Changes in the ear canal microbiota of dogs with otitis externa.

AIMS: Otitis externa (OE), one of the most common ear diseases in dogs, is caused by bacterial pathogens such as Staphylococcus sp. To understand the network of microbial communities in the canine ear canal affected with OE, we performed a cross-sectional study using next-generation sequencing. METHODS AND RESULTS: Ear swab samples were collected from 23 OE-affected and 10 healthy control dogs, and the 16S rRNA gene sequenced using Illumina MiSeq. The otic microbiota in the OE-affected dogs showed significantly decreased alpha diversity compared to controls. The community composition also differed in the affected group, with significantly higher relative abundance of the phylum Firmicutes and the genus Staphylococcus (P = 0·01 and 0·04 respectively). Contrary to our expectations, the severity of the disease did not impact the otic microbiota in OE-affected dogs. CONCLUSIONS: The ear canal microbiota of OE-affected dogs is distinct from that of healthy dogs, irrespective of disease status. SIGNIFICANCE AND IMPACT OF THE STUDY: This study, one of the few detailed analyses of the otic microbiota, can provide practical information for the appropriate treatment of canine OE.

Long-term clinical impact of serum albumin in coronary artery disease patients with preserved renal function.

BACKGROUND AND AIMS: Low serum albumin level is reportedly associated with worse clinical outcomes in patients with chronic kidney disease (CKD). However, associations between decreased serum albumin level and outcomes in non-CKD patients with coronary artery disease (CAD) remain unclear. Therefore, we aimed to evaluate the prognostic value of serum albumin concentrations in stable CAD patients with preserved renal function. METHODS AND RESULTS: We studied 1316 patients with CAD and preserved renal function (estimated glomerular filtration rate ≥60 mL/min/1.73 m2) who underwent their first PCI between 2000 and 2011 and had data available for pre-procedural serum albumin. Patients were assigned to quartiles based on pre-procedural albumin concentrations. The incidence of major adverse cardiac events (MACE), including all-cause death and non-fatal myocardial infarction, was evaluated. Mean albumin concentration was 4.1 ± 0.4 g/dL. During the median follow-up of 7.5 years, 181 events occurred (13.8%). Kaplan-Meier curves revealed that patients with decreased serum albumin concentrations showed a higher event rate for MACE (log-rank, p < 0.0001). Using the highest tertiles (>4.3 g/dL) as reference, adjusted hazard ratios were 1.97 (95% CI, 1.12-3.55), 1.77 (95% CI, 0.99-3.25), and 1.19 (95% CI, 0.68-2.15) for serum albumin concentrations of <3.9, 3.9-4.0, and 4.1-4.3 g/dL, respectively. Decreased serum albumin concentration was associated with MACE even after adjusting for other independent variables (HR, 2.21 per 1-g/dL decrease; 95% CI, 1.37-3.56, p = 0.001). CONCLUSION: Decreased serum albumin concentration independently predicted worse long-term prognosis in non-CKD patients after PCI. Pre-procedural serum albumin concentration could offer a useful predictor for patients with CAD and preserved renal function.

Increased ratio of FoxP3+ regulatory T cells/CD3+ T cells in skin lesions in drug-induced hypersensitivity syndrome/drug rash with eosinophilia and systemic symptoms.

BACKGROUND: Drug-induced hypersensitivity syndrome/drug rash with eosinophilia with systemic symptoms (DIHS/DRESS) is a severe drug eruption accompanied by multiorgan disorders. Several unique aspects of DIHS/DRESS, including herpesvirus reactivation, liver dysfunction and hypogammaglobulinaemia, have similarities to graft-versus-host disease (GVHD). AIM: In this study, we focused on the dynamics of regulatory T cells (Tregs) infiltrating into the skin lesions of DIHS/DRESS and GVHD. METHODS: Skin biopsies were taken from patients with DIHS/DRESS, GVHD, or maculopapular drug eruption. Tregs were detected using immunostaining with anti-FoxP3. RESULTS: The ratio of FoxP3+ T cells to CD3+ T cells was significantly higher in the skin lesions of patients with DIHS/DRESS than in those of patients with GVHD, and was positively correlated with the number of days from disease onset in the acute phase. CONCLUSIONS: The dynamics of Tregs in skin lesions are different between DIHS/DRESS and GVHD, despite there being many similarities between these conditions.

