Structure and partial amino acid sequence of calf thymus DNA topoisomerase II: comparison with other type II enzymes.
The partial amino acid sequence of p140 calf thymus DNA topoisomerase II was determined by analysis of cyanogen bromide peptides. Five peptides were aligned and shared extensive homology with sequences derived from cDNA clones for the human topoisomerase II isoenzyme forms. Less homology was seen with the Drosophila, yeast and bacterial type II enzymes. Calf and human enzymes shared epitopes allowing isolation of a cDNA clone to human topoisomerase II isoenzyme alpha. Our results indicate that calf thymus p140 topoisomerase II is an active N-terminal proteolytic fragment of the native p180 enzyme and demonstrate that mammalian type II enzymes exhibit close sequence similarity.


Association of physical activity with vascular endothelial function and intima-media thickness.
Impairment of vascular endothelial function and increased intima-media thickness (IMT) are important early steps in atherogenesis. Longitudinal data on the effect of physical activity on endothelial function and IMT in healthy adolescents are lacking. We investigated prospectively the association of leisure-time physical activity with endothelial function (brachial artery flow-mediated dilatation; FMD) and aortic IMT in adolescents. FMD and IMT were measured with ultrasonography at 13 (n=553), 15 (n=531), and 17 (n=494) years of age in adolescents participating in a longitudinal atherosclerosis prevention study (Special Turku Coronary Risk Factor Intervention Project for Children). Mean aortic IMT, maximum FMD, and total FMD response (area under the dilatation curve 40 to 180 seconds after hyperemia) were calculated. Leisure-time physical activity was assessed with a questionnaire, and metabolic equivalent (MET) hours per week of leisure-time physical activity were calculated by multiplying weekly mean exercise intensity, duration, and frequency. Leisure-time physical activity was directly associated with endothelial function (P for maximum FMD=0.0021, P for total FMD response=0.0036) and inversely with IMT (P=0.011) after adjustment for age, sex, body mass index, high-density lipoprotein/total cholesterol, systolic blood pressure, and C-reactive protein and regarding FMD brachial artery diameter. Sedentary adolescents who increased their leisure-time physical activity from <5 to >5 (IMT) or >30 (maximum FMD) MET h/wk between 13 and 17 years of age had an increased maximum FMD (P=0.031) and decreased progression of IMT (P=0.047) compared with adolescents who remained sedentary. IMT progression was attenuated in persistently active adolescents compared with those who became sedentary (P=0.0072). Physical activity is favorably associated with endothelial function and IMT in adolescents. Importantly, a moderate increase in physical activity is related to decreased progression of IMT. A physically active lifestyle seems to prevent the development of subclinical atherosclerotic vascular changes in healthy adolescents. Clinical Trial Registration- URL: http://www.clinicaltrials.gov. Unique identifier: NCT00223600 (STRIP19902010).


Isolated Fe and Co dual active sites on nitrogen-doped carbon for a highly efficient oxygen reduction reaction.
We successfully prepared Fe and Co isolated single atoms on metal-organic framework derived nitrogen-doped carbon (FeCo-ISAs/CN) by an adsorption-calcination strategy. The obtained FeCo-ISAs/CN exhibited top-level catalytic reactivity for the alkaline oxygen reduction reaction (ORR) with a half-wave potential of 0.920 V, which was 70 mV more positive than that of commercial Pt/C. Moreover, the catalyst was durable and showed negligible activity decay in the alkaline ORR during 5000 voltage cycles.


Duration of therapy recommended for bacteraemic illness varies widely amongst clinicians.
The optimum duration of antimicrobial therapy would eradicate infection whilst minimising potential adverse drug effects to the patient. Australian and New Zealand infectious diseases (ID) and ICU specialists were surveyed regarding their recommended duration of antibiotic treatment for five common bacteraemic syndromes. A total of 239 clinicians responded to the survey (15.5% ICU and 84.5% ID). Overall, the most common reported durations were 7 (33.7%), 10 (25.9%) and 14 (26.0%) days, with 46% of responses recommending ≤7 days. Most respondents (>75% for each characteristic) would not modify duration based on host characteristics such as patient age or co-morbidities. ID physicians recommended longer durations than ICU physicians for all five syndromes (ID, median 10, IQR 7-14, range 1-28 days; ICU, median 7, IQR 5-10, range 2-21 days). Across all respondents, the median (IQR) duration for each syndrome was: CVC-BSI, 7 (7-10) days; bacteraemic pneumonia, 7 (7-10) days; bacteraemic UTI, 10 (7-14) days; bacteraemic IAI, 7 (7-12) days; and bacteraemic SSTI, 10 (7-14) days. Marked variation exists amongst clinicians' recommended duration of antibiotic treatment for BSI. A proportion of clinicians recommend therapy of ≤7 days at present (33.3-59.7% across scenarios). Patient characteristics are not strongly considered in the decision on therapy duration. This survey was undertaken as preparatory work for initiation of the BALANCE study, an ongoing randomised trial comparing 7 days with 14 days of therapy for BSI, providing an evidence base to inform best clinical treatment for this patient population.


