Urinary steroid profiling using liquid chromatography-tandem mass spectrometry for the diagnosis of canine Cushing's syndrome.

Serum cortisol measurements by chemiluminescence enzyme immunoassay (CLEIA) are widely used to diagnose hypercortisolism (HC) or Cushing's syndrome in dogs. However, they are associated with problems such as the need for multiple blood collections under stressful conditions or cross-reactivity between hormones. Therefore, a less invasive and more accurate diagnostic method is required. This study aimed to develop a urinary steroid profile analysis method using liquid chromatography-tandem mass spectrometry (LC/MS/MS) and to evaluate its clinical usefulness. Sixty-five healthy dogs and 38 dogs with suspected HC were included in the study. Using LC/MS/MS, the levels of 11 steroid hormones in the urine were determined. We established the upper limit of the reference interval for each urinary steroid-to-creatinine ratio and evaluated their diagnostic performances. The levels of the five steroid hormones were significantly higher in the 14 dogs with HC than in the 24 dogs with mimicking HC and 65 healthy dogs. The urinary corticosterone-to-creatinine ratio showed the highest diagnostic accuracy (area under the curve, 0.96). A significant correlation was seen between urinary cortisol concentrations measured by LC/MS/MS and CLEIA (rs = 0.88, P <0.001), although the CLEIA measurements were significantly higher than the LC/MS/MS measurements (P <0.001). LC/MS/MS-based urinary steroid profiles are a promising tool for diagnosing canine HC.

Evaluation of responses to immunosuppressive therapy in dogs with suspected non-regenerative immune-mediated anaemia: 11 cases (2012-2018).

OBJECTIVES: We aimed to determine the response time to immunosuppressive therapy and time required to achieve a 5% increase in haematocrit among dogs with non-regenerative immune-mediated anaemia. MATERIALS AND METHODS: Client-owned dogs diagnosed with non-regenerative immune-mediated anaemia in Hokkaido University Veterinary Teaching Hospital between December 2012 and May 2018 were enrolled. The first treatment regimen included prednisolone (2 mg/kg/day) and ciclosporin (up to 10 mg/kg/day) for 8 weeks. Dogs that did not respond to the first regimen proceeded to the second regimen comprising prednisolone and mycophenolate mofetil (15 mg/kg, twice a day). Reticulocyte count and haematocrit were monitored every 1 to 2 weeks. Treatment response was defined as an absolute reticulocyte count more than 60×103 /µL or increasing haematocrit. RESULTS: During the study period, 23 dogs fulfilled the inclusion criteria for non-regenerative immune-mediated anaemia. Twelve dogs were excluded from this study for various reasons and response to therapy was evaluated in the remaining 11 dogs. Treatment responses were observed in 8 of 11 dogs, and the median time to response was 39.5 days (range 8 to 92 days). Two responders were unable to continue the first treatment regimen and were switched to the second regimen owing to anorexia and nausea, possibly induced by ciclosporin; withdrawal of ciclosporin improved their symptoms. The time required to achieve a 5% increase in haematocrit was assessed in the other six dogs, with a median of 55.5 days (range 8 to 135 days). CLINICAL SIGNIFICANCE: Here we report the response to a standardised treatment protocol in dogs with non-regenerative immune-mediated anaemia. Knowledge of potential side effects and expected therapeutic outcomes may be of use for veterinary practitioners treating this condition.

Prognostic value of left atrial stiffness estimated using echocardiography in dogs with myxomatous mitral valve disease.

INTRODUCTION: No studies have investigated the prognostic value of left atrial stiffness (LASt) estimated using echocardiography in dogs with myxomatous mitral valve disease (MMVD). ANIMALS: Seventy-two dogs had MMVD and 46 dogs were healthy dogs. MATERIALS AND METHODS: Clinical retrospective cohort study. The survival information of MMVD dogs that underwent echocardiographic examination was obtained. The peak velocities of early diastolic transmitral flow (E) and mitral annular motion as determined by pulsed wave Doppler (E') were determined. The left atrial reservoir strain (εS) was determined by two-dimensional speckle tracking echocardiography of the left atrium. The LASt was estimated by the formula: E/E'/εS. The 95% prediction interval of LASt was computed from the data of the healthy dogs. RESULTS: Seventeen dogs having MMVD died of cardiac-related causes with 55 MMVD dogs censored. The MMVD dogs with LASt > its 95% upper prediction limit (LASt > 0.56; n = 26; median survival time, 484 days; 95% confidence interval, 283 days-indeterminable) had shorter survival times (P<0.001) than those with LASt ≤ its 95% upper prediction limit (LASt ≤ 0.56; n = 46; median survival time, >1112 days; 95% confidence interval, indeterminable). Multivariable Cox's proportional hazard analysis demonstrated that the ratio of the left atrial dimension to the aortic annulus dimension and LASt were independent predictors of cardiac-related death among conventional echocardiographic indices, εS and LASt in MMVD dogs. CONCLUSIONS: In dogs with MMVD, increased LASt estimated using echocardiography is an independent predictor of cardiac-related death, and LASt can be more useful for prognostication than εS.

