Surface, Interface, and Temperature Effects on the Phase Separation and Nanoparticle Self Assembly of Bi-Metallic Ni0.5Ag0.5: A Molecular Dynamics Study.
Classical molecular dynamics (MD) simulations were used to investigate how free surfaces, as well as supporting substrates, affect phase separation in a NiAg alloy. Bulk samples, droplets, and droplets deposited on a graphene substrate were investigated at temperatures that spanned regions of interest in the bulk NiAg phase diagram, i.e., miscible and immiscible liquid, liquid-crystal, and crystal-crystal regions. Using MD simulations to cool down a bulk sample from 3000 K to 800 K, it was found that phase separation below 2400 K takes place in agreement with the phase diagram. When free surface effects were introduced, phase separation was accompanied by a core-shell transformation: spherical droplets created from the bulk samples became core-shell nanoparticles with a shell made mostly of Ag atoms and a core made of Ni atoms. When such droplets were deposited on a graphene substrate, the phase separation was accompanied by Ni layering at the graphene interface and Ag at the vacuum interface. Thus, it should be possible to create NiAg core-shell and layer-like nanostructures by quenching liquid NiAg samples on tailored substrates. Furthermore, interesting bimetallic nanoparticle morphologies might be tuned via control of the surface and interface energies and chemical instabilities of the system.


Seamless gene tagging by endonuclease-driven homologous recombination.
Gene tagging facilitates systematic genomic and proteomic analyses but chromosomal tagging typically disrupts gene regulatory sequences. Here we describe a seamless gene tagging approach that preserves endogenous gene regulation and is potentially applicable in any species with efficient DNA double-strand break repair by homologous recombination. We implement seamless tagging in Saccharomyces cerevisiae and demonstrate its application for protein tagging while preserving simultaneously upstream and downstream gene regulatory elements. Seamless tagging is compatible with high-throughput strain construction using synthetic genetic arrays (SGA), enables functional analysis of transcription antisense to open reading frames and should facilitate systematic and minimally-invasive analysis of gene functions.


The stem cell-specific long noncoding RNA HOXA10-AS in the pathogenesis of KMT2A-rearranged leukemia.
HOX genes are highly conserved, and their precisely controlled expression is crucial for normal hematopoiesis. Accordingly, deregulation of HOX genes can cause leukemia. However, despite of intensive research on the coding HOX genes, the role of the numerous long noncoding RNAs (lncRNAs) within the HOX clusters during hematopoiesis and their contribution to leukemogenesis are incompletely understood. Here, we show that the lncRNA HOXA10-AS, located antisense to HOXA10 and mir-196b in the HOXA cluster, is highly expressed in hematopoietic stem cells (HSCs) as well as in KMT2A-rearranged and NPM1 mutated acute myeloid leukemias (AMLs). Using short hairpin RNA- and locked nucleic acid-conjugated chimeric antisense oligonucleotide (LNA-GapmeR)-mediated HOXA10-AS-knockdown and CRISPR/Cas9-mediated excision in vitro, we demonstrate that HOXA10-AS acts as an oncogene in KMT2A-rearranged AML. Moreover, HOXA10-AS knockdown severely impairs the leukemic growth of KMT2A-rearranged patient-derived xenografts in vivo, while high HOXA10-AS expression can serve as a marker of poor prognosis in AML patients. Lentiviral expression of HOXA10-AS blocks normal monocytic differentiation of human CD34+ hematopoietic stem and progenitor cells. Mechanistically, we show that HOXA10-AS localizes in the cytoplasm and acts in trans to induce NF-κB target genes. In total, our data imply that the normally HSC-specific HOXA10-AS is an oncogenic lncRNA in KMT2A-r AML. Thus, it may also represent a potential therapeutic target in KMT2A-rearranged AML.


A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks.
Finding the components of cellular circuits and determining their functions systematically remains a major challenge in mammalian cells. Here, we introduced genome-wide pooled CRISPR-Cas9 libraries into dendritic cells (DCs) to identify genes that control the induction of tumor necrosis factor (Tnf) by bacterial lipopolysaccharide (LPS), a key process in the host response to pathogens, mediated by the Tlr4 pathway. We found many of the known regulators of Tlr4 signaling, as well as dozens of previously unknown candidates that we validated. By measuring protein markers and mRNA profiles in DCs that are deficient in known or candidate genes, we classified the genes into three functional modules with distinct effects on the canonical responses to LPS and highlighted functions for the PAF complex and oligosaccharyltransferase (OST) complex. Our findings uncover new facets of innate immune circuits in primary cells and provide a genetic approach for dissection of mammalian cell circuits.


The role of insulin and IGF-1 signaling in longevity.
There are many theories of aging and parameters that influence lifespan, including genetic instability, telomerase activity and oxidative stress. The role of caloric restriction, metabolism and insulin and insulin-like growth factor-1 signaling in the process of aging is especially well conserved throughout evolution. These latter factors interact with each other, the former factors and histone deacetylases of the SIR family in a complex interaction to influence lifespan.


Human embryonic stem cells: biology and clinical implications.
Embryonic stem cells (ESCs) are derived from the inner cell mass of the preimplantation stage embryo and are capable of prolonged symmetrical self-renewal (both daughter cells remain escs) as well as differentiation into derivatives of all three embryonic germ layers. ESCs therefore have the potential to provide an unlimited supply of transplantable cells to replace or regenerate damaged or diseased tissues. However, several barriers must be overcome before successful clinical trials are possible: for example, pure populations of the desired cell type need to be selected and expanded in clinically relevant numbers, and a method for preventing immunological rejection of the transplanted cells without long-term immunosuppressive therapy is also required. In this review, we highlight recent developments in human ESC derivation and expansion, outline current understanding of the signalling pathways underlying stem cell renewal, and discuss challenging problems related to the selective differentiation and immune properties of human ESCs.


Mechanism of activation and selectivity in a ligand-gated ion channel: structural and functional studies of GluR2 and quisqualate.
Glutamate is the major excitatory neurotransmitter in the mammalian brain. The (S)-2-amino-3-(3-hydroxy-5-methyl-4-isoxazole)propionic acid (AMPA)-subtype glutamate receptor, a ligand-gated ion channel, mediates most of the fast excitatory synaptic transmission in the mammalian central nervous system. Here we present electrophysiological, biochemical, and crystallographic data on the interactions between quisqualate and the GluR2 receptor ion channel and its corresponding ligand binding core. Quisqualate is a high-affinity, full agonist which like AMPA and glutamate elicits maximum peak current responses, and stabilizes the ligand binding core in a fully closed conformation, reinforcing the concept that full agonists produce similar conformational changes [Armstrong, N., and Gouaux, E. (2000) Neuron 28, 165-181]. Nevertheless, the mechanism of quisqualate binding is different from that of AMPA but similar to that of glutamate, illustrating that quisqualate is a faithful glutamate analogue. A detailed comparison of the three agonist complexes reveals distinct binding mechanisms, particularly in the region of a hydrophobic pocket that is proximal to the anionic gamma-substituents, and demonstrates the importance of agonist-water-receptor interactions. The hydrophobic pocket, which is predicted to vary in chemical character between receptor subtypes, probably plays an important role in determining receptor subtype specificity.


Proteomic analysis of shoot tissue during photoperiod induced growth cessation in V. riparia Michx. grapevines.
Growth cessation, cold acclimation and dormancy induction in grapevines and other woody perennial plants native to temperate continental climates is frequently triggered by short photoperiods. The early induction of these processes by photoperiod promotes winter survival of grapevines in cold temperate zones. Examining the molecular processes, in particular the proteomic changes in the shoot, will provide greater insight into the signaling cascade that initiates growth cessation and dormancy induction. To begin understanding transduction of the photoperiod signal, Vitis riparia Michx. grapevines that had grown for 35 days in long photoperiod (long day, LD, 15 h) were subjected to either a continued LD or a short photoperiod (short day, SD, 13 h) treatment. Shoot tips (4-node shoot terminals) were collected from each treatment at 7 and 28 days of LD and SD for proteomic analysis via two-dimensional (2D) gel electrophoresis. Protein profiles were characterized in V. riparia shoot tips during active growth or SD induced growth cessation to examine physiological alterations in response to differential photoperiod treatments. A total of 1054 protein spots were present on the 2D gels. Among the 1054 proteins, 216 showed differential abundance between LD and SD (>/= two-fold ratio, p-value </= 0.05). After 7 days, 39 protein spots were more abundant in LD and 30 were more abundant in SD. After 28 days, 93 protein spots were more abundant in LD and 54 were more abundant in SD. MS/MS spectrometry was performed to determine the functions of the differentially abundant proteins. The proteomics analysis uncovered a portion of the signal transduction involved in V. riparia grapevine growth cessation and dormancy induction. Different enzymes of the Calvin-Benson cycle and glutamate synthetase isoforms were more abundant either in LD or SD treatments. In LD tissues the significantly differentially more abundant proteins included flavonoid biosynthesis and polyphenol enzymes, cinnamyl alcohol dehydrogenase, and TCP-1 complexes. In the SD tissue photorespiratory proteins were more abundant than in the LD. The significantly differentially more abundant proteins in SD were involved in ascorbate biosynthesis, photosystem II and photosystem I subunits, light harvesting complexes, and carboxylation enzymes.


