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This thesis examines and compares two natural resource management regimes, those of the Australian State of Tasmania and the sovereign state of Taiwan, with a focus upon their respective terrestrial natural reserve systems. Recommendations for future improvements are made for both islands. Taiwan is an island about half the size of Tasmania, yet the former has a population more than 48 times greater than the latter. The two island ecosystems are similar in some respects, but the contrasts are more marked than the similarities. It would be beneficial for both islands to share their experiences of natural resource management. This study undertakes such a comparison with a view to facilitating exchange of knowledge in the field of environmental management. Despite its dramatically smaller population, Tasmania's terrestrial natural resource management is more highly developed than Taiwan's in some respects. For example, the New Public Management (NPM) model has been employed as a framework for regime reform in Tasmania, but not in Taiwan. There is, nevertheless, room for improvement in planning and practice on both islands. The Tasmanian government structure provides a more integrated approach to natural resource management, especially with regard to its nature reserve system, and Taiwan could learn from this in planning for the future. The successful Landcare movement and accumulated treaty-derived conservation experience, in, for example, World Heritage Area and Ramsar site management, are appropriate for adaptation in Taiwan to foster community involvement and prepare itself for the transition to involvement in international affairs. On the other hand, the integrated environmental education coordination across governmental agencies in Taiwan, although not yet implemented, could be considered as a future approach in Tasmania.

Landcare education is part of environmental education and unique to Australia. This paper examines Landcare as an approach to Australian environmental education experience and looks at the practice of environmental education in Australia from the perspective of a Chinese student from Taiwan. This study should therefore benefit both Australia and the Republic of China (Taiwan). The study focuses on the Landcare for Teachers Program which is a promising teacher training course which will help educators to start or enhance their teaching in Landcare. The skills acquired by participants are also applicable to other subject areas and non-teaching activities. Evaluation is necessary before the program can successfully be extended from Tasmania to other Australian States and Territories, and be considered for use in other countries. An evaluation of the Landcare for Teachers Program was made using a questionnaire survey along with interviews of some of the participants and key persons including seminar leaders. Findings are based upon the questionnaire, interviews, and related documentary and statistical analysis. The results of the evaluation have allowed recommendations to be made for the Landcare for Teachers program and the potential for developing a similar program in the Republic of China to be assessed. The major findings are that the Landcare for Teachers Program has a very positive response from participants, more than 90% satisfaction rate, which is an indication of its success; however, the course content lacks a clear philosophical direction, such as might be derived from the environmental ethic of, for example, the Australian Aboriginal culture. In addition, some course topics need more careful design to cater for the differing needs of primary and secondary school teachers, taking account of their different training backgrounds and teaching subject areas.

During COVID-19, the anatomy faculty and students at Schulich School of Medicine and Dentistry observed strengths and weaknesses in their transition to online learning. A "four-prong" approach to teaching anatomy was developed. Asynchronous content modules were tailored to specific learning objectives, virtual labs were implemented to work through case-based applications, "live from the lab" review sessions provided the opportunity for interaction and integration, and finally, limited face-to-face laboratory sessions provided an opportunity for supervised consolidation with cadaveric specimens. Our approach may be used by other institutions to enhance anatomical education and student engagement.

Introduction: Perivascular fibrosis, characterized by increased amount of connective tissue around vessels, is a hallmark for vascular disease. Angiotensin II (Ang II) contributes to vascular disease via promoting T-cell activation and end-organ damage. Despite recent data suggesting the role of T cells in the progression of perivascular fibrosis, the underlying mechanisms are poorly understood. Objective: Kruppel-like Factor 10 is a transcription factor expressed in T cell subsets. We sought to investigate the role of KLF10 in CD4+ T cells in regulating vascular damage in an AngII mouse model. Methods: CD4-targeted KLF10 deficient (TKO) and CD4-Cre (WT) mice were generated and subjected to 28 days of Ang II infusion. Endpoint characterization included fibrotic organ transcriptomic analysis, multi-organ histology, flow cytometry, cytokine analysis, myograph vasoreactivity, and cardiac echocardiography. Results: TKO mice showed enhanced perivascular fibrosis compared to WT mice by histological analysis in the aorta, heart, and kidney. TKO mice had vessels with enhanced vasoconstriction to phenylephrine, hearts with impaired global longitudinal strain by echo, and kidneys with elevated albumin/creatinine ratio. However, no change was found in blood pressure between TKO and WT. Plasma IL-9 and IL-15 were increased in TKO mice. IL-9 mRNA and secreted protein were also increased in activated TKO CD4+ T cells, as well as in Ang II treated WT-CD4+ T cells in vitro . Mechanistic studies revealed that KLF10 regulated Il9 transcription through interaction with promoter region of Il9 by ChIP assay. Notably, injection of anti-IL9 antibody reversed perivascular fibrosis in Ang II infused TKO mice. Conclusion: CD4+ T cell deficiency of Klf10 exacerbated perivascular fibrosis and multi-organ dysfunction in response to Ang II via upregulation of IL-9. Klf10 or IL-9 in T cells might represent novel therapeutic targets for the treatment of vascular or fibrotic diseases.