Chito-oligosaccharide reduces diarrhea incidence and attenuates the immune response of weaned pigs challenged with Escherichia coli K88.
Seventy-two barrows (Landrace × Large White, initial BW of 4.9 ± 0.3 kg and 17 ± 3 d old) were used to determine if dietary chito-oligosaccharides can replace antibiotics as a means to reduce signs associated with infection in weaned pigs challenged with Escherichia coli. Pigs were assigned to 1 of 4 treatments in a randomized complete block design using 6 pens per treatment with 3 pigs per pen. The treatments consisted of pigs fed the unsupplemented corn-soybean meal diet challenged or unchallenged with E. coli K88 and pigs fed the same diet supplemented with 160 mg of chito-oligosaccharides or 100 mg of cyadox/kg and challenged with E. coli K88. On d 7, 1 group of pigs fed the unsupplemented diet, as well as all pigs fed diets containing chito-oligosaccharides or cyadox, were orally dosed with 30 mL of an alkaline broth containing E. coli K88. Another group of pigs fed the unsupplemented diet was orally dosed with 30 mL of sterilized alkaline broth. Fecal consistency was visually assessed each morning from d 7 to 14. Blood samples were collected at 0, 24, 48, and 168 h postinfection. On d 14 postchallenge, all pigs were killed to evaluate intestinal morphology and determine E. coli concentrations in the intestine. During the postchallenge period (wk 2), unsupplemented pigs challenged with E. coli had decreased (P < 0.05) BW gain, feed intake, fecal consistency, villus height, villus height:crypt depth ratio, and plasma IGF-1, and increased (P < 0.05) diarrhea incidence, E. coli counts in the intestine, plasma interleukin-1β, plasma IL-10, and IGA-positive cells in the jejunal and ileal lamina propria, compared with unchallenged pigs. Supplementation with cyadox largely mitigated these effects. Although chito-oligosaccharide reduced the incidence of diarrhea, the growth performance of E. coli-challenged pigs supplemented with chito-oligosaccharide was not better than that of unsupplemented pigs challenged with E. coli. Therefore, chito-oligosaccharide, at the amount used in this experiment, does not seem to be an effective substitute for antibiotics as a growth promoter for newly weaned pigs challenged with E. coli.


Contributions of individual mechanisms to fluoroquinolone resistance in 36 Escherichia coli strains isolated from humans and animals.
Twenty-eight human isolates of Escherichia coli from Argentina and Spain and eight veterinary isolates received from the Ministry of Agriculture Fisheries and Foods in the United Kingdom required 2 to > 128 micrograms of ciprofloxacin per ml for inhibition. Fragments of gyrA and parC encompassing the quinolone resistance-determining region were amplified by PCR, and the DNA sequences of the fragments were determined. All isolates contained a mutation in gyrA of a serine at position 83 (Ser83) to an Leu, and 26 isolates also contained a mutation of Asp87 to one of four amino acids: Asn (n = 14), Tyr (n = 6), Gly (n = 5), or His (n = 1). Twenty-four isolates contained a single mutation in parC, either a Ser80 to Ile (n = 17) or Arg (n = 2) or a Glu84 to Lys (n = 3). The role of a mutation in gyrB was investigated by introducing wild-type gyrB (pBP548) into all isolates; for three transformants MICs of ciprofloxacin were reduced; however, sequencing of PCR-derived fragments containing the gyrB quinolone resistance-determining region revealed no changes. The analogous region of parE was analyzed in 34 of 36 isolates by single-strand conformational polymorphism analysis and sequencing; however, no amino acid substitutions were discovered. The outer membrane protein and lipopolysaccharide profiles of all isolates were compared with those of reference strains, and the concentration of ciprofloxacin accumulated (with or without 100 microM carbony cyanide m-chlorophenylhydrazone [CCCP] was determined. Twenty-two isolates accumulated significantly lower concentrations of ciprofloxacin than the wild-type E. coli isolate; nine isolates accumulated less then half the concentration. The addition of CCCP increased the concentration of ciprofloxacin accumulated, and in all but one isolate the percent increase was greater than that in the control strains. The data indicate that high-level fluoroquinolone resistance in E. coli involves the acquisition of mutations at multiple loci.


