Molecular and physical characterization of the complex pericentromeric heterochromatin of the vole species Microtus thomasi.
A new repeated DNA from Microtus thomasi, Mth-Alu2.2, was cloned and characterized and is presented here for the first time. Digestion of genomic DNA from M. thomasi with AluI restriction enzyme revealed a 2.2-kb repetitive DNA sequence with a high AT content (69%). This sequence consists of a tandemly repeated nonanucleotide of the consensus sequence CACAATGTA, which constitutes approximately 93-95% of the total unit length. The location of the Mth-Alu2.2 sequence in the karyotype was determined by FISH, demonstrating strong hybridization signals in the pericentromeric regions of all chromosomes and in the heterochromatin blocks of several X chromosome variants. In addition, the distribution of the 4 pericentromeric repeat sequences Msat-160, Mth-Alu900, Mth-Alu2.2, and interstitial telomeric repeats was analyzed by in situ hybridization in M. thomasi, in order to shed light on the complex composition of the chromosomal pericentromeric regions in this species. The order and organization of these sequences in the pericentromeric regions are conserved, with slight variations in both the degree of overlapping and the amount of each repeated DNA in the chromosomes. Specifically, Mth-Alu2.2 is localized in the terminal regions of the chromosomes, with Msat-160 occupying the immediately inner region, partially intermixed with Mth-Alu2.2. The sequence Mth-Alu900 is found in internal positions below Msat-160, and the interstitial telomeric repeats are located close to the long-arm euchromatin of the chromosomes.


Nonlinear pulse propagation in a quantum dot laser.
We investigate the nonlinear propagation of an ultra-short, 150 fs, optical pulse along the waveguide of a quantum dot (QD) laser operating above threshold. We demonstrate that among the various nonlinear processes experienced by the propagating pulse, four-wave mixing (FWM) between the pulse and the two oscillating counter-propagating cw fields of the laser is the dominant one. FWM has two important consequences. One is the creation of a spectral hole located in the vicinity of the cw oscillating frequency. The width of the spectral hole is determined by an effective carrier and gain relaxation time. The second is a modification of the shape of the trailing edge of the pulse. The wave mixing involves first and second order processes which result in a complicated interaction among several fields inside the cavity, some of which are cw while the others are time varying, all propagating in both directions. The nonlinear pulse propagation is analyzed using two complementary theoretical approaches. One is a semi-analytical model which considers only the wave mixing interaction between six field components, three of which propagate in each direction (two cw fields and four time-varying signals). This model predicts the deformation of the tail of the output signal by a secondary idler wave, produced in a cascaded FWM process, which co-propagates with the original injected pulse. The second approach is a finite-difference time-domain simulation, which considers also additional nonlinear effects, such as gain saturation and self-phase modulation. The theoretical results are confirmed by a series of experiments in which the time dependent amplitude and phase of the pulse after propagation are measured using the cross-frequency-resolved optical gating technique.


Electrochemical synthesis of luminescent MoS2 quantum dots.
Size-controlled synthesis of luminescent quantum dots of MoS2 (≤2 layers) with narrow size distribution, ranging from 2.5 to 6 nm, from their bulk material using a unique electrochemical etching of bulk MoS2 is demonstrated. Excitation-dependent photoluminescence emission is observed in the MoS2 QDs. "As-synthesized" MoS2 QDs also exhibit excellent electrocatalytic activity towards hydrogen evolution reactions.


Clinical role of atrial arrhythmias in patients with arrhythmogenic right ventricular dysplasia.
The clinical role of atrial fibrillation/atrial flutter (AF-AFl) and variables predicting these arrhythmias are not well defined in patients with arrhythmogenic right ventricular dysplasia (ARVD). We hypothesized that transthoracic echocardiography (TTE) and 12-lead electrocardiography (ECG) would be helpful in predicting AF-AFl in these patients. METHODS AND RESULTS: ECGs and TTEs of 90 patients diagnosed with definite or borderline ARVD (2010 Task Force Criteria) were analyzed. Data were compared in (1) patients with AF-AFl and (2) all other patients. A total of 18 (20%) patients experienced AF-AFl during a median follow-up of 5.8 years (interquartile range 2.0-10.4). Kaplan-Meier analysis revealed reduced times to AF-AFl among patients with echocardiographic RV fractional area change <27% (P<0.001), left atrial diameter ≥24.4 mm/m(2)(parasternal long-axis, P=0.001), and right atrial short-axis diameter ≥22.1 mm/m(2)(apical 4-chamber view, P=0.05). From all ECG variables, P mitrale conferred the highest hazard ratio (3.37, 95% confidence interval 0.92-12.36, P=0.067). Five patients with AF-AFl experienced inappropriate implantable cardioverter-defibrillator (ICD) shocks compared with 4 without AF-AFl (36% vs. 9%, P=0.03). AF-AFl was more prevalent in heart-transplant patients and those who died of cardiac causes (56% vs. 16%, P=0.014). AF-AFl is associated with inappropriate ICD shocks, heart transplantation, and cardiac death in patients with ARVD. Evidence of reduced RV function and atrial dilation helps to identify the ARVD patients at increased risk for AF-AFl.


Low prevalence of lipid lowering drug use in older men with established coronary heart disease.
To determine the prevalence and correlates of lipid lowering drug use among older British men with established coronary heart disease (CHD). Cross sectional survey within a cohort study (British regional heart study) carried out at 20 years of follow up in 1998-2000. General practices in 24 British towns. 3689 men aged 60-75 years (response rate 76%). Diagnoses of myocardial infarction and angina based on detailed review of general practice records. Lipid lowering drug use and blood cholesterol concentrations ascertained at 20 year follow up examination. Among 286 men with definite myocardial infarction, 102 (36%) were taking a lipid lowering drug (93 (33%) a statin); among 360 men with definite angina without myocardial infarction, 84 (23%) were taking a lipid lowering drug (78 (21%) a statin). Most men with documented CHD who were not receiving a lipid lowering drug had a total cholesterol concentration of 5.0 mmol/l or more (87% of those with myocardial infarction, 82% with angina). Fewer than half of men with CHD receiving a statin had a total cholesterol concentration below 5.0 mmol/l (45% of those with myocardial infarction and 47% of those with angina). Only one third of the men taking a statin were receiving trial validated dosages. Among men with CHD, a history of revascularisation, more recent diagnosis, and younger age at diagnosis were associated with a higher probability of receiving lipid lowering drug treatment. Among patients with established CHD, the prevalence of lipid lowering drug use remains low and statin regimens suboptimal. Major improvements in secondary prevention are essential if the benefits of statins are to be realised.


Consequences of unlocking the cardiac myosin molecule in human myocarditis and cardiomyopathies.
Myocarditis, often initiated by viral infection, may progress to autoimmune inflammatory heart disease, dilated cardiomyopathy and heart failure. Although cardiac myosin is a dominant autoantigen in animal models of myocarditis and is released from the heart during viral myocarditis, the characterization, role and significance of anti-cardiac myosin autoantibodies is poorly defined. In our study, we define the human cardiac myosin epitopes in human myocarditis and cardiomyopathies and establish a mechanism to explain how anti-cardiac myosin autoantibodies may contribute to heart disease. We show that autoantibodies to cardiac myosin in sera from myocarditis and dilated cardiomyopathies in humans targeted primarily epitopes in the S2 hinge region of cardiac myosin. In addition, anti-cardiac myosin antibodies in sera or purified IgG from myocarditis and cardiomyopathy targeted the beta-adrenergic receptor and induced antibody-mediated cAMP-dependent protein kinase A (PKA) cell signaling activity in heart cells. Antibody-mediated PKA activity in sera was abrogated by absorption with anti-human IgG. Antibody-mediated cell signaling of PKA was blocked by antigen-specific inhibition by human cardiac myosin or the beta-adrenergic receptor but not the alpha adrenergic receptor or bovine serum albumin. Propranolol, a beta blocker and inhibitor of the beta-adrenergic receptor pathway also blocked the antibody-mediated signaling of the beta-adrenergic receptor and PKA. The data suggest that IgG antibody against human cardiac myosin reacts with the beta-adrenergic receptor and triggers PKA signaling in heart cells. In summary, we have identified a new class of crossreactive autoantibodies against human cardiac myosin and the beta-adrenergic receptor in the heart. In addition, we have defined disease specific peptide epitopes in the human cardiac myosin rod S2 region in human myocarditis and cardiomyopathy as well as a mechanistic role of autoantibody in the pathogenesis of disease.


Relaxation filtered hyperfine (REFINE) spectroscopy: a novel tool for studying overlapping biological electron paramagnetic resonance signals applied to mitochondrial complex I.
A simple strategy to separate overlapping electron paramagnetic resonance (EPR) signals in biological systems is presented. Pulsed EPR methods (inversion- and saturation-recovery) allow the determination of the T(1) spin-lattice relaxation times of paramagnetic centers. T(1) may vary by several orders of magnitude depending on the species under investigation. These variations can be employed to study selectively individual species from a spectrum that results from an overlap of two species using an inversion-recovery filtered (IRf) pulsed EPR technique. The feasibility of such an IRf field-swept technique is demonstrated on model compounds (alpha,gamma-bisphenylene-beta-phenylallyl-benzolate, BDPA, and 2,2,6,6-tetramethyl-piperidine-1-oxyl, TEMPO) and a simple strategy for the successful analysis of such mixtures is presented. Complex I is a multisubunit membrane protein of the respiratory chain containing several iron-sulfur (FeS) centers, which are observable with EPR spectroscopy. It is not possible to investigate the functionally important FeS cluster N2 separately because this EPR signal always overlaps with the other FeS signals. This cluster can be studied selectively using the IRf field-swept technique and its EPR spectrum is in excellent agreement with previous cw-EPR data from the literature. In addition, the possibility to separate the hyperfine spectra of two spectrally overlapping paramagnetic species is demonstrated by applying this relaxation filter together with hyperfine spectroscopy (REFINE). For the first time, the application of this filter to a three-pulse electron spin-echo envelope modulation (ESEEM) pulse sequence is demonstrated to selectively observe hyperfine spectra on a system containing two paramagnetic species. Finally, REFINE is used to assign the observed nitrogen modulation in complex I to an individual iron-sulfur cluster.


