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Objective: To determine multidimensional impulsivity levels across different early stages of α-synucleinopathy., Methods: This cross-sectional study investigated motor and decisional impulsivity levels using a panel of computerized tasks among drug-naïve parkinsonism patients, isolated/idiopathic rapid eye movement sleep behavior disorder (iRBD) patients and their first-degree relatives (iRBD-FDRs), and control participants. Trait impulsivity and impulse control behaviors were assessed by self-reported questionnaires., Results: A total of 27 drug-naïve parkinsonism patients, 157 iRBD patients, 66 iRBD-FDRs, and 82 control participants were recruited. Parkinsonism and iRBD patients had fewer numbers of extracted beads in beads task 1 and 2 (both p < 0.001), and a higher rate of irrational choice in task 1 (p = 0.046) before making decisions, and fewer numbers of pumps of unexploded blue balloons in the balloon analog risk task (p = 0.004) than control participants, indicating a higher level of reflection impulsivity and a lower level of risk taking, respectively. iRBD patients had more no-go errors in the go/no-go task than control participants (p adjusted  = 0.036), suggesting a higher level of motor impulsivity. iRBD-FDRs with dream-enactment behaviors had fewer numbers of extracted beads (p = 0.047) in beads task 2 than FDRs without dream-enactment behaviors, suggesting a possible higher level of reflection impulsivity., Interpretation: A complex construct of altered impulsivity with decreased risk taking, but increased reflection and motor impulsivity, has already occurred at the prodromal and early stages of α-synucleinopathy, which have implications for underlying pathophysiology and clinical management of α-synucleinopathy, especially for impulse control behaviors upon dopaminergic drug treatment. ANN NEUROL 2024;95:544-557., (© 2023 The Authors. Annals of Neurology published by Wiley Periodicals LLC on behalf of American Neurological Association.)

Objective: Age at onset of neurodegenerative disease has significant implications in differentiating disease profiles. We aimed to determine whether age at onset could identify clinical and neurodegenerative profiles in patients with isolated/idiopathic rapid eye movement sleep behavior disorder (iRBD) - a prodromal stage of α-synucleinopathies., Methods: In this retrospective cohort study, the time of the first episode of dream-enactment behaviors that the patient/bed-partners recalled at the time of the patient's first visit to sleep clinic was collected. The distribution of age at onset was examined and patients were dichotomized into early- and late-onset groups based on the intersection point of underlying two Gaussian distributions of onset age., Results: A total of 241 patients were included. The intersection of underlying two Gaussian models of onset age was 64.6 years, yielding 168 early- (median onset age: 58.0 years, range: 38.0-64.0) and 73 late-onset patients (median onset age: 70.0 years, range: 65.0-82.0). Among them, 154 of early- and 68 late-onset patients were followed-up. Late-onset patients had milder RBD symptoms, but worse sleep, cognition, olfactory and motor functions, and a higher risk of phenoconversion (adjusted hazard ratio (aHR) = 2.2, 95% confidence interval (CI) = 1.2-3.9), especially to probable dementia with Lewy bodies (DLB) (aHR = 8.9, 95% CI = 3.0-26.2), than early-onset patients., Conclusions: Late-onset iRBD was associated with a higher level of neurodegenerative markers and a quicker phenoconversion, especially to probable DLB. Age at onset of iRBD could help identify clinical features and predict prognosis of iRBD., Competing Interests: Declaration of competing interest Dr. Li Zhou and Dr. Bei Huang were supported by the Faculty Postdoctoral Fellowship Scheme of the Chinese University of Hong Kong. Dr. Joey WY Chan received personal fee from Eisai. Co.,Ltd for joining an expert panel. Prof. Yun Kwok Wing received personal fees from Eisai Co., Ltd for lecture, travel support from Lundbeck HK Limited, which are outside the submitted work. Other co-authors report no disclosures., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Positive affect is associated with more adaptive responses to psychological stress. However, few studies have examined the association between gratitude, a specific type of positive affect, with physiological responses to acute psychological stress. The current study aimed to replicate and extend on previous work examining the associations between state and trait gratitude and cardiovascular stress reactivity in 324 (59.9% female, 67.0% Caucasian, 17.9% Hispanic) healthy participants. State gratitude was measured at the beginning of the laboratory session using the Gratitude Adjective Checklist-Three Items. Trait gratitude was measured using the Gratitude Questionnaire-Six Items. Blood pressure and heart rate reactions to an acute mental arithmetic task were measured. In regression models that adjusted for baseline cardiovascular activity, body mass index, sex, depressive symptomology, performance on the acute mental arithmetic task, and state positive affect, state gratitude was associated with lower systolic blood pressure reactivity. There were no associations between trait gratitude and any of the cardiovascular variables. Results support previous work demonstrating that state, but not trait, gratitude is related to cardiovascular stress reactivity. Higher levels of state gratitude immediately preceding a stressful encounter may be protective., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Pluripotent stem cell (PSC) differentiation in vitro represents a powerful and tractable model to study mammalian development and an unlimited source of cells for regenerative medicine. Within hematology, in vitro PSC hematopoiesis affords novel insights into blood formation and represents an exciting potential approach to generate hematopoietic and immune cell types for transplantation and transfusion. Most studies to date have focused on in vitro hematopoiesis from mouse PSCs and human PSCs. However, differences in mouse and human PSC culture protocols have complicated the translation of discoveries between these systems. We recently developed a novel chemical media formulation, expanded potential stem cell medium (EPSCM), that maintains mouse PSCs in a unique cellular state and extraembryonic differentiation capacity. Herein, we describe how EPSCM can be directly used to stably maintain human PSCs. We further demonstrate that human PSCs maintained in EPSCM can spontaneously form embryoid bodies and undergo in vitro hematopoiesis using a simple differentiation protocol, similar to mouse PSC differentiation. EPSCM-maintained human PSCs generated at least two hematopoietic cell populations, which displayed distinct transcriptional profiles by RNA-sequencing (RNA-seq) analysis. EPSCM also supports gene targeting using homologous recombination, affording generation of an SPI1 (PU.1) reporter PSC line to study and track in vitro hematopoiesis. EPSCM therefore provides a useful tool not only to study pluripotency but also hematopoietic cell specification and developmental-lineage commitment., (Copyright © 2019 ISEH -- Society for Hematology and Stem Cells. Published by Elsevier Inc. All rights reserved.)