Therapeutic Effect of Cabazitaxel and Blood-Brain Barrier opening in a Patient-Derived Glioblastoma Model.
Treatment of glioblastoma and other diseases in the brain is especially challenging due to the blood-brain barrier, which effectively protects the brain parenchyma. In this study we show for the first time that cabazitaxel, a semi-synthetic derivative of docetaxel can cross the blood-brain barrier and give a significant therapeutic effect in a patient-derived orthotopic model of glioblastoma. We show that the drug crosses the blood-brain barrier more effectively in the tumor than in the healthy brain due to reduced expression of p-glycoprotein efflux pumps in the vasculature of the tumor. Surprisingly, neither ultrasound-mediated blood-brain barrier opening (sonopermeation) nor drug formulation in polymeric nanoparticles could increase either accumulation of the drug in the brain or therapeutic effect. This indicates that for hydrophobic drugs, sonopermeation of the blood brain barrier might not be sufficient to achieve improved drug delivery. Nonetheless, our study shows that cabazitaxel is a promising drug for the treatment of brain tumors.


Combined treatment strategies for microtubule stabilizing agent-resistant tumors.
Resistance to microtubule-stabilizing agents is a major hurdle for successful cancer therapy. We investigated combined treatment of microtubule-stabilizing agents (MSAs) with inhibitors of angiogenesis to overcome MSA resistance. Treatment regimens of clinically relevant MSAs (patupilone and paclitaxel) and antiangiogenic agents (everolimus and bevacizumab) were investigated in genetically defined MSA-resistant lung (A549EpoB40) and colon adenocarcinoma (SW480) tumor xenografts in nude mice (CD1-Foxn1<nu>, ICRnu; 5-14 per group). Tumor growth delays were calculated by Kaplan-Meier analysis with Holm-Sidak tests. All statistical tests were two-sided. Inhibition of mTOR-kinase by everolimus only minimally reduced the proliferative activity of β tubulin-mutated lung adenocarcinoma cells alone and in combination with the MSA patupilone, but everolimus inhibited expression and secretion of vascular endothelial growth factor (VEGF) from these cells. mTOR-kinase inhibition strongly sensitized tumor xenografts derived from these otherwise MSA-resistant tumor cells to patupilone. Tumors treated with the combined modality of everolimus and patupilone had statistically significantly reduced tumor volume and stronger tumor growth delay (16.2 ± 1.01 days) than control- (7.7 ± 0.3 days, P = .004), patupilone- (10 ± 0.97 days, P = .009), and everolimus-treated (10.6 ± 1.4 days, P = .014) tumors. A combined treatment modality with bevacizumab also resensitized this MSA-refractory tumor model to patupilone. Treatment combination also strongly reduced microvessel density, corroborating the relevance of VEGF targeting for the known antivasculature-directed potency of MSA alone in MSA-sensitive tumor models. Resensitization to MSAs was also probed in P glycoprotein-overexpressing SW480-derived tumor xenografts. Different bevacizumab regimens also sensitized this otherwise-resistant tumor model to clinically relevant MSA paclitaxel. A treatment combination of MSAs with antiangiogenic agents is potent to overcome tumor cell-linked MSA resistance and should be considered as strategy for MSA-refractory tumor entities.


A Comprehensive NGS Data Analysis of Differentially Regulated miRNAs, piRNAs, lncRNAs and sn/snoRNAs in Triple Negative Breast Cancer.
Cancer is the second leading cause of death in the United States and is a major public health concern worldwide. Basic, clinical and epidemiological research is leading to improved cancer detection, prevention, and outcomes. Recent technological advances have allowed unbiased and comprehensive screening of genome-wide gene expression. Small non-coding RNAs (sncRNAs) have been shown to play an important role in biological processes and could serve as a diagnostic, prognostic and therapeutic biomarker for specific diseases. Recent findings have begun to reveal and enhance our understanding of the complex architecture of sncRNA expression including miRNAs, piRNAs, lncRNAs, sn/snoRNAs and their relationships with biological systems. We used publicly available small RNA sequencing data that was derived from 24 triple negative breast cancers (TNBC) and 14 adjacent normal tissue samples to remap various types of sncRNAs. We found a total of 55 miRNAs were aberrantly expressed (p<0.005) in TNBC samples (8 miRNAs upregulated; 47 downregulated) compared to adjacent normal tissues whereas the original study reported only 25 novel miRs. In this study, we used pathway analysis of differentially expressed miRNAs which revealed TGF-beta signaling pathways to be profoundly affected in the TNBC samples. Furthermore, our comprehensive re-mapping strategy allowed us to discover a number of other differentially expressed sncRNAs including piRNAs, lncRNAs, sn/snoRNAs, rRNAs, miscRNAs and nonsense-mediated decay RNAs. We believe that our sncRNA analysis workflow is extremely comprehensive and suitable for discovery of novel sncRNAs changes, which may lead to the development of innovative diagnostic and therapeutic tools for TNBC.