The Cytotoxicity and Synergistic Potential of Aspirin and Aspirin Analogues Towards Oesophageal and Colorectal Cancer.
Oesophageal cancer (OC) is a deadly cancer because of its aggressive nature with survival rates that have barely improved in decades. Epidemiologic studies have shown that low-dose daily intake of aspirin can decrease the incidence of OC. The toxicity of aspirin and aspirin derivatives to OC and a CRC cell line were investigated in the presence and absence of platins. The data in this study show the effects of a number of aspirin analogues and aspirin on OC cell lines that originally presented as squamous cell carcinoma (SSC) and adenocarcinoma (ADC). The aspirin analogues fumaryldiaspirin (PN517) and the benzoylsalicylates (PN524, PN528 and PN529), were observed to be more toxic against the OC cell lines than aspirin. Both quantitative and qualitative apoptosis experiments reveal that these compounds largely induce apoptosis, although some necrosis was evident with PN528 and PN529. Failure to recover following the treatment with these analogues emphasized that these drugs are largely cytotoxic in nature. The OE21 (SSC) and OE33 (ADC) cell lines were more sensitive to the aspirin analogues compared to the Flo-1 cell line (ADC). A non-cancerous oesophageal primary cells NOK2101, was used to determine the specificity of the aspirin analogues and cytotoxicity assays revealed that analogues PN528 and PN529 were selectively toxic to cancer cell lines, whereas PN508, PN517 and PN524 also induced cell death in NOK2101. In combination index testing synergistic interactions of the most promising compounds, including aspirin, with cisplatin, oxaliplatin and carboplatin against the OE33 cell line and the SW480 colorectal cancer (CRC) cell line were investigated. Compounds PN517 and PN524, and to a lesser extent PN528, synergised with cisplatin against OE33 cells. Cisplatin and oxaliplatin synergised with aspirin and PN517 when tested against the SW480 cell line. These findings indicate the potential and limitations of aspirin and aspirin analogues as chemotherapeutic agents against OC and CRC when combined with platins.


Faces in early visual environments are persistent not just frequent.
The regularities in very young infants' visual worlds likely have out-sized effects on the development of the visual system because they comprise the first-in experience that tunes, maintains, and specifies the neural substrate from low-level to higher-level representations and therefore constitute the starting point for all other visual learning. Recent evidence from studies using head cameras suggests that the frequency of faces available in early infant visual environments declines over the first year and a half of life. The primary question for the present paper concerns the temporal structure of face experiences: Is frequency the key exposure dimension distinguishing younger and older infants' face experiences, or is it the duration for which faces remain in view? Our corpus of head-camera images collected as infants went about their daily activities consisted of over a million individually coded frames sampled at 0.2 Hz from 232 h of infant-perspective scenes, recorded from 51 infants aged 1 month to 15 months. The major finding from this corpus is that very young infants (1-3 months) not only have more frequent face experiences but also more temporally persistent ones. The repetitions of the same very few face identities presenting up-close and frontal views are exaggerated in more persistent runs of the same face, and these persistent runs are more frequent for the youngest infants. The implications of early experiences consisting of extended repeated exposures of up-close frontal views for visual learning are discussed.


Simulation of realistic retinoscopic measurement.
Realistic simulation of ophthalmic measurements on normal and diseased eyes is presented. We use clinical data of ametropic and keratoconus patients to construct anatomically accurate three-dimensional eye models and simulate the measurement of a streak retinoscope with all the optical elements. The results show the clinical observations including the anomalous motion in high myopia and the scissors reflex in keratoconus. The demonstrated technique can be applied to other ophthalmic instruments and to other and more extensively abnormal eye conditions. It provides promising features for medical training and for evaluating and developing ocular instruments.


A germline oncogenic MITF mutation and tumor susceptibility.
MITF (Microphthalmia-associated transcription factor) is a lineage specific transcription factor that plays a critical role in melanocyte homeostasis and whose deregulation has been shown to contribute to melanoma disease. A germline mutation in MITF, impairing SUMOylation and predisposing to cutaneous malignant melanoma, was recently identified. Interestingly, an association of the MITF mutation with coexisting melanoma and renal cell carcinoma was also shown. Collectively, these data suggest that MITF has an important oncogenic function in tumorigenesis of multiple tissues/melanocytes and kidney cells.


The adhesiveness of native platelets and its prevention.
THE FOLLOWING METHODS WERE USED TO MEASURE THE ADHESION TO VARIOUS SURFACES OF PLATELETS IN WHOLE BLOOD OR PLASMA: 1, two measurements gave an estimate in vivo of platelet adhesion to cut human capillaries; 2, platelets adhering to damaged cells in vitro were counted directly; 3, a highly reproducible method for estimating platelet adhesion to glass was devised; 4, the manner in which platelets adhere to each other (aggregation) was also studied. Platelet adhesion to all these surfaces was found to be dependent upon calcium and independent of all clotting factors except that platelet aggregation is probably dependent upon thrombin. A number of drugs-mostly antimalarials, antihistaminics, and local anaesthetics-in suitable concentration inhibited adhesion. They probably form a fixed, orientated layer on glass and possibly on cells and make these surfaces unattractive to a platelet. They also stick reversibly to live cells (including platelets) altering their permeability, and they may make platelets less adhesive. Consequently the possibility of using antiadhesive drugs therapeutically to inhibit thrombus formation was considered.


Hexokinase 2 as a novel selective metabolic target for rheumatoid arthritis.
Recent studies indicate that glucose metabolism is altered in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS). Hexokinases (HKs) catalyse the first step in glucose metabolism, and HK2 constitutes the principal HK inducible isoform. We hypothesise that HK2 contributes to the synovial lining hypertrophy and plays a critical role in bone and cartilage damage. HK1 and HK2 expression were determined in RA and osteoarthritis (OA) synovial tissue by immunohistochemistry. RA FLS were transfected with either HK1 or HK2 siRNA, or infected with either adenovirus (ad)-GFP, ad-HK1 or ad-HK2. FLS migration and invasion were assessed. To study the role of HK2 in vivo, 108 particles of ad-HK2 or ad-GFP were injected into the knee of wild-type mice. K/BxN serum transfer arthritis was induced in HK2F/F mice harbouring Col1a1-Cre (HK2Col1), to delete HK2 in non-haematopoietic cells. HK2 is particular of RA histopathology (9/9 RA; 1/8 OA) and colocalises with FLS markers. Silencing HK2 in RA FLS resulted in a less invasive and migratory phenotype. Consistently, overexpression of HK2 resulted in an increased ability to migrate and invade. It also increased extracellular lactate production. Intra-articular injection of ad-HK2 in normal knees dramatically increased synovial lining thickness, FLS activation and proliferation. HK2 was highly expressed in the synovial lining after K/BxN serum transfer arthritis. HK2Col1 mice significantly showed decreased arthritis severity, bone and cartilage damage. HK2 is specifically expressed in RA synovial lining and regulates FLS aggressive functions. HK2 might be an attractive selective metabolic target safer than global glycolysis for RA treatment.


Threats to graduate medical education funding and the need for a rational approach: a statement from the alliance for academic internal medicine.
With the current focus on reducing the federal budget deficit, funding for graduate medical education (GME) has come under scrutiny, particularly those monies labeled as indirect medical education payments; these are intended to cover ill-defined costs inherent to training programs, such as increased lengths of stay, additional testing, and patients with more complex conditions. Although there are cogent arguments that indirect medical education expenses have decreased over the past 25 years, there is also a reasonable expectation that direct medical education expenses, such as those related to resident salaries, faculty involvement, administration of programs, and overhead costs, have increased. Our current system of GME financing is complex and cumbersome and relies almost exclusively on government support. This article examines the adequacy of current funding, focusing on the economics of the entire system rather than individual hospitals, states, or regions. It also recommends reexamining GME financing and considering options that ensure appropriate levels of government support and participation of other health care insurers to adequately fund GME.


Continuous-wave ultraviolet generation at 320 nm by intracavity frequency doubling of red-emitting Praseodymium lasers.
We describe a new approach for the generation of coherent ultraviolet radiation. Continuous-wave ultraviolet light at 320 nm has been obtained by intracavity frequency doubling of red-emitting Praseodymium lasers. Lasing at the 640-nm fundamental wavelength in Pr:LiYF(4) and Pr:BaY(2)F(8) was realized by employing an optically pumped semiconductor laser at 480 nm as pump source.Using LiB(3)O(5) as nonlinear medium, ~19 mW of ultraviolet radiation with ~9% optical efficiency with respect to absorbed power was reached for both laser crystals; the visible-to-ultraviolet conversion efficiency was 26% and 35% for Pr:LiYF(4) and Pr:BaY(2)F(8), respectively.


Biochanin A attenuates myocardial ischemia/reperfusion injury through the TLR4/NF-κB/NLRP3 signaling pathway.
Myocardial ischemia/reperfusion (Ml/R) injury is a leading cause of damage in cardiac tissues, with high rates of mortality and disability. Biochanin A (BCA) is a main constituent of Trifolium pratense L. This study was intended to explore the effect of BCA on Ml/R injury and explore the potential mechanism. In vivo MI/R injury was established by transient coronary ligation in Sprague-Dawley rats. Triphenyltetrazolium chloride staining (TTC) was used to measure myocardial infarct size. ELISA assay was employed to evaluate the levels of myocardial enzyme and inflammatory cytokines. Western blot assay was conducted to detect related protein levels in myocardial tissues. BCA significantly ameliorated myocardial infarction area, reduced the release of myocardial enzyme levels including aspartate transaminase (AST), creatine kinase (CK-MB) and lactic dehydrogenase (LDH). It also decreased the production of inflammatory cytokines (IL-1β, IL-18, IL-6 and TNF-α) in serum of Ml/R rats. Further mechanism studies demonstrated that BCA inhibited inflammatory reaction through blocking TLR4/NF-kB/NLRP3 signaling pathway. The present study is the first evidence demonstrating that BCA attenuated Ml/R injury through suppressing TLR4/NF-kB/NLRP3 signaling pathway-mediated anti-inflammation pathway.