Cell death signalling is competitively but coordinately regulated by repressor-type and activator-type ethylene response factors in tobacco (Nicotiana tabacum) plants.

Ethylene response factors (ERFs) comprise one of the largest transcription factor families in many plant species. Tobacco (Nicotiana tabacum) ERF3 (NtERF3) and other ERF-associated amphiphilic repression (EAR) motif-containing ERFs are known to function as transcriptional repressors. NtERF3 and several repressor-type ERFs induce cell death in tobacco leaves and are also associated with a defence response against tobacco mosaic virus (TMV). We investigated whether transcriptional activator-type NtERFs function together with NtERF3 in the defence response against TMV infection by performing transient ectopic expression, together with gene expression, chromatin immunoprecipitation (ChIP) and promoter analyses. Transient overexpression of NtERF2 and NtERF4 induced cell death in tobacco leaves, albeit later than that induced by NtERF3. Fusion of the EAR motif to the C-terminal end of NtERF2 and NtERF4 abolished their cell death-inducing ability. The expression of NtERF2 and NtERF4 was upregulated at the early phase of N gene-triggered hypersensitive response (HR) against TMV infection. The cell death phenotype induced by overexpression of wild-type NtERF2 and NtERF4 was suppressed by co-expression of an EAR motif-deficient form of NtERF3. Furthermore, ChIP and promoter analyses suggested that NtERF2, NtERF3 and NtERF4 positively or negatively regulate the expression of NtERF3 by binding to its promoter region. Overall, our results revealed the cell death-inducing abilities of genes encoding activator-type NtERFs, including NtERF2 and NtERF4, suggesting that the HR-cell death signalling via the repressor-type NtERF3 is competitively but coordinately regulated by these NtERFs.

A novel multi-target strategy to attenuate the progression of Parkinson's disease by diamine hybrid AGE/ALE inhibitor.

Instead of a conventional 'one-drug-one-target approach', this article presents a novel multi-target approach with a concept of trapping simultaneously as many detrimental factors as possible involved in the progression of Parkinson's disease. These factors include reactive carbonyl species, reactive oxygen species, Fe3+/Cu2+ and ortho-quinones (o-quinone), in particular. Different from the known multi-target strategies for Parkinson's disease, it is a sort of 'vacuum cleaning' strategy. The new agent consists of reactive carbonyl species scavenging moiety and reactive oxygen species scavenging and metal chelating moiety linked by a spacer. Provided that the capacity of scavenging o-quinones is demonstrated, this type of agent can further broaden its potential therapeutic profile. In order to support this new hypothetical approach, a number of simple in vitro experiments are proposed.

Changes in fears and worries related to COVID-19 during the pandemic among current employees in Japan: a 5-month longitudinal study.

OBJECTIVES: This study investigates and describes the time course of fears and worries about COVID-19 among current employees during this outbreak. STUDY DESIGN: This was a longitudinal study. METHODS: This study was a part of the Employee Cohort Study in Japan. The study comprised 4120 individuals from February 2019. A baseline survey in March 2020, a 2-month follow-up survey in May 2020, and a 5-month follow-up survey in August 2020 were conducted. Questions surveyed respondents' global fear and worry and six items related to COVID-19. A mixed model for repeated measures of an analysis of variance was used. RESULTS: A total of 1421 respondents completed the baseline survey. At 2- and 5-month follow-ups, 1032 and 1181 respondents completed surveys, respectively. Of those, 64 and 33 individuals who were temporarily laid off or on leave were recorded as missing values. Global fear and worry about COVID-19 significantly increased from March to August 2020. Fears of personal or family infection, limiting one's activities and national and local government policies also significantly increased with time. In contrast, fears of lack of knowledge and difficulty of obtaining hygiene products significantly decreased. CONCLUSION: To conduct efficient risk communication during a pandemic, knowing the concerns of the populace, providing correct information and a sufficient supply of products, and setting clear guidelines are essential.

Predictors of Cerebral Aneurysm Rupture after Coil Embolization: Single-Center Experience with Recanalized Aneurysms.

BACKGROUND AND PURPOSE: Recanalization after coil embolization is widely studied. However, there are limited data on how recanalized aneurysms rupture. Herein, we describe our experience with the rupture of recanalized aneurysms and discuss the type of recanalized aneurysms at greatest rupture risk. MATERIALS AND METHODS: A total of 426 unruptured aneurysms and 169 ruptured aneurysms underwent coil embolization in our institution between January 2009 and December 2017. Recanalization occurred in 38 (8.9%) of 426 unruptured aneurysms (unruptured group) and 37 (21.9%) of 169 ruptured aneurysms (ruptured group). The Modified Raymond-Roy classification on DSA was used to categorize the recanalization type. Follow-up DSA was scheduled until 6 months after treatment, and follow-up MRA was scheduled yearly. If recanalization was suspected on MRA, DSA was performed. RESULTS: In the unruptured group, the median follow-up term was 74.0 months. Retreatment for recanalization was performed in 18 aneurysms. Four of 20 untreated recanalized aneurysms (0.94% of total coiled aneurysms) ruptured. In untreated recanalized aneurysms, class IIIb aneurysms ruptured significantly more frequently than class II and IIIa (P = .025). In the ruptured group, the median follow-up term was 28.0 months. Retreatment for recanalization was performed in 16 aneurysms. Four of 21 untreated recanalized aneurysms (2.37% of total coiled aneurysms) ruptured. Class IIIb aneurysms ruptured significantly more frequently than class II and IIIa (P = .02). CONCLUSIONS: The types of recanalization after coil embolization may be predictors of rupture. Coiled aneurysms with class IIIb recanalization should undergo early retreatment because of an increased rupture risk.