Background : Inadequately controlled sternotomy pain after cardiac surgery can lead to delayed recovery and patient suffering. Preoperative intravenous methadone is effective for reducing both postoperative pain and opioid consumption. Despite ease of administration, the effects of preoperative oral methadone are not well described in the literature. Aims : This pilot study investigated the effect of preoperative oral methadone on pain scores, analgesia requirements, and opioid-induced side effects. Methods : A randomized double-blind placebo-controlled model was used with sampling of patients undergoing sternotomy for isolated coronary artery bypass graft (CABG) surgery (ClinicalTrials.gov registration no. NCT02774499). Participants were randomized to receive oral methadone (0.3 mg/kg) or oral placebo prior to entering the operating room. The primary outcome was pain scores on a 0-10 Verbal Rating Scale. Secondary outcomes included morphine requirements using patient-controlled analgesia (PCA), time to extubation, level of sedation, and side effects such as nausea, vomiting, pruritus, hypoventilation, and hypoxia over a 72-h monitoring time. Results : Twenty-one patients completed the study. Oral methadone did not reduce pain scores in the methadone group ( P = 0.08). However, postoperative morphine requirement during the first 24 h was reduced by a mean of 23 mg in the methadone group (mean difference, -23; 99% confidence interval [CI], 37-13 mg; P < 0.005). No reduction in pain scores or PCA morphine was observed beyond 24 h postoperatively. There was no difference in incidence of opioid-related side effects between groups throughout the postoperative period. Conclusions : Though preoperative oral methadone did not reduce pain scores, morphine requirements were reduced in the first 24 h post-CABG., Competing Interests: No potential conflict of interest was reported by the authors., (© 2019 The Author(s). Published with license by Taylor & Francis Group, LLC.)

Mouse embryonic stem cells derived from the epiblast contribute to the somatic lineages and the germline but are excluded from the extra-embryonic tissues that are derived from the trophectoderm and the primitive endoderm upon reintroduction to the blastocyst. Here we report that cultures of expanded potential stem cells can be established from individual eight-cell blastomeres, and by direct conversion of mouse embryonic stem cells and induced pluripotent stem cells. Remarkably, a single expanded potential stem cell can contribute both to the embryo proper and to the trophectoderm lineages in a chimaera assay. Bona fide trophoblast stem cell lines and extra-embryonic endoderm stem cells can be directly derived from expanded potential stem cells in vitro. Molecular analyses of the epigenome and single-cell transcriptome reveal enrichment for blastomere-specific signature and a dynamic DNA methylome in expanded potential stem cells. The generation of mouse expanded potential stem cells highlights the feasibility of establishing expanded potential stem cells for other mammalian species.

Innate lymphoid cells (ILCs) functionally resemble T lymphocytes in cytotoxicity and cytokine production but lack antigen-specific receptors, and they are important regulators of immune responses and tissue homeostasis. ILCs are generated from common lymphoid progenitors, which are subsequently committed to innate lymphoid lineages in the α-lymphoid progenitor, early innate lymphoid progenitor, common helper innate lymphoid progenitor and innate lymphoid cell progenitor compartments. ILCs consist of conventional natural killer cells and helper-like cells (ILC1, ILC2 and ILC3). Despite recent advances, the cellular heterogeneity, developmental trajectory and signalling dependence of ILC progenitors are not fully understood. Here, using single-cell RNA-sequencing (scRNA-seq) of mouse bone marrow progenitors, we reveal ILC precursor subsets, delineate distinct ILC development stages and pathways, and report that high expression of programmed death 1 (PD-1 hi ) marked a committed ILC progenitor that was essentially identical to an innate lymphoid cell progenitor. Our data defined PD-1 hi IL-25R hi as an early checkpoint in ILC2 development, which was abolished by deficiency in the zinc-finger protein Bcl11b but restored by IL-25R overexpression. Similar to T lymphocytes, PD-1 was upregulated on activated ILCs. Administration of a PD-1 antibody depleted PD-1 hi ILCs and reduced cytokine levels in an influenza infection model in mice, and blocked papain-induced acute lung inflammation. These results provide a perspective for exploring PD-1 and its ligand (PD-L1) in immunotherapy, and allow effective manipulation of the immune system for disease prevention and therapy.