Atypical antipsychotic metabolism and excretion.
The metabolic/biotransformation pathways of atypical antipsychotics (aripiprazole, clozapine, iloperidone, olanzapine, paliperidone, quetiapine, risperidone, and ziprasidone) have been characterized and reviewed. However, comparisons of excretory pathways remain unexplored. To analyze the excretion profile of atypical antipsychotic agents and compare the overall magnitude of metabolism (changed vs. unchanged drug) and route of excretion (feces vs. urine). Secondary objectives include providing: 1) dosing information in hepatic and renal impairment, and 2) context of the specific enzymes and pathways involved in each agents' biotransformation. Published literature and each manufacturer's radiolabeled drug absorption, distribution, metabolism and excretion data and U.S. prescribing information were reviewed. With the exception of paliperidone, atypical antipsychotics undergo extensive metabolism (i.e., <or=50% of dose recovered unchanged). Quetiapine undergoes the greatest overall metabolism (<1% of the dose recovered unchanged) and paliperidone the least (59% recovered unchanged in the urine). Between-agent differences exist in the extent of cytochrome P450 (CYP450) metabolism and the specific isozymes involved. After administration of a radioactive dose, fecal elimination of unchanged drug plus metabolites ranged from 11% (paliperidone) to 71% (ziprasidone) and renal elimination ranged from 21% (ziprasidone) to 80% (paliperidone). Understanding the differences in the elimination profiles of atypical antipsychotics agents may permit better-informed drug and dose selection in special populations such as those with comorbid conditions (e.g. hepatitis, diabetes, end-stage renal disease) or pharmacogenetic variability; or at risk for drug-drug interactions. The use of patient tailored drug and dose-selection may result in greater treatment efficacy and a reduction in adverse events.


[Neuropeptides in dermatologic therapy].
The presence of neuropeptides and their specific receptors has been detected in the skin and the epithelial tissues. They are involved in innervation, immunomodulation, glandular secretion, control of cellular proliferation and regulation of blood flow. The fact that they act in so many different ways means that neuropeptides and their agonists and antagonists are now being regarded as potential therapeutic agents in dermatologic diseases. Among the substances which act as antagonists, particular attention should be paid to capsaicin, which has therapeutic potential for three types of indication: peripheral neurologic pain, affections with a neurogenic inflammatory component and pruriginous dermatosis; peptide T with therapeutic potential for psoriasis; and spantide, which might prove useful in dermatoses related to substance P. The influence of topical corticosteroids on the mechanism of action of neuropeptides can explain its efficacy in the treatment of many dermatoses. Among the agonists, the possibility of taking advantage of the vasodilatory activity of the calcitonin gene-related peptide is being considered in Raynaud's disease and erectile disfunction of the penis; and the immunomodulatory and anti-inflammatory action of the alpha-melanocyte stimulating hormone is being studied as a potential means of controlling inflammatory dermatoses of immunological origin.


Cone-beam composite-circling scan and exact image reconstruction for a quasi-short object.
Here we propose a cone-beam composite-circling mode to solve the quasi-short object problem, which is to reconstruct a short portion of a long object from longitudinally truncated cone-beam data involving the short object. In contrast to the saddle curve cone-beam scanning, the proposed scanning mode requires that the X-ray focal spot undergoes a circular motion in a plane facing the short object, while the X-ray source is rotated in the gantry main plane. Because of the symmetry of the proposed mechanical rotations and the compatibility with the physiological conditions, this new mode has significant advantages over the saddle curve from perspectives of both engineering implementation and clinical applications. As a feasibility study, a backprojection filtration (BPF) algorithm is developed to reconstruct images from data collected along a composite-circling trajectory. The initial simulation results demonstrate the correctness of the proposed exact reconstruction method and the merits of the proposed mode.


Effects of Laser Photobiomodulation and Ozone Therapy on Palatal Epithelial Wound Healing and Patient Morbidity.
The aim of this study was to compare the effects of laser photobiomodulation (PBM) and topical ozone therapy on the reepithelialization of palatal donor site wounds through clinical assessment and computer-aided image analysis and to assess the patient morbidity following free gingival graft (FGG) surgeries. Thirty-six patients requiring FGG were randomly allocated into three groups: laser group (n = 12), ozone group (n = 12), and control group (n = 12). Epithelialization was evaluated by applying 3% hydrogen peroxide (H2O2) to the wound area and also measured by using digital image analysis (ImageJ). Bland-Altman plots were used for assessing agreement between H2O2 and ImageJ measurements. Parameters in relation to patient morbidity were assessed by using visual analog scale (VAS) on the first 3, 7, 14, and 30 days postoperatively. At day 14, statistically significant smaller wounds were observed with digital image analysis in the ozone group as compared with the control group (p = 0.034). However, intergroup comparison of the remaining wound area evaluated by the clinician using the H2O2 method did not reveal any significant differences (p > 0.05). Nonetheless, according to Bland-Altman analysis, the lower and upper limits showed a moderate agreement between the two measurement methods. The mean VAS sores exhibiting postoperative discomfort was observed to be significantly higher in the control group compared with the laser group (p = 0.002) and ozone group (p < 0.001) at day 7. Adjunctive ozone therapy could have a significantly beneficial effect on the acceleration of palatal wound healing following FGG procedures. Both PBM and ozone treatment modalities reduced postoperative discomfort as compared with spontaneous healing.


Roles of Cyclooxygenase, Prostaglandin E2 and EP Receptors in Mucosal Protection and Ulcer Healing in the Gastrointestinal Tract.
Endogenous prostaglandins (PGs), produced from arachidonic acid by the two isoforms of cyclooxygenase (COX), play a pivotal role in maintaining mucosal integrity by modulating various functions of the gastrointestinal (GI) tract, and PGE2 is most effective in these actions. The PGE2 receptor is classified into 4 specific G-protein coupled subtypes, EP1-EP4, and their distribution accounts for the multiple effects of this prostanoid. PGE2 prevents acid-reflux esophagitis and indomethacin-induced gastric lesions through EP1 receptors, while endogenous PGs protect the stomach against cold restraint stress mediated by mainly PGI2/IP receptors and partly EP4 receptors. PGE2 also exhibits a protective effect against acid-induced duodenal damage and indomethacin-induced small intestinal lesions mediated by EP3/EP4 receptors; these effects in the stomach, duodenum, or small intestine are associated functionally with inhibition of gastric contraction (EP1), stimulation of duodenal HCO3 - secretion (EP3/EP4), or suppression of bacterial invasion due to the inhibition of intestinal motility (EP4) as well as stimulation of mucus secretion (EP3/EP4), respectively. PGE2 also prevents ischemiainduced enteritis and dextran sulfate sodium-induced colitis mediated by EP4 receptors, and the protective mechanisms may be related to the stimulation of mucus secretion and the down-regulation of immune response, respectively. Furthermore, PGE2 shows a healing-promoting effect on gastric ulcers and small intestinal lesions through the up-regulated expression of vascular endothelial growth factor (VEGF) and stimulation of angiogenesis via the activation of EP4 receptors. Finally, COX-1 is mainly responsible for the production of endogenous PGs involved in mucosal protection, while COX-2 is mainly responsible for those involved in the healing of gastric ulcers or small intestinal lesions. These findings contribute to future development of new strategies for the treatment of GI diseases.


Development of a fatal noncompressible truncal hemorrhage model with combined hepatic and portal venous injury in normothermic normovolemic swine.
Noncompressible truncal hemorrhage and brain injury currently account for most early mortality of warfighters on the battlefield. There is no effective treatment for noncompressible truncal hemorrhage, other than rapid evacuation to a surgical facility. The availability of an effective field treatment for noncompressible truncal hemorrhage could increase the number of warfighters salvaged from this frequently-lethal scenario. Our intent was to develop a porcine model of noncompressible truncal hemorrhage with a ∼ 50% one-hour mortality so that we could develop new treatments for this difficult problem. Normovolemic normothermic domestic swine (barrows, 3 months old, 34-36 kg) underwent one of three injury types through a midline incision: 1) central stellate injury (N = 6); 2) excision of a portal vein branch distal to the main PV trunk (N = 6); or 3) hemi-transection of the left lateral lobe of the liver at its base (N = 10). The one-hour mortality of these injuries was 0, 82, and 40%, respectively; the final mean arterial pressure was 65, 24, and 30 mm Hg, respectively; and the final hemoglobin was 8.3, 2.3, and 3.6 g/dL, respectively. Hemi-transection of the left lateral lobe of the liver appeared to target our desired mortality rate better than the other injury mechanisms.


Is air transport faster? A comparison of air versus ground transport times for interfacility transfers in a regional referral system.
Helicopter transport is often considered in an effort to minimize time to critical interventions, such as cardiac catheterization or arterial thrombolysis in stroke patients. However, for interfacility transports, the extra time considerations for helicopter preparation, takeoff, and time to get to the transferring hospital may not offset the slower transport times for local ground ambulances. The purpose of this study was to compare transport times for helicopter with traditional ground ambulance for interfacility transfers within a regional referral system. All patients transported from an outside hospital to the intensive care unit of the University of Wisconsin were eligible for this study. Equal numbers of patients transferred by ground and by helicopter from each facility were sequentially selected. The following intervals were compared: time from call to dispatch, time from dispatch to arrival at the referring hospital, time at the referring hospital, transport time to the receiving hospital, and total transport time. One hundred forty-five patients were included in this study, transferred from 20 hospitals within the UW referral system. Dispatch times and time at the referring hospital were location independent, and each was shorter for ground transport. Ground dispatch times were 5+/-6 minutes, whereas for helicopter transport dispatch times were 17+/-8 (P<.001). Times at the referral hospital were on average longer for helicopter transport (31+/-11 minutes for air ambulance, 25+/-13 for ground; P=.008). Other intervals were location dependent. Arrivals were much more variable for ground transport, reflecting the fact that private ambulance services may have to travel some distance to reach the referring hospital (helicopter 18+/-8 minutes, ground 19+/-18 minutes). As expected, transport time from the referring hospital was shorter for helicopter transport. From each of the hospitals, average helicopter transport was as fast as the best ground transport. Helicopter transport was faster than ground transport for interfacility transfer of patients from all hospitals studied in our referral system. Under optimal dispatch and transport conditions, the time difference from several hospitals was minimal. For stable patients for whom the only issue is time to critical procedure, it may be reasonable for those hospitals to try ground transport first if timely service is available to transport in that way. Even for those hospitals, helicopter transport should be considered for these patients if ground transport is not optimally available, as well as for patients in whom minimizing time outside of the hospital is a significant consideration, or when transport of such patients impacts emergency medical services availability to the community for a significant time.