We report normal-incidence planar GeSn resonant-cavity-enhanced photodetectors (RCE-PDs) with a lateral p - i - n homojunction configuration on a silicon-on-insulator (SOI) platform for short-wave infrared (SWIR) integrated photonics. The buried oxide of the SOI platform and the deposited S i O 2 layer serve as the bottom and top reflectors, respectively, creating a vertical cavity for enhancing the optical responsivity. The planar p - i - n diode structure is favorable for complementary-metal-oxide-semiconductor-compatible, large-scale integration. With the bandgap reduction enabled by the 4.2% Sn incorporation into the GeSn active layer, the photodetection range extends to 1960 nm. The promising results demonstrate that the developed planar GeSn RCE-PDs are potential candidates for SWIR integrated photonics.

A young child presented with severe ventricular dysfunction and troponin leak in the setting of Coronavirus-19 disease (COVID-19). He developed intermittent, self-resolving, and hemodynamically insignificant episodes of complete heart block (CHB), which were diagnosed on telemetry and managed conservatively. This report is the first description of COVID-19 induced transient CHB in a child., Graphical abstract

Neonates with critical left heart obstruction and intact atrial septum (IAS) or restrictive atrial septum (RAS) are at risk for hypoxia within hours of birth and remain a group at high risk for mortality.Prenatally diagnosed fetuses with critical left heart obstruction and IAS or RAS with follow-up from January 1, 2005, to February 14, 2017, were included. Primary outcome was a composite measure of severe neonatal illness (pH 7.15, venous pH 7.10, bicarbonate 16 mmol/L, lactic acid 5 mmol/L, or median oxygen saturation 60% within 2 hours of birth).Of 68 live born fetuses, 52 (76.5%) had hypoplastic left heart syndrome, 14 (20.5%) had critical aortic stenosis, and two (3%) had complex anatomy with mitral stenosis/atresia. There were 27 (39.7%) fetuses with IAS and 41 (60.3%) with RAS. Severe neonatal illness was present in 36 (52.9%). The strongest discriminators for severe neonatal illness were a pulmonary vein A:R VTI ≤ 2.7 (P 0.001, AUC 0.93) and larger pulmonary vein diameter (P = 0.025, AUC 0.77). A:R VTI ≤ 2.7 predicted death or transplant (log-rank P = 0.03).In neonates with hypoplastic left heart syndrome and IAS or RAS, A:R VTI ≤ 2.7 is predictive of severe neonatal instability. This threshold can help guide resource planning, delivery management, and improve fetal intervention criteria.

Publicly available data from the Pediatric Heart Network's Single Ventricle Reconstruction Trial was analyzed to determine the prevalence, timing, risk factors for, and impact of second- and third-degree heart block (HB) on outcomes in patients who underwent stage 1 palliation (S1P) for hypoplastic left heart syndrome (HLHS).The presence and date of onset of post-S1P HB occurring within the first year of life, potential risk factors for HB, and factors known to predict poor outcomes after S1P were extracted. Multivariable logistic and Cox regression analyses were performed to identify risk factors for HB and to determine the effect of HB on 3-year transplantation-free survival.Among the 549 patients in the cohort, 33 (6%) developed HB after S1P. The median interval between S1P and HB was 8 days (interquartile range, 0-133 days). Regression analysis showed that tricuspid valve repair during S1P and obstruction of pulmonary venous drainage requiring pre-S1P intervention were independently associated with HB (adjusted odds ratio [aOR], 11.6, 95% confidence interval [CI] 3.3-40; P .001 and aOR, 5.1; 95% CI, 1.3-20.6; P = .02, respectively). Transplantation-free survival at 3 years was lower for those with HB (39% vs 65%; P = .004). HB remained associated with transplantation-free survival after controlling for known risk factors (adjusted hazard ratio, 3.1; 95% CI, 1.9-5.0; P .001). Nine children (27%) had a pacemaker implanted, and 7 of these children (78%) died or underwent heart transplantation.HB after S1P is rare but heralds a poor outcome. Careful monitoring of these patients is recommended given their significantly increased risks of death and heart transplantation.