Embryonic stem cell (ESC) culture conditions are important for maintaining long-term self-renewal, and they influence cellular pluripotency state. Here, we report single cell RNA-sequencing of mESCs cultured in three different conditions: serum, 2i, and the alternative ground state a2i. We find that the cellular transcriptomes of cells grown in these conditions are distinct, with 2i being the most similar to blastocyst cells and including a subpopulation resembling the two-cell embryo state. Overall levels of intercellular gene expression heterogeneity are comparable across the three conditions. However, this masks variable expression of pluripotency genes in serum cells and homogeneous expression in 2i and a2i cells. Additionally, genes related to the cell cycle are more variably expressed in the 2i and a2i conditions. Mining of our dataset for correlations in gene expression allowed us to identify additional components of the pluripotency network, including Ptma and Zfp640, illustrating its value as a resource for future discovery., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Hematopoietic stem cells (HSCs) are a rare cell type with the ability of long-term self-renewal and multipotency to reconstitute all blood lineages. HSCs are typically purified from the bone marrow using cell surface markers. Recent studies have identified significant cellular heterogeneities in the HSC compartment with subsets of HSCs displaying lineage bias. We previously discovered that the transcription factor Bcl11a has critical functions in the lymphoid development of the HSC compartment., Results: In this report, we employ single-cell transcriptomic analysis to dissect the molecular heterogeneities in HSCs. We profile the transcriptomes of 180 highly purified HSCs (Bcl11a (+/+) and Bcl11a (-/-)). Detailed analysis of the RNA-seq data identifies cell cycle activity as the major source of transcriptomic variation in the HSC compartment, which allows reconstruction of HSC cell cycle progression in silico. Single-cell RNA-seq profiling of Bcl11a (-/-) HSCs reveals abnormal proliferative phenotypes. Analysis of lineage gene expression suggests that the Bcl11a (-/-) HSCs are constituted of two distinct myeloerythroid-restricted subpopulations. Remarkably, similar myeloid-restricted cells could also be detected in the wild-type HSC compartment, suggesting selective elimination of lymphoid-competent HSCs after Bcl11a deletion. These defects are experimentally validated in serial transplantation experiments where Bcl11a (-/-) HSCs are myeloerythroid-restricted and defective in self-renewal., Conclusions: Our study demonstrates the power of single-cell transcriptomics in dissecting cellular process and lineage heterogeneities in stem cell compartments, and further reveals the molecular and cellular defects in the Bcl11a-deficient HSC compartment.

The transcription activator-like effectors (TALEs) and the RNA-guided clustered regularly interspaced short palindromic repeat (CRISPR) associated protein (Cas9) utlilize distinct molecular mechanisms in targeting site recognition. The two proteins can be modified to carry additional functional domains to regulate expression of genomic loci in mammalian cells. In this study, we have compared the two systems in activation and suppression of the Oct4 and Nanog loci by targeting their enhancers. Although both are able to efficiently activate the luciferase reporters, the CRISPR/dCas9 system is much less potent in activating the endogenous loci and in the application of reprogramming somatic cells to iPS cells. Nevertheless, repression by CRISPR/dCas9 is comparable to or even better than TALE repressors. We demonstrated that dCas9 protein binding results in significant physical interference to binding of native transcription factors at enhancer, less efficient active histone markers induction or recruitment of activating complexes in gene activation. This study thus highlighted the merits and drawbacks of transcription regulation by each system. A combined approach of TALEs and CRISPR/dCas9 should provide an optimized solution to regulate genomic loci and to study genetic elements such as enhancers in biological processes including somatic cell reprogramming and guided differentiation., (© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Recent researches have identified multiple transcription factors as permissible reprogramming factors to pluripotency and lineage switching. The current standard strategy by ectopic factor overexpression however has intrinsic limitations in studying the reprogramming mechanism. There is a growing interest in engineering novel chimeric reprogramming factors and applying designer transcription factors technology to improve reprogramming efficiency and dissect the process of endogenous pluripotency network reactivation. Here, we provide a concise review on the latest progress in studying cellular reprogramming by transcription factor engineering., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The modular DNA recognition code of the transcription-activator-like effectors (TALEs) from plant pathogenic bacterial genus Xanthomonas provides a powerful genetic tool to create designer transcription factors (dTFs) targeting specific DNA sequences for manipulating gene expression. Previous studies have suggested critical roles of enhancers in gene regulation and reprogramming. Here, we report dTF activator targeting the distal enhancer of the Pou5f1 (Oct4) locus induces epigenetic changes, reactivates its expression, and substitutes exogenous OCT4 in reprogramming mouse embryonic fibroblast cells (MEFs) to induced pluripotent stem cells (iPSCs). Similarly, dTF activator targeting a Nanog enhancer activates Nanog expression and reprograms epiblast stem cells (EpiSCs) to iPSCs. Conversely, dTF repressors targeting the same genetic elements inhibit expression of these loci, and effectively block reprogramming. This study indicates that dTFs targeting specific enhancers can be used to study other biological processes such as transdifferentiation or directed differentiation of stem cells.