The effect of microstructure of octacalcium phosphate on the bone regenerative property.
The present study was designed to investigate whether the microstructure of synthetic octacalcium phosphate (OCP) affects its intrinsic bone regenerative properties as a scaffold and its conversion process into hydroxyapatite (HA). Our previous studies indicated that an agregate of OCP crystals, consisting of randomly oriented plate-like crystals, are capable of enhancing both osteoblastic cell differentiation in vitro and bone regeneration. While the transformation of OCP into HA has been considered in relation to the stimulatory capacity of OCP in bone regeneration, little is known about the effect of the microstructure of OCP granules on these capabilities. Two types of OCP granules, with identical diameters (300-500 microm) but composed of crystals with distinct crystal dimensions (4.0 and 26.6 microm length), were prepared (hereafter referred to as fine OCP granules [F-OCP] and coarse OCP granules [C-OCP], respectively). The intergranule distances and the porosity, including the intergranule spaces, were 108.5 microm and 93.7% for F-OCP, and 67.5 microm and 95.7% for C-OCP, as estimated by mercury intrusion. The OCP granules were implanted in mouse critical-sized calvarial defects for up to 14 days. Histological examination demonstrated that osteoblastic cells aligned on the surface of F-OCP at day 7 and formed new bone around the granules up to day 14. On the other hand, cells around C-OCP were sparse at day 7, and resulted in only slight bone formation around the granules at day 14. X-ray diffraction showed that both OCP granules tended to be converted to an apatite structure with similar conversion velocity by the implantation. Adhesion of mouse bone marrow stromal ST-2 cells was markedly inhibited on C-OCP compared to F-OCP in vitro. These results suggested that the microstructure consisting of plate-like crystals of OCP controls cell adhesion on the crystal surfaces and their resultant bone regenerative properties as well as the physicochemical effect associated with the transitory nature of OCP previously reported.


Evidence for activation of circulating inactive renin by the human kidney.
1. In eight patients with essential hypertension (EHT) and six patients with renovascular hypertension (RVHT) peripheral venous enzymatically active and inactive renin values were followed after acute stimulation of renin release by the vasodilating agent diazoxide (300 mg intravenously). Active renin rose during the first hour after diazoxide and remained high during the following 15 h, but inactive renin fell during the first hour and rose thereafter. Peripheral venous active and inactive renin were not different from arterial values both before and after diazoxide. 2. Sixteen patients with EHT received propranolol, 80 mg, four times a day. Six of them had a first injection of diazoxide the day before propranolol was started and a second one after 10--14 days of propranolol treatment. Peripheral vein active renin was lowered by propranolol, but inactive renin was raised. Both the diazoxide-induced rapid rise of active renin and the fall of inactive renin observed in untreated patients were absent during treatment with propranolol. 3. In four patients with EHT and seven patients with RVHT renal vein sampling was performed before and 30 min after diazoxide. Increased release of active renin from kidneys that were not markedly contracted was associated with a fall of the renal vein to peripheral vein ratio of inactive renin to a value less than one. 4. It is concluded that under certain circumstances stimulated release of active renin is associated with removal of inactive renin from the circulation by the kidney. This may in fact be due to intrarenal transformation of circulating inactive renin into its active counterpart. The findings suggest that a beta-adrenoreceptor might be involved in this activation process.


Effects of Pegvisomant and Pasireotide LAR on Vertebral Fractures in Acromegaly Resistant to First-generation SRLs.
Osteopathy is an emerging complication of acromegaly. In somatostatin receptor ligands (SRL)-resistant patients, pegvisomant (PegV) and pasireotide LAR (Pasi) are used for acromegaly treatment, but their effect on skeletal health is still not defined. In a longitudinal retrospective international study, we evaluated incidence of radiological vertebral fractures (VFs) in 55 patients with acromegaly resistant to first-generation SRL. At study entry, prevalent VFs occurred in 23 patients (41.8%). Biochemical acromegaly control was reached in 66.7% of patients on PegV and in 66.7% of patients on Pasi. During the follow-up, incident VFs (iVFs) were detected in 16 patients (29.1%). Occurrence of iVFs was associated with prevalent VFs (P = .002), persistence of active acromegaly (P = .01) and higher value of insulin-like growth factor 1 (IGF-1) during follow-up (P = .03). Among patients with active disease at last visit, iVFs occurred less frequently in patients on treatment with Pasi (25%) compared to PegV (77.8% P = .04), independently of the IGF-1 values (P = .90). In patients who reached biochemical control, 22.7% on PegV and 12.5% on Pasi had iVFs (P = .40). Among both treatment groups, the presence of pre-existent VFs was the main determinant for iVFs. Our data show for the first time that patients with biochemically active disease treated with Pasi had lower risk of iVFs versus those treated with PegV. It also confirms that the presence of pre-existent VFs was the main determinant for iVFs. Additional studies on larger populations and with longer follow-up are needed to confirm our data and disclose the mechanisms underlying our findings.


Cholecystokinin and leptin: their influence upon the eating behaviour and nutrient intake of dialysis patients.
We have used serial visual analogue scores to demonstrate disturbances of the appetite profile in dialysis patients. This is potentially important as dialysis patients are prone to malnutrition yet have a lower nutrient intake than controls. Appetite disturbance may be influenced by accumulation of appetite inhibitors such as leptin and cholecystokinin (CCK) in dialysis patients. Fasting blood samples were drawn from 43 controls, 50 haemodialysis (HD) and 39 peritoneal dialysis (PD) patients to measure leptin and CCK. Hunger and fullness scores were derived from profiles compiled using hourly visual analogue scores. Nutrient intake was derived from 3 day dietary records. Fasting CCK was elevated for PD (6.73 +/- 4.42 ng/l vs control 4.99 +/- 2.23 ng/l, P < 0.05; vs HD 4.43 +/- 2.15 ng/l, P < 0.01). Fasting CCK correlated with the variability of the hunger (r = 0.426, P = 0.01) and fullness (r = 0.52, P = 0.002) scores for PD. There was a notable relationship with the increase in fullness after lunch for PD (r = 0.455, P = 0.006). When well nourished PD patients were compared with their malnourished counterparts, CCK was higher in the malnourished group (P = 0.004). Leptin levels were higher for the dialysis patients than controls (HD and PD, P < 0.001) with pronounced hyperleptinaemia evident in some PD patients. Control leptin levels demonstrated correlation with fullness scores (e.g. peak fullness, r = 0.45, P = 0.007) but the dialysis patients did not. PD nutrient intake (energy and protein intake, r = -0.56, P < 0.0001) demonstrated significant negative correlation with leptin. Increased CCK levels appear to influence fullness and hunger perception in PD patients and thus may contribute to malnutrition. Leptin does not appear to affect perceived appetite in dialysis patients but it may influence nutrient intake in PD patients via central feeding centres.


Predictable dynamic program of timing of DNA replication in human cells.
The organization of mammalian DNA replication is poorly understood. We have produced high-resolution dynamic maps of the timing of replication in human erythroid, mesenchymal, and embryonic stem (ES) cells using TimEX, a method that relies on gaussian convolution of massive, highly redundant determinations of DNA copy-number variations during S phase to produce replication timing profiles. We first obtained timing maps of 3% of the genome using high-density oligonucleotide tiling arrays and then extended the TimEX method genome-wide using massively parallel sequencing. We show that in untransformed human cells, timing of replication is highly regulated and highly synchronous, and that many genomic segments are replicated in temporal transition regions devoid of initiation, where replication forks progress unidirectionally from origins that can be hundreds of kilobases away. Absence of initiation in one transition region is shown at the molecular level by single molecule analysis of replicated DNA (SMARD). Comparison of ES and erythroid cells replication patterns revealed that these cells replicate about 20% of their genome in different quarters of S phase. Importantly, we detected a strong inverse relationship between timing of replication and distance to the closest expressed gene. This relationship can be used to predict tissue-specific timing of replication profiles from expression data and genomic annotations. We also provide evidence that early origins of replication are preferentially located near highly expressed genes, that mid-firing origins are located near moderately expressed genes, and that late-firing origins are located far from genes.


In Vivo Performance of Innovative Polyelectrolyte Matrices for Hot Melt Extrusion of Amorphous Drug Systems.
Hot melt extrusion of amorphous systems has become a pivotal technology to cope with challenges of poorly water-soluble drugs. Previous research showed that small molecular additives with targeted molecular interactions enabled introduction of a polyelectrolyte matrix into hot melt extrusion that would otherwise not be possible to process due to the unfavorable properties upon heating of the pure polymer. Carboxymethyl cellulose sodium (NaCMC) with lysine or alternatively meglumine led to modified polymeric matrices that showed adequate processability by hot melt extrusion and yielded stable amorphous formulations. The investigated formulations, including fenofibrate as a model drug, were characterized by attenuated total reflectance Fourier transform infrared spectroscopy, differential scanning calorimetry, and viscosity measurements after aqueous dispersion. Further biopharmaceutical assessment started with biorelevant nonsink dissolution testing followed by a pharmacokinetic in vivo study in rats. The in vitro assessment showed superiority of the lysine-containing formulation in the extent of in vitro supersaturation and overall drug release. In accordance with this, the in vivo study also demonstrated increased exposure of the amorphous formulations and in particular for the system containing lysine. In summary, the combination of polyelectrolytes with interacting additives presents a promising opportunity for the formulation of poorly water-soluble drugs.


The accuracy of diagnostic coding for acute kidney injury in England - a single centre study.
Acute kidney injury (AKI) is an independent risk factor for mortality and is responsible for a significant burden of healthcare expenditure, so accurate measurement of its incidence is important. Administrative coding data has been used for assessing AKI incidence, and shows an increasing proportion of hospital bed days attributable to AKI. However, the accuracy of coding for AKI and changes in coding over time have not been studied in England. We studied a random sample of admissions from 2005 and 2010 where ICD-10 code N17 (acute renal failure) was recorded in the administrative coding data at one acute NHS Foundation Trust in England. Using the medical notes and computerised records we examined the demographic and clinical details of these admissions. Against a 6.3% (95% CI 4.8-7.9%) increase in all non-elective admissions, we found a 64% increase in acute renal failure admissions (95% CI 41%-92%, p < 0.001) in 2010 compared to 2005. Median age was 78 years (IQR 72-87), 11-25% had a relevant pre-admission co-morbidity and 64% (55-73%) were taking drugs known to be associated with AKI. Over both years, 95% (91-99%) of cases examined met the Kidney Disease: Improving Global Outcomes criteria for AKI. Patients with hospital admissions where AKI has been coded are elderly with multiple co-morbidities. Our results demonstrate a high positive predictive value of coding data for a clinical diagnosis of AKI, with no suggestion of marked changes in coding of AKI between 2005 and 2010.