Outcomes of Casualties Without Airway Trauma Undergoing Prehospital Airway Interventions: A Department of Defense Trauma Registry Study.
Airway obstruction is the second leading cause of preventable death on the battlefield. Most airway obstruction occurs secondary to traumatic disruptions of the airway anatomical structures. However, casualties may require airway interventions for other indications (e.g., depressed mental status). We describe casualties undergoing airway intervention in the prehospital, combat setting without apparent upper airway trauma. We used a series of emergency department procedure codes to identify patients within the Department of Defense Trauma Registry (DODTR) from January 2007 to August 2016. This is a subgroup analysis of those patients with a documented prehospital airway intervention and no apparent airway trauma as defined by abbreviated injury scale of 0 for body regions 1 (head/neck) and 2 (face). Our predefined search codes captured 28222 DODTR subjects of whom 409 (1.4%) met criteria for study inclusion. Subjects included members of host nation forces (34%) and civilians (30%). Most subjects sustained injuries in Afghanistan (82%). Explosive (57%) and gunshot wounds (36%) were the most frequent mechanisms of injury. Median injury severity scores were 17. The most common anatomical locations of injuries for included subjects included extremities (53%) and thorax (29%). A majority of subjects underwent intubation (89%); comparatively few casualties underwent placement of a nasopharyngeal airway (2%) or supraglottic airway (2%). The proportion of subjects surviving to hospital discharge was 80% and was highest among subjects undergoing intubation (82%). In this subgroup analysis of casualties without apparent upper airway trauma, survival rates were lower when compared to our previous report. Higher quality data are necessary to better understand the resuscitation needs of this critically ill subset of combat casualties.


A comparison of costs of universal versus targeted lead screening for young children.
The purposes of this study were to compare universal blood lead screening for young children versus targeting by a risk assessment questionnaire and to examine the cost implications of each approach. Costs reflect the total number of blood tests required and cost of specimen collection, handling, and testing per elevated case. The setting included the metropolitan areas of Minneapolis and St. Paul, Minnesota. Children (N=9603) from 17 community organizations had blood tests. In addition, each child's parent or guardian completed a questionnaire assessing potential risk for lead poisoning. Four different screening approaches are presented. Each screening approach is presented with associated costs of overall screening and cost per child identified at blood levels of >/=10 microg/dl (N=1140) and >/=15 microg/dl (N=317). Based on the screening strategy selected and an estimate of $17 per blood test, total screening costs ranged from $91,596 to $165,945. The cost per child identified with elevated lead levels ranged from $361 to $523 at >/=15 and $105 to $146 at >/=10. Nine to 13% of children would not have been detected by policies other than universal screening. A geographically based approach was able to detect 90% of children with elevated blood levels at two-thirds the cost of universal screening. Blood tests would be taken for all children living within city limits. Those residing elsewhere would be tested only if answers to questionnaire items pertaining to age of housing, prior history of lead poisoning, or eating paint chips indicated risk. The new CDC guidelines suggest that screening be based on an assessment of housing, population demographics, and community risk and resources. This paper presents such an assessment.


An integrated model of environmental effects on growth, carbohydrate balance, and mortality of Pinus ponderosa forests in the southern Rocky Mountains.
Climate-induced tree mortality is an increasing concern for forest managers around the world. We used a coupled hydrologic and ecosystem carbon cycling model to assess temperature and precipitation impacts on productivity and survival of ponderosa pine (Pinus ponderosa). Model predictions were evaluated using observations of productivity and survival for three ponderosa pine stands located across an 800 m elevation gradient in the southern Rocky Mountains, USA, during a 10-year period that ended in a severe drought and extensive tree mortality at the lowest elevation site. We demonstrate the utility of a relatively simple representation of declines in non-structural carbohydrate (NSC) as an approach for estimating patterns of ponderosa pine vulnerability to drought and the likelihood of survival along an elevation gradient. We assess the sensitivity of simulated net primary production, NSC storage dynamics, and mortality to site climate and soil characteristics as well as uncertainty in the allocation of carbon to the NSC pool. For a fairly wide set of assumptions, the model estimates captured elevational gradients and temporal patterns in growth and biomass. Model results that best predict mortality risk also yield productivity, leaf area, and biomass estimates that are qualitatively consistent with observations across the sites. Using this constrained set of parameters, we found that productivity and likelihood of survival were equally dependent on elevation-driven variation in temperature and precipitation. Our results demonstrate the potential for a coupled hydrology-ecosystem carbon cycling model that includes a simple model of NSC dynamics to predict drought-related mortality. Given that increases in temperature and in the frequency and severity of drought are predicted for a broad range of ponderosa pine and other western North America conifer forest habitats, the model potentially has broad utility for assessing ecosystem vulnerabilities.


Developments in the treatment of candidiasis: more choices and new challenges.
The incidence of oesophageal candidiasis, candidaemia and disseminated candidiasis has increased dramatically. In addition to the amphotericin B formulations and fluconazole, the echinocandins anidulafungin, caspofungin and micafungin and the newer triazoles posaconazole and voriconazole are in the last stages of development and are becoming available for the management of candidiasis. This review presents these new agents and addresses their role in the treatment of candidiasis. All new antifungal agents exhibit potent activity against Candida spp. and echinocandins are fungicidal against most Candida spp. but appear to be less potent against certain species, such as Candida parapsilosis and C. guilliermondii. Systemic antifungal therapy can now be individualised based on the severity of the infection, comorbid conditions and the Candida spp. causing the infection. Studies are needed to investigate the possible development of resistance and the efficacy of these antifungal agents against the more resistant Candida spp.


Cyclovergence evoked by up-down acceleration along longitudinal axis in humans.
We present results of a study of torsional eye movements evoked by earth-vertical accelerations along the subject's longitudinal axis. The earth-vertical stimulus leads to a gravito-inertial acceleration vector that changes magnitude but not direction. It can therefore be viewed as a dynamic change of the gravity level. Up-down oscillations induced relatively symmetric cyclovergence (0.6-2.2 degrees peak-to-peak). Eyes intorted/extorted for higher/lower effective gravity. The phase of this modulation was small relative to chair acceleration. We contrast this behaviour to the dynamics of cycloversion in response to interaural acceleration, which shows a considerably larger phase lag. This strikingly different dynamics suggest a different processing of otolith signals during interaural and longitudinal stimulation.


Tocotrienol rich fraction supplementation improved lipid profile and oxidative status in healthy older adults: A randomized controlled study.
Vitamin E supplements containing tocotrienols are now being recommended for optimum health but its effects are scarcely known. The objective was to determine the effects of Tocotrienol Rich Fraction (TRF) supplementation on lipid profile and oxidative status in healthy older individuals at a dose of 160 mg/day for 6 months. Sixty-two subjects were recruited from two age groups: 35-49 years (n = 31) and above 50 years (n = 31), and randomly assigned to receive either TRF or placebo capsules for six months. Blood samples were obtained at 0, 3rd and 6th months. HDL-cholesterol in the TRF-supplemented group was elevated after 6 months (p < 0.01). Protein carbonyl contents were markedly decreased (p < 0.001), whereas AGE levels were lowered in the > 50 year-old group (p < 0.05). Plasma levels of total vitamin E particularly tocopherols were significantly increased in the TRF-supplemented group after 3 months (p < 0.01). Plasma total tocotrienols were only increased in the > 50 year-old group after receiving 6 months of TRF supplementation. Changes in enzyme activities were only observed in the > 50 year-old group. SOD activity was decreased after 3 (p < 0.05) and 6 (p < 0.05) months of TRF supplementation whereas CAT activity was decreased after 3 (p < 0.01) and 6 (p < 0.05) months in the placebo group. GPx activity was increased at 6 months for both treatment and placebo groups (p < 0.05). The observed improvement of plasma cholesterol, AGE and antioxidant vitamin levels as well as the reduced protein damage may indicate a restoration of redox balance after TRF supplementation, particularly in individuals over 50 years of age.


Measuring correlated electronic and vibrational spectral dynamics using line shapes in two-dimensional electronic-vibrational spectroscopy.
Two-dimensional electronic-vibrational (2DEV) spectroscopy is an experimental technique that shows great promise in its ability to provide detailed information concerning the interactions between the electronic and vibrational degrees of freedom in molecular systems. The physical quantities 2DEV is particularly suited for measuring have not yet been fully determined, nor how these effects manifest in the spectra. In this work, we investigate the use of the center line slope of a peak in a 2DEV spectrum as a measure of both the dynamic and static correlations between the electronic and vibrational states of a dye molecule in solution. We show how this center line slope is directly related to the solvation correlation function for the vibrational degrees of freedom. We also demonstrate how the strength with which the vibration on the electronic excited state couples to its bath can be extracted from a set of 2DEV spectra. These analytical techniques are then applied to experimental data from the laser dye 3,3'-diethylthiatricarbocyanine iodide in deuterated chloroform, where we determine the lifetime of the correlation between the electronic transition frequency and the transition frequency for the backbone C = C stretch mode to be ∼1.7 ps. Furthermore, we find that on the electronic excited state, this mode couples to the bath ∼1.5 times more strongly than on the electronic ground state.


MRI-determined total volumes of visceral and subcutaneous abdominal and trunk adipose tissue are differentially and sex-dependently associated with patterns of estimated usual nutrient intake in a northern German population.
Visceral (VAT) and subcutaneous abdominal (SAAT) and trunk (STRAT) adipose tissue (AT) have been suggested to be differentially influenced by diet. We investigated whether and to what extent usual patterns of nutrient intake are associated with VAT, SAAT, and STRAT compared with nondietary predictors in northern German adults (n = 583). AT volumes were quantified by magnetic resonance imaging. Nutrient intake was estimated by a 112-item food-frequency questionnaire linked to the German Food Code and Nutrient Database. Exploratory nutrient patterns were derived by principal components analysis (PCA) and partial least-squares regression (PLS) of 87 nutrients. Cross-sectional associations between nutrient patterns, single nutrients, or total energy intake and AT compartments were analyzed by multiple linear regression. Next to sex and age, respectively, which were important nondietary predictors and accounted for more of the variation in VAT (∼13% and ∼4%) than in SAAT or STRAT (both 4-7% and <1%), variation in VAT (16.8% or 17.6%) was explained to a greater extent by 9 or 2 nutrient patterns derived by principal components analysis or partial least-squares regression, respectively, than was variation in SAAT (10.6% or 8.2%) or STRAT (11.5% or 8.6%). Whereas VAT (16.6%) was primarily explained by nutrient quality, SAAT (6.9%) and STRAT (7.4%) were mainly explained by total energy intake. VAT was positively associated with nutrients characteristic of animal (except for dairy) products, including arachidonic acid (standardized β: 0.25; 95% CI: 0.15, 0.34; P < 0.0001), but negatively with dietary fiber, including polypentoses (standardized β: -0.17; 95% CI: -0.24, -0.09; P < 0.0001), and nutrients found in milk. The direction and strength of many associations, however, depended strongly on sex and adjustment for BMI. VAT may be particularly associated with sex-specific interplays of nutrients found in animal products and fiber, whereas SAAT and STRAT are associated with total energy intake.