Background: The diagnosis of coronary artery disease (CAD) in women remains a challenge, given their lower prevalence of obstructive disease and the suboptimal performance of traditional noninvasive tests (exercise electrocardiography and stress myocardial perfusion imaging). Coronary computed tomographic angiography (CTA) is a validated method for detection and exclusion of obstructive coronary artery stenosis., Objectives: We compared the diagnostic accuracy of coronary CTA between men and women without known CAD with the use of invasive coronary angiography (ICA) as the reference standard., Methods: We prospectively evaluated 230 subjects with chest pain at 16 sites who were clinically referred for ICA. ICAs were evaluated for coronary stenosis according to quantitative coronary angiography., Results: Subjects (136 men and 94 women; mean ± age, 57 ± 10 years) underwent both CTA and ICA. For a patient-based model for stenosis >50%, sensitivity, specificity, positive and negative predictive values in men versus women were 96%, 78%, 69%, 100% and 90%, 88%, 47%, 99%, respectively. Subgroup analyses were performed for age and lifestyle risk factors. For stenosis > 50% in patients < 55 years, specificity in men versus women was 88% versus 95%, whereas for patients > 55 years, specificity in men versus women was 68% versus 82% (P < 0.05)., Conclusions: Coronary CTA found comparable diagnostic accuracy for women in comparison with men for the detection of obstructive coronary stenosis at both thresholds of 50% and 70%., (Copyright © 2012 Society of Cardiovascular Computed Tomography. Published by Elsevier Inc. All rights reserved.)

Introduction: During a 4-week rheumatology elective at our institution, opportunities for internal medicine residents to perform arthrocentesis were limited, particularly for sites other than the knee. Consequently, residents were inadequately prepared and had less self-confidence to perform such procedures. To overcome these educational deficiencies, an arthrocentesis workshop was developed. We report our quality improvement data that was collected during the first year of workshop implementation., Methods: We devised a structured half-day arthrocentesis workshop for rheumatology fellows as well as rotating internal medicine residents. This program consisted of a one hour lecture immediately followed by a hands-on workshop that used mannequin models for 5 anatomic sites. A self-assessment questionnaire and medical knowledge test were administered before and after each session. The accuracy of the self-assessment questionnaire was analyzed by comparing responses to an external objective measure of knowledge in the same content area. Finally, an optional postworkshop survey addressed resident satisfaction., Results: Thirty-eight trainees participated in the workshop between July 2006 and June 2007. There were statistically significant improvements in self-confidence in 9 content areas (P < 0.0002), cognitive testing (P < 0.0001) and in self-assurance of procedural skill at all anatomic sites. A high degree of discordance was found between the perceived level of competence and the actual performance on the medical knowledge test during the preworkshop analysis. In contrast, the postworkshop analysis displayed modestly higher concordance. All residents completing a postworkshop survey believed that it was a useful exercise, and 96% stated that they would change their practice habits., Conclusion: The arthrocentesis workshop provided a solid foundation from which trainees can learn key concepts of joint injection, increase their self-confidence and refine their motor skills. The accuracy of resident self-reported confidence is poor and should therefore be used only to complement other means of competency assessment and medical knowledge acquisition.

Carbon nanotubes and graphene are candidate materials for nanoscale electronic devices. Both materials show weak acoustic phonon scattering and long mean free paths for low-energy charge carriers. However, high-energy carriers couple strongly to optical phonons, which leads to current saturation and the generation of hot phonons. A non-equilibrium phonon distribution has been invoked to explain the negative differential conductance observed in suspended metallic nanotubes, while Raman studies have shown the electrical generation of hot G-phonons in metallic nanotubes. Here, we present a complete picture of the phonon distribution in a functioning nanotube transistor including the G and the radial breathing modes, the Raman-inactive zone boundary K mode and the intermediate-frequency mode populated by anharmonic decay. The effective temperatures of the high- and intermediate-frequency phonons are considerably higher than those of acoustic phonons, indicating a phonon-decay bottleneck. Most importantly, inclusion of scattering by substrate polar phonons is needed to fully account for the observed electronic transport behaviour.