Hydroxypyridinone-Diamine Hybrids as Potential Neuroprotective Agents in the PC12 Cell-Line Model of Alzheimer's Disease.

There is an urgent need to propose effective treatments for Alzheimer's disease (AD). Although the origin of the disease is poorly understood, several therapeutic options have been proposed. The new therapeutic approaches targeting biometal-mediated neurodegenerative pathways appear to be interesting ones. As a continuation of our preceding studies, two novel series of advanced glycation endproducts (AGE)/advanced lipid peroxidation endproducts (ALE) inhibitors have been developed as multifunctional scavengers. This extended work allowed us to highlight the new hydroxypyridinone-diamine hybrid IIa-3 bearing a C4 alkyl linker between the two pharmacophores. This derivative exhibited preserved potent capacities to trap reactive carbonyl species (vicinal diamine function) as well as reactive oxygen species and transition metals (hydroxypyridinone moiety) in comparison with previously described lead compound 1. In addition, its good predicted absorption, distribution, metabolism and excretion (ADME) properties were correlated with a better efficacy to inhibit in vitro methylglyoxal-induced apoptosis in neuronal-like PC12 cells. This new promising agent revealed improved druglikeness and ability to prevent biometal-mediated oxidative and carbonyl stress amplification involved in AD pathogenesis.

Right ventricular function and dyssynchrony measured by echocardiography in dogs with precapillary pulmonary hypertension.

INTRODUCTION: Assessment of the right ventricular (RV) function by echocardiography is important in dogs with pulmonary hypertension (PH). Few reports are available on RV function and dyssynchrony in dogs, especially in the context of precapillary PH. ANIMALS: The study included 79 client-owned dogs: 25 dogs with precapillary PH and 54 control dogs. METHODS: Dogs with precapillary PH were prospectively enrolled between December 2013 and February 2017. The echocardiographic indices of RV function, including RV strain and the dyssynchrony index by speckle-tracking echocardiography, were measured. Multivariate analysis was used to determine independent predictors of the RV dyssynchrony index. RESULTS: Dogs with precapillary PH showed RV dilation, hypertrophy and right atrial dilation. Impaired echocardiographic indices of RV function, including RV strain, were observed. In addition, RV dyssynchrony occurred in dogs with precapillary PH. Multivariate analysis revealed that tricuspid regurgitation velocity and RV dilation were independent predictors of the RV dyssynchrony index. CONCLUSIONS: Echocardiographic RV function indices are impaired in dogs with precapillary PH. In addition, RV dilation and elevated systolic pulmonary arterial pressure estimated by echocardiography are associated with RV dyssynchrony.

The Repeatability and Left Atrial Strain Analysis Obtained via Speckle Tracking Echocardiography in healthy Dogs.

INTRODUCTION: In left atrial (LA) strain-derived two-dimensional speckle tracking echocardiography, the reference intervals in healthy dogs can provide useful information to evaluate the LA function in dogs with heart disease. ANIMALS: Six laboratory beagles and 120 privately owned dogs without cardiac diseases were recruited. MATERIALS AND METHODS: The LA strain and strain rate (SR) and echocardiographic indices were obtained in dogs who underwent standard echocardiography and offline analysis for LA strain and SR measurement by speckle tracking echocardiography. RESULTS: The intra-observer within-day variations of strain variables showed adequate repeatability (coefficient of variation <20%). The mean values of strain were 25.37 for the LA reservoir function, 11.06 for the LA conduit function, and 14.17 for the LA booster-pump function; the strain was significantly correlated with the LA fractional volume change at each phasic function. The left atrial longitudinal strain during early ventricular diastole showed moderate correlation with the peak velocity of early diastolic transmitral flow (r = 0.5560) and ratio of peak velocity of early diastolic transmitral flow to peak velocity of late transmitral flow (r = 0.5515). In multiple regression analysis, only age was significantly related to the strain/SR and volumetric change indices, indicating conduit function. CONCLUSIONS: Left atrial speckle tracking echocardiographic analysis provided useful information to assess the LA function in healthy dogs. The influencing factors on strain and SR variables including the age, body weight, and heart rate should be considered in interpretation of these parameters in a clinical setting.