Clinical Practice Patterns of Radiotherapy in Patients with Hepatocellular Carcinoma: A Korean Radiation Oncology Group Study (KROG 14-07).
The aim of this study was to examine patterns of radiotherapy (RT) in Korean patients with hepatocellular carcinoma (HCC) according to the evolving guideline for HCC established by the Korean Liver Cancer Study Group-National Cancer Center (KLCSG-NCC). We reviewed 765 patients with HCC who were treated with RT between January 2011 and December 2012 in 12 institutions. The median follow-up period was 13.3 months (range, 0.2 to 51.7 months). Compared with previous data between 2004 and 2005, the use of RT as a first treatment has increased (9.0% vs. 40.8%). Increased application of intensity-modulated RT resulted in an increase in radiation dose (fractional dose, 1.8 Gy vs. 2.5 Gy; biologically effective dose, 53.1 Gy10 vs. 56.3 Gy10). Median overall survival was 16.2 months, which is longer than that reported in previous data (12 months). In subgroup analysis, treatments were significantly different according to stage (p < 0.001). Stereotactic body RT was used in patients with early HCC, and most patients with advanced stage were treated with three-dimensional conformal RT. Based on the evolving KLCSG-NCC practice guideline for HCC, clinical practice patterns of RT have changed. Although RT is still used mainly in advanced HCC, the number of patients with good performance status who were treated with RT as a first treatment has increased. This change in practice patterns could result in improvement in overall survival.


The impact of mistranslation on phenotypic variability and fitness.
Phenotypic variation is widespread in natural populations, and can significantly alter population ecology and evolution. Phenotypic variation often reflects underlying genetic variation, but also manifests via non-heritable mechanisms. For instance, translation errors result in about 10% of cellular proteins carrying altered sequences. Thus, proteome diversification arising from translation errors can potentially generate phenotypic variability, in turn increasing variability in the fate of cells or of populations. However, the link between protein diversity and phenotypic variability remains unverified. We manipulated mistranslation levels in Escherichia coli, and measured phenotypic variability between single cells (individual-level variation), as well as replicate populations (population-level variation). Monitoring growth and survival, we find that mistranslation indeed increases variation across E. coli cells, but does not consistently increase variability in growth parameters across replicate populations. Interestingly, although any deviation from the wild-type (WT) level of mistranslation reduces fitness in an optimal environment, the increased variation is associated with a survival benefit under stress. Hence, we suggest that mistranslation-induced phenotypic variation can impact growth and survival and has the potential to alter evolutionary trajectories.


Posttranslational modifications required for cell surface localization and function of the fungal adhesin Aga1p.
Adherence of fungal cells to host substrates and each other affects their access to nutrients, sexual conjugation, and survival in hosts. Adhesins are cell surface proteins that mediate these different cell adhesion interactions. In this study, we examine the in vivo functional requirements for specific posttranslational modifications to these proteins, including glycophosphatidylinositol (GPI) anchor addition and O-linked glycosylation. The processing of some fungal GPI anchors, creating links to cell wall beta-1,6 glucans, is postulated to facilitate postsecretory traffic of proteins to cell wall domains conducive to their functions. By studying the yeast sexual adhesin subunit Aga1p, we found that deletion of its signal sequence for GPI addition eliminated its activity, while deletions of different internal domains had various effects on function. Substitution of the Aga1p GPI signal domain with those of other GPI-anchored proteins, a single transmembrane domain, or a cysteine capable of forming a disulfide all produced functional adhesins. A portion of the cellular pool of Aga1p was determined to be cell wall resident. Aga1p and the alpha-agglutinin Agalpha1p were shown to be under glycosylated in cells lacking the protein mannosyltransferase genes PMT1 and PMT2, with phenotypes manifested only in MATalpha cells for single mutants but in both cell types when both genes are absent. We conclude that posttranslational modifications to Aga1p are necessary for its biogenesis and activity. Our studies also suggest that in addition to GPI-glucan linkages, other cell surface anchorage mechanisms, such as transmembrane domains or disulfides, may be employed by fungal species to localize adhesins.


A computational model for reference-frame synthesis with applications to motion perception.
As discovered by the Gestaltists, in particular by Duncker, we often perceive motion to be within a non-retinotopic reference frame. For example, the motion of a reflector on a bicycle appears to be circular, whereas, it traces out a cycloidal path with respect to external world coordinates. The reflector motion appears to be circular because the human brain subtracts the horizontal motion of the bicycle from the reflector motion. The bicycle serves as a reference frame for the reflector motion. Here, we present a general mathematical framework, based on vector fields, to explain non-retinotopic motion processing. Using four types of non-retinotopic motion paradigms, we show how the theory works in detail. For example, we show how non-retinotopic motion in the Ternus-Pikler display can be computed.


High heterogeneity in methods used for the laboratory confirmation of pertussis diagnosis among European countries, 2010: integration of epidemiological and laboratory surveillance must include standardisation of methodologies and quality assurance.
Despite extensive childhood immunisation, pertussis remains one of the world's leading causes of vaccine preventable deaths. The current methods used for laboratory diagnosis of pertussis include bacterial culture, polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) serology. We conducted a questionnaire survey to identify variations in the laboratory methods and protocols used among participating countries included in the European surveillance network for vaccine-preventable diseases(EUVAC.NET). In February 2010, we performed the survey using a web-based questionnaire and sent it to the country experts of 25 European Union countries,and two European Economic Area (EEA) countries,Norway and Iceland. The questionnaire consisted of 37 questions which covered both general information on surveillance methods and detailed laboratory methods used. A descriptive analysis was performed.Questionnaires were answered by all 27 contacted countries. Nineteen countries had pertussis reference laboratories at the national level; their functions varied from performing diagnosis to providing technical advice for routine microbiology laboratories. Culture,PCR and serology were used in 17, 18 and 20 countries,respectively. For PCR, nine laboratories used insertion sequence IS481 as the target gene, which is present in multiple copies in the Bordetella pertussis genome and thus has a greater sensitivity over single copy targets, but has been proved not to be specific for B.pertussis. Antibodies directed against pertussis toxin(PT) are specific for B. pertussis infections. For ELISA serology, only 13 countries' laboratories used purified PT as coating antigen and 10 included World Health Organization (WHO) or Food and Drug Administration (FDA) reference sera in their tests. This present survey shows that methods used for laboratory confirmation of pertussis differ widely among European countries and that there is a great heterogeneity of the reference laboratories and functions. To evaluate the effects of different pertussis immunisation programmes in Europe, standardisation and harmonisation of the laboratory methods are needed.


Effect of laughter on salivary endocrinological stress marker chromogranin A.
We investigated the effect of laughter on salivary endocrinological stress marker chromogranin A (CgA). In saliva samples collected from 11 healthy males before and after watching a comic film or a non-humorous control film, salivary CgA levels were determined by enzyme-linked immunosorbent assay (ELISA). Samples taken after watching the comic film showed increased levels of CgA. This tendency was more pronounced in individuals with lower initial levels of stress. The control samples showed no significant change in CgA levels. Stress score, subjectively evaluated using a visual analog scale, decreased significantly after watching the comic film. These findings suggest that, in addition to a stress relief effect, laughter can bring about feeling uplifted or fulfilled.


Rapid engraftment after allogeneic transplantation of density-enriched peripheral blood CD34+ cells in patients with advanced hematologic malignancies.
Acute graft versus host disease (GVHD) remains a major cause of morbidity and mortality after allogeneic hematopoietic cell transplantation. Preclinical studies have suggested that a T-cell subset with a CD4-/CD8- double-negative (DN) T-cell phenotype is capable of suppressing GVHD. Double-negative T cells can be mobilized into the peripheral blood with granulocyte colony-stimulating factor (G-CSF) and enriched by density centrifugation. The current study was performed to study the feasibility and safety of applying a density gradient separation technique for enrichment of CD34+ and DN T cells, while depleting CD4+ and CD8+ single-positive (SP) T cells from peripheral blood progenitor cells (PBPCs) for the purpose of allogeneic transplantation. Twenty-five patients with advanced hematologic malignancies were treated with a myeloablative preparative regimen consisting of fractionated total body irradiation, etoposide, and cyclophosphamide. Human leukocyte antigen identical donors were mobilized with G-CSF PBPC collected by apheresis. The apheresis product was applied to a single-step density gradient, and the low-density cell population was collected. The low-density cell population was infused as the sole source of allogeneic cells after myeloablative therapy. Graft versus host disease prophylaxis consisted of cyclosporine with or without prednisone. CD34 cell recovery was efficient with a median 72% yield, providing for a median CD34+ cell dose of 6.5 x 10(6)/kg (range,1.0- 13.9 x 10(6)/kg). CD3+CD4+ or CD3+CD8+ SP T cells were depleted by a median of 94.4% (range, 58.8- 99.2%), and the ratio of CD34+:SP T cells increased 10-fold. Double-negative T cells were depleted by 92% (range, 18.8- 99.4%), thus the ratio of DN:SP T cells increased less than 2-fold in 71% of apheresis samples tested. Hematopoietic engraftment was rapid, and there was no occurrence of graft failure in examinable patients. Median time to absolute neutrophil count greater than 0.5 x 10(9)/L and platelet count greater than 20 x 10(9)/L was 10.5 and 12 days, respectively. The incidence of Grade 2-4 acute GVHD was 26% (95% confidence interval [CI], 6-45%), although not all patients were examinable due to an unexpectedly high nonrecurrence mortality that at Day 180 was 62% (95% CI, 40-83%). These data suggest that T-cell subset manipulation via density gradient separation is a safe procedure and allowed rapid hematopoietic recovery. Selective enrichment of a donor DN T-cell subset was observed in only a few and was not associated with a reduced incidence of GVHD. However, the low-density selected cells still resulted in GVHD, and there was a high treatment-related mortality.