Synthetic zinc finger nuclease design and rapid assembly.
Engineered zinc finger nucleases (ZFNs) are a tool for genome manipulation that are of great interest to scientists in many fields. To meet the needs of researchers wishing to employ ZFNs, an inexpensive, rapid assembly procedure would be beneficial to laboratories that do not have access to the proprietary reagents often required for ZFN production. Using freely available sequence data derived from the Zinc Finger Targeter database, we developed a protocol for synthesis and directed insertion of user-defined ZFNs into a versatile plasmid expression system. This oligonucleotide-based isothermal DNA assembly protocol was used to determine whether we could generate functional nucleases capable of endogenous gene editing. We targeted the human α-l-iduronidase (IDUA) gene on chromosome 4, mutations of which result in the severe lysosomal storage disease mucopolysaccharidosis type I. In approximately 1 week we were able to design, assemble, and test six IDUA-specific ZFNs. In a single-stranded annealing assay five of the six candidates we tested performed at a level comparable to or surpassing previously reported ZFNs. One of the five subsequently showed nuclease activity at the endogenous genomic IDUA locus. To our knowledge, this is the first demonstration of in silico-designed, oligonucleotide-assembled, synthetic ZFNs, requiring no specialized templates or reagents that are capable of endogenous human gene target site activity. This method, termed CoDA-syn (context-dependent assembly-synthetic), should facilitate a more widespread use of ZFNs in the research community.


Fluorescence correlation spectroscopy of fast chain dynamics within denatured protein L.
Intramolecular dynamics in the denatured state of a protein are of importance for protein folding. Native-like contact formation within the ensemble of denatured conformations of a protein may guide its transformation towards the native conformation. The efficiency of folding is thus dependent on the diffusion of chain fragments, which facilitates contact formation. Herein we investigate intramolecular diffusion of denatured molecules of the small two-state-folding protein L with fluorescence correlation spectroscopy (FCS). We utilize the specific quenching of the fluorescence of the oxazine dye Atto655 labeling a cysteine at position 64 (the C-terminus of the protein) by the side chain of a tryptophan at position 47. FCS measurements allow us to probe processes ranging in timescales from tens of nanoseconds to seconds. Two fast photophysical processes can be distinguished in the fluorescence correlation curves. The slower of the two is found to be due to triplet dynamics, while the faster process is attributed to the quenching of the Atto655 by the tryptophan upon transient ground-state complex formation. We study the dependence of the intrachain dynamics of the denatured protein on the concentration of the denaturant guanidinium chloride (GdmCl), and extract complex association and dissociation rates. While the dissociation rate does not depend on the denaturant, the association rate decreases as denaturant concentration is increased from 3 to 7 M GdmCl. This decrease in contact formation rate tracks the expansion of denatured protein L, measured in our previous work. Thus, the intramolecular diffusion coefficient calculated from the results is found to be essentially independent of the denaturant concentration over this range, even as the protein expands by more than 20%.


Structure, sequon recognition and mechanism of tryptophan C-mannosyltransferase.
C-linked glycosylation is essential for the trafficking, folding and function of secretory and transmembrane proteins involved in cellular communication processes. The tryptophan C-mannosyltransferase (CMT) enzymes that install the modification attach a mannose to the first tryptophan of WxxW/C sequons in nascent polypeptide chains by an unknown mechanism. Here, we report cryogenic-electron microscopy structures of Caenorhabditis elegans CMT in four key states: apo, acceptor peptide-bound, donor-substrate analog-bound and as a trapped ternary complex with both peptide and a donor-substrate mimic bound. The structures indicate how the C-mannosylation sequon is recognized by this CMT and its paralogs, and how sequon binding triggers conformational activation of the donor substrate: a process relevant to all glycosyltransferase C superfamily enzymes. Our structural data further indicate that the CMTs adopt an unprecedented electrophilic aromatic substitution mechanism to enable the C-glycosylation of proteins. These results afford opportunities for understanding human disease and therapeutic targeting of specific CMT paralogs.


Capacity building in nutrition science: revisiting the curricula for medical professionals.
The current nutrition education curricula for students in U.S. medical schools, and schools of other health professions, such as nursing and oral health, do not provide enough opportunity to gain knowledge of the interactions among micro- and macronutrients, their role in maintaining optimal body functions, factors that interfere with these interactions, or, importantly, how to integrate this knowledge into medical practice. There is a need to better prepare healthcare professionals for identifying nutrition risk and managing hospitalized patients, especially those with chronic conditions, using an interprofessional, team-based approach. A major goal of this report is to revisit current nutrition training programs for physicians and other healthcare professionals in order to explore opportunities for providing healthcare providers with the essential tools of preventative and therapeutic nutrition intervention strategies. The issues addressed include whether a consensus exists on how to integrate basic and applied nutrition into the general healthcare professional curriculum, and if so, at which stages of training and at what depth should these integrations occur; how nutrition education is dealt with and achieved throughout all the health professions; and whether current nutrition education models are sufficient. To help address these issues, the report will review current nutrition education practices-their strengths and weaknesses-as well as evaluate promising new initiatives, and offer proposals for new directions for nutrition education training of future generation of medical practitioners.


Health effects caused by noise: evidence in the literature from the past 25 years.
Traffic noise is the most important source of environmental annoyance. According to the Environmental Expert Council of Germany, severe annoyance persistent over prolonged periods of time is to be regarded as causing distress. Previously, extraaural noise effects were mostly assessed using a paradigm in which the sound level played the major role. On the basis of this paradigm the relatively low sound level of environmental noise was not considered to be a potential danger to health. In contrast to this numerous empirical results have shown long-term noise-induced health risks. Therefore a radical change of attitude - a change of paradigm - is necessary. For an immediate triggering of protective reactions (fight/flight or defeat reactions) the information conveyed by noise is very often more relevant than the sound level. It was shown recently that the first and fastest signal detection is mediated by a subcortical area - the amygdala. For this reason even during sleep the noise from aeroplanes or heavy goods vehicles may be categorised as danger signals and induce the release of stress hormones. In accordance with the noise stress hypothesis chronic stress hormone dysregulations as well as increases of established endogenous risk factors of ischaemic heart diseases have been observed under long-term environmental noise exposure. Therefore, an increased risk of myocardial infarction is to be expected. The results of individual studies on this subject in most cases do not reach statistical significance. However, according to the Environmental Expert Council, these studies show a consistent trend towards an increased cardiovascular risk if the daytime immission level exceeds 65 dB(A). Most of the previous studies on the extraaural effects of occupational noise have been invalidated by exposure misclassifications. In future studies on health effects of noise a correct exposure assessment is one of the most important preconditions.


Identification and Quantification of Transient Receptor Potential Vanilloid 1 (TRPV1) in Equine Articular Tissue.
Joint pain and osteoarthritis (OA) are some of the most common causes of lameness in horses, and most of the available treatments focus on symptomatic relief without a disease-modifying effect. TRPV1 is a potential target for treating joint diseases, including OA, and the present study aims to investigate if the TRPV1 receptor is present in equine articular tissue and determine whether the number of receptors is upregulated in joint inflammation. Metacarpo/metatarsophalangeal (MCP/MTP) joints from 15 horses euthanised for reasons unrelated to this study were included. Based on synovial fluid analysis, macroscopic evaluation, and magnetic resonance imaging (MRI), joints were divided into two groups: healthy joints and joints with pathology. ELISA analysis was performed on synovial tissue harvested from all joints. TPRV1 was found in all joints. The mean concentration of TRPV1 compared to total protein in healthy joints (8.4 × 10-7 ng/mL) and joints with pathology (12.9 × 10-7 ng/mL) differed significantly (p = 0.01, t-test with Welch correction). Quantitative real-time reverse transcriptase PCR analysis was performed on RNA isolates from synovial tissue from all joints. TRPV1 mRNA expression ratio normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in healthy joints (0.16 (SD: 0.19)) and joints with pathology (0.24 (SD: 0.14)) did not differ significantly (p = 0.43, t-test with Welch correction). mRNA expression of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-) was very low for both groups. In conclusion, TRPV1 was detected both on mRNA and the protein level, with a higher expression of TRPV1 in samples from joints with pathology. Future studies will determine the clinical potential of equine TRPV1 as a target in the management of joint pain and inflammation.


Analysis of cytokine-induced NO-dependent apoptosis using RNA interference or inhibition by 1400W.
RNA interference has been used to silence gene expression and evaluate the contribution of a gene product to cell function. Here, we investigated conditions under which expression of an inducible protein, nitric oxide synthase 2 (NOS2), is decreased by RNA interference. Cytokine treatment of insulin-producing RINm5F cells results in NOS2 induction and cell death. Conditions used here favoured cytokine-induced apoptosis, for the first time--rather than necrosis, previously recorded. In RINm5F cells, transfected with NOS2-specific small interfering RNA followed by a 12 h exposure to cytokines, there was a significant reduction in NOS2 protein, nitrite, and apoptosis. There were no changes in these three parameters when experiments were carried out with unrelated vimentin siRNA. To interpret the NOS2-siRNA result further, we compared it with complete pharmacological inhibition of nitric oxide (NO) production by the NOS2 competitive inhibitor, 1400W, which lowered apoptosis by only 50% in the RINm5F cells. We conclude that the use of NOS2-specific siRNA has resulted in the subsequent lowering of expression of a cytokine-inducible protein whose function can be quantified. siRNA results have compared favourably with use of a pharmacological inhibitor of NOS2, in revealing the subtle, partial contribution of cytokine-induced NO to apoptosis induction in these insulin-producing cells.


Rate limit of protein elastic response is tether dependent.
The elastic restoring force of tissues must be able to operate over the very wide range of loading rates experienced by living organisms. It is surprising that even the fastest events involving animal muscle tissues do not surpass a few hundred hertz. We propose that this limit is set in part by the elastic dynamics of tethered proteins extending and relaxing under a changing load. Here we study the elastic dynamics of tethered proteins using a fast force spectrometer with sub-millisecond time resolution, combined with Brownian and Molecular Dynamics simulations. We show that the act of tethering a polypeptide to an object, an inseparable part of protein elasticity in vivo and in experimental setups, greatly reduces the attempt frequency with which the protein samples its free energy. Indeed, our data shows that a tethered polypeptide can traverse its free-energy landscape with a surprisingly low effective diffusion coefficient D(eff) ~ 1,200 nm(2)/s. By contrast, our Molecular Dynamics simulations show that diffusion of an isolated protein under force occurs at D(eff) ~ 10(8) nm(2)/s. This discrepancy is attributed to the drag force caused by the tethering object. From the physiological time scales of tissue elasticity, we calculate that tethered elastic proteins equilibrate in vivo with D(eff) ~ 10(4)-10(6) nm(2)/s which is two to four orders magnitude smaller than the values measured for untethered proteins in bulk.