Safety of supplemental endplate screws in thoracic pedicle hook fixation.
The AO Universal Spine System thoracic pedicle hook design includes a fixation screw that passes obliquely through the inferior facet into the pedicle to engage in the posterior portion of the superior vertebral body endplate. This endplate screw provides additional purchase at the hook-bone interface. To determine the safety of this fixation system the authors reviewed the operative notes, radiographs, and outcomes of patients who underwent placement of endplate screws. Thirty-six patients (16 male and 20 female patients) who required posterior thoracic instrumentation for spinal deformity (11 cases), neoplasm (15 cases), and traumatic injury (10 cases) were included in this study. One hundred sixty-four endplate screws were placed (mean 4.3/patient) to augment pedicle hooks for posterior thoracic instrumentation. The number of instrumented levels ranged from seven to 16. The positions of the screws in relation to the pedicle, neural foramen, spinal canal, and endplate were evaluated by assessing plain radiographs and computerized tomography scans (10 cases). Eighty-two screws (56%) were in ideal position. Lateral pedicle wall perforation occurred with 51 screws (35%). Three screws violated the medial wall and nine screws violated the superior or inferior walls of the pedicle. There were no clinical sequelae associated with any of the malpositioned screws. Adequate follow-up radiographic data were not available in five patients. The mean follow-up duration was 19.8 months (range 3-61 months). Two patients required revision surgery at 3 months and 18 months, respectively, because of hook/endplate screw displacement. There was also one case of an endplate screw fracture without hook displacement that was discovered during subsequent revision surgery. The remainder of the endplate screws and associated pedicles hooks maintained their original positions. There was no case of spinal cord, nerve root, pulmonary, or vascular injury. The placement of supplemental endplate screws in conjunction with thoracic pedicle hooks can be conducted safely.


Fabry-Perot etalon aperture requirements for direct detection Doppler wind lidar from Earth orbit.
The design of Fabry-Perot etalons for direct detection Doppler wind lidar from a satellite is considered for two direct detection methods, fringe imaging (multichannel) and double edge. The area solid-angle product of the etalon for each technique is derived and shown to be inherently larger, for a given etalon aperture, for the fringe imager than for the double-edge Doppler analyzer. Modeling of the Doppler measurement accuracy of a spaceflight direct detection wind lidar shows that a very large optical aperture, 2 m or more, is necessary. Optical throughput matching to a 2-m collector requires, for the fringe-imaging Doppler analyzer, an etalon with 60 mm aperture, whereas the double-edge technique would require two etalons of 200 mm aperture, or a split-aperture etalon of 400 mm working aperture. Because the two direct detection methods have been shown to have practically identical intrinsic sensitivities (measurement accuracies per unit signal), this difference in etalon dimensions may be a significant selection consideration.


Emerging gene therapy products for RPGR-associated X-linked retinitis pigmentosa.
Mutations in the RPGR gene are responsible for one of the most prevalent and severe types of retinitis pigmentosa. Gene therapy has shown great promise to treat inherited retinal diseases, and currently, four RPGR gene therapy vectors are being evaluated in clinical trials. This manuscript reviews the gene therapy products that are in development for X-linked retinitis pigmentosa caused by mutations in RPGR, and the challenges that scientists and clinicians have faced. The development of a gene therapy product for RPGR-associated retinal degeneration has been a great challenge due to the incomplete understanding of the underlying genetics and mechanism of action of RPGR, and on the other hand, due to the instability of the RPGR gene. Three of the four gene therapy vectors currently in clinical trials include a codon-optimized version of the human RPGR sequence, and the other vector contains a shortened version of the human RPGR. To date, the only Phase I/II results published in a peer-reviewed journal demonstrate a good safety profile and an improvement in the visual field using a codon optimized version of RPGRORF15.


Hemodynamic and metabolic correlates of perinatal white matter injury severity.
Although the spectrum of perinatal white matter injury (WMI) in preterm infants is shifting from cystic encephalomalacia to milder forms of WMI, the factors that contribute to this changing spectrum are unclear. We hypothesized that the variability in WMI quantified by immunohistochemical markers of inflammation could be correlated with the severity of impaired blood oxygen, glucose and lactate. We employed a preterm fetal sheep model of in utero moderate hypoxemia and global severe but not complete cerebral ischemia that reproduces the spectrum of human WMI. Since there is small but measurable residual brain blood flow during occlusion, we sought to determine if the metabolic state of the residual arterial blood was associated with severity of WMI. Near the conclusion of hypoxia-ischemia, we recorded cephalic arterial blood pressure, blood oxygen, glucose and lactate levels. To define the spectrum of WMI, an ordinal WMI rating scale was compared against an unbiased quantitative image analysis protocol that provided continuous histo-pathological outcome measures for astrogliosis and microgliosis derived from the entire white matter. A spectrum of WMI was observed that ranged from diffuse non-necrotic lesions to more severe injury that comprised discrete foci of microscopic or macroscopic necrosis. Residual arterial pressure, oxygen content and blood glucose displayed a significant inverse association with WMI and lactate concentrations were directly related. Elevated glucose levels were the most significantly associated with less severe WMI. Our results suggest that under conditions of hypoxemia and severe cephalic hypotension, WMI severity measured using unbiased immunohistochemical measurements correlated with several physiologic parameters, including glucose, which may be a useful marker of fetal response to hypoxia or provide protection against energy failure and more severe WMI.