We demonstrate silicon-based p - n - p floating-base GeSn heterojunction phototransistors with enhanced optical responsivity for efficient short-wave infrared (SWIR) photodetection. The narrow-bandgap GeSn active layer sandwiched between the p - G e collector and n - G e base effectively extends the photodetection range in the SWIR range, and the internal gain amplifies the optical response by a factor of more than three at a low driving voltage of 0.4 V compared to that of a reference GeSn p - i - n photodetector (PD). We anticipate that our findings will be leveraged to realize complementary metal-oxide-semiconductor-compatible, sensitive, low driving voltage SWIR PDs in a wide range of applications.

Extracorporeal membrane oxygenation (ECMO) is life saving for many critically ill children with congenital heart disease (CHD). However, limited information is available about their ensuing neurodevelopmental (ND) outcomes. We describe early ND outcomes in a cohort of children supported with ECMO for cardiac indications. Twenty-eight patients supported with ECMO at age < 36 months underwent later ND testing at 12-42 months of age using the Bayley Scales of Infant and Toddler Development, Third Edition (Bayley-III). ND scores were compared with normative means and with ND outcomes of a matched cohort of 79 children with CHD undergoing cardiac surgery but not requiring ECMO support. Risk factors for worse ND outcomes were identified using multivariable linear regression models. Cardiac ECMO patients had ND scores at least one standard deviation below the normative mean in the gross motor (61%), language (43%), and cognitive (29%) domains of the Bayley-III. Cardiac ECMO patients had lower scores on the motor, language, and cognitive domains as compared to the matched non-ECMO group and clinically important (1/2 SD) differences in the motor domain persisted after controlling for primary caregiver education and number of cardiac catheterizations. Risk factors of worse ND outcomes among cardiac ECMO patients in more than one developmental domain, included older age at first cannulation and more cardiac catheterization and cardiac surgical procedures prior to ND assessment. Overall, children supported on ECMO for cardiac indications have significant developmental delays and warrant close ND follow-up.

We report an investigation of strain-dependent Raman frequency shift in Ge 0.917 Sn 0.083 epilayer where the Sn composition is near the indirect-to-direct band gap transition point. The strain is modulated by ex-situ rapid thermal annealing and the amount of strain relaxation is determined by X-ray diffraction measurement. The results show that the Raman shift of Ge–Ge longitudinal optical phonon is linearly dependent on the in-plane strain of the GeSn layers with a coefficient b = − 299.3 cm − 1 . Such a relationship enables characterization of GeSn epilayers using the readily accessible Raman spectroscopy.

Background: Type-2 diabetes (T2D) is associated with both reduced and preserved ejection fraction heart failure. Obese db/db mice and Goto Kakizaki (GK) rats represents animals models of T2D that develop cardiac dysfunction similar to human diabetic cardiomyopathy, in which dominant early findings are of diastolic (and not systolic) dysfunction. Circulating extracellular vesicles (EV) contain microRNAs (miR) that can be transferred to recipient cells to modulate their function. We explored whether analysis of EV content from animals models of T2D would inform on the pathophysiology, diagnosis and therapeutic targets of cardiac dysfunction. Hypothesis: EV from animal models of T2D will have altered miR content that contributes to the pathophysiology of diabetic cardiomyopathy. Methods & Results: miR qPCR arrays on circulating EV isolated from plasma of db/db mice reveal several miR (-7, -15, -25, -30e, -148a, -150, -195) modulated during disease progression. These changes in miR content occur prior to echocardiographic evidence of diastolic dysfunction, including global longitudinal strain and strain rate. Among circulating EV miR from the GK rat model, miR-30 was also upregulated (1.42 fold, p=0.03) compared to Wistar rat. In GK rat left ventricle, and in H9C2 rat cardiac myoblast cultured in 25 mM high glucose media, mass spectrometry revealed proteins that were overexpressed in the diabetic heart including oxidative phosphorylation, glycolysis, fatty acid degredation and the citrate cycle. Using a bioinformatics approach, we next identified metabolic pathways affected by miR-30. Based on these findings, in vivo therapy with antagomiR and mimics of miR-30 are underway to test causality and reversibility of the observed cardiomyopathy. Conclusion: EV from animal models of T2D have altered miR content, including miR-30. We also identify alterations in the expression of a network of metabolism genes in the heart, which are implicated in diabetic cardiomyopathy. If causality is supported by experiments that enhance or block miR-30 expression in these models of disease, we will have identified a novel biomarker and therapeutic target for diabetic cardiomyopathy.