Fine-scale genetic structure in a wild bird population: the role of limited dispersal and environmentally based selection as causal factors.
Individuals are typically not randomly distributed in space; consequently ecological and evolutionary theory depends heavily on understanding the spatial structure of populations. The central challenge of landscape genetics is therefore to link spatial heterogeneity of environments to population genetic structure. Here, we employ multivariate spatial analyses to identify environmentally induced genetic structures in a single breeding population of 1174 great tits Parus major genotyped at 4701 single-nucleotide polymorphism (SNP) loci. Despite the small spatial scale of the study relative to natal dispersal, we found multiple axes of genetic structure. We built distance-based Moran's eigenvector maps to identify axes of pure spatial variation, which we used for spatial correction of regressions between SNPs and various external traits known to be related to fitness components (avian malaria infection risk, local density of conspecifics, oak tree density, and altitude). We found clear evidence of fine-scale genetic structure, with 21, seven, and nine significant SNPs, respectively, associated with infection risk by two species of avian malaria (Plasmodium circumflexum and P. relictum) and local conspecific density. Such fine-scale genetic structure relative to dispersal capabilities suggests ecological and evolutionary mechanisms maintain within-population genetic diversity in this population with the potential to drive microevolutionary change.


Solving Community SARS-CoV-2 Testing With Telehealth: Development and Implementation for Screening, Evaluation and Testing.
Telehealth has emerged as a crucial component of the SARS-CoV-2 pandemic emergency response. Simply stated, telehealth is a tool to provide health care from a distance. Jefferson Health has leveraged its acute care telehealth platform to screen, order testing, and manage patients with COVID-19-related concerns. This study aims to describe the expansion and results of using a telehealth program to increase access to care while minimizing additional potential exposures during the early period of the COVID-19 pandemic. Screening algorithms for patients with SARS-CoV-2-related complaints were created, and 150 new clinicians were trained within 72 hours to address increased patient demand. Simultaneously, Jefferson Health created mobile testing sites throughout eastern Pennsylvania and the southern New Jersey region. Visit volume, the number of SARS-CoV-2 tests ordered, and the number of positive tests were evaluated, and the volume was compared with preceding time periods. From March 8, 2020, to April 11, 2020, 4663 patients were screened using telehealth, representing a surge in visit volume. There were 1521 patients sent to mobile testing sites, and they received a telephone call from a centralized call center for results. Of the patients who were tested, nearly 20% (n=301) had a positive result. Our model demonstrates how using telehealth for a referral to central testing sites can increase access to community-based care, decrease clinician exposure, and minimize the demand for personal protective equipment. The scaling of this innovation may allow health care systems to focus on preparing for and delivering hospital-based care needs.


Patients experiencing local anaesthesia and hip surgery.
The aim of this study was to show what the experience of local anaesthesia and a surgical situation meant to patients. Many patients who receive local anaesthesia will remain awake during surgery. The very fact of staying awake causes psychological and psychosocial needs. These needs must be met and it is therefore important to identify them. Seven patients (aged 61-79) experiencing local anaesthesia and hip surgery (hip replacement surgery or repair of fractured hip) were interviewed. An interpretive phenomenological method developed by Benner was used to extract the experience of what it means to be in local anaesthesia and surgery. One paradigm case is used to illustrate the results. Results show that the well-being and comfort of patients is compromised by challenges such as severe pain and long waits, which may be experienced as endless and which leave the patient thinking of nothing else. By contrast, the experience of trust helps the patient to feel control even in situations where the treatment is hard to grasp. The results highlight that local anaesthesia and surgery force patients to overcome and handle experiences of pain, trust and distrust as well as feelings of alienation and unreality. The results illustrate the patient's needs, as seen from the patient's perspective. A shared experience makes it possible to understand, and thus to recognize, the required qualitative care to facilitate and to help the patient to remain in control and face the psychological challenges presented by anaesthesia care and surgical situations.


Predicting the risk of nephrotoxicity in patients receiving colistimethate sodium: a multicentre, retrospective, cohort study.
With increasing rates of infections caused by MDR Gram-negative organisms, clinicians resort to older agents such as colistimethate sodium (CMS) despite a significant risk of nephrotoxicity. Several risk factors for CMS-associated nephrotoxicity have been reported, but they have yet to be validated. We compared the performance of published mathematical models in predicting the risk of CMS-associated nephrotoxicity. In a multicentre, retrospective, cohort study, adult patients (≥18 years of age) were evaluated from five large academic medical centres in the USA. Patients with normal renal function (baseline serum creatinine ≤1.5 mg/dL) who received intravenous CMS for ≥72 h were followed for up to 30 days. The development of nephrotoxicity was as defined by the RIFLE criteria. Each published model was conditioned using patient-specific variables to predict the risk of nephrotoxicity. The predictive performance of the models was evaluated using the observed-to-expected (O/E) ratio. The most significant cut-off threshold for stratifying patients into high and low risk of nephrotoxicity was identified using classification and regression tree analysis. A total of 106 patients were examined (mean age 53.3 ± 14.9 years, 66% male); the overall observed nephrotoxicity rate was 52.8%. We identified a simple model demonstrating reasonable overall nephrotoxicity risk assessment [O/E ratio of 1.07 (95% CI = 0.81-1.39)] and high sensitivity (92.9%) in predicting nephrotoxicity development in patients on CMS therapy. We identified a model that could be incorporated into patient management strategies to reduce the risk of nephrotoxicity in patients requiring CMS therapy.


Common variants of the liver fatty acid binding protein gene influence the risk of type 2 diabetes and insulin resistance in Spanish population.
The main objective was to evaluate the association between SNPs and haplotypes of the FABP1-4 genes and type 2 diabetes, as well as its interaction with fat intake, in one general Spanish population. The association was replicated in a second population in which HOMA index was also evaluated. 1217 unrelated individuals were selected from a population-based study [Hortega study: 605 women; mean age 54 y; 7.8% with type 2 diabetes]. The replication population included 805 subjects from Segovia, a neighboring region of Spain (446 females; mean age 52 y; 10.3% with type 2 diabetes). DM2 mellitus was defined in a similar way in both studies. Fifteen SNPs previously associated with metabolic traits or with potential influence in the gene expression within the FABP1-4 genes were genotyped with SNPlex and tested. Age, sex and BMI were used as covariates in the logistic regression model. One polymorphism (rs2197076) and two haplotypes of the FABP-1 showed a strong association with the risk of DM2 in the original population. This association was further confirmed in the second population as well as in the pooled sample. None of the other analyzed variants in FABP2, FABP3 and FABP4 genes were associated. There was not a formal interaction between rs2197076 and fat intake. A significant association between the rs2197076 and the haplotypes of the FABP1 and HOMA-IR was also present in the replication population. The study supports the role of common variants of the FABP-1 gene in the development of type 2 diabetes in Caucasians.


FUS pathology defines the majority of tau- and TDP-43-negative frontotemporal lobar degeneration.
Through an international consortium, we have collected 37 tau- and TAR DNA-binding protein 43 (TDP-43)-negative frontotemporal lobar degeneration (FTLD) cases, and present here the first comprehensive analysis of these cases in terms of neuropathology, genetics, demographics and clinical data. 92% (34/37) had fused in sarcoma (FUS) protein pathology, indicating that FTLD-FUS is an important FTLD subtype. This FTLD-FUS collection specifically focussed on aFTLD-U cases, one of three recently defined subtypes of FTLD-FUS. The aFTLD-U subtype of FTLD-FUS is characterised clinically by behavioural variant frontotemporal dementia (bvFTD) and has a particularly young age of onset with a mean of 41 years. Further, this subtype had a high prevalence of psychotic symptoms (36% of cases) and low prevalence of motor symptoms (3% of cases). We did not find FUS mutations in any aFTLD-U case. To date, the only subtype of cases reported to have ubiquitin-positive but tau-, TDP-43- and FUS-negative pathology, termed FTLD-UPS, is the result of charged multivesicular body protein 2B gene (CHMP2B) mutation. We identified three FTLD-UPS cases, which are negative for CHMP2B mutation, suggesting that the full complement of FTLD pathologies is yet to be elucidated.


Fluconazole Susceptibility in Cryptococcus gattii Is Dependent on the ABC Transporter Pdr11.
Cryptococcus gattii isolates from the Pacific Northwest have exhibited higher fluconazole MICs than isolates from other sites. The mechanism of fluconazole resistance in C. gattii is unknown. We sought to determine the role of the efflux pumps Mdr1 and Pdr11 in fluconazole susceptibility. Using biolistic transformation of the parent isolate, we created a strain lacking Mdr1 (mdr1Δ) and another strain lacking Pdr11 (pdr11Δ). Phenotypic virulence factors were assessed by standard methods (capsule size, melanin production, growth at 30 and 37 °C). Survival was assessed in an intranasal murine model of cryptococcosis. Antifungal MICs were determined by the M27-A3 methodology. No differences in key virulence phenotypic components were identified. Fluconazole susceptibility was unchanged in the Mdr1 knockout or reconstituted isolates. However, fluconazole MICs decreased from 32 μg/ml for the wild-type isolate to <0.03 μg/ml for the pdr11Δ strain and reverted to 32 μg/ml for the reconstituted strain. In murine models, no difference in virulence was observed between wild-type, knockout, or reconstituted isolates. We conclude that Pdr11 plays an essential role in fluconazole susceptibility in C. gattii. Genomic and expression differences between resistant and susceptible C. gattii clinical isolates should be assessed further in order to identify other potential mechanisms of resistance.