Terahertz near-field microscopy with subwavelength spatial resolution based on photoconductive antennas.
Imaging and sensing applications based on pulsed terahertz radiation have opened new possibilities for scientific and industrial applications. Many exploit the unique features of the terahertz (THz) spectral region, where common packaging materials are transparent and many chemical compounds show characteristic absorptions. Because of their diffraction limit, THz far-field imaging techniques lack microscopic resolution and, if subwavelength features have to be resolved, near-field techniques are required. Here, we present a THz near-field microscopy approach based on photoconductive antennas as the THz emitter and as a near-field probe. Our system allows us to measure amplitude, phase, and polarization of the electric fields in the vicinity of a sample with a spatial resolution on the micrometer scale (approximately lambda/20). Using a dielectric (plant leaf) and a metallic structure (microwire) as examples, we demonstrate the capabilities of our approach.


Role of vitamin b(6) deficiency in the nitrogen balance of streptozotocin-diabetic rats.
ABSTRACT Male Wistar rats with streptozotocin-induced diabetes received a control diet and a pyridoxine-deficient diet. The animals were divided at random into four groups: control rats (CR), control diabetic rats (CDR), diabetic rats receiving a pyridoxine-free diet (DRB6), and diabetic rats receiving saline solution and no insulin treatment (DRSS). The experiment lasted 45 days. During the first 15 days the animals were observed for the development of diabetes and during the remaining 30 days they received the respective diets. The absence of vitamin B(6) did not influence the glycemia levels at the end of the experiment or the weight evolution of the animals. The rats that did not receive pyridoxine (DRB6) only showed a reduction in GPT activity (17.79 U/mL) compared to the other groups. The DRB6 group presented a significantly lower (p <0.05) nitrogen balance during each period (2.38 +/- 0.44 g N/7 days) compared to the CDR group (3.28 +/- 0.56 g N/7 days). The DRSS group presented similar or significantly higher values (2.81 +/- 0.77 g N/7 days) compared to the CDR group. Pyridoxine-deficient diabetic rats treated with insulin suffered important changes in the utilization of dietary proteins, as observed by nitrogen balance and enzyme activity studies.


A chiral salen-based MOF catalytic material with high thermal, aqueous and chemical stabilities.
A highly stable chiral salen-based metal-organic framework [(Cu4I4)2L4]·20DMF·3CH3CN (1) [L = (R,R)-N,N'-bis(3-tert-butyl-5-(4-pyridyl)salicylidene)-1,2-diphenylethylenediamine nickel(ii)] has been synthesized and characterized by single crystal X-ray diffraction and other physicochemical methods. 1 exhibits a rare 8-fold interpenetrated 3D framework constructed by a 4-connecting Cu4I4 cluster and a 2-coordinating L ligand. Remarkably, in spite of 8-fold interpenetration, 1 still possesses two types of 1D chiral hydrophobic channels with pore window sizes of 6.77 × 8.64 Å2 and 6.09 × 10.96 Å2 along the crystallographic a axis. All Ni(salen) moieties of L lie inside the 1D channels and the empty coordination sites of Ni2+ are oriented to the cavities. PXRD and N2 adsorption measurements confirmed that 1 is extremely stable under high temperature (>400 °C), in water vapor (90% relative humidity), in acid/base aqueous solution (pH 0-14), and in saturated NaOH solution at 100 °C, as well as in 30 wt% H2O2 and 70 wt% tert-butyl hydroperoxide solution. 1 was proved to be an excellent recycled heterogeneous catalyst for the conversion of simulated industrial CO2 (that is, involving tiny amounts of water vapor and other acidic gases) with epoxides into cyclic carbonates under mild conditions for the first time. The synthesis of β-hydroxy-1,2,3-triazoles from the same epoxides, alkyne and sodium azide was also catalyzed by 1 in aqueous solution with high yield. Interestingly, the cycloaddition reaction of CO2 to bulky epoxides shows a decrease in the activity with an increase in the alkyl chain length of the substrate because of confinement of the channel size of 1, showing size-dependent selectivity. The plausible catalytic mechanisms for these two reactions have also been proposed.


Education in the workplace for the physician: clinical management states as an organizing framework.
Medical educators are interested in approaches to making selected relevant knowledge available in the context of problem-based care. This is of value both during the process of care and as a means of organizing information for offline self-study. Four trends in health information technology are relevant to achieving the goal and can be expected to play a growing role in the future. First, health care enterprises are developing approaches for access to information resources related to the care of a patient, including clinical data and images but also communication tools, referral and other logistic tools, decision support, and educational materials. Second, information for patients and methods for patient-doctor interaction and decision making are becoming available. Third, computer-based methods for representation of practice guidelines are being developed to support applications that can incorporate their logic. Finally, considering patients as being in particular "clinical management states" (or CMSs) for specific problems, approaches are being developed to use guidelines as a kind of "predictive" framework to enable development of interfaces for problem-based clinical encounters. The guidelines for a CMS can be used to identify the kinds of resources specifically needed for clinical encounters of that type. As the above trends converge to produce problem-specific environments, professional specialty organizations and continuing medical education course designers will need to focus energies on organizing and updating medical knowledge to make it available in CMS-specific contexts.


6 months versus 12 months of adjuvant trastuzumab in early breast cancer (PHARE): final analysis of a multicentre, open-label, phase 3 randomised trial.
In 2013, the interim analysis of the Protocol for Herceptin as Adjuvant therapy with Reduced Exposure (PHARE) trial could not show that 6 months of adjuvant trastuzumab was non-inferior to 12 months. Here, we report the planned final analysis based on the prespecified number of occurring events. PHARE is an open-label, phase 3, non-inferiority randomised trial of patients with HER2-positive early breast cancer comparing 6 months versus 12 months of trastuzumab treatment concomitant with or following standard neoadjuvant or adjuvant chemotherapy. The study was undertaken in 156 centres in France. Eligible patients were women aged 18 years or older with non-metastatic, operable, histologically confirmed adenocarcinoma of the breast and either positive axillary nodes or negative axillary nodes but a tumour of at least 10 mm. Participants must have received at least four cycles of a chemotherapy for this breast cancer and have started receiving adjuvant trastuzumab-treatment. Eligible patients were randomly assigned to either 6 months or 12 months of trastuzumab therapy duration between the third and sixth months of adjuvant trastuzumab. The randomisation was stratified by concomitant or sequential treatment with chemotherapy, oestrogen receptor status, and centre. The primary objective was non-inferiority in the intention-to-treat population in the 6-month group in terms of disease-free survival with a prespecified hazard margin of 1·15. This trial is registered with ClinicalTrials.gov, number NCT00381901. 3384 patients were enrolled and randomly assigned to either 12 months (n=1691) or 6 months (n=1693) of adjuvant trastuzumab. One patient in the 12-month group and three patients in the 6-month group were excluded, so 1690 patients in each group were included in the intention-to-treat analysis. At a median follow-up of 7·5 years (IQR 5·3-8·8), 704 events relevant to disease-free survival were observed (345 [20·4%] in the 12-month group and 359 [21·2%] in the 6-month group). The adjusted hazard ratio for disease-free survival in the 12-month group versus the 6-month group was 1·08 (95% CI 0·93-1·25; p=0·39). The non-inferiority margin was included in the 95% CI. No differences in effects pertaining to trastuzumab duration were found in any of the subgroups. After the completion of trastuzumab treatment, rare adverse events occurred over time and the safety analysis remained similar to the previously published report. In particular, we found no change in the cardiac safety comparison, and only three additional cases in which the left ventricular ejection fraction decreased to less than 50% have been reported in the 12-month group. The PHARE study did not show the non-inferiority of 6 months versus 12 months of adjuvant trastuzumab. Hence, adjuvant trastuzumab standard duration should remain 12 months. The French National Cancer Institute.


Life span extension by calorie restriction depends on Rim15 and transcription factors downstream of Ras/PKA, Tor, and Sch9.
Calorie restriction (CR), the only non-genetic intervention known to slow aging and extend life span in organisms ranging from yeast to mice, has been linked to the down-regulation of Tor, Akt, and Ras signaling. In this study, we demonstrate that the serine/threonine kinase Rim15 is required for yeast chronological life span extension caused by deficiencies in Ras2, Tor1, and Sch9, and by calorie restriction. Deletion of stress resistance transcription factors Gis1 and Msn2/4, which are positively regulated by Rim15, also caused a major although not complete reversion of the effect of calorie restriction on life span. The deletion of both RAS2 and the Akt and S6 kinase homolog SCH9 in combination with calorie restriction caused a remarkable 10-fold life span extension, which, surprisingly, was only partially reversed by the lack of Rim15. These results indicate that the Ras/cAMP/PKA/Rim15/Msn2/4 and the Tor/Sch9/Rim15/Gis1 pathways are major mediators of the calorie restriction-dependent stress resistance and life span extension, although additional mediators are involved. Notably, the anti-aging effect caused by the inactivation of both pathways is much more potent than that caused by CR.


Dimension-controlled ion-pairing assemblies based on π-electronic charged species.
This feature article summarizes the recent progress in the study of ion-pairing assemblies based on π-electronic ion pairs, including anion complexes of π-electronic molecules. Recently, ionic self-assembly (or ion-pairing assembly) has been widely investigated and utilized in the fabrication of various functional materials including soft materials, surface morphologies and organic-inorganic hybrid materials. It is significantly important to precisely design and synthesize charged π-electronic systems for obtaining highly organized assemblies, but the development of π-electronic ion pairs with an aim to fabricate soft materials has hardly been examined. This is because in many cases it is difficult to arrange the desired charged species due to their synthetic difficulties. In view of this scenario, tailor-made π-electronic ion pairs consisting of ion complexes of π-electronic molecules and their counterions can be used as the building blocks for ion-pairing assemblies. In particular, dimension-controlled ion-pairing assemblies can be formed based on π-electronic charged species, which have potential for use as functional nanostructured materials. In this feature article, we overview the basic synthetic strategies and recent examples of ion-pairing assemblies comprising π-electronic systems.