Objective: The purpose of this study was to define the optimal Doppler criteria for the diagnosis of inferior mesenteric artery (IMA) stenosis in patients with suspected chronic mesenteric ischemia (CMI)., Methods: A retrospective review of 205 dedicated color and pulsed Doppler sonographic studies of mesenteric arteries was performed in 205 patients. All studies were performed in patients with suspected CMI. Correlative angiography was available in 50 patients., Results: The IMA was visualized in 176 of 205 Doppler sonographic examinations (86%) and in 92% of the correlative studies. The visualization rate for the detection of a patent IMA by Doppler sonography in this series was 90%. The ranges of the peak systolic velocity (PSV), end-diastolic velocity (EDV), and mesenteric-aortic velocity ratio (MAR) in the nonstenotic IMA were 70 to 200 cm/s, 0 to 33 cm/s, and 0.7 to 3.7, respectively. The ranges of the PSV, EDV, and MAR in IMA stenosis were 200 to 485 cm/s, 0 to 177 cm/s, and 0.69 to 8.1. The threshold values for severe IMA stenosis by logistic regression analysis (n = 42) were as follows: PSV, greater than 200 cm/s; EDV, greater than 25 cm/s; and MAR, greater than 2.5, with sensitivities of 90%, 40%, and 80%; specificities of 97%, 91%, and 88%; positive predictive values (PPVs) of 90%, 57%, and 67%; negative predictive values (NPVs) of 97%, 83%, and 93%; and accuracy of 95%, 79%, and 86%, respectively., Conclusions: We found that a PSV of greater than 200 cm/s was the best criterion for the diagnosis of IMA stenosis. The sensitivity, specificity, PPV, NPV, and accuracy for the PSV were 90%, 97%, 90%, 97%, and 95%, respectively.

We measure the temperature distribution in a biased single-layer graphene transistor using Raman scattering microscopy of the 2D-phonon band. Peak operating temperatures of 1050 K are reached in the middle of the graphene sheet at 210 kW cm(-2) of dissipated electric power. The metallic contacts act as heat sinks, but not in a dominant fashion. To explain the observed temperature profile and heating rate, we have to include heat flow from the graphene to the gate oxide underneath, especially at elevated temperatures, where the graphene thermal conductivity is lowered due to umklapp scattering. Velocity saturation due to phonons with about 50-60 meV energy is inferred from the measured charge density via shifts in the Raman G-phonon band, suggesting that remote scattering (through field coupling) by substrate polar surface phonons increases the energy transfer to the substrate and at the same time limits the high-bias electronic conduction of graphene.

We investigate poly(ethylene imine) and diazonium salts as stable, complementary dopants on graphene. Transport in graphene devices doped with these molecules exhibits asymmetry in electron and hole conductance. The conductance of one carrier is preserved, while the conductance of the other carrier decreases. Simulations based on nonequilibrium Green's function formalism suggest that the origin of this asymmetry is imbalanced carrier injection from the graphene electrodes caused by misalignment of the electrode and channel neutrality points.

Background: The specific detection of a minor population of mutant DNA molecules requires methods of high specificity and sensitivity. While the single-allele base extension reaction (SABER) was shown to be useful for the detection of certain beta-thalassemia mutations, we encountered problems with false positivity during development of SABER for the noninvasive prenatal diagnosis of the hemoglobin E (HbE) disease. Systematic optimization resulted in an alternative protocol, the allele-specific base extension reaction (ASBER)., Methods: An artificial model was established by mixing genomic DNA of HbE carriers and normal individuals. Effects of terminator concentration and annealing temperature on the nonspecificity of SABER were then studied. The use of a single relevant terminator and the other 3 types of dideoxynucleotide as competing terminators were also compared in the development of the ASBER protocol. Thirteen cases of HbE-susceptible pregnancies were tested to compare the SABER and the ASBER protocols., Results: Decreasing the single relevant terminator concentration and increasing the annealing temperature in SABER were found to improve specificity. The use of the other 3 types of dideoxynucleotide as competing terminators was shown to offer better detection sensitivity than a single terminator in ASBER. Genotyping results were all correctly determined by ASBER, except one false-negative detection (sensitivity: 80%, specificity: 100%)., Conclusions: An alternative mass spectrometry-based protocol for noninvasive prenatal diagnosis, ASBER, has been successfully developed to allow the detection of a minor DNA population with a point mutation.

We show that the Raman frequency associated with the vibrational mode at approximately 1,580 cm(-1) (the G mode) in both metallic and semiconducting carbon nanotubes shifts in response to changes in the charge density induced by an external gate field. These changes in the Raman spectra provide us with a powerful tool for probing local doping in carbon nanotubes in electronic device structures, or charge carrier densities induced by environmental interactions, on a length scale determined by the light diffraction limit. The G mode shifts to higher frequency and narrows in linewidth in metallic carbon nanotubes at large fields. This behaviour is analogous to that observed recently in graphene. In semiconducting carbon nanotubes, on the other hand, induced changes in the charge density only shift the phonon frequency, but do not affect its linewidth. These spectral changes are quantitatively explained by a model that involves the renormalization of the carbon nanotube phonon energy by the electron-phonon interaction as the carrier density in the carbon nanotube is changed.