In Vitro Reconstitution of OxyA Enzymatic Activity Clarifies Late Steps in Vancomycin Biosynthesis.
Studies on the biosynthesis of glycopeptide antibiotics have provided many insights into the strategies that Nature employs to build architecturally strained molecules. A key structural feature of vancomycin, the founding member of this class, is a set of three aromatic cross-links that are introduced via yet unknown mechanisms. Previous reports have identified three cytochrome P450 enzymes involved in this process and demonstrated enzymatic activity for OxyB, which installs the first aromatic cross-link. However, the activities of the remaining two P450 enzymes have not been recapitulated. Herein, we show that OxyA generates the second bis-aryl ether bond in vancomycin and that it exhibits strict substrate specificity toward the chlorinated, OxyB-cross-linked product. No OxyA product is detected with the unchlorinated substrate. Together with previous results, these data suggest that chlorination occurs after OxyB- but before OxyA-catalyzed cross-link formation. Our results have important implications for the chemo-enzymatic synthesis of vancomycin and its analogs.


Occupational therapy: The key to unlocking locked-up occupations during the COVID-19 pandemic.
Occupations refer to the everyday activities that people do as individuals, in families and with communities to occupy time and bring meaning and purpose to life. It is not always limited to just paid employment. Occupations of the global population have been adversely affected in one way or the other because of this COVID-19 pandemic. Four different key sects of occupations were majorly affected. These are the occupations of those who are or were COVID-positive, occupations of healthy individuals affected by COVID-19/lockdown, occupations of the population highly susceptible and vulnerable of contracting COVID-19 and occupations having a direct impact on global market, supply chain or economy. These occupations were locked up due to the pandemic lockdown. Occupational therapists can scientifically analyse occupations and help formulate exit strategies for the lockdown. They are experts who understand and study the different ways of measuring participation in occupation to develop innovative strategies and therapeutic interventions to facilitate individuals' engagement in occupations. They can unravel the pragmatic strategies for preventing transmission (physical distancing, hand hygiene, personal protective equipment usage and decontamination) despite engaging in occupations safely and effectively. Nourishing this niche and essential science is pertinent, not just in this pandemic context but also against a backdrop of health and social care research, policy, practice and education for the future.


Beta-cell deterioration determines the onset and rate of progression of secondary dietary failure in type 2 diabetes mellitus: the 10-year follow-up of the Belfast Diet Study.
Secondary failure of plasma glucose control following initial successful response to diet therapy may be due to dietary indiscretion, or to progression of the intrinsic diabetic condition. We report a 10-year prospective natural history study of 432 newly diagnosed diabetic patients aged 40-69 years undertaken to assess the effect of intensive dietary management, where patients were transferred to insulin, or oral hypoglycaemic therapy (tolbutamide, metformin) by predetermined criteria of weight and plasma glucose. Secondary failure to diet therapy occurred in 41 patients in years 2-4, 67 patients in years 5-7, and 51 patients in years 8-10; 173 patients remained on diet alone until death or the end of the study. Continuation on diet alone was associated with a lower ongoing fasting plasma glucose, greater beta-cell function assessed by an oral glucose tolerance test at 6 months, and increasing age. The rate of rise of fasting plasma glucose was inversely related to the duration of successful dietary therapy, but mean weight remained constant in all groups while on diet alone. The ongoing fall in beta-cell function assessed by HOMA modelling closely mirrored the progressive rise in fasting plasma glucose: there was no change in mean insulin sensitivity in any of the groups.


Parasite fauna of bream Abramis brama and roach Rutilus rutilus from a man-made waterway and a freshwater habitat in northern Germany.
Fifty specimens each of bream Abramis brama and roach Rutilus rutilus were examined for metazoan parasite fauna and trichodinid ciliates; 25 specimens of each species were collected from the Kiel Canal, a man-made waterway, and a nearby freshwater lake, the Dieksee. This is the first detailed parasitological examination of A. brama and R. rutilus at these locations: 30 parasite species were found, comprising 4 protozoans, 4 myxozoans, 5 digeneans, 3 monogeneans, 2 cestodes, 6 nematodes, 2 acanthocephalans, 3 crustaceans and 1 hirudinean. The crustacean Caligus lacustris occurred in both habitats while 2 other crustacean species, 2 acanthocephalans and 1 hirudinean were recorded exclusively for the lake habitat. Larval as well as adult stages of the different parasite species were found, indicating that both fish species act as intermediate and final hosts in both habitats. The Kiel Canal (total of 17 parasite species) showed a lower parasite species richness for A. brama and R. rutilus (14 and 10 parasite species, respectively) than the lake (25 parasite species). A. brama had a higher parasite richness (22 species) than R. rutilus (16 species) in the lake habitat. Most parasites collected were of freshwater origin. Consequently, the observed infection pattern of both fish species in the waterway is mainly influenced by the limited salinity tolerance of freshwater parasites, which are negatively affected even by a salinity of 2.3 to 4.5. In the central Kiel Canal, neither fish species was infected with marine parasites of low host specifity. These parasites are either limited by the low salinity at this sampling site (<4.5 to 6.0) or they cannot enter the canal due to the environmental conditions prevailing in this artificial brackish water habitat. Thus, the canal may comprise a natural barrier preventing the distribution of North Sea parasites into the Baltic Sea. However, the brackish water Baltic Sea nematodes Paracuaria adunca and Cosmocephalus obvelatus were found in R. rutilus from the canal, demonstrating the ability of some parasite species to invade and extend their range of distribution through this man-made shipping route from the Baltic to the North Sea.


Engaging, recruiting, and retaining black men who have sex with men in research studies: don't underestimate the importance of staffing--lessons learned from HPTN 061, the BROTHERS study.
HIV/AIDS in the United States continues to primarily impact men who have sex with men (MSM), with disproportionately high rates among black MSM. The purpose of this study was to identify factors that may influence engagement and retention of black MSM in HIV research. This was a qualitative evaluation of study implementation within a multisite, prospective, observational study (HIV Prevention Trials Network 061, BROTHERS) that enrolled 1553 black MSM in 6 cities throughout the United States. Data collection for this evaluation included a written, structured survey collected from each of the sites describing site characteristics including staff and organizational structure, reviews of site standard operating procedures, and work plans; semistructured key informant interviews were conducted with site coordinators to characterize staffing, site-level factors facilitating or impeding effective community engagement, study recruitment, and retention. Data from completed surveys and site standard operating procedures were collated, and notes from key informant interviews were thematically coded for content by 2 independent reviewers. Several key themes emerged from the data, including the importance of inclusion of members of the community being studied as staff, institutional hiring practices that support inclusive staffing, cultivating a supportive working environment for study implementation, and ongoing relationships between research institutions and community. This study underscores the importance of staffing in implementing research with black MSM. Investigators should consider how staffing and organizational structures affect implementation during study design and when preparing to initiate study activities. Ongoing monitoring of community engagement can inform and improve methods for engagement and ensure cultural relevance while removing barriers for participation.


Sexual behaviours, knowledge and attitudes regarding safe sex, and prevalence of non-viral sexually transmitted infections among attendees of youth clinics in St. Petersburg, Russia.
Adolescents and young adults are at increased risk of sexually transmitted infections (STIs). Knowledge of STI prevalence and risk factors are essential tools to elaborate preventive strategies. However, internationally reported studies on epidemiology of STIs among the youth in Russia are mainly lacking. To ascertain sexual behaviours, knowledge and attitudes about safe sex and prevalence and correlates with STIs in attendees of youth clinics in St. Petersburg, Russia. A total of 301 women and 131 men, who self-referred for STI testing, completed a questionnaire and were screened for Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, and Trichomonas vaginalis using nucleic acid amplification tests. The overall STI prevalence was 16.9%, and similar in the female patients and male patients (15.6% and 19.8% respectively). C. trachomatis, N. gonorrhoeae, M. genitalium and T. vaginalis were detected in 13%, 2.5%, 4.6% and 1.2% of the attendees respectively. The men displayed riskier sexual behaviours and worse knowledge and attitudes regarding safe sex compared to the women, with the most distinguishing features being younger age at first intercourse (P < 0.0005), higher numbers of sex partners during lifetime (P = 0.001) and latest 6 months (P < 0.0005), more frequently consuming alcohol (P < 0.0005), poorer knowledge of STI/HIV prevention measures (P < 0.0005), and less positive attitudes towards safe sex (P = 0.001). However, no significant predictors of STI positivity were found in the men. In the women, the strongest predictors of STI positivity were young age (15-19 years) and multiple sex partners (≥ 2) during latest 6 months. The overall prevalence of STIs among users of STI services at youth clinics in St. Petersburg was high. Comprehensive epidemiological data on STI prevalence and sexual behaviour correlates are necessary to initiate new and strengthen existing STI prevention programmes for the youth, in Russia as well as in many other settings.