In this paper, the current standard method and other commonly used engineering practices for linear or linearized engineering failure data analysis are critically reviewed first, and the existing issues are also indicated. To overcome these issues, a linear data analysis method based on a new equilibrium mechanism is subsequently presented. Based on the equilibrium mechanism, three possible ideal data patterns are identified and the corresponding best curve fitting approaches are proposed. Compared to the existing methods, the equilibrium method not only provides quantitative solutions of fit parameters, but also gives an obvious physical meaning and, therefore, is more intuitive in quickly and correctly identifying data patterns, subsequent data preprocessing, and evaluating goodness-of-fit. The equilibrium method is further applied to fatigue and creep data analyses to demonstrate its applicability.

Vascular calcification is a common feature of chronic kidney disease, cardiovascular disease, and aging. Such abnormal calcium deposition occurs in medial and/or intimal layers of blood vessels as well as in cardiac valves. Once considered a passive and inconsequential finding, the presence of calcium deposits in the vasculature is widely accepted as a predictor of increased morbidity and mortality. Recognition of the importance of vascular calcification in health is driving research into mechanisms that govern its development, progression, and regression. Diverse, but highly interconnected factors, have been implicated, including disturbances in lipid metabolism, oxidative stress, inflammatory cytokines, and mineral and hormonal balances, which can lead to formation of osteoblast-like cells in the artery wall. A tight balance of procalcific and anticalcific regulators dictates the extent of disease. In this review, we focus on the main regulatory circuits modulating vascular cell calcification.

Doxorubicin-induced cardiotoxicity (CTX) occurs in 10 to 15% of patients. As such, there is an important need to improve our understanding of the relevance of cardiac mechanics in predicting CTX. The overall objective of our study was to use machine learning algorithms to identify new patterns in

We describe the fabrication of a screen printed disposable electrode system with integrated elements to allow for control of the working electrode temperature. The device has fully functional working, counter, and reference electrodes with underlying thermocouple and heating elements to allow feedback control of temperature. In our experiments, we determined that the accuracy of the temperature measurements on the electrode was within 0.5 °C over the temperature range of liquid water using a single universal calibration, and we demonstrated that the control system could maintain a setpoint temperature with a root mean squared error of 0.35 °C based on the indicated temperature. We used these electrodes to determine the temperature dependence of the diffusivity of ferricyanide ([Fe(CN)6]3−) ion using linear sweep voltammetry. The diffusion coefficient determined at 25 °C (0.85 × 10−5 cm2 s−1) was similar to other reported values in the literature, although our observed activation energy for diffusion (32.1 kJ mol−1) was significantly higher. Our observed temperature coefficient of −1.63 mV K−1 for the redox potential of the [Fe(CN)6]3−/[Fe(CN)6]4− couple is reasonably close to other reported values. The system shows promise for use in other disposable electrochemical diagnostic systems in which temperature control plays a key role, especially where perturbations or variations in ambient temperature or electrolyte rheological properties make feedback control necessary.

Tuberous sclerosis complex (TSC) is an autosomal dominant, tumor predisposition disorder characterized by significant neurodevelopmental brain lesions, such as tubers and subependymal nodules. The neuropathology of TSC is often associated with seizures and intellectual disability. To learn about the developmental perturbations that lead to these brain lesions, we created a mouse model that selectively deletes the Tsc2 gene from radial glial progenitor cells in the developing cerebral cortex and hippocampus. These Tsc2 mutant mice were severely runted, developed post-natal megalencephaly and died between 3 and 4 weeks of age. Analysis of brain pathology demonstrated cortical and hippocampal lamination defects, hippocampal heterotopias, enlarged dysplastic neurons and glia, abnormal myelination and an astrocytosis. These histologic abnormalities were accompanied by activation of the mTORC1 pathway as assessed by increased phosphorylated S6 in brain lysates and tissue sections. Developmental analysis demonstrated that loss of Tsc2 increased the subventricular Tbr2-positive basal cell progenitor pool at the expense of early born Tbr1-positive post-mitotic neurons. These results establish the novel concept that loss of function of Tsc2 in radial glial progenitors is one initiating event in the development of TSC brain lesions as well as underscore the importance of Tsc2 in the regulation of neural progenitor pools. Given the similarities between the mouse and the human TSC lesions, this model will be useful in further understanding TSC brain pathophysiology, testing potential therapies and identifying other genetic pathways that are altered in TSC.