The photovoltage produced by local illumination at the Schottky contacts of carbon nanotube field-effect transistors varies substantially with gate voltage. This is particularly pronounced in ambipolar nanotube transistors where the photovoltage switches sign as the device changes from p-type to n-type. The detailed transition through the insulating state can be recorded by mapping the open-circuit photovoltage as a function of excitation position. These photovoltage images show that the band-bending length can grow to many microns when the device is depleted. In our palladium-contacted devices, the Schottky barrier for electrons is much higher than that for holes, explaining the higher p-type current in the transistor. The depletion width is 1.5 mum near the n-type threshold and smaller than our resolution of 400 nm near the p-type threshold. Internal photoemission from the metal contact to the carbon nanotube and thermally assisted tunneling through the Schottky barrier are observed in addition to the photocurrent that is generated inside the carbon nanotube.

Since the discovery of circulating nucleic acids in plasma in 1948, many diagnostic applications have emerged. For example, diagnostic and prognostic potentials of circulating tumour-derived DNA have been demonstrated for many types of cancer. The parallel development of fetal-derived DNA detection in maternal plasma has opened up the possibility of non-invasive prenatal diagnosis and monitoring of many pregnancy-associated disorders. In this regard, non-invasive fetal rhesus blood group genotyping has already been translated to clinical practice. Other applications of circulating DNA in traumatology and transplant monitoring have also been reported. The more recent discoveries of circulating tumour-derived RNA and fetal-derived RNA have proven to be equally important as their DNA counterparts. Successful prenatal diagnosis of Down's syndrome by fetal RNA analysis has recently been reported. However, the definite origin and release mechanisms of circulating nucleic acids have remained incompletely understood, with cell death being suggested to be associated with such nucleic acid release. Pre-analytical standardisation will become increasingly relevant when comparing data from different laboratories. In conclusion, studies of circulating nucleic acids have promised exciting developments in molecular diagnostics in the years to come.

Carbon nanotube field-effect transistors (CNTFETs) produce band gap derived infrared emission under both ambipolar and unipolar transport conditions. We demonstrate here that heterogeneities/defects in the local environment of a CNTFET perturb the local potentials and, as a result, the characteristic bias dependent motion of the ambipolar light emission. Such defects can also introduce localized infrared emission due to impact excitation by carriers accelerated by a voltage drop at the defect. The correlation of the change in the motion of the ambipolarlight emission and of the stationary electroluminescence with the electrical characteristics of the CNTFETs shows that stationaryelectroluminescence can identify "environmental defects" in carbon nanotubes and help evaluate their influence on electrical transport and device operation. A number of different defects are studied involving local dielectric environment changes (partially polymer-covered nanotubes), nanotube-nanotube contacts in looped nanotubes, and nanotube segments close to the electronic contacts. Random defects due to local charging are also observed.

Current antagonists for the parathyroid hormone (PTH)/PTH-related protein (PTHrP) receptor (PTHR) are N-terminally truncated or N-terminally modified analogs of PTH(1-34) or PTHrP(1-34) and are thought to bind predominantly to the N-terminal extracellular (N) domain of the receptor. We hypothesized that ligands that bind only to PTHR region comprised of the extracellular loops and seven transmembrane helices (the juxtamembrane or J domain) could also antagonize the PTHR. To test this, we started with the J domain-selective agonists [Gln(10),Ala(12),Har(11),Trp(14),Arg(19) (M)]PTH(1-21), [M]PTH(1-15), and [M]PTH(1-14), and introduced substitutions at positions 1-3 that were predicted to dissociate PTHR binding and cAMP signaling activities. Strong dissociation was observed with the tri-residue sequence diethylglycine (Deg)(1)-para-benzoyl-l-phenylalanine (Bpa)(2)-Deg(3). In HKRK-B7 cells, which express the cloned human PTHR, [Deg(1,3),Bpa(2),M]PTH(1-21), [Deg(1,3),Bpa(2),M]PTH(1-15), and [Deg(1,3),Bpa(2),M]PTH(1-14) fully inhibited (IC(50)s = 100-700 nm) the binding of (125)I-[alpha-aminoisobutyric acid(1,3),M]PTH(1-15) and were severely defective for stimulating cAMP accumulation. In ROS 17/2.8 cells, which express the native rat PTHR, [Deg(1,3),Bpa(2),M]PTH(1-21) and [Deg(1,3),Bpa(2),M]PTH(1-15) antagonized the cAMP-agonist action of PTH(1-34), as did PTHrP(5-36) (IC(50)s = 0.7 microm, 2.6 microm, and 36 nm, respectively). In COS-7 cells expressing PTHR-delNt, which lacks the N domain of the receptor, [Deg(1,3),Bpa(2), M]PTH(1-21) and [Deg(1,3),Bpa(2),M]PTH(1-15) inhibited the agonist actions of [alpha-aminoisobutyric acid(1,3)]PTH(1-34) and [M]PTH(1-14) (IC(50)s approximately 1 microm), whereas PTHrP(5-36) failed to inhibit. [Deg(1,3),Bpa(2),M]PTH(1-14) inhibited the constitutive cAMP-signaling activity of PTHR-tether-PTH(1-9), in which the PTH(1-9) sequence is covalently linked to the PTHR J domain, as well as that of PTHR(cam)H223R. Thus, the J-domain-selective N-terminal PTH fragment analogs can function as antagonists as well as inverse agonists for the PTHR. The new ligands described should be useful for further studies of the ligand binding and activation mechanisms that operate in the critical PTHR J domain.