Importance of chromophore environment on the near-infrared absorption of polymeric waveguides.
The near-infrared absorption of two chromophore functionalized polymers and combinations of seventeen different guest chromophores in seven different organic polymer matrices were investigated to assess the effect of chromophore structure and environment on absorption. The near-infrared absorption losses were found to be dramatically larger by as much as 2-3 orders of magnitude in polymer matrices than in solution. Furthermore, the absorption of the long-wavelength tail appears to be related to the glass transition temperature of the polymer matrix that contains the chromophore. These results are interpreted in terms of inhomogeneous broadening.


Physiologic actions and molecular expression of the renin-angiotensin system in the diabetic rat.
Clinical, experimental, biochemical, and molecular biologic studies all invoke an important role for the renin-angiotensin system (RAS) in the pathogenesis of diabetic complications. Studies of pharmacologic interruption of the RAS with angiotensin-converting enzyme (ACE) inhibition have implicated this hormonal system in the progression of diabetic nephropathy, both experimentally and clinically. Preliminary evidence also suggests a beneficial effect of angiotensin II (Ang II) receptor antagonists. The relative roles of the systemic vs. intrarenal RAS in the pathogenesis of diabetic nephropathy have recently been evaluated. Though plasma renin is generally low, it is not yet clear whether RAS component processing is normal in diabetes; there may be subtle changes in Ang II metabolism which sustain relatively higher plasma Ang II levels. Furthermore, the intrarenal RAS may not be suppressed. Renal renin levels tend to be disproportionately elevated, as compared to plasma renin values. Renal Ang II levels are normal, and renal mRNAs for RAS components have been variable, though not suppressed. In general, lack of RAS suppression (despite plasma volume and increased exchangeable sodium) may indicate inappropriate activity of the RAS in diabetes. Indeed, disproportionate activity of the intrarenal RAS may be a proximate cause of the observed suppression of the systemic RAS. RAS-mediated injury may occur via stimulation of a number of sclerosing mediators, and there is evidence that hyperglycemia acts synergistically with Ang II to promote cellular injury. Together, these recent investigations lend further support to the notion that the RAS plays an important role in diabetic nephropathy, and are beginning to shed light on the mechanisms of progressive renal injury.


Hydrogenation of titanocene and zirconocene bis(trimethylsilyl)acetylene complexes.
Reactions following the addition of dihydrogen under maximum atmospheric pressure to bis(trimethylsilyl)acetylene (BTMSA) complexes of titanocenes, [(η5-C5H5-nMen)2Ti(η2-BTMSA)] (n = 0, 1, 3, and 4) (1A-1D), and zirconocenes, [(η5-C5H5-nMen)2Zr(η2-BTMSA)] (n = 2-5) (4A-4D), proceeded in diverse ways and, depending on the metal, afforded different products. The former complexes lost, in all cases, their BTMSA ligand via its hydrogenation to bis-1,2-(trimethylsilyl)ethane when reacted at 80 °C for a prolonged reaction time. For n = 0, 1, and 3, the titanocene species formed in situ dimerised via the formation of fulvalene ligands and two bridging hydride ligands, giving known green dimeric titanocenes (2A-2C). For n = 4, a titanocene hydride [(η5-C5HMe4)2TiH] (2D) was formed, similarly to the known [(η5-C5Me5)2TiH] (2E) for n = 5; however, in contrast to this example, 2D in the absence of dihydrogen spontaneously dehydrogenated to the known Ti(iii)-Ti(iii) dehydro-dimer [{Ti(η5-C5HMe4)(μ-η1:η5-C5Me4)}2] (3B). This complex has now been fully characterised via spectroscopic methods, and was shown through EPR spectroscopy to attain an intramolecular electronic triplet state. The zirconocene-BTMSA complexes 4A-4D reacted uniformly with one hydrogen molecule to give Zr(iv) zirconocene hydride alkenyls, [(η5-C5H5-nMen)2ZrH{C(SiMe3)[double bond, length as m-dash]CH(SiMe3)}] (n = 2-5) (5A-5D). These were identified through their 1H and 13C NMR spectra, which show features typical of an agostically bonded proton, [double bond, length as m-dash]CH(SiMe3). Compounds 5A-5D formed equilibria with the BTMSA complexes 4A-4D depending on hydrogen pressure and temperature.


OPA1 (dys)functions.
Mitochondrial morphology varies according to cell type and cellular context from an interconnected filamentous network to isolated dots. This morphological plasticity depends on mitochondrial dynamics, a balance between antagonistic forces of fission and fusion. DRP1 and FIS1 control mitochondrial outer membrane fission and Mitofusins its fusion. This review focuses on OPA1, one of the few known actors of inner membrane dynamics, whose mutations provoke an optic neuropathy. Since its first identification in 2000 the characterization of the functions of OPA1 has made rapid progress thus providing numerous clues to unravel the pathogenetic mechanisms of ADOA-1.