ObjectivesNeonates undergoing cardiac surgery are at high risk for adverse outcomes. B-type natriuretic peptide is used as a biomarker in patients with cardiac disease, but the predictive value of B-type natriuretic peptide after cardiac surgery in neonates has not been evaluated. Therefore, the objective of this study was to determine the predictive value of perioperative B-type natriuretic peptide levels for postoperative outcomes in neonates undergoing cardiac surgery.MethodsPlasma B-type natriuretic peptide determinations were made before and 2, 12, and 24 hours after surgery in 36 consecutive neonates. B-type natriuretic peptide levels and changes in perioperative B-type natriuretic peptide were evaluated as predictors of postoperative outcome.ResultsB-type natriuretic peptide levels at 24 hours were lower than preoperative levels (24-h/pre B-type natriuretic peptide ratio < 1) in 29 patients (81%) and higher (24-h/pre B-type natriuretic peptide ratio ≥ 1) in 7 patients (19%). A 24-hour/pre B-type natriuretic peptide level of 1 or greater was associated with an increased incidence of low cardiac output syndrome (100% vs 34%, P = .002) and fewer ventilator-free days (17 ± 13 days vs 26 ± 3 days, P = .002), and predicted the 6-month composite end point of death, an unplanned cardiac operation, or cardiac transplant (57% vs 3%, P = .003). A 24-hour/pre B-type natriuretic peptide level of 1 or greater had a sensitivity of 80% and a specificity of 90% for predicting a poor postoperative outcome (P = .003).ConclusionIn neonates undergoing cardiac surgery, an increase in B-type natriuretic peptide 24 hours after surgery predicts poor postoperative outcome.

This report summarizes the genome-wide effects of ACTH on transcript accumulation in mouse adrenal Y1 cells and the relative contributions of the cAMP-, protein kinase C- and Ca(2+)-dependent signaling pathways to these actions of the hormone. ACTH affected the accumulation of 1386 transcripts, a much larger number than previously appreciated. The cAMP signaling pathway accounted for approximately 56% of the ACTH effects whereas the protein kinase C- and Ca(2+)-dependent pathways made smaller contributions to ACTH action. Approximately 38% of the ACTH-affected transcripts could not be assigned to these signaling pathways and thus represent candidates for regulation via other mechanisms. The set of ACTH-regulated transcripts included clusters with functions in steroid metabolism, cell proliferation and alternative splicing. Collectively, our results suggest that Y1 adrenal cells undergo extensive remodeling upon prolonged stimulation with ACTH. The functional implications of ACTH on alternative splicing are explored.

ACTH regulates the steroidogenic capacity, size, and structural integrity of the adrenal cortex through a series of actions involving changes in gene expression; however, only a limited number of ACTH-regulated genes have been identified, and these only partly account for the global effects of ACTH on the adrenal cortex. In this study, a National Institute on Aging 15K mouse cDNA microarray was used to identify genome-wide changes in gene expression after treatment of Y1 mouse adrenocortical cells with ACTH. ACTH affected the levels of 1275 annotated transcripts, of which 46% were up-regulated. The up-regulated transcripts were enriched for functions associated with steroid biosynthesis and metabolism; the down- regulated transcripts were enriched for functions associated with cell proliferation, nuclear transport and RNA processing, including alternative splicing. A total of 133 different transcripts, i.e. only 10% of the ACTH-affected transcripts, were represented in the categories above; most of these had not been described as ACTH-regulated previously. The contributions of protein kinase A and protein kinase C to these genome-wide effects of ACTH were evaluated in microarray experiments after treatment of Y1 cells and derivative protein kinase A-defective mutants with pharmacological probes of each pathway. Protein kinase A-dependent signaling accounted for 56% of the ACTH effect; protein kinase C-dependent signaling accounted for an additional 6%. These results indicate that ACTH affects the expression profile of Y1 adrenal cells principally through cAMP- and protein kinase A- dependent signaling. The large number of transcripts affected by ACTH anticipates a broader range of actions than previously appreciated.

A collection of 4457 Saccharomyces cerevisiae mutants deleted for nonessential genes was screened for mutants with increased or decreased mobilization of the gypsylike retroelement Ty3. Of these, 64 exhibited increased and 66 decreased Ty3 transposition compared with the parental strain. Genes identified in this screen were grouped according to function by using GOnet software developed as part of this study. Gene clusters were related to chromatin and transcript elongation, translation and cytoplasmic RNA processing, vesicular trafficking, nuclear transport, and DNA maintenance. Sixty-six of the mutants were tested for Ty3 proteins and cDNA. Ty3 cDNA and transposition were increased in mutants affected in nuclear pore biogenesis and in a subset of mutants lacking proteins that interact physically or genetically with a replication clamp loader. Our results suggest that nuclear entry is linked mechanistically to Ty3 cDNA synthesis but that host replication factors antagonize Ty3 replication. Some of the factors we identified have been previously shown to affect Ty1 transposition and others to affect retroviral budding. Host factors, such as these, shared by distantly related Ty retroelements and retroviruses are novel candidates for antiviral targets.

