presented with oedema around the neck, including pus and cutaneous necrosis, 2 days after starting firocoxib treatment and placement of a cervical collar for intervertebral disc disease. Cytology of the pus revealed predominantly mature neutrophils with fewer macrophages and lymphocytes, indicating sterile inflammation. Although a skin biopsy could have provided more diagnostic information, it was not performed at presentation. Firocoxib treatment was discontinued, and immunosuppressive therapy including cyclosporine was initiated, which significantly alleviated the skin lesions. The dog recovered fully in 7 weeks. The final diagnosis was a possible cutaneous adverse drug reaction to firocoxib based on history, clinical signs and response to therapy.

Malarone® induced pancreatitis and alopecia in a dog: a case report.

BACKGROUND: Malarone® is a drug used for the treatment of malaria in humans. This drug is also particularly effective in the treatment of canine Babesia gibsoni infections. Malarone® is rarely used in dogs, and its adverse effects have not been widely reported. Its mechanism of action is related to the inhibition of cytochrome b and electron transport in the cell. This is the first known report of the development of acute pancreatitis and alopecia in a dog following the administration of Malarone®. CASE PRESENTATION: A 3-year-old, intact, female Maltese was referred to our clinic with intermittent vomiting and sudden, generalized alopecia. Two months previously, the dog had been prescribed Malarone® for the treatment of a suspected B. gibsoni infection. The dog was evaluated using hematology, radiography, ultrasonography, a PCR for Babesia detection, and a canine pancreatic lipase immunoreactivity (cPLI) assay. The result of the PCR test was negative, whereas the cPLI assay yielded a positive result. Dermatologic examination revealed bacterial infection with hair cycle arrest. CONCLUSIONS: Based on these findings, drug-induced acute pancreatitis and alopecia with superficial pyoderma were diagnosed. Malarone® may induce severe adverse reactions in dogs. Therefore, careful monitoring for adverse effects is required when using Malarone® in dogs.

Human salivary gland cells express bradykinin receptors that modulate the expression of proinflammatory cytokines.

Bradykinin is an important peptide modulator that affects the function of neurons and immune cells. However, there is no evidence of the bradykinin receptors and their functions in human salivary glands. Here we have identified and characterized bradykinin receptors on human submandibular gland cells. Both bradykinin B1 and B2 receptors are expressed on human submandibular gland cells, A253 cells, and HSG cells. Bradykinin increased the intracellular Ca2+ concentration ([Ca2+ ]i ) in a concentration-dependent manner. Interestingly, a specific agonist of the B1 receptor did not have any effect on [Ca2+ ]i in HSG cells, whereas specific agonists of the B2 receptor had a Ca2+ mobilizing effect. Furthermore, application of the B1 receptor antagonist, R715, did not alter the bradykinin-mediated increase in cytosolic Ca2+ , whereas the B2 receptor antagonist, HOE140, showed a strong inhibitory effect, which implies that bradykinin B2 receptors are functional in modulating the concentration of cytosolic Ca2+ . Bradykinin did not affect a carbachol-induced rise of [Ca2+ ]i and did not modulate translocation of aquaporin-5. However, bradykinin did promote the expression of proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), implying the role of bradykinin in salivary gland inflammation. These data suggest that bradykinin receptors are involved in Ca2+ signaling in human submandibular gland cells and serve a unique role, which is separate from that of other salivary gland G protein-coupled receptors.

Low levels of methyl ß-cyclodextrin disrupt GluA1-dependent synaptic potentiation but not synaptic depression.

Methyl-ß-cyclodextrin (MßCD) is a reagent that depletes cholesterol and disrupts lipid rafts, a type of cholesterol-enriched cell membrane microdomain. Lipid rafts are essential for neuronal functions such as synaptic transmission and plasticity, which are sensitive to even low doses of MßCD. However, how MßCD changes synaptic function, such as N-methyl-d-aspartate receptor (NMDA-R) activity, remains unclear. We monitored changes in synaptic transmission and plasticity after disrupting lipid rafts with MßCD. At low concentrations (0.5 mg/mL), MßCD decreased basal synaptic transmission and miniature excitatory post-synaptic current without changing NMDA-R-mediated synaptic transmission and the paired-pulse facilitation ratio. Interestingly, low doses of MßCD failed to deplete cholesterol or affect α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPA-R) and NMDA-R levels, while clearly reducing GluA1 levels selectively in the synaptosomal fraction. Low doses of MßCD decreased the inhibitory effects of NASPM, an inhibitor for GluA2-lacking AMPA-R. MßCD successfully decreased NMDA-R-mediated long-term potentiation but did not affect the formation of either NMDA-R-mediated or group I metabotropic glutamate receptor-dependent long-term depression. MßCD inhibited de-depression without affecting de-potentiation. These results suggest that MßCD regulates GluA1-dependent synaptic potentiation but not synaptic depression in a cholesterol-independent manner.

Rescue of fragile X syndrome phenotypes in Fmr1 KO mice by the small-molecule PAK inhibitor FRAX486.

Fragile X syndrome (FXS) is the most common inherited form of autism and intellectual disability and is caused by the silencing of a single gene, fragile X mental retardation 1 (Fmr1). The Fmr1 KO mouse displays phenotypes similar to symptoms in the human condition--including hyperactivity, repetitive behaviors, and seizures--as well as analogous abnormalities in the density of dendritic spines. Here we take a hypothesis-driven, mechanism-based approach to the search for an effective therapy for FXS. We hypothesize that a treatment that rescues the dendritic spine defect in Fmr1 KO mice may also ameliorate autism-like behavioral symptoms. Thus, we targeted a protein that regulates spines through modulation of actin cytoskeleton dynamics: p21-activated kinase (PAK). Our results demonstrate that a potent small molecule inhibitor of group I PAKs reverses dendritic spine phenotypes in Fmr1 KO mice. Moreover, this PAK inhibitor--which we call FRAX486--also rescues seizures and behavioral abnormalities such as hyperactivity and repetitive movements, thereby supporting the hypothesis that a drug treatment that reverses the spine abnormalities can also treat neurological and behavioral symptoms. Finally, a single administration of FRAX486 is sufficient to rescue all of these phenotypes in adult Fmr1 KO mice, demonstrating the potential for rapid, postdiagnostic therapy in adults with FXS.