Prognostic utility of heart-type fatty acid binding protein in patients with acute coronary syndromes.
Heart-type fatty acid binding protein (H-FABP) is a cytosolic protein that is released rapidly from the cardiomyocyte in response to myocardial injury. Although it has been investigated as an early marker of acute myocardial infarction, its prognostic utility in acute coronary syndromes has not been established. We measured H-FABP in 2287 patients with acute coronary syndromes from the OPUS-TIMI 16 trial. H-FABP was elevated (> 8 ng/mL) in 332 patients (14.5%). Patients with an elevated H-FABP were more likely to suffer death (hazard ratio [HR], 4.1; 95% CI, 2.6 to 6.5), recurrent myocardial infarction (HR, 1.6; 95% CI, 1.0 to 2.5), congestive heart failure (HR, 4.5; 95% CI, 2.6 to 7.8), or the composite of these end points (HR, 2.6; 95% CI, 1.9 to 3.5) through the 10-month follow-up period. H-FABP predicted the risk of the composite end point both in patients who were troponin I negative (HR, 2.1; 95% CI, 1.3 to 3.4) and in those who were troponin I positive (HR, 3.3; 95% CI, 2.0 to 5.3). In a Cox proportional-hazards model that adjusted for baseline variables, including demographics, clinical characteristics, creatinine clearance, ST deviation, index diagnosis, and troponin I, elevated H-FABP remained a significant predictor of the composite end point (HR, 1.9; 95% CI, 1.3 to 2.7), as well as the individual end points of death (HR, 2.7; 95% CI, 1.5 to 4.9) and CHF (HR, 2.4; 95% CI, 1.2 to 5.0). In a multimarker approach, H-FABP, troponin I, and B-type natriuretic peptide provided complementary information. Elevation of H-FABP is associated with an increased risk of death and major cardiac events in patients presenting across the spectrum of acute coronary syndromes and is independent of other established clinical risk predictors and biomarkers.


Analysis of Heme Iron Coordination in DGCR8: The Heme-Binding Component of the Microprocessor Complex.
DGCR8 is the RNA-binding partner of the nuclease Drosha. Their complex (the "Microprocessor") is essential for processing of long, primary microRNAs (pri-miRNAs) in the nucleus. Binding of heme to DGCR8 is essential for pri-miRNA processing. On the basis of the split Soret ultraviolet-visible (UV-vis) spectrum of ferric DGCR8, bis-thiolate sulfur (cysteinate, Cys(-)) heme iron coordination of DGCR8 heme iron was proposed. We have characterized DGCR8 heme ligation using the Δ276 DGCR8 variant and combined electron paramagnetic resonance (EPR), magnetic circular dichroism (MCD), electron nuclear double resonance, resonance Raman, and electronic absorption spectroscopy. These studies indicate DGCR8 bis-Cys heme iron ligation, with conversion from bis-thiolate (Cys(-)/Cys(-)) axial coordination in ferric DGCR8 to bis-thiol (CysH/CysH) coordination in ferrous DGCR8. Pri-miRNA binding does not perturb ferric DGCR8's optical spectrum, consistent with the axial ligand environment being separated from the substrate-binding site. UV-vis absorption spectra of the Fe(II) and Fe(II)-CO forms indicate discrete species exhibiting peaks with absorption coefficients substantially larger than those for ferric DGCR8 and that previously reported for a ferrous form of DGCR8. Electron-nuclear double resonance spectroscopy data exclude histidine or water as axial ligands for ferric DGCR8 and favor bis-thiolate coordination in this form. UV-vis MCD and near-infrared MCD provide data consistent with this conclusion. UV-vis MCD data for ferrous DGCR8 reveal features consistent with bis-thiol heme iron coordination, and resonance Raman data for the ferrous-CO form are consistent with a thiol ligand trans to the CO. These studies support retention of DGCR8 cysteine coordination upon reduction, a conclusion distinct from those of previous studies of a different ferrous DGCR8 isoform.


Melanotic neuroectodermal tumor in a newborn.
Melanotic neuroectodermal tumor of infancy (MNTI) is an uncommon, fast-growing, pigmented neoplasm of neural crest origin; it primarily affects the maxilla of the infants during the first year of life. Approximately 200 instances of this tumor are reported in the medical literature. We present a case of newborn with MNTI on the left maxillary alveolar ridge. The patient underwent complete surgical resection without maxillectomy. There has been no recurrence in a follow-up period of 1 year. The diagnostic features and management alternatives of MNTI are discussed.