Distinct effects of XBF-1 in regulating the cell cycle inhibitor p27(XIC1) and imparting a neural fate.
XBF-1 is an anterior neural plate-specific, winged helix transcription factor that affects neural development in a concentration-dependent manner. A high concentration of XBF-1 results in suppression of endogenous neuronal differentiation and an expansion of undifferentiated neuroectoderm. Here we investigate the mechanism by which this expansion is achieved. Our findings suggest that XBF-1 converts ectoderm to a neural fate and it does so independently of any effects on the mesoderm. In addition, we show that a high dose of XBF-1 promotes the proliferation of neuroectodermal cells while a low dose inhibits ectodermal proliferation. Thus, the neural expansion observed after high dose XBF-1 misexpression is due both to an increase in the number of ectodermal cells devoted to a neural fate and an increase in their proliferation. We show that the effect on cell proliferation is likely to be mediated by p27(XIC1), a cyclin-dependent kinase (cdk) inhibitor. We show that p27(XIC1) is expressed in a spatially restricted pattern in the embryo, including the anterior neural plate, and when misexpressed it is sufficient to block the cell cycle in vivo. We find that p27(XIC1 )is transcriptionally regulated by XBF-1 in a dose-dependent manner such that it is suppressed or ectopically induced by a high or low dose of XBF-1, respectively. However, while a low dose of XBF-1 induces ectopic p27(XIC1 )and ectopic neurons, misexpression of p27(XIC1 )does not induce ectopic neurons, suggesting that the effects of XBF-1 on cell fate and cell proliferation are distinct. Finally, we show that p27(XIC1 )is suppressed by XBF-1 in the absence of protein synthesis, suggesting that at least one component of p27(XIC1 )regulation by XBF-1 may be direct. Thus, XBF-1 is a neural-specific transcription factor that can independently affect both the cell fate choice and the proliferative status of the cells in which it is expressed.


Self-collection of oral epithelial cell DNA under instruction from epidemiologic interviewers.
Oral epithelial cells provide an easily accessible source of germline DNA. Two methods for collection were compared in a 1992-1995 case-control study of oral cancer in Puerto Rico. One group of subjects (55 controls without oral cancer) collected oral rinse samples at home or work under the direction of a nonmedically trained interviewer ("self-collection"); the other group (94 controls) participated in a clinic-based collection, which also included blood and urine samples, conducted by a medical technician ("clinic collection"). Participation was higher for self-collection (98.2%) than for clinic collection (70.7%) (p < 0.001). DNA yields ranged from 2.0 to 204.5 microg (median, 25.9 microg) and did not differ by collection method, although yields varied by interviewer among self-collected samples (p = 0.02). Success rates for polymerase chain reaction amplification of the ADH3, NAT1, and multiplex CYP1A1/GSTT1/GSTM1 genotyping assays ranged from 76.4% (NAT1) to 98.2% (ADH3) for self-collected samples and were similar to those for clinic-collected samples (87.2-97.9%). Failure to amplify was associated with low DNA content (p = 0.015). Similar results were observed among cases (91 self-collected, 66 clinic collected), except that DNA yields did not vary by interviewer and a larger fraction (10.2%) of samples contained less than 5 microg of DNA, perhaps because of disease-related oral impairment. Self-collection of oral epithelial DNA samples appears satisfactory and efficient for many epidemiologic studies.


Contagion in Mass Killings and School Shootings.
Several past studies have found that media reports of suicides and homicides appear to subsequently increase the incidence of similar events in the community, apparently due to the coverage planting the seeds of ideation in at-risk individuals to commit similar acts. Here we explore whether or not contagion is evident in more high-profile incidents, such as school shootings and mass killings (incidents with four or more people killed). We fit a contagion model to recent data sets related to such incidents in the US, with terms that take into account the fact that a school shooting or mass murder may temporarily increase the probability of a similar event in the immediate future, by assuming an exponential decay in contagiousness after an event. We find significant evidence that mass killings involving firearms are incented by similar events in the immediate past. On average, this temporary increase in probability lasts 13 days, and each incident incites at least 0.30 new incidents (p = 0.0015). We also find significant evidence of contagion in school shootings, for which an incident is contagious for an average of 13 days, and incites an average of at least 0.22 new incidents (p = 0.0001). All p-values are assessed based on a likelihood ratio test comparing the likelihood of a contagion model to that of a null model with no contagion. On average, mass killings involving firearms occur approximately every two weeks in the US, while school shootings occur on average monthly. We find that state prevalence of firearm ownership is significantly associated with the state incidence of mass killings with firearms, school shootings, and mass shootings.


The neural mediators of kindness-based meditation: a theoretical model.
Although kindness-based contemplative practices are increasingly employed by clinicians and cognitive researchers to enhance prosocial emotions, social cognitive skills, and well-being, and as a tool to understand the basic workings of the social mind, we lack a coherent theoretical model with which to test the mechanisms by which kindness-based meditation may alter the brain and body. Here, we link contemplative accounts of compassion and loving-kindness practices with research from social cognitive neuroscience and social psychology to generate predictions about how diverse practices may alter brain structure and function and related aspects of social cognition. Contingent on the nuances of the practice, kindness-based meditation may enhance the neural systems related to faster and more basic perceptual or motor simulation processes, simulation of another's affective body state, slower and higher-level perspective-taking, modulatory processes such as emotion regulation and self/other discrimination, and combinations thereof. This theoretical model will be discussed alongside best practices for testing such a model and potential implications and applications of future work.


Systematic review of the efficacy and safety of shock wave therapy for lateral elbow pain.
To determine the efficacy and safety of extracorporeal shock wave therapy (ESWT) for lateral elbow pain. Systematic review of randomized controlled trials using Cochrane Collaboration methodology. Nine placebo-controlled trials (1006 participants) and one trial of ESWT versus steroid injection (93 participants) were included. The 9 placebo-controlled trials reported conflicting results, although 11 of 13 pooled analyses found no significant benefit of ESWT over placebo, e.g., weighted mean difference for improvement in pain (on a 100-point scale) from baseline to 4-6 weeks (pooled analysis of 3 trials, 446 participants) was -9.42 (95% CI -20.70 to 1.86). Two pooled results favored ESWT, e.g., relative risk of treatment success (at least 50% improvement in pain with resisted wrist extension at 12 weeks) for ESWT in comparison to placebo (pooled analysis of 2 trials, 192 participants) was 2.2 (95% CI 1.55 to 3.12). However, this finding was not supported by the results of 4 other trials that were unable to be pooled. Steroid injection was more effective than ESWT at 3 months after the end of treatment assessed by a reduction of pain of 50% from baseline [21/25 (84%) vs 29/48 (60%); p < 0.05]. Minimal adverse effects of ESWT were reported. Based upon systematic review of 9 placebo-controlled trials, there is "platinum" level evidence that ESWT provides little or no benefit in terms of pain and function in lateral elbow pain. There is "silver" level evidence based upon one trial that steroid injection may be more effective than ESWT.


Benefits and harms of citrate locking solutions for hemodialysis catheters: a systematic review and meta-analysis.
Citrate has theoretical advantages over heparin for locking hemodialysis central venous catheters (CVCs), but the comparative effectiveness of these agents is not clear. 1) To compare the benefits and harms of citrate versus heparin locking solutions among patients undergoing hemodialysis through CVCs; 2) to appraise methodological quality of the supporting evidence. CENTRAL, MEDLINE, EMBASE, CINAHL, ISI Web of Science, and nephrology conference abstracts. We included randomized, parallel arm clinical trials that enrolled adult patients (>18 years) receiving chronic hemodialysis through CVCs using a citrate locking solution. We excluded studies in which citrate was combined with other agents, such as antibiotics. We used the GRADE approach to systematic reviews and quality appraisal. Two reviewers performed data extraction independently and in duplicate. We pooled count data using generic inverse variance with random-effects models, and used fixed-effect models when only two studies were available for pooling. Subgroups included low (≤5%) vs. higher (≥30%) citrate. We screened 600 citations. Forty-one proceeded to full-text screen; 5 met inclusion criteria. Studies included between 19 and 291 participants (Median N = 61) followed for a total of 174.6 catheter-years; 2 were multi-centred trials. Three studies assessed all-cause mortality; the pooled relative risk for death was 0.71 (95% CI = 0.42-1.24; p = 0.21; I(2) = 0%). The rate ratio for bacteremic episodes was 0.54 (95% CI = 0.23-1.29; p = 0.16; I(2) = 65%) while the rate ratio for bleeding was 0.48 (95% CI = 0.3-0.75; p = 0.001;I I(2) = 5%). Rates of catheter exchange/replacement, all-cause hospitalization and in-situ thrombolysis were not significantly different between groups in any of the pooled analyses. Risk of bias within pooled studies was low. Outcome definitions varied across studies. Imprecision due to small sample sizes and low event rates reduce our overall confidence in the pooled effect estimates. Benefits and harms of citrate vs. heparin locking solutions remain unclear; larger studies and standardization of outcome measurement and reporting are warranted. Protocol Registration Number: CRD42013004781.


Culture in whales and dolphins.
Studies of animal culture have not normally included a consideration of cetaceans. However, with several long-term field studies now maturing, this situation should change. Animal culture is generally studied by either investigating transmission mechanisms experimentally, or observing patterns of behavioural variation in wild populations that cannot be explained by either genetic or environmental factors. Taking this second, ethnographic, approach, there is good evidence for cultural transmission in several cetacean species. However, only the bottlenose dolphin (Tursiops) has been shown experimentally to possess sophisticated social learning abilities, including vocal and motor imitation; other species have not been studied. There is observational evidence for imitation and teaching in killer whales. For cetaceans and other large, wide-ranging animals, excessive reliance on experimental data for evidence of culture is not productive; we favour the ethnographic approach. The complex and stable vocal and behavioural cultures of sympatric groups of killer whales (Orcinus orca) appear to have no parallel outside humans, and represent an independent evolution of cultural faculties. The wide movements of cetaceans, the greater variability of the marine environment over large temporal scales relative to that on land, and the stable matrilineal social groups of some species are potentially important factors in the evolution of cetacean culture. There have been suggestions of gene-culture coevolution in cetaceans, and culture may be implicated in some unusual behavioural and life-history traits of whales and dolphins. We hope to stimulate discussion and research on culture in these animals.


Antioxidant activities of aqueous extracts from 12 Chinese edible flowers in vitro and in vivo.
The antioxidant function of edible flowers have attracted increasing interest. However, information is lacking on the impact of edible flowers on oxidative injury including hypoxia-re-oxygenation and hyperlipidemia. The antioxidant activities of aqueous extracts from 12 Chinese edible flowers were assessed in four different antioxidant models, including total antioxidant capacity (TAC), oxygen radical absorbance capacity (ORAC), scavenging hydroxyl radical capacity (SHRC) and scavenging superoxide anion radical capacity (SSARC). Subsequently, the potential antioxidant effects on rat cardiac microvascular endothelial cells (rCMEC) treated with hypoxia-re-oxygenation and hyperlipidemia rats induced by high-fat diet were also evaluated. The highest TAC, ORAC, SHRC and SSARC were Lonicera japonica Thunb., Rosa rugosa Thunb., Chrysanthemum indicum L. and Rosa rugosa Thunb., respectively. Most aqueous extracts of edible flowers exhibited good antioxidant effects on injury of rCMEC induced by hypoxia-re-oxygenation. In addition, the aqueous extracts of Lonicera japonica Thunb., Carthamus tinctorius L., Magnolia officinalis Rehd. et Wils., Rosmarinus officinalis L. and Chrysanthemum morifolium Ramat. could suppress the build-up of oxidative stress by increasing serum superoxide dismutase, glutathion peroxidase, and reducing malonaldehyde concentration in hyperlipidemia rats. These findings provided scientific support for screening edible flowers as natural antioxidants and preventative treatments for oxidative stress-related diseases.