The upper airway in sleep-disordered breathing: UA in SDB.

Sleep disordered breathing (SDB) is a common condition and could be a risk factor for cardiovascular morbidity and mortality. However, the pathogenesis of SDB remains to be elucidated. In general, SDB is divided into two forms, obstructive and central sleep apnea (OSA and CSA, respectively). OSA results from the sleep-related collapse of the upper airway (UA) in association with multiple factors like race, gender, obesity and UA dimensions. CSA primarily results from a fall in PaCO2 to a level below the apnea threshold during sleep through the reflex inhibition of central respiratory drive. It has been reported that UA alterations (i.e., collapse or dilation) can be observed in CSA. This review highlights the roles of the UA in the pathogenesis and pathophysiology of SDB.

Involvement of autoimmunity to REG, a regeneration factor, in patients with primary Sjögren's syndrome.

The regenerating gene (Reg) was isolated originally as a gene specifically over-expressed in regenerating pancreatic islets and constitute a growth factor family. Reg gene product (Reg) is important in the pathophysiology of various human inflammatory diseases. Recently, the possible involvement of human REG in the regeneration of salivary ductal epithelial cells of patients with primary Sjögren's syndrome (SS) was reported. However, the expression of the REG family genes in minor salivary glands (MSG) and the occurrence of anti-REG Iα autoantibodies in SS patients were obscured. In this study, we examined the expression of REG family genes in the MSG of SS and screened anti-REG Iα autoantibodies in SS. The mRNA levels of REG family genes in MSG were quantified using real-time reverse transcription-polymerase chain reaction (RT-PCR) and REG Iα expression in the MSG was analysed by immunohistochemistry. The mRNA level of REG Iα in the MSG of SS patients was significantly higher than that of control. REG Iα protein was expressed highly in SS ductal epithelial cells. Anti-REG Iα autoantibodies in the sera were found in 11% of SS. All the MSG in the anti-REG Iα autoantibody-positive group showed REG Iα expression, whereas only 40% showed REG Iα expression in the anti-REG Iα autoantibody-negative group. The anti-REG Iα autoantibody-positive group showed significantly lower saliva secretion and a higher ratio of grade 4 (by Rubin-Holt) in sialography. These data suggest strongly that autoimmunity to REG Iα might play a role in the degeneration of MSG ductal epithelial cells in primary SS.

Phase I/II trial of a biweekly combination of S-1 plus docetaxel in patients with previously treated non-small cell lung cancer (KRSG-0601).

BACKGROUND: Combination of S-1, an oral fluorouracil derivative, plus docetaxel against non-small cell lung cancer (NSCLC) showed promising efficacy but clinically problematic emesis. A phase I/II study utilising a new schedule for this combination was conducted. METHODS: A biweekly regimen of docetaxel on day 1 with oral S-1 on days 1-7 was administered to previously treated NSCLC patients. Doses of docetaxel/S-1 were escalated to 30/80, 35/80, and 40/80 mg m(-2), respectively, and its efficacy was investigated at the recommended dose below maximum tolerated dose (MTD). RESULTS: In phase I study employing 13 patients, dose-limiting toxicities were febrile neutropenia and treatment delay, with the respective MTDs for docetaxel 40 mg m(-2)/S-1 80 mg m(-2). In the phase II study, 34 patients were treated with docetaxel 35 mg m(-2)/S-1 80 mg m(-2) for a median cycle of 6. The response and disease control rates were 34.3% (95% confidence interval (CI), 18.6-50.0%) and 62.9% (95% CI, 46.8-72.9%), respectively. Median progression-free survival was 150.5 days. Haematologic grade 4 toxicities were observed in neutropenia (11.8%) and thrombocytopenia (2.9%). Regarding non-haematologic toxicities, including emesis, there were no grade 3/4 side effects. CONCLUSION: Combination of 1-week administration of S-1 with biweekly docetaxel is safe and active for NSCLC.