Understanding and exploiting nanoparticles' intimacy with the blood vessel and blood.
While the blood vessel is seldom the target tissue, almost all nanomedicine will interact with blood vessels and blood at some point of time along its life cycle in the human body regardless of their intended destination. Despite its importance, many bionanotechnologists do not feature endothelial cells (ECs), the blood vessel cells, or consider blood effects in their studies. Including blood vessel cells in the study can greatly increase our understanding of the behavior of any given nanomedicine at the tissue of interest or to understand side effects that may occur in vivo. In this review, we will first describe the diversity of EC types found in the human body and their unique behaviors and possibly how these important differences can implicate nanomedicine behavior. Subsequently, we will discuss about the protein corona derived from blood with foci on the physiochemical aspects of nanoparticles (NPs) that dictate the protein corona characteristics. We would also discuss about how NPs characteristics can affect uptake by the endothelium. Subsequently, mechanisms of how NPs could cross the endothelium to access the tissue of interest. Throughout the paper, we will share some novel nanomedicine related ideas and insights that were derived from the understanding of the NPs' interaction with the ECs. This review will inspire more exciting nanotechnologies that had accounted for the complexities of the real human body.


Programmed Cell Death-1 Inhibitor-Induced Type 1 Diabetes Mellitus.
Pembrolizumab (Keytruda; Merck Sharp & Dohme) is a humanized IgG4 monoclonal antibody used in cancer immunotherapy. It targets the programmed cell death-1 (PD-1) receptor, which is important in maintaining self-tolerance. However, immune checkpoint blockade is associated with a risk for immune-related adverse events (irAEs) potentially affecting the endocrine organs. Type 1 diabetes mellitus is a rare irAE of PD-1 inhibitors, occurring in 0.2% of cases. Systematic search of four databases (MEDLINE, Embase, Web of Science, and Cochrane Library) using the search terms "diabetes" or "ketoacidosis" and "pembrolizumab," "nivolumab," "PD-1 inhibitor," or "immunotherapy." Included were articles published in English between 1 January 2012 and 1 January 2018. The search was supplemented by bibliographic searches of the complete reference lists of all included papers. We provide an overview of all published cases (n = 42) of PD-1 inhibitor-induced type 1 diabetes mellitus to date, including a well-characterized case of islet cell antibody and glutamic acid decarboxylase antibody-positive diabetes mellitus, in a patient with a diabetes-prone HLA genotype. She presented with diabetic ketoacidosis during pembrolizumab therapy for a metastatic uveal melanoma. Furthermore, we discuss potential pathogenic mechanisms, clinical presentation, prognostic markers (β-cell antibodies and HLA type), treatment, and a screening protocol. Because the use of immunotherapy will increase, it is essential that all clinicians are aware of diabetic ketoacidosis as a rare and life-threatening side effect of immunotherapy. Blood glucose monitoring during anti-PD-1 therapy is necessary.


Watch the target! Effects in the affective misattribution procedure become weaker (but not eliminated) when participants are motivated to provide accurate responses to the target.
Previous research showed that priming effects in the affective misattribution procedure (AMP) are unaffected by direct warnings to avoid an influence of the primes. The present research examined whether a priming influence is diminished by task procedures that encourage accurate judgments of the targets. Participants were motivated to categorize the affective meaning of nonsense targets accurately by being made to believe that a true word was presented in each trial and by providing feedback on (allegedly) incorrect responses. This condition produced robust priming effects. Priming was however reduced and less reliable relative to more typical AMP conditions in which participants guessed the meaning of openly presented nonsense targets. Affective judgments of nonsense targets were not affected by advance knowledge of the response mapping during the priming phase, which argues against a response-priming explanation of AMP effects. These findings show that affective primes influence evaluative judgments even in conditions in which the motivation to provide accurate responses is high and a priming of motor responses is not possible. Priming effects were however weaker with high accuracy motivation, suggesting that a focus on accurate judgments is an effective strategy to control for an unwanted priming influence in the AMP.


Analysis of interval-censored data from circular migrant and non-migrant sexual partnerships using the EM algorithm.
In epidemiological studies where subjects are seen periodically on follow-up visits, interval-censored data occur naturally. The exact time the change of state (such as HIV seroconversion) occurs is not known exactly, only that it occurred sometime within a specific time interval. Methods of estimation for interval-censored data are readily available when data are independent. However, methods for correlated interval-censored data are not well developed. This paper considers an approach for estimating the parameters when data are interval-censored and correlated within sexual partnerships. We consider the exact event times for interval-censored observations as unobserved data, only known to be between two time points. Dependency induced by sexual partnerships is modelled as frailties assuming a gamma distribution for frailties and an exponential distribution on the time to infection. This formulation facilitates application of the expectation-maximization (EM) algorithm. Maximization process maximizes the standard survival frailty model. Results show high degree of heterogeneity between sexual partnerships. Intervention strategies aimed at combating the spread of HIV and other sexually transmitted infections (STI)s should treat sexual partnerships as social units and fully incorporate the effects of migration in their strategies.