Neuroacanthocytosis. A clinical, haematological and pathological study of 19 cases.
Nineteen cases are described, including 12 cases from three different families and 7 nonfamilial cases, in which multisystem neurological disease was associated with acanthocytosis in peripheral blood and normal plasma lipoproteins. Mild acanthocytosis can easily be overlooked, and scanning electron microscopy may be helpful. Some neurologically asymptomatic relatives with significant acanthocytosis were identified during family screening, including some who were clinically affected. The mean age of onset was 32 (range 8-62) yrs and the clinical course was usually progressive but there was marked phenotypic variation. Cognitive impairment, psychiatric features and organic personality change occurred in over half the cases, and more than one-third had seizures. Orofaciolingual involuntary movements and pseudobulbar disturbance commonly caused dysphagia and dysarthria that was sometimes severe, but biting of the lips or tongue was rarely seen. Chorea was seen in almost all symptomatic cases but dystonia, tics, involuntary vocalizations and akinetic-rigid features also occurred. Two cases had no movement disorder at all. Computerized tomography often demonstrated cerebral atrophy. Caudate atrophy was seen less commonly, and nonspecific focal and symmetric signal abnormalities from the caudate or lentiform nuclei were seen by magnetic resonance imaging in 3 out of 4 cases. Depression or absence of tendon reflexes was noted in 13 cases and neurophysiological abnormalities often indicated an axonal neuropathy. Sural nerve biopsies from 3 cases showed evidence of a chronic axonal neuropathy with prominent regenerative activity, predominantly affecting the large diameter myelinated fibres. Serum creatine kinase activity was increased in 11 cases but without clinical evidence of a myopathy. Postmortem neuropathological examination in 1 case revealed extensive neuronal loss and gliosis affecting the corpus striatum, pallidum, and the substantia nigra, especially the pars reticulata. The cerebral cortex appeared spared and the spinal cord showed no evidence of anterior horn cell loss. Two examples of the McLeod phenotype, an X-linked abnormality of expression of Kell blood group antigens, were identified in a single family and included 1 female. The genetics of neuroacanthocytosis are unclear and probably heterogeneous, but the available pedigree data and the association with the McLeod phenotype suggest that there may be a locus for this disorder on the short arm of the X chromosome.


Spatial characteristics of white matter abnormalities in schizophrenia.
There is considerable evidence implicating brain white matter (WM) abnormalities in the pathophysiology of schizophrenia; however, the spatial localization of WM abnormalities reported in the existing studies is heterogeneous. Thus, the goal of this study was to quantify the spatial characteristics of WM abnormalities in schizophrenia. One hundred and fourteen patients with schizophrenia and 138 matched controls participated in this multisite study involving the Universities of Iowa, Minnesota, and New Mexico, and the Massachusetts General Hospital. We measured fractional anisotropy (FA) in brain WM regions extracted using 3 different image-processing algorithms: regions of interest, tract-based spatial statistics, and the pothole approach. We found that FA was significantly lower in patients using each of the 3 image-processing algorithms. The region-of-interest approach showed multiple regions with lower FA in patients with schizophrenia, with overlap at all 4 sites in the corpus callosum and posterior thalamic radiation. The tract-based spatial statistic approach showed (1) global differences in 3 of the 4 cohorts and (2) lower frontal FA at the Iowa site. Finally, the pothole approach showed a significantly greater number of WM potholes in patients compared to controls at each of the 4 sites. In conclusion, the spatial characteristics of WM abnormalities in schizophrenia reflect a combination of a global low-level decrease in FA, suggesting a diffuse process, coupled with widely dispersed focal reductions in FA that vary spatially among individuals (ie, potholes).


Objectives and Design of the Russian Acute Coronary Syndrome Registry (RusACSR).
The Russian Acute Coronary Syndrome Registry (RusACSR) is a retrospective, continuous, nationwide, Web-based registry of patients with acute coronary syndromes (ACS). The RusACSR is a database that uses a secure Web-based interface for data entry by individual users. Participation in the RusACSR is voluntary. Any clinical center that provides health care to ACS patients can take part in the RusACSR. The RusACSR enrolls ACS patients who have undergone care in Russian hospitals from February 2008 to the present. Key data elements and methods of data analysis in the RusACSR are presented in this article. Up to 2015, 213 clinical centers from 36 regions of Russia had participated in the RusACSR. Currently, the database contains data on more than 250 000 ACS patients who underwent care from 2008 to 2015. Some current problems are highlighted in this article. The RusACSR is a perspective project for different epidemiologic studies in Russian ACS patients.


Dissociation between decoding and reasoning about mental states in patients with theory of mind reasoning impairments.
Theory of mind (ToM) reasoning may involve a multiplicity of processes, including an initial stage, where cues relevant for social processes are detected and decoded, and a mentalizing stage, where the decoded information is used to reason about mental states. Here we report that the processing of lower-order facial cues relevant to social judgments can be relatively spared in patients with impaired ToM reasoning. We discuss the implications for understanding the mechanisms underlying social judgments in brain-lesioned patients.


Comparison of conventional viral cultures with direct fluorescent antibody stains for diagnosis of community-acquired respiratory virus infections in hospitalized children.
Because of the widespread availability of rapid viral antigen testing, many institutions never adopted a routine practice of ordering viral cultures to detect community-acquired respiratory viruses (CRVs). The ease of performing complete viral studies in our on site laboratory allowed us to assess the clinical implications of the absence of conventional culture results in previously healthy hospitalized children with CRV infections. From June 1997 through May 2000, the results of direct immunofluorescence assay (DFA) of 1069 nasopharyngeal swab (NP) specimens were compared with simultaneously inoculated conventional tube cell cultures for detection of CRVs. In addition the medical records of 140 previously healthy infants and children hospitalized for management of lower respiratory tract infections caused by culture-proved CRVs were reviewed. Viruses were isolated or detected by DFA or viral culture or both in 468 (30%) of the 1557 NP samples evaluated. The most common CRV isolated was respiratory syncytial virus (49%), followed by parainfluenza viruses (15%), influenza A viruses (14%), rhinoviruses (8%), adenoviruses (4%), enteroviruses (4%) and influenza B viruses (1%). Of the 1069 NP specimens for which both viral culture and rapid antigen testing were performed, 190 specimens were DFA-positive and culture-positive, 7 specimens were DFA-positive and culture-negative, 35 specimens were DFA-negative and culture-positive and 837 specimens were DFA-negative and culture-negative. The overall sensitivity, specificity, positive predictive value and negative predictive value of DFA were 84, 99, 96 and 96%, respectively. Of the 140 hospitalized patients with culture-proved viral cultures (89 respiratory syncytial virus, 22 influenza A, 20 parainfluenza virus and 9 adenovirus), the mean duration of hospital stay was 3.6 days, and the mean time for viral cultures to become positive was 7.7 days (P < 0.001, signed rank test). One hundred twenty (86%) viral cultures did not become positive until after the patient had been discharged from the hospital. In no case was the clinical decision regarding the patient's treatment or discharge from the hospital based on the results of viral culture. We conclude that positive viral cultures have no impact on clinical decision making and management of healthy children during hospitalization for illness attributable to community-acquired respiratory viruses.


C-reactive protein levels and atrial fibrillation after cardiac surgery in women.
The aim of this study was to evaluate whether risk for postoperative atrial fibrillation in women is related to preexisting inflammation as detected by plasma C-reactive protein (CRP) concentrations. We further sought to assess the importance of atrial fibrillation for outcome after cardiac surgery in women. The CRP was measured before coronary artery bypass grafting and (or) valvular surgery using cardiopulmonary bypass in 141 women. Univariate and multivariate analyses were used to evaluate for differences in CRP levels between women with and without atrial fibrillation, and to assess for the importance of the arrhythmia and postoperative outcomes. Atrial fibrillation developed in 46 (33%) women. Neither CRP concentrations (median +/- standard error, 13.3 +/- 2.5 mg/L vs 11.7 +/- 1.4 mg/L, p = 0.847), nor the frequency of elevated levels (defined as > upper 95% confidence interval or >19.2 mg/L) (19% vs 21%, p = 0.807) differed between women with or without atrial fibrillation. Patient age and previous stroke, but not CRP levels, were independently associated with atrial fibrillation. Women with atrial fibrillation were more likely to have low cardiac output syndrome (p = 0.018), stroke (p = 0.031), longer duration of hospitalization in the intensive care unit (p = 0.012) and on the postoperative (p = 0.0008) ward, and they were more likely to require an extended care facility after surgery (p = 0.046). In contrast to findings from studies that have included mostly men, preoperative CRP concentrations are not associated with risk for atrial fibrillation after cardiac surgery for women. Postoperative atrial fibrillation in women is associated with increased risk for stroke, longer hospitalization, and extended care facility admission.


Egg-shaped core/shell α-Mn2O3@α-MnO2 as heterogeneous catalysts for decomposition of phenolics in aqueous solutions.
Novel uniform ellipsoid α-Mn2O3@α-MnO2 core/shell (McMs) nanocomposites were prepared via a hydrothermal process with a shape-control protocol followed by calcination at different temperatures. The properties of the composites were characterized by a number of techniques such as thermogravimetric analysis (TGA), X-ray diffraction (XRD), N2 adsorption, and scanning electron microscopy (SEM). The core/shell materials were much effective in heterogeneous oxone(®) activation to generate sulfate and hydroxyl radicals for degradation of aqueous phenol. The McMs composites demonstrated catalytic activity for 100% phenol decomposition in short duration varying between 20 and 120 min, much higher than that of homogeneous Mn(2+) system with 95% phenol degradation in 120 min. They also showed a higher activity than single-phase α-Mn2O3 or α-MnO2. The catalytic activity of phenol degradation depends on temperature, oxone(®) concentration, phenol concentration, and catalyst loading. The catalysts also showed a stable activity in several cycles. Kinetic study demonstrated that phenol degradation reactions follow a first order reaction on McMs catalysts giving activation energies at 32.1-68.8 kJ/mol. With the detection of radicals by electron paramagnetic resonance (EPR), the generation mechanism was proposed.


Establishment of human retinal microvascular endothelial cells with extended life-span.
To generate and characterize a telomerase-immortalized human retinal microvascular endothelial cell (HREC) line. This cell line may be utilized as an in vitro model to study the molecular basis of several diseases of the human retina. Primary retinal neuronal cells were isolated and transfected with plasmid encoding full-length human telomerase reverse transcriptase (hTERT). Transfected cells were selected and characterized to determine telomerase activity, karyotype, proliferative capacity and functionality. HREC-hTERT cells appear morphologically similar to primary endothelial cells and have an extended in vitro life-span. HREC-hTERT cells express the progenitor/stem cell marker nestin. They have active telomerase and a high proliferative capacity. These cells also maintain a diploid karyotype. The HREC-hTERT cells showed high colony-formation capacity and plating efficiency compared to the primary cells. These cells are capable of differentiation into neuronal and glial cell phenotypes and the differentiated cells express the astrocyte marker glial fibrillary acidic protein (GFAP) and the neuronal marker microtubule-associated protein-2 (MAP2), respectively. The in vitro life-span of human retinal neuronal endothelial cells can be extended by ectopic expression of hTERT without altering the genetic stability and functionality of these cells. These cells will be a valuable tool to further our understanding on the role of HRECs in the human blood-retinal-barrier and in angiogenesis and neovascularization.