Antihypertensive effect of a fixed-dose combination of losartan/hydrochlorothiazide in patients with uncontrolled hypertension: a multicenter study.

BACKGROUND: Achieving adequate blood pressure (BP) control often requires more than one antihypertensive agent. The purpose of this study was to determine whether a fixed-dose formulation of losartan (LOS) plus hydrochlorothiazide (HCTZ) (LOS/HCTZ) is effective in achieving a greater BP lowering in patients with uncontrolled hypertension. METHODS: The study was a prospective, multicenter, observational trial exploring the antihypertensive effect of a single tablet of LOS 50 mg/HCTZ 12.5 mg. A total of 228 patients whose BP had previously been treated with more than one antihypertensive agents without having achieved BP goal below 130/80 mmHg enrolled in the study. RESULTS: A significant decrease in systolic and diastolic BP was observed in both clinic and home measurement after switching from the previous treatment to LOS/HCTZ. There was a significant decrease in both B-type natriuretic peptide (BNP) and urinary albumin creatinine (Cr) excretion ratio (ACR), especially in patients with elevated values. In contrast, there was a significant increase in serum Cr concentration in conjunction with a decrease in estimated glomerular filtration rate (eGFR). Overall serum uric acid (UA) concentration increased, whereas in patients with hyperuricemia there was a significant reduction in this value. CONCLUSION: Switching to LOS/HCTZ provides a greater reduction in clinic and home BP in patients with uncontrolled hypertension. This combination therapy may lead to cardio-, reno protection and improve UA metabolism.

Rotationally correlated reactivity in the CH (v = 0, J, F(i)) + O2 → OH (A) + CO reaction.

The rotational-state-selected CH (v = 0, J, F(i)) beam has been prepared by using an electric hexapole and applied to the crossed beam reaction of CH (v = 0, J, F(i)) + O(2) → OH (A) + CO at different O(2) beam conditions. The rotational state selected reactive cross sections of CH (RSSRCS-CH) turn out to depend remarkably on the rotational state distribution of O(2) molecules at a collision energy of ∼ 0.19 eV. The reactivity of CH molecules in the N = 1 rotational states (namely ∣J = 1∕2, F(2)> and ∣J = 3∕2, F(1)> states, N designates the angular momentum excluding spin) becomes strongly enhanced upon a lowering of the rotational temperature of the O(2) beam. The RSSRCS-CH in these two rotational states correlate linearly with the population of O(2) molecule in the specific K(O(2)) frame rotation number states: CH(|J = 1/2,F(2)>) with O(2)(|K(O(2)) = 1>);CH(|J = 3/2,F(1)>) with O(2)(|K(O(2)) = 3>). These linear correlations mean that the rotational-state-selected CH molecules are selectively reactive upon the incoming O(2) molecules in a specific rotational state; here, we use the term "rotationally correlated reactivity" to such specific reactivity depending on the combination of the rotational states between two molecular reactants. In addition, the steric asymmetry in the oriented CH (∣J = 1∕2, F(2), M = 1∕2>) + O(2) (|K(O(2)) = 1>) reaction turns out to be negligible (< ±1%). This observation supports the reaction mechanism as theoretically predicted by Huang et al. [J. Phys. Chem. A 106, 5490 (2002)] that the first step is an intermediate formation with no energy barrier in which C-atom of CH molecule attacks on one O-atom of O(2) molecule at a sideways configuration.

Multi-dimensional steric effect for XeI* (B) formation in the oriented Xe* ((3)P(2), M(J) = 2) + oriented CH(3)I reaction.