We spatially resolve the infrared light emission from ambipolar carbon-nanotube field-effect transistors with long-channel lengths. Electrons and holes are injected from opposite contacts into a single nanotube molecule. The ambipolar domain, where electron and hole currents overlap, forms a microscopic light emitter within the carbon nanotube. We can control its location by varying gate and drain voltages. At high electric fields, additional stationary spots appear due to defect-assisted Zener tunneling or impact ionization. The laterally resolved measurement provides valuable insight into the transistor behavior, complementary to electronic device characteristics.

Polarized infrared optical emission was observed from a carbon nanotube ambipolar field-effect transistor (FET). An effective forward-biased p-n junction, without chemical dopants, was created in the nanotube by appropriately biasing the nanotube device. Electrical measurements show that the observed optical emission originates from radiative recombination of electrons and holes that are simultaneously injected into the undoped nanotube. These observations are consistent with a nanotube FET model in which thin Schottky barriers form at the source and drain contacts. This arrangement is a novel optical recombination radiation source in which the electrons and holes are injected into a nearly field-free region. Sucha source may form the basis for ultrasmall integrated photonic devices.

Unlabelled: Single aortic clamp (SAC) versus partial occluding clamp (POC) technique for the construction of proximal anastomosis has been suggested to provide better cerebral protection during coronary artery bypass grafting (CABG). The aim of this study was to assess this hypothesis in a prospective randomized trial., Methods: Two hundred sixty-eight consecutive patients underwent CABG at a single institution. All patients were randomized to either SAC (Group S) or POC (Group P) for the construction of the proximal anastomosis. Myocardial protection consisted of multidose antegrade cold blood cardioplegia with topical cooling. The operations were performed using standard cardiopulmonary bypass support and moderate systemic hypothermia (29 to 32 degrees C). The incidences of neurological events, perioperative myocardial infarction (MI), and mortality were prospectively evaluated., Results: The two groups were similar in mean age, gender, urgency of operation, and number of bypasses. Group S patients had a significantly longer cross-clamp (61 +/- 21 minutes [S] vs 44 +/- 13.8 minutes [P], p < 0.05) and bypass times (85 +/- 25 minutes [S] vs 74 +/- 19.7 minutes [P], p < 0.05). There were no differences in the number of perioperative MIs (Group S = 3 [2.3%]; Group P = 2 [1.5%], p = 0.50) or mortality (Group S = 2 [1.5%]; Group P = 3 [2.2%], p = 0.50). Two patients randomized to POC were switched to SAC intraoperatively because of severe calcification of the ascending aorta. In Group P, there were two strokes (1.5%) and two (1.5%) postoperative confusions versus none in Group S (relative risk = 2.0, p < 0.05, respectively)., Conclusion: The SAC technique improved cerebral protection without any adverse effect on myocardial protection and postoperative outcome in patients undergoing CABG.

Recent data suggest that the binding of parathyroid hormone (PTH)-(1-34) to the PTH-1 receptor (P1R) involves a high-affinity interaction between the C-terminal (15-34) domain of the ligand and the amino-terminal extracellular (N) domain of the receptor and a low-affinity interaction between the N-terminal (1-14) portion of PTH and the juxtamembrane (J) region of the receptor, with the latter interaction giving rise to signal transduction. We investigated whether residues C-terminal of position 14 in PTH(1-34) contribute to the J component of the interaction mechanism by comparing the capacity of PTH analogues N-terminally modified to improve J domain affinity and C-terminally truncated at position 14, 20, or 34 to stimulate cAMP formation in COS-7 cells transiently transfected with P1R-delNt, a P1R construct that lacks most of the N domain. In these cells, the potency of [M]PTH(1-34) (M = Ala(1,3,12),Gln(10),Har(11),Trp(14),Arg(19)) was 120-fold greater than that of [M]PTH(1-14) (EC(50)s = 3.0 +/- 0.8 and 360 +/- 90 nM, respectively) but was equal to that of [M]PTH(1-20) (EC(50) = 2.3 +/- 0.3 nM). Reverting the Arg(19) substitution of [M]PTH(1-20) to the native Glu reduced cAMP signaling potency on P1R-delNt by 12-fold (EC(50) of [M]PTH(1-20)-Glu(19) = 27 +/- 4 nM), and it decreased the analog's capacity to inhibit the binding of the J domain-selective radioligand, (125)I-[Aib(1,3),Nle(8),M,Tyr(21)]ratPTH(1-21), to the full-length P1R stably expressed in LLC-PK1 cells by 40-fold. The Glu(19) --> Arg modification, however, did not affect the capacity of PTH(15-31) to inhibit the binding of the N domain-selective radioligand (125)I-bPTH(3-34) to the full-length receptor. The overall data suggest that residues (15-20) of PTH, and particularly residue 19, contribute to the capacity of the N-terminal portion of the ligand to interact with the juxtamembrane region of the receptor. The NMR data presented in the accompanying manuscript suggests that this role could involve intramolecular effects on secondary structure in the N-terminal portion of the ligand.