NeuroD Factors Discriminate Mineralocorticoid From Glucocorticoid Receptor DNA Binding in the Male Rat Brain.
In the limbic brain, mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs) both function as receptors for the naturally occurring glucocorticoids (corticosterone/cortisol) but mediate distinct effects on cellular physiology via transcriptional mechanisms. The transcriptional basis for specificity of these MR- vs GR-mediated effects is unknown. To address this conundrum, we have identified the extent of MR/GR DNA-binding selectivity in the rat hippocampus using chromatin immunoprecipitation followed by sequencing. We found 918 and 1450 nonoverlapping binding sites for MR and GR, respectively. Furthermore, 475 loci were co-occupied by MR and GR. De novo motif analysis resulted in a similar binding motif for both receptors at 100% of the target loci, which matched the known glucocorticoid response element (GRE). In addition, the Atoh/NeuroD consensus sequence was found in co-occurrence with all MR-specific binding sites but was absent for GR-specific or MR-GR overlapping sites. Basic helix-loop-helix family members Neurod1, Neurod2, and Neurod6 showed hippocampal expression and were hypothesized to bind the Atoh motif. Neurod2 was detected at rat hippocampal MR binding sites but not at GR-exclusive sites. All three NeuroD transcription factors acted as DNA-binding-dependent coactivators for both MR and GR in reporter assays in heterologous HEK293 cells, likely via indirect interactions with the receptors. In conclusion, a NeuroD family member binding to an additional motif near the GRE seems to drive specificity for MR over GR binding at hippocampal binding sites.


Hippocampal volume reduction in 22q11.2 deletion syndrome.
Hippocampal volume reduction and decreased memory skills form a characteristic neurofunctional alteration observed in schizophrenia. Individuals affected with 22q11.2 deletion syndrome (22q11DS), while exhibiting memory deficits throughout development, are also at high risk for developing schizophrenia. The present study sought to investigate hippocampal volume reduction as separate of global grey matter reduction in a large, independent sample of individuals with 22q11DS. Volumetric data from structural magnetic resonance imaging was obtained for 43 individuals affected with 22q11DS, aged 6-39 years of age, as well as for 40 healthy individuals matched for age and gender. Drawing of the amygdala was included to enhance the delineation of the hippocampus, and circumscription of both the amygdala and the hippocampus were executed using an increased resolution matrix. After controlling for total grey volume reductions observed in affected individuals, a significant decrease in hippocampus volume was observed in the 22q11DS group, driven by significant bilateral volumetric reduction of the body of the hippocampus. These results are discussed in reference to memory and cerebral alterations already reported in 22q11DS. Further, the specific implications of hippocampus body volume reduction are outlined in light of its anatomical relationships and its function in memory. Finally, reduction of hippocampal volume in 22q11DS is examined in the context of psychiatric risk status associated to the deletion.


Association of Functional, Cognitive, and Psychological Measures With 1-Year Mortality in Patients Undergoing Major Surgery.
More older adults are undergoing major surgery despite the greater risk of postoperative mortality. Although measures, such as functional, cognitive, and psychological status, are known to be crucial components of health in older persons, they are not often used in assessing the risk of adverse postoperative outcomes in older adults. To determine the association between measures of physical, cognitive, and psychological function and 1-year mortality in older adults after major surgery. Retrospective analysis of a prospective cohort study of participants 66 years or older who were enrolled in the nationally representative Health and Retirement Study and underwent 1 of 3 types of major surgery. Major surgery, including abdominal aortic aneurysm repair, coronary artery bypass graft, and colectomy. Our outcome was mortality within 1 year of major surgery. Our primary associated factors included functional, cognitive, and psychological factors: dependence in activities of daily living (ADL), dependence in instrumental ADL, inability to walk several blocks, cognitive status, and presence of depression. We adjusted for other demographic and clinical predictors. Of 1341 participants, the mean (SD) participant age was 76 (6) years, 737 (55%) were women, 99 (7%) underwent abdominal aortic aneurysm repair, 686 (51%) coronary artery bypass graft, and 556 (42%) colectomy; 223 (17%) died within 1 year of their operation. After adjusting for age, comorbidity burden, surgical type, sex, race/ethnicity, wealth, income, and education, the following measures were significantly associated with 1-year mortality: more than 1 ADL dependence (29% vs 13%; adjusted hazard ratio [aHR], 2.76; P = .001), more than 1 instrumental ADL dependence (21% vs 14%; aHR, 1.32; P = .05), the inability to walk several blocks (17% vs 11%; aHR, 1.64; P = .01), dementia (21% vs 12%; aHR, 1.91; P = .03), and depression (19% vs 12%; aHR, 1.72; P = .01). The risk of 1-year mortality increased within the increasing risk factors present (0 factors: 10.0%; 1 factor: 16.2%; 2 factors: 27.8%). In this older adult cohort, 223 participants (17%) who underwent major surgery died within 1 year and poor function, cognition, and psychological well-being were significantly associated with mortality. Measures in function, cognition, and psychological well-being need to be incorporated into the preoperative assessment to enhance surgical decision-making and patient counseling.