Multi-dimensional steric effect for the XeI* (B) formation in the oriented Xe* ((3)P(2), M(J) = 2) + oriented CH(3)I reaction has been observed as a function of the mutual configuration between the molecular orientation and the atomic alignment in the collision frame. The molecular steric opacity function has been determined as a function of the atomic orbital alignment. The large molecular orientation dependence (i.e., the largest reactivity at the I-end and the large difference in the reaction probability between the I-end and the CH(3)-end) and the large molecular alignment dependence (the poor reactivity at the sideway) is recognized for each atomic orbital alignment. In addition, a clear correlation between the molecular orientation and the atomic orbital alignment is recognized (i.e., the L(Z)' = 0 atomic orbital alignment is favorable for the molecular axis direction, while the |L(Z)'| = 1 atomic orbital alignment is favorable for the sideway direction).

Multidimensional steric effect for the XeBr* (B, C) formation in the oriented Xe*((3)P2, M(J) = 2) + oriented CF3Br reaction.

Steric effect for the XeBr(*) (B, C) formation in the oriented Xe(*)((3)P(2), M(J) = 2) + oriented CF(3)Br reaction has been observed as a function of the mutual configuration between the molecular orientation and the atomic orientation in the collision frame. Molecular steric opacity function has been determined as a function of the atomic orbital alignment (L(Z)(')) in the collision frame. The L(Z)(') selectivity in the molecular steric opacity function is different between the XeBr(*) (B, C) channels: For the XeBr(*) (C) channel, the L(Z)(') = 0 alignment is favorable at the molecular axis direction and the absolute value(L(Z)(')) = 1 alignment is favorable at the sideway direction, whereas for the XeBr(*) (B) channel, the L(Z)(') = 0 alignment is favorable at the sideway direction and the absolute value(L(Z)(')) = 1 alignment is favorable at the molecular axis direction. However, the shape of the steric opacity function for the XeBr(*) (B) channel at the L(Z)(') = 0 (and absolute value(L(Z)(')) = 1) alignment is similar to that for the XeBr(*) (C) channel at the absolute value(L(Z)(')) = 1 (and L(Z)(') = 0) alignment, respectively: A large molecular orientation dependence (i.e., the largest reactivity at the Br-end with the small molecular alignment dependence) is recognized for the XeBr(*) (B) channel at the L(Z)(') = 0 alignment and for the XeBr(*) (C) channel at the absolute value(L(Z)(')) = 1 alignment, whereas a large molecular alignment dependence (i.e., the largest reactivity at the Br-end with the poor reactivity at the sideway) is recognized for the XeBr(*) (B) channel at the absolute value(L(Z)(')) = 1 alignment and for the XeBr(*) (C) channel at the L(Z)(') = 0 alignment. We propose the indirect mechanism for the dark channels (Xe + Br + CF(3)) via the back-electron transfer from the CF(3) segment (or dissociating CF(3)...Br(-)) to Xe(+) as the origin of the significant molecular alignment dependence in the molecular steric opacity function.

Getting back to basics during a public health emergency: a framework to prepare and respond to infectious disease public health emergencies.

This paper describes a simple framework developed by the World Health Organization, used to convey the concept of comprehensive and integrated public health response structures and to identify core public health capacity development needs. The framework highlights five core components of a response: surveillance, healthcare response, public health intervention, communication and command. This paper notes that to mount an effective public health response, each component requires sufficient capacity to meet demand, and effective relationships and mechanisms need to be established between components that support coordination, communication and collaboration.

Mammalian transgenesis by intracytoplasmic sperm injection.

Coinjection of unfertilized mouse oocytes with sperm heads and exogenous DNA encoding either a green fluorescent protein (GFP) or beta-galactosidase reporter produced 64 to 94 percent transgene-expressing embryos, reflecting DNA-sperm head association before coinjection. Nonselective transfer to surrogate mothers of embryos in the GFP series generated about 20 percent offspring expressing the integrated transgene. These data indicate that exogenous DNA can reproducibly be delivered into an oocyte by microinjected spermatozoa and suggest an adaptable method of transgenesis.

The Culdotomy FourS Two U procedure for transvaginal access to the peritoneal cavity.