Structure-activity relationship (SAR) studies of 2-[3-di(and tri)fluoromethyl-5-arylpyrazol-1-yl]-5-methanesulfonylpyridine derivatives for canine COX enzymes are described. This led to the identification of 12a as a lead candidate for further progression. The in vitro and in vivo activity of 12a for the canine COX-2 enzyme as well as its in vivo efficacy and pharmacokinetic properties in dog are highlighted.

Müllerian-inhibiting substance (MIS), a member of the transforming growth factor-beta family of cytokines that signal through a heteromeric complex of single-transmembrane serine/threonine kinase receptors, is required for Müllerian duct regression and normal reproductive tract development in the male embryo. However, the continued expression of MIS at high levels in males until puberty and its induction in females after birth suggested other roles for MIS. Additionally, Leydig cell development and steroidogenic capacity and ovarian follicle recruitment were abnormal in MIS-knockout or MIS-overexpressing mice. We have shown that MIS inhibits the cAMP-induced expression of cytochrome P450 C17alpha-hydroxylase/C17-20 lyase (Cyp17) mRNA both in vitro and in vivo. Our current efforts are to understand the molecular mechanisms regulating both MIS type II receptor (MISRII) expression and its signaling in rodent Leydig cell lines. MISRII expression in R2C cells requires both steroidogenic factor-1 and an unknown protein to bind to its proximal promoter in the context of 1.6 kb 5'-flanking DNA. When bound by MIS, signaling by the receptor in MA-10 cells blocks the protein kinase A-mediated induction of Cyp17 expression by a cAMP regulatory element-binding protein independent mechanism. We continue to investigate the molecular mechanisms of MISRII expression and possible interactions between MIS-regulated SMAD activation and cAMP signaling. These studies will provide a better understanding of the role played by MIS during postnatal life.

Background To guide treatment of infectious diseases, clinicians need sensitive, specific, and rapid diagnostics. We aim to incorporate complementary methods of microbial sequencing and host-response profiling to improve the diagnosis of patients at risk for acute infections. Methods We enrolled 200 adult patients with systemic inflammatory response syndrome (SIRS) at the Stanford Emergency Department. Physicians with specialty training in infectious diseases conducted retrospective two-physician chart review to establish likely admission diagnoses. Blood samples were tested with a previously described 18-gene host-response integrated antibiotics decision model (IADM) that distinguishes noninfectious SIRS, bacterial infections and viral infections. Plasma samples were tested with shotgun metagenomic next-generation sequencing (NGS) and viral sequencing with VirCapSeq. A novel statistical algorithm was developed to identify contaminant organism sequences in NGS data. Results The physician chart review classified 99 patients (49%) as infected, 69 (35%) possibly infected and 32 (16%) non-infected. Compared with chart review, the IADM distinguished bacterial from viral infections with an area under curve of 0.85 (95% confidence interval 0.77–0.93). NGS results to date confirmed positive blood cultures in seven of nine patients, with two of four blood culture-positive E. coli patients turning up negative on NGS due to E. coli contamination. NGS also confirmed positive cultures from other sites in two of six patients with negative blood cultures. Preliminary VirCapSeq data from 23 patients confirmed positive viral tests in five of six patients with Hepatitis C, BK Virus, Cytomegalovirus and Epstein–Barr Virus infections. VirCapSeq did not identify a causative agent in the plasma of 11 patients with confirmed respiratory viral infection and intestinal Norovirus infection, and six patients with idiopathic illness. Interestingly, VirCapSeq found viral reactivation in 8 of 12 immunocompromised patients. Conclusion The diagnosis of suspected infections may be enhanced by integrating host-response and microbial data alongside clinical judgment. Our results and large cohort lay the foundation to demonstrate the utility of this approach and in which patients these tools may be most useful. Disclosures T. E. Sweeney, Inflammatix, Inc: Employee and Shareholder, Salary; T. Briese, Roche: Columbia University has licensed VirCapSeq to Roche, Licensing agreement or royalty; W. I. Lipkin, Roche: Columbia University has licensed VirCapSeq to Roche., Licensing agreement or royalty; P. Khatri, Inflammatix, Inc.: Co-founder, Scientific Advisor and Shareholder, Licensing agreement or royalty and ownership stock; D. A. Relman, Karius: Consultant, Stock options; Arc Bio LLC: Consultant, Stock options