We report a rare case of severe hemolytic anemia following repair of a congenital heart defect without the use of prosthetic material. A review of the literature, diagnosis, and management are described. Although this is an unusual complication following congenital heart surgery, a high index of suspicion must be maintained and a possible mechanical cause should be sought and corrected.

The concept of myocardial sinusoids has been described in the literature during the past 60 years. They have been the basis of several revascularization procedures, such as the "Vineberg" procedure and more recently transmural laser revascularization. This article will review the historical evolution as well as the validity of the concept of "myocardial sinusoid."

There has been an upsurge of research on myocardial preconditioning because of its potential clinical application in areas such as cardiology, cardiac surgery, and transplantation. From a broad biological standpoint, a conceptual framework may help in both promoting understanding and suggesting future research paths. The living organism's tendency toward developing evolutionarily advantageous strategies has led to a fight or flight response, for which the authors consider preconditioning a component. Added to preconditioning, the production of stress proteins and altered myocardial states (especially that of "hibernating myocardium") can be seen as a series of biological strategies developed and maintained during evolution. An increased understanding of the mechanisms involved in the body's self-defense strategies should lead to better approaches, those in which we can help the cells, including those comprising the myocardium, to preserve themselves.

A monoclonal antibody, F3H7, was generated by immunizing mice with a synthetic peptide corresponding to residues 63-84 of the B*2705 allele of the HLA-B27 antigens. The reactive epitope and the contact residues on the peptide were localized by ELISA using a large panel of overlapping peptides as well as peptides with substituted amino acids. Residues corresponding to R75, D77 and L78 on the HLA-B27 protein appeared to be critical. The clarity of these results indicate that this is a potentially useful approach to the study of HLA class I epitopes.

With reference sera and homozygous typing cells (HTCs) of 3rd Asia-Oceania Histocompatibility Workshop Conference, 56 healthy unrelated subjects in Shanghai were typed for HLA-A, B, C, DR, DQ, and Dw. This paper presents the results of HLA-Dw typing, its relationship to serological class II antigens, and the distribution of Dw in the population. The polymorphism patterns of Chinese Dw specificities were quite different from those in Caucasoids and Japanese. The predominant Dw phenotypes detected in Shanghai Chinese were Dw 2, Dw 3, DKT 2, Dw 7 c, (Dw7 + Dw 17) and Dw 23 (DB 5). And significant correlations were observed between Dw 1 and DR 1, Dw 2 and DR 2, Dw 3 and DR 3, Dw 7 c and DR 7, DB 7 and DRw 8, as well as Dw 23 and DR 9. SMY 129, a novel Dw specificity defined by local HTCs and co-studied by the laboratories joined for Dw typing in 3rd AOHWC showed its correlation with DR 5. Nevertheless, more than fifty percent of Dw specificities could not be assigned in the four correspondent designated serological antigens, DR 2, DR 5, DRw 8 and DR 9, respectively, which, together with other blank Dw specificities, gave a total blank Dw gene frequency as high as 43.2% in the population. It was suggested by further analysis that novel Dw specificities might be identified more effectively if efforts would be concentrated on DR 5 and DR 9, two antigen families which, in some way, might represent the characteristics of HLA system in Chinese. Besides, certain HTC-defined antigens, e.g. Dw 3 and the DR 4-related Dw specificities, have been revealed to be in linkage disequilibrium with other DR antigens in addition with the correspondent designated ones, resulting in some unique haplotype combinations in Shanghai Chinese. It seems to us that the particular patterns of polymorphisms of serum- and cell-defined HLA class II antigens would be helpful to elucidate the mechanisms by which certain diseases are in association with HLA in Chinese in a different manner as compared with that in Caucasoids.

In vitro and in vivo complex formations of polymyxin B and lipopolysaccharides (LPS) from resistant and sensitive cells of Serratia marcescens were studied by polyacrylamide gel electrophoresis in sodium dodecyl sulfate and electron microscopy. In vitro treatment of LPS from resistant cells with polymyxin B gave two populations of spherical complexes of differnt molecular weights as determined electrophoretically. Similar treatment of LPS from sensitive cells resulted in dissociation of the LPS-protein and subsequent complexing with the LPS moiety into stable spheres. In vivo treatment of resistant cells with polymyxin B resulted in LPS-polymyxin B complexes which were comparatively smaller and existed in two morphological forms; spheres and linear ribbons. LPS from the sensitive cells were degraded extensively into small rods and an amorphous mass by the in vivo polymyxin B treatment. In both systems, the electrophoretic results consistently matched the electron microscopic evidences for complex formation of LPS with polymyxin B. It is suggested that the disruptive effects of polymyxin B on LPS in the outer membrane of S. marcescens may be the explanation for the change in permeability barrier in the resistant cells and disorganization of the outer membrane and subsequent death in the sensitive cells. Furthermore, the ability of the LPS to complex with the polymyxin B molecules in resistant cells may be the basis of their resistance to the antibiotic.