A reliable method of transvaginal access is needed for natural orifice transluminal endoscopic surgery. We introduce a new culdotomy procedure, using a technique for the creation of space in the cul-de-sac, transvaginal ultrasound, and a newly developed umbrella Hakko needle. An artificially developed, saline-containing space in the cul-de-sac was punctured by an umbrella Hakko needle from the vagina under the guidance of ultrasound. The vaginal walls on both sides of the needle were incised with an electric scalpel. In five cases with benign gynecological tumors, culdotomy was successfully performed. Operating time was less than 10 minutes and blood loss was less than 10 mL. There were no culdotomy-associated complications. This procedure, named Culdotomy FourS Two U, is a simple, safe, and reliable method for entry into the cul-de-sac in transvaginal gynecological surgeries and may have future applications in transluminal endoscopic surgery through the vagina.

Collision energy dependence of the rotational-state-resolved cross section in the CH (v = 0, J, F(i)) + O(2) --> OH(A) + CO reaction.

A velocity variable rotational-state-selected CH (v = 0, J, F(i)) beam has been prepared by using an electric hexapole and applied to the CH (v = 0, J, F(i)) + O(2) --> OH(A) + CO reaction. The CH rotational-state-resolved reaction cross sections have been determined under the beam-cell condition at the collision energy range of 0.06-0.18 eV. The N = 2 rotational states are 2-3 times more reactive than the other states (N = 1, 3). In addition, we observed a noticeable difference in the collision energy dependence of the cross section between the CH rotational states. The reaction cross section for the N = 2 states has a gentle negative dependence on collision energy, while, the reaction cross section for the N = 1 states has a positive dependence on collision energy.

Steric effect in the energy transfer reaction of oriented Kr (3P2, MJ = 2) + CO.

Atomic alignment effect for the formations of CO (a' (3)Sigma(+)) and CO (d (3)Delta) in the energy transfer reaction of oriented Kr ((3)P(2), M(J) = 2) + CO has been measured at a collision energy of 0.07 eV. The emission intensities of CO (a' (3)Sigma(+)) and CO (d (3)Delta) were similarly highly enhanced when the electron angular momentum of Kr ((3)P(2)) is aligned perpendicular to the relative velocity vector. We observed the analogous atomic alignment effect between the CO (a' (3)Sigma(+)) and CO (d (3)Delta) formations. That is, the |M(J)'| = 2 magnetic substate in the collision frame is significantly less reactive than the other M(J)' states. In addition, the large difference of the cross section (sigma(Sigma)/sigma(Pi) approximately 2.0) between the Sigma- and Pi-configuration of the unpaired 4p orbital of Kr ((3)P(2)) is recognized.

Correlation between the atomic alignment and the alignment of XeX* (B, C) rotation in the reactions of oriented Xe ((3)P(2), M(J) = 2) + halogen (X)-containing molecules.

The polarization of the XeX* (B) and XeX* (C) emissions in the reactions of oriented Xe* ((3)P(2), M(J) = 2) + halogen (X)-containing molecules (CCl(4), CF(3)Br, CF(3)I, CH(3)I, NF(3)) has been measured as a function of each magnetic M(J)' substate in the collision frame. The parallel polarization of the XeX* (B, C) emissions to the relative velocity vector is commonly observed for all magnetic M(J)' substates. The correlation between the atomic alignment (M(J)') and the M(J)'-dependent alignment (A(M(J)')) of the XeX* (B, C) rotation is found to be extremely different between the XeX* (B) and XeX* (C) channels: For XeX* (B), A(M(J)') is highest for the M(J)' = 0 state, except CCl(4), whereas the |M(J)'| = 2 states give the highest A(M(J)') for XeX* (C). Alternatively, the correlation between the configuration (L(Z)') of the inner 5p orbital and the L(Z)'-dependent alignment (A(L(Z)')) of the XeX* (B, C) rotations is revealed. The collision with |L(Z)'| = 1 causes a similar positive alignment A(L(Z)') for the XeX* (B) and XeX* (C) channels. The alignment A(L(Z)') at the collision with |L(Z)'| = 0 is extremely different between the XeX* (B) and XeX* (C) channels. The collision with L(Z)' = 0 induces no alignment of XeX* (C) except CF(3)I, that is, A(L(Z)'=0) approximately 0, whereas it induces the higher positive A(L(Z)') of XeX* (B). The different M(J)' dependence on the alignment of the XeX* (B, C) rotation between the XeX* (B) and XeX* (C) channels can be recognized as the change of reaction mechanism due to the difference in the favorable impact parameter for each M(J)' state between the XeX* (B) and XeX* (C) channels, which reflects the Omega' conservation in the course of ion-pair (Xe(+)-RX(-)) formation.