Residency training subjects healthy young people to frequent fluctuation in sleep-wake cycles. Although shift work is known to increase cardiovascular risk, little is known about the physiology of phase-shifted sleep in residency. This study uses 24-hr ambulatory blood pressure monitoring to investigate diurnal BP variation in medical residents working alternating day/night periods. We hypothesized that diurnal variation of BP is dependent upon sleep/wake status, not upon day/night hours. Residents were recruited from the Johns Hopkins Medicine Residency Program during the 2013-2014 academic year. Thirty-six individuals participated while working in three clinical environments (intensive care unit, general medicine wards, outpatient clinic), and during various sleep-wake schedules (night shift, 24-hr overnight call, day shift). BP was measured every 30 min during waking hours and every 2 hr during sleep. Residents self-reported during which hours they slept. Ninety-eight separate 24-hr periods of data were collected (Table 1). There is no difference in mean 24-hr SBP, awake SBP/DBP, or asleep SBP/DBP between those working different schedules. However, there are statistically significant differences in day SBP/DBP and night SBP/DBP between participants working different schedules. In all schedules, participants showed at least a 9 mmHg drop in SBP between awake and asleep . Diurnal variation of BP in this population is present, and dependent upon sleep/wake status, not day/night hours. Further investigation into the correlation between this reversal of diurnal BP variation and cardiovascular outcomes in shift workers is warranted.

Background Changes in energy substrate metabolism are first responders to hemodynamic stress in the heart. We have previously shown that hexose‐6‐phosphate levels regulate mammalian target of rapamycin ( mTOR ) activation in response to insulin. We now tested the hypothesis that inotropic stimulation and increased afterload also regulate mTOR activation via glucose 6‐phosphate (G6P) accumulation. Methods and Results We subjected the working rat heart ex vivo to a high workload in the presence of different energy‐providing substrates including glucose, glucose analogues, and noncarbohydrate substrates. We observed an association between G6P accumulation, mTOR activation, endoplasmic reticulum ( ER ) stress, and impaired contractile function, all of which were prevented by pretreating animals with rapamycin ( mTOR inhibition) or metformin ( AMPK activation). The histone deacetylase inhibitor 4‐phenylbutyrate, which relieves ER stress, also improved contractile function. In contrast, adding the glucose analogue 2‐deoxy‐ d ‐glucose, which is phosphorylated but not further metabolized, to the perfusate resulted in mTOR activation and contractile dysfunction. Next we tested our hypothesis in vivo by transverse aortic constriction in mice. Using a micro‐ PET system, we observed enhanced glucose tracer analog uptake and contractile dysfunction preceding dilatation of the left ventricle. In contrast, in hearts overexpressing SERCA2a, ER stress was reduced and contractile function was preserved with hypertrophy. Finally, we examined failing human hearts and found that mechanical unloading decreased G6P levels and ER stress markers. Conclusions We propose that glucose metabolic changes precede and regulate functional (and possibly also structural) remodeling of the heart. We implicate a critical role for G6P in load‐induced mTOR activation and ER stress.

Highlights: ► Anabolic effects of PTH were tested in hyperlipidemic mice overexpressing PON1. ► Expression of antioxidant regulatory genes was induced in PON1 overexpression. ► Bone resorptive activity was reduced in PON1 overexpressing hyperlipidemic mice. ► PON1 restored responsiveness to intermittent PTH in bones of hyperlipidemic mice. -- Abstract: Hyperlipidemia blunts anabolic effects of intermittent parathyroid hormone (PTH) on cortical bone, and the responsiveness to PTH are restored in part by oral administration of the antioxidant ApoA-I mimetic peptide, D-4F. To evaluate the mechanism of this rescue, hyperlipidemic mice overexpressing the high-density lipoprotein-associated antioxidant enzyme, paraoxonase 1 (Ldlr{sup −/−}PON1{sup tg}) were generated, and daily PTH injections were administered to Ldlr{sup −/−}PON1{sup tg} and to littermate Ldlr{sup −/−} mice. Expression of bone regulatory genes was determined by realtime RT-qPCR, and cortical bone parameters of the femoral bones by micro-computed tomographic analyses. PTH-treated Ldlr{sup −/−}PON1{sup tg} mice had significantly greater expression of PTH receptor (PTH1R), activating transcription factor-4 (ATF4), and osteoprotegerin (OPG) in femoral cortical bone, as well as significantly greater cortical bone mineral content, thickness, and area in femoral diaphyses compared with untreated Ldlr{sup −/−}PON1{sup tg} mice. In contrast, in control mice (Ldlr{sup −/−}) without PON1 overexpression, PTH treatment did notmore » induce these markers. Calvarial bone of PTH-treated Ldlr{sup −/−}PON1{sup tg} mice also had significantly greater expression of osteoblastic differentiation marker genes as well as BMP-2-target and Wnt-target genes. Untreated Ldlr{sup −/−}PON1{sup tg} mice had significantly greater expression of PTHR1 than untreated Ldlr{sup −/−} mice, whereas sclerostin expression was reduced. In femoral cortical bones, expression levels of transcription factors, FoxO1 and ATF4, were also elevated in the untreated, control Ldlr{sup −/−}PON1{sup tg} mice, suggesting enhancement of cellular protection against oxidants. These findings suggest that PON1 restores responsiveness to PTH through effects on oxidant stress, PTH receptor expression, and/or Wnt signaling.« less