An interactive and intuitive visualisation method for X-ray computed tomography data of biological samples in 3D Portable Document Format.
3D imaging approaches based on X-ray microcomputed tomography (microCT) have become increasingly accessible with advancements in methods, instruments and expertise. The synergy of material and life sciences has impacted biomedical research by proposing new tools for investigation. However, data sharing remains challenging as microCT files are usually in the range of gigabytes and require specific and expensive software for rendering and interpretation. Here, we provide an advanced method for visualisation and interpretation of microCT data with small file formats, readable on all operating systems, using freely available Portable Document Format (PDF) software. Our method is based on the conversion of volumetric data into interactive 3D PDF, allowing rotation, movement, magnification and setting modifications of objects, thus providing an intuitive approach to analyse structures in a 3D context. We describe the complete pipeline from data acquisition, data processing and compression, to 3D PDF formatting on an example of craniofacial anatomical morphology in the mouse embryo. Our procedure is widely applicable in biological research and can be used as a framework to analyse volumetric data from any research field relying on 3D rendering and CT-biomedical imaging.


Conformation types of ubiquitin [M+8H]8+ Ions from water:methanol solutions: evidence for the N and A States in aqueous solution.
Ion mobility and mass spectrometry measurements are used to examine the gas-phase populations of [M+8H](8+) ubiquitin ions formed upon electrospraying 20 different solutions from 100:0 to 5:95 water:methanol that are maintained at pH = 2.0. Over this range of solution conditions, mobility distributions for the +8 charge state show substantial variations. Here we develop a model that treats the combined measurements as one data set. By varying the relative abundances of a discrete set of conformation types, it is possible to represent distributions obtained from any solution. For solutions that favor the well-known A-state ubiquitin, it is possible to represent the gas-phase distributions with seven conformation types. Aqueous conditions that favor the native structure require four more structural types to represent the distribution. This analysis provides the first direct evidence for trace amounts of the A state under native conditions. The method of analysis presented here should help illuminate how solution populations evolve into new gas-phase structures as solvent is removed. Evidence for trace quantities of previously unknown states under native solution conditions may provide insight about the relationship of dynamics to protein function as well as misfolding and aggregation phenomena.


Expression of angiotensin-converting enzyme mRNA gene in the kidneys of patients with glomerulonephrites.
A little is known about the behavior of the renin-angiotensin system (RAS) in glomerulo-nephritis (GN), although it is activated in other models of injury. To study renal angiotensin-converting enzyme (ACE) messenger ribonucleic acid (mRNA) gene expression in patients with GN to determine its role in the disease process and other factors that may influence the course of the disease and the prognosis, e.g. treatment with ACE inhibitor (ACEI) drugs, we studied 20 patients with GN allocated to two groups: ten patients received an ACEI drug and ten patients did not receive ACEI in addition to a control group of ten healthy subjects. Routine and special laboratory investigation, histopathological studies and quantitative polymerase chain reaction analysis for renal ACE mRNA were done for both the study and the control groups. There was a statistically significant increase in ACE mRNA gene expression in the GN groups than in control group, but no statistically significant difference in ACE mRNA gene expression between the patients group that received and the group that did not receive ACEI. A significant correlation was found between the ACE mRNA gene expression and the mean blood pressure, serum creatinine, blood urea nitrogen and 24-h urinary protein. In conclusion, a higher level of ACE mRNA gene expression in patients suffering from GN may suggest a role of the RAS in the process of GN, perhaps contributing to glomerular hypertrophy and matrix overproduction. The use of ACEI drugs possibly slows the rate of progression of renal failure and plays a role in controlling the pathophysiology.


Transient hepatic attenuation differences at biphasic spiral CT examinations.
Transient hepatic attenuation difference (THAD) is a perfusion disorder that can sometimes be observed in hepatic arterial phase of a biphasic spiral CT examination. The involved site of liver by THAD appears as an area of high attenuation on the hepatic arterial phase image and returns to normal attenuation on portal venous phase image. The knowledge of the shape, distribution and causes of THAD is essential to differentiate the THAD from other pathologies such as neoplasm.


Coordination of Cell Cycle Progression and Mitotic Spindle Assembly Involves Histone H3 Lysine 4 Methylation by Set1/COMPASS.
Methylation of histone H3 lysine 4 (H3K4) by Set1 complex/COMPASS is a hallmark of eukaryotic chromatin, but it remains poorly understood how this post-translational modification contributes to the regulation of biological processes like the cell cycle. Here, we report a H3K4 methylation-dependent pathway in Saccharomyces cerevisiae that governs toxicity toward benomyl, a microtubule destabilizing drug. Benomyl-sensitive growth of wild-type cells required mono- and dimethylation of H3K4 and Pho23, a PHD-containing subunit of the Rpd3L complex. Δset1 and Δpho23 deletions suppressed defects associated with ipl1-2 aurora kinase mutant, an integral component of the spindle assembly checkpoint during mitosis. Benomyl resistance of Δset1 strains was accompanied by deregulation of all four tubulin genes and the phenotype was suppressed by tub2-423 and Δtub3 mutations, establishing a genetic link between H3K4 methylation and microtubule function. Most interestingly, sine wave fitting and clustering of transcript abundance time series in synchronized cells revealed a requirement for Set1 for proper cell-cycle-dependent gene expression and Δset1 cells displayed delayed entry into S phase. Disruption of G1/S regulation in Δmbp1 and Δswi4 transcription factor mutants duplicated both benomyl resistance and suppression of ipl1-2 as was observed with Δset1 Taken together our results support a role for H3K4 methylation in the coordination of cell-cycle progression and proper assembly of the mitotic spindle during mitosis.


Increased risk of brain injury in IVF babies.
The aim of this paper was to assess brain injury occurrence among in vitro fertilization (IVF) babies. We examined all babies born in our hospital in the triennium 2004-2006, comparing the presence of brain injuries between IVF babies and the rest of the population. In IVF group (180 babies), brain injury was present in 4 babies, while in the rest of population (n=3602) it was present in 23 babies (P=0.042, RR: 3.18). IVF babies have a higher risk of being born with a birthweight less than 2 500 grams (P<0.0001; RR: 5.133). When we considered only babies born with a birth weight less than 2 500 grams, the difference of brain injury between the two groups was not significant. In IVF babies, brain injury occurred more frequently than in the rest of population. This is probably due to a higher rate of premature births and low birth weight in IVF population. Anyway, this data should be disclosed to future parents to make an informed decision.


The assembly of the export apparatus (YscR,S,T,U,V) of the Yersinia type III secretion apparatus occurs independently of other structural components and involves the formation of an YscV oligomer.
YscV (FlhA in the flagellum) is an essential component of the inner membrane (IM) export apparatus of the type III secretion injectisome. It contains eight transmembrane helices and a large C-terminal cytosolic domain. YscV was expressed at a significantly higher level than the other export apparatus components YscR, YscS, YscT, and YscU, and YscV-EGFP formed bright fluorescent spots at the bacterial periphery, colocalizing in most cases with YscC-mCherry. This suggested that YscV is the only protein of the export apparatus that oligomerizes. Oligomerization required the cytosolic domain of YscV, as well as YscR, -S, -T, but no other Ysc protein, indicating that an IM platform can assemble independently from the membrane-ring forming proteins YscC, -D, -J. However, in the absence of YscC, -D, -J, this IM platform moved laterally at the bacterial surface. YscJ, but not YscD could be recruited to the IM platform in the absence of the secretin YscC. As YscJ was shown earlier to be also recruited by the outer membrane (OM) platform made of YscC and YscD, we infer that assembly of the injectisome proceeds through the independent assembly of an IM and an OM platform that merge through YscJ.


Interactions between intracellular domains as key determinants of the quaternary structure and function of receptor heteromers.
G protein-coupled receptor (GPCR) heteromers are macromolecular complexes with unique functional properties different from those of its individual protomers. Little is known about what determines the quaternary structure of GPCR heteromers resulting in their unique functional properties. In this study, using resonance energy transfer techniques in experiments with mutated receptors, we provide for the first time clear evidence for a key role of intracellular domains in the determination of the quaternary structure of GPCR heteromers between adenosine A(2A), cannabinoid CB(1), and dopamine D(2) receptors. In these interactions, arginine-rich epitopes form salt bridges with phosphorylated serine or threonine residues from CK1/2 consensus sites. Each receptor (A(2A), CB(1), and D(2)) was found to include two evolutionarily conserved intracellular domains to establish selective electrostatic interactions with intracellular domains of the other two receptors, indicating that these particular electrostatic interactions constitute a general mechanism for receptor heteromerization. Mutation experiments indicated that the interactions of the intracellular domains of the CB(1) receptor with A(2A) and D(2) receptors are fundamental for the correct formation of the quaternary structure needed for the function (MAPK signaling) of the A(2A)-CB(1)-D(2) receptor heteromers. Analysis of MAPK signaling in striatal slices of CB(1) receptor KO mice and wild-type littermates supported the existence of A(1)-CB(1)-D(2) receptor heteromer in the brain. These findings allowed us to propose the first molecular model of the quaternary structure of a receptor heteromultimer.