Atomic alignment effects for the formation of excimers RgX* (B, C) in the reaction of oriented Rg ((3)P2, M(J) = 2) (Rg = Xe, Kr, Ar) + halogen(X)-containing molecules.

Atomic alignment effects for the formation of excimers, RgX* (B, Omega = 1/2) and RgX* (C, Omega = 3/2), in the reaction of Rg ((3)P(2)) (Rg = Xe, Kr, Ar) with halogen (X)-containing molecules (RX) (CH(3)I, CF(3)I, CF(3)Br, NF(3), CHBr(3), CHCl(3), CCl(3)F, and CCl(4)) have been measured by using an oriented Rg ((3)P(2), M(J) = 2) atomic beam at a collision energy of approximately 0.07 eV. The emission intensities for RgX* (B, C) have been measured as a function of the magnetic orientation field direction in the collision frame. The reactant (RX) dependence of the atomic alignment effect is extremely different between the RgX* (B) and the RgX* (C) channels. For RgX* (C), an analogous atomic alignment effect is commonly observed despite the difference of RX and Rg. In contrast, for RgX* (B), the atomic alignment effect shows a diverse dependence on RX and Rg.

Multidimensional steric effects for the XeI* (B, C) formations in the oriented Xe* (3P(2),M(J) = 2) + oriented CF3I reaction.

Steric effects for the XeI(*) (B) and XeI(*) (C) formations in the oriented Xe(*) ((3)P(2),M(J)=2)+oriented CF(3)I reaction have been observed as a function of the mutual configuration between molecular orientation and atomic alignment in the collision frame. The mutual configuration exercises the significant influences on the stereoanisotropy for both the reactivity and the branching to the XeI(*) (B) and XeI(*) (C) channels.

Long-time exposure of mouse embryos to the sperm produces high levels of reactive oxygen species in culture medium and relates to poor embryo development.

Small amounts of reactive oxygen species (ROS), metabolites of oxygen, are necessary for sperm-fertilizing capability. However, in excessive levels, their role in infertility has been extensively studied. The conventional in vitro fertilization (IVF) method employs a prolonged co-incubation of gametes for 16-18 h to reach fertilization. However, it has been shown that this long period might create high levels of ROS. We aimed at finding out whether ROS increases in vitro during prolonged incubation with fertilized oocytes and whether high level of ROS relates to poor embryo development. To confirm if levels of ROS relate to length of time, we measured the ROS levels in fertilization medium (FM), which contained mouse embryos exposed to spermatozoa. To evaluate the contribution of sperm in production of ROS, we measured the ROS in the medium with only sperm. The measurements were performed by chemiluminescence assay using luminol as a probe after 4 and 18 h of incubation separately. The ROS levels were significantly increased after 18 h as compared with 4 h (p < 0.0001). Moreover, ROS in the medium with only sperm was also increased after 18 h (p < 0.0001), demonstrating that they were generated either by spermatozoa or as a result of possible reaction of sperm with medium during prolonged incubation. In addition, we compared embryo development after 2, 4, 6, 8, 10, 12 and 18 h of incubation. The number of degenerated embryos exposed to sperm for 12 and 18 h was significantly higher than those exposed for 4 or 6 h (p < 0.01). These results demonstrate that ROS concentrations appear to be related to the length of incubation time, and their excessive levels have a negative effect on embryo development. We suggest reducing incubation time to at least 4 h.