To describe patterns of care for pediatric patients with advanced heart disease who experience in-hospital death.Retrospective single-institution medical record review.A tertiary care pediatric hospital.All patients younger than 21 years who died in the inpatient setting between January 1, 2007, and December 31, 2009, with primary cardiac diagnoses or who had ever received a cardiology consult (N=468). After excluding patients with significant noncardiac primary diagnoses, 111 children formed the analytic sample.In-hospital deaths of children with heart disease during a 3-year period.Median age at death was 4.8 months (age range,1 day to 20.5 years), with 84 deaths (75.7%) occurring before age 1 year. Median length of terminal hospital stay was 22 days (range, 1-199 days). Diagnoses included 84 patients (75.7%) with congenital heart disease, 10 (9.0%)with cardiomyopathy/myocarditis, 9 (8.1%) with pulmonary hypertension, and 8 (7.2%) with heart transplants.Sixty-two patients (55.9%) had received cardio-pulmonary resuscitation during their last hospital admission. At the end of life, 21 children (18.9%) had gastrostomy tubes and 26 (23.4%) had peritoneal drains.Most patients (91.9%) received ventilation, with half also receiving mechanical circulatory support. Eighty-three patients (74.8%) experienced additional end-organ failure. Classified by mode of death, 76 patients (68.5%) had disease-directed support withdrawn, 28 (25.2%) died during resuscitation, and 7 (6.3%) died while receiving comfort care after birth. Eighty-three percent of parents were present at the time of death.Infants and children who die of advanced heart disease frequently succumb in the intensive care setting with multisystem organ failure and exposure to highly technical care.

Asrac Caegor 27. Cogeal Carolog Soluos earcreseao Numer 1135-124Auhors Henry Cheng, David Wypij, Peter Laussen, David Bellinger, Christian D. Stopp, Jane Newburger, Barry Kussman, Childrens Hospital Boston, Boston, MA, USABackground: Cereral loo low veloc (CV) measure rascraal Doppler soograph has prove sgh o cereral perfuso paes uergog surgcal repar of cogeal hear sease. However, a relaoshp ewee CV a evelopmeal oucome has o ee esalshe.Methods: Ifas

In embryogenesis, structural patterns, such as vascular branching, may form via a reaction-diffusion mechanism in which activator and inhibitor morphogens guide cells into periodic aggregates. We previously found that vascular mesenchymal cells (VMCs) spontaneously aggregate into nodular structures and that morphogen pairs regulate the aggregation into patterns of spots and stripes. To test the effect of a focal change in activator morphogen on VMC pattern formation, we created a focal zone of high cell density by plating a second VMC layer within a cloning ring over a confluent monolayer. After 24 h, the ring was removed and pattern formation monitored by phase-contrast microscopy. At days 2–8, the patterns progressed from uniform distributions to swirl, labyrinthine and spot patterns. Within the focal high-density zone (HDZ) and a narrow halo zone, cells aggregated into spot patterns, whilst in the outermost zone of the plate, cells formed a labyrinthine pattern. The area occupied by aggregates was significantly greater in the outermost zone than in the HDZ or halo. The rate of pattern progression within the HDZ increased as a function of its plating density. Thus, focal differences in cell density may drive pattern formation gradients in tissue architecture, such as vascular branching.

In this study, we describe a disposable electrode system on a polycarbonate substrate intended for DNA-based detection of the bacterial plant pathogen Ralstonia solanacearum. The electrode system was prepared by using a laser engraver to expose and isolate areas of gold film on gold compact discs to make working, auxiliary, and reference electrodes. Silver and silver chloride were subsequently coated onto the reference electrode by electrochemical treatment. The CD electrode system performed comparably to commercial screen-printed electrodes when used for analysis of ferricyanide by cyclic voltammetry, and also showed superior electrode surface smoothness. Current efforts are focused on coupling DNA hybridization probes to these electrode systems to detect loop-mediated isothermal amplification (LAMP) amplicons of R. solanacearum.