A Computer-Aided Diagnosis System With EEG Based on the P3b Wave During an Auditory Odd-Ball Task in Schizophrenia.
To design a Computer-aided diagnosis (CAD) system using an optimized methodology over the P3b wave in order to objectively and accurately discriminate between healthy controls (HC) and schizophrenic subjects (SZ). We train, test, analyze, and compare various machine learning classification approaches optimized in terms of the correct classification rate (CCR), the degenerated Youden's index (DYI) and the area under the receiver operating curve (AUC). CAD system comprises five stages: electroencephalography (EEG) preprocessing, feature extraction, seven electrode groupings, discriminant feature selection, and binary classification. With two optimal combinations of electrode grouping, filtering, feature selection algorithm, and classification machine, we get either a mean CCR = 93.42%, specificity = 0.9673, sensitivity = 0.8727, DYI = 0.9188, and AUC = 0.9567 (total-15 Hz-J5-MLP), or a mean CCR = 92.23%, specificity = 0.9499, sensitivity = 0.8838, DYI = 0.9162, and AUC = 0.9807 (right hemisphere-35 Hz-J5-SVM), which to our knowledge are higher than those available to date. We have verified that a more restrictive low-pass filtering achieves higher CCR as compared to others at higher frequencies in the P3b wave. In addition, results validate previous hypothesis about the importance of the parietal-temporal region, associated with memory processing, allowing us to identify powerful {feature,electrode} pairs in the diagnosis of schizophrenia, achieving higher CCR and AUC in classification of both right and left Hemispheres, and parietal-temporal EEG signals, like, for instance, the {PSE, P4} pair (J5 and mutual information feature selection). Diagnosis of schizophrenia is made thoroughly by psychiatrists but as any human-based decision that has a subjective component. This CAD system provides the human expert with an objective complimentary measure to help him in diagnosing schizophrenia.


[In vitro susceptibilities to levofloxacin and various antibacterial agents of 12,866 clinical isolates obtained from 72 centers in 2010].
Postmarketing surveillance of levofloxacin (LVFX) has been conducted continuously since 1992. The present survey was performed to investigate in vitro susceptibility of recent clinical isolates in Japan to 30 selected antibacterial agents, focusing on fluoroquinolones (FQs). The common respiratory pathogens Streptococcus pyogenes, Streptococcus pneumoniae, Moraxella catarrhalis, and Haemophilus influenzae continue to show a high susceptibility to FQs. In contrast, widely-prevailing resistance to macrolides was markedly noted among S pneumoniae and S. pyogenes. Regarding H. influenzae, the prevalence of beta-lactamase-negative ampicillin-resistant isolates has been increasing year by year (25.8% in 2002, 40.0% in 2004, 50.1% in 2007, and 57.9% in 2010). Enterobacteriaceae showed high susceptibility to FQs, however, prevalence of LVFX-resistant Escherichia coli, including intermediate resistance, was 29.3%, showing an increase over time. Nevertheless, the increase in the prevalence of LVFX-resistant E. coli isolates has slowed since 2007 (8.2% in 2000, 11.8% in 2002, 18.8% in 2004, 26.2% in 2007, and 29.3% in 2010), suggesting the influence of LVFX 500 mg tablets since its approval in 2009. Another Enterobacteriaceae member, Klebsiella pneumoniae, showed low resistance to FQs, in contrast with E. coli. In methicillin-resistant Staphylococcus aureus (MRSA), the percentage of FQ-susceptible isolates was low, at 51.6% for susceptibility to sitafloxacin, and at only around 10% for susceptibility to other FQs. However, methicillin-susceptible S. aureus (MSSA) isolates were highly susceptible to FQs, with the percentage ranging from 88.5% to 99.1%. The prevalence of FQs-resistant isolates in methicillin-resistant coagulase-negative staphylococci was higher than that in methicillin-susceptible coagulase-negative staphylococci, although it was lower than the prevalence of FQ-resistance in MRSA. The prevalence of FQs-resistant Pseudomonas aeruginosa isolates derived from urinary tract infections (UTIs) was 15.4-21.3%, higher than the prevalence of 6.1-12.3% in P. aeruginosa isolates from respiratory tract infections (RTIs). While this trend was consistent with the results of previous surveillance, gradual decreases were noted in the prevalence of FQ-resistant P. aeruginosa isolates derived from UTIs. The prevalence of multidrug-resistant P. aeruginosa was 2.3% among isolates derived from UTIs and 0.3% among isolates from RTIs, a decrease from the results of 2007. Acinetobacter spp. showed high susceptibility to FQs. Imipenem-resistant Acinetobacter baumannii, which is currently an emerging issue, was detected at a prevalence of 2.4% (13 isolates). Neisseria gonorrhoeae showed a high resistance of 81.3-82.5%, to FQs. Ceftriaxone (CTRX) continued to show 100% susceptibility until 2007, but the present survey revealed the advent of resistance to CTRX in some clinical isolates. The result of the present survey indicated that although methicillin-resistant staphylococci, Enterococcus faecium, P. aeruginosa from UTIs, N. gonorrhoeae, and E. coli showed resistance of about 20% or more (19.5-89.2%) against the FQs which have been used clinically for over 17 years, the trends observed were similar to the results of previous surveillance. While FQ resistance has been prevailing in E. coli, E. coli still shows more than 70% susceptibility to FQs. The other bacterial species maintained high susceptibility rates of greater than 80%, against FQs.


Fifteen compelling open questions in plant cell biology.
As scientists, we are at least as excited about the open questions-the things we do not know-as the discoveries. Here, we asked 15 experts to describe the most compelling open questions in plant cell biology. These are their questions: How are organelle identity, domains, and boundaries maintained under the continuous flux of vesicle trafficking and membrane remodeling? Is the plant cortical microtubule cytoskeleton a mechanosensory apparatus? How are the cellular pathways of cell wall synthesis, assembly, modification, and integrity sensing linked in plants? Why do plasmodesmata open and close? Is there retrograde signaling from vacuoles to the nucleus? How do root cells accommodate fungal endosymbionts? What is the role of cell edges in plant morphogenesis? How is the cell division site determined? What are the emergent effects of polyploidy on the biology of the cell, and how are any such "rules" conditioned by cell type? Can mechanical forces trigger new cell fates in plants? How does a single differentiated somatic cell reprogram and gain pluripotency? How does polarity develop de-novo in isolated plant cells? What is the spectrum of cellular functions for membraneless organelles and intrinsically disordered proteins? How do plants deal with internal noise? How does order emerge in cells and propagate to organs and organisms from complex dynamical processes? We hope you find the discussions of these questions thought provoking and inspiring.


Spatial clusters of life expectancy and association with cardiovascular disease mortality and cancer mortality in the contiguous United States: 1980-2014.
The average life expectancy varies greatly from county to county in USA and there are also spatial variations in the county mortality rates for cardiovascular disease (CVD) and cancer, the top two causes of death. An association between these two groups of diseases has not been identified by cluster analysis previously. The main objective in this study was to investigate and quantify the associations between mortality due to CVD, cancer mortality and life expectancy based on US county data between 1980 and 2014. Regression analysis was used to adjust life expectancy for the mortality due to CVD and that due to cancer. In addition to the spatial life expectancy trends, we also studied existing trends over time with the software JOINPOINT to see how life expectancy is influenced by changes in mortality due to CVD and cancer mortality. The study setting was the 48 contiguous US states, while participants were 3,100 counties and their populations of all ages during the period 1980-2014. The main outcomes are spatial clusters of unusually low or high levels of life expectancy in addition to identifying which county level life expectancy locations were significantly associated with mortality due to CVD and/or cancer. Life expectancy has been improving steadily from 1980 to 2014, but the rate of increase per year (indicated by variation of the trend slope) changed significantly at five joinpoints, the latest of which occurred in 2010 when the slope changed from 0.29 (1980-1982) to 0.03 (2010-2014). Our results reveal that there are significant, purely spatial clusters in some geographical areas where life expectancy rates are significantly higher (or lower) than in the rest of the contiguous US. It is also shown that there is a significant association between the life expectancy level and the corresponding CVD mortality, and there is also a significant association between life expectancy level and the corresponding overall cancer mortality. The general trends (regression slopes) over time for the USA in life expectancy mortality, CVD mortality and cancer mortality have changed significantly after 2009-2010.


Comparative and functional genomics provide insights into the pathogenicity of dermatophytic fungi.
Millions of humans and animals suffer from superficial infections caused by a group of highly specialized filamentous fungi, the dermatophytes, which exclusively infect keratinized host structures. To provide broad insights into the molecular basis of the pathogenicity-associated traits, we report the first genome sequences of two closely phylogenetically related dermatophytes, Arthroderma benhamiae and Trichophyton verrucosum, both of which induce highly inflammatory infections in humans. 97% of the 22.5 megabase genome sequences of A. benhamiae and T. verrucosum are unambiguously alignable and collinear. To unravel dermatophyte-specific virulence-associated traits, we compared sets of potentially pathogenicity-associated proteins, such as secreted proteases and enzymes involved in secondary metabolite production, with those of closely related onygenales (Coccidioides species) and the mould Aspergillus fumigatus. The comparisons revealed expansion of several gene families in dermatophytes and disclosed the peculiarities of the dermatophyte secondary metabolite gene sets. Secretion of proteases and other hydrolytic enzymes by A. benhamiae was proven experimentally by a global secretome analysis during keratin degradation. Molecular insights into the interaction of A. benhamiae with human keratinocytes were obtained for the first time by global transcriptome profiling. Given that A. benhamiae is able to undergo mating, a detailed comparison of the genomes further unraveled the genetic basis of sexual reproduction in this species. Our results enlighten the genetic basis of fundamental and putatively virulence-related traits of dermatophytes, advancing future research on these medically important pathogens.


The association of early post-resuscitation hypotension with discharge survival following targeted temperature management for pediatric in-hospital cardiac arrest.
Approximately 40% of children who have an in-hospital cardiac arrest (IHCA) in the US survive to discharge. We aimed to evaluate the impact of post-cardiac arrest hypotension during targeted temperature management following IHCA on survival to discharge. This is a secondary analysis of the therapeutic hypothermia after pediatric cardiac arrest in-hospital (THAPCA-IH) trial. "Early hypotension" was defined as a systolic blood pressure less than the fifth percentile for age and sex for patients not treated with extracorporeal membrane oxygenation (ECMO) or a mean arterial pressure less than fifth percentile for age and sex for patients treated with ECMO during the first 6 h of temperature intervention. The primary outcome was survival to hospital discharge. Of 299 children, 142 (47%) patients did not receive ECMO and 157 (53%) received ECMO. Forty-two of 142 (29.6%) non-ECMO patients had systolic hypotension. Twenty-three of 157 (14.7%) ECMO patients had mean arterial hypotension. After controlling for confounders of interest, non-ECMO patients who had early systolic hypotension were less likely to survive to hospital discharge (40.5% vs. 72%; adjusted OR [aOR] 0.34; 95%CI, 0.12-0.93). There was no difference in survival to discharge by blood pressure groups for children treated with ECMO (30.4% vs. 49.3%; aOR = 0.60; 95%CI, 0.22-1.63). In this secondary analysis of the THAPCA-IH trial, in patients not treated with ECMO, systolic hypotension within 6 h of temperature intervention was associated with lower odds of discharge survival. Blood pressure groups in patients treated with ECMO were not associated with survival to discharge.