Resveratrol induces growth arrest and apoptosis through activation of FOXO transcription factors in prostate cancer cells.
Resveratrol, a naturally occurring phytopolyphenol compound, has attracted extensive interest in recent years because of its diverse pharmacological characteristics. Although resveratrol possesses chemopreventive properties against several cancers, the molecular mechanisms by which it inhibits cell growth and induces apoptosis have not been clearly understood. The present study was carried out to examine whether PI3K/AKT/FOXO pathway mediates the biological effects of resveratrol. Resveratrol inhibited the phosphorylation of PI3K, AKT and mTOR. Resveratrol, PI3K inhibitors (LY294002 and Wortmannin) and AKT inhibitor alone slightly induced apoptosis in LNCaP cells. These inhibitors further enhanced the apoptosis-inducing potential of resveratrol. Overexpression of wild-type PTEN slightly induced apoptosis. Wild type PTEN and PTEN-G129E enhanced resveratrol-induced apoptosis, whereas PTEN-G129R had no effect on proapoptotic effects of resveratrol. Furthermore, apoptosis-inducing potential of resveratrol was enhanced by dominant negative AKT, and inhibited by wild-type AKT and constitutively active AKT. Resveratrol has no effect on the expression of FKHR, FKHRL1 and AFX genes. The inhibition of FOXO phosphorylation by resveratrol resulted in its nuclear translocation, DNA binding and transcriptional activity. The inhibition of PI3K/AKT pathway induced FOXO transcriptional activity resulting in induction of Bim, TRAIL, p27/KIP1, DR4 and DR5, and inhibition of cyclin D1. Similarly, resveratrol-induced FOXO transcriptional activity was further enhanced when activation of PI3K/AKT pathway was blocked. Over-expression of phosphorylation deficient mutants of FOXO proteins (FOXO1-TM, FOXO3A-TM and FOXO4-TM) induced FOXO transcriptional activity, which was further enhanced by resveratrol. Inhibition of FOXO transcription factors by shRNA blocked resveratrol-induced upregulation of Bim, TRAIL, DR4, DR5, p27/KIP1 and apoptosis, and inhibition of cyclin D1 by resveratrol. These data suggest that FOXO transcription factors mediate anti-proliferative and pro-apoptotic effects of resveratrol, in part due to activation of extrinsic apoptosis pathway.


Calibration of Elasto-Magnetic Sensors on In-Service Cable-Stayed Bridges for Stress Monitoring.
The recent developments in measurement technology have led to the installation of efficient monitoring systems on many bridges and other structures all over the world. Nowadays, more and more structures have been built and instrumented with sensors. However, calibration and installation of sensors remain challenging tasks. In this paper, we use a case study, Adige Bridge, in order to present a low-cost method for the calibration and installation of elasto-magnetic sensors on cable-stayed bridges. Elasto-magnetic sensors enable monitoring of cable stress. The sensor installation took place two years after the bridge construction. The calibration was conducted in two phases: one in the laboratory and the other one on site. In the laboratory, a sensor was built around a segment of cable that was identical to those of the cable-stayed bridge. Then, the sample was subjected to a defined tension force. The sensor response was compared with the applied load. Experimental results showed that the relationship between load and magnetic permeability does not depend on the sensor fabrication process except for an offset. The determination of this offset required in situ calibration after installation. In order to perform the in situ calibration without removing the cables from the bridge, vibration tests were carried out for the estimation of the cables' tensions. At the end of the paper, we show and discuss one year of data from the elasto-magnetic sensors. Calibration results demonstrate the simplicity of the installation of these sensors on existing bridges and new structures.


Using sensorimotor integrative treatment with mentally retarded adults.
Thirty-four mentally retarded, institutionalized adults were subjects in a study of the effects of sensorimotor integrative treatment procedures on sensorimotor development. The 25 subjects in the experimental group received group sensorimotor integrative treatment in addition to regular institutional programming; the 9 subjects in the control group received only the regular institutional programming. The Sensorimotor Performance Analysis was used to test sensorimotor performance of all subjects four times during the 12-month study. Results showed that both the experimental group and the control group made gains in sensorimotor development, with no significant differences between groups. The findings indicate that treatment procedures for improving sensorimotor development of the mentally retarded adult need further examination so that appropriate modalities can be developed. Periodic reassessment of the client following completion of treatment is suggested because of possible delays in improvement of sensorimotor development.


Graphene oxide/nucleic-acid-stabilized silver nanoclusters: functional hybrid materials for optical aptamer sensing and multiplexed analysis of pathogenic DNAs.
Hybrid systems consisting of nucleic-acid-functionalized silver nanoclusters (AgNCs) and graphene oxide (GO) are used for the development of fluorescent DNA sensors and aptasensors, and for the multiplexed analysis of a series of genes of infectious pathogens. Two types of nucleic-acid-stabilized AgNCs are used: one type includes the red-emitting AgNCs (616 nm) and the second type is near-infrared-emitting AgNCs (775 nm). Whereas the nucleic-acid-stabilized AgNCs do not bind to GO, the conjugation of single-stranded nucleic acid to the DNA-stabilized AgNCs leads to the adsorption of the hybrid nanostructures to GO and to the fluorescence quenching of the AgNCs. By the conjugation of oligonucleotide sequences acting as probes for target genes, or as aptamer sequences, to the nucleic-acid-protected AgNCs, the desorption of the probe/nucleic-acid-stabilized AgNCs from GO through the formation of duplex DNA structures or aptamer-substrate complexes leads to the generation of fluorescence as a readout signal for the sensing events. The hybrid nanostructures are implemented for the analysis of hepatitis B virus gene (HBV), the immunodeficiency virus gene (HIV), and the syphilis (Treponema pallidum) gene. Multiplexed analysis of the genes is demonstrated. The nucleic-acid-AgNCs-modified GO is also applied to detect ATP or thrombin through the release of the respective AgNCs-labeled aptamer-substrate complexes from GO.


New Opportunities in Synchrotron X-ray Crystallography.
Several high-intensity synchrotron x-ray sources have been constructed over the past few years in the United States, West Germany, Great Britain, Japan, France, Italy, and the Soviet Union. Crystallographers have begun to use these facilities for experiments that take advantage of the characteristics of synchrotron radiation, namely, a broad distribution of wavelengths, high intensity, low divergence, strong polarization, and a pulsed time structure. In addition to more familiar diffraction experiments on single crystals and powdered samples, new types of crystallographic studies, for example, energy-dispersive and surface diffraction studies, have progressed rapidly with more general accessibility of synchrotron sources. These high-intensity sources allow diffraction experiments to be performed on very small crystals or on large biological molecules, and permit weak magnetic scattering to be detected Anomalous dispersion experiments can exploit he ability to vary the wavelength of the radiation, and the pulsed time structure of the beam makes possible fast time-resolved experiments. Because of the availability of synchrotron x-radiation, these and other kinds of experiments will be in the forefront of crystallographic research for the next several years.


Interleukin 17-Mediated Host Defense against Candida albicans.
Candida albicans is part of the normal microbiota in most healthy individuals. However, it can cause opportunistic infections if host defenses are breached, with symptoms ranging from superficial lesions to severe systemic disease. The study of rare congenital defects in patients with chronic mucocutaneous candidiasis led to the identification of interleukin-17 (IL-17) as a key factor in host defense against mucosal fungal infection. Experimental infections in mice confirmed the critical role of IL-17 in mucocutaneous immunity against C. albicans. Research on mouse models has also contributed importantly to our current understanding of the regulation of IL-17 production by different cellular sources and its effector functions in distinct tissues. In this review, we highlight recent findings on IL-17-mediated immunity against C. albicans in mouse and man.


Antagonistic regulation of actin dynamics and cell motility by TRPC5 and TRPC6 channels.
The Rho family of small guanosine triphosphatases (Rho GTPases: RhoA, Cdc42, and Rac1) regulates many aspects of cell behavior, including actin dynamics and cell migration. The generation of calcium ion (Ca(2+)) microdomains is critical in promoting cell migration because they control the localized activity of Rho GTPases. We identified receptor-activated TRPC5 and TRPC6 (transient receptor potential canonical type 5 and 6) channels as antagonistic regulators of actin remodeling and cell motility in fibroblasts and kidney podocytes. We show that TRPC5 is in a molecular complex with Rac1, whereas TRPC6 is in a molecular complex with RhoA. TRPC5-mediated Ca(2+) influx induces Rac1 activation, thereby promoting cell migration, whereas TRPC6-mediated Ca(2+) influx increases RhoA activity, thereby inhibiting cell migration. Our data unveil antagonistic Ca(2+) influx pathways as a conserved signaling mechanism for the integrated regulation of cell migration.


[Prevalence, clinical-endoscopic characteristics and predictive factors of Barrett's Esophagus in endoscopic screening for gastric cancer].
Determine the prevalence, clinical-epidemiological characteristics and predictive factors for Barrets Esophagus (BE). An analytical and transversal study was conducted, followed by a case-control study nested in a population participating in a gastric cancer screening campaign, from January to June 2008 at the Central Hospital of the Peruvian Air Force (FAP). A BE case was defined due to the endoscopic presence of columnar-appearing mucosa confirmed by intestinal metaplasia at biopsy. Demographic, clinical and endoscopic variables were evaluated, and bivariate and multivariate studies were conducted to identify predictive risk factors, using patients with gastroesophagic reflux and control patients of the study population as control groups. Out of 2273 patients studied, 11 (0.48%) patients with an average age of 52.2 +/- 7.7 years (43-69 year range) suffered from BE, out of which 81.8% were male. The 81.8% were symptomatic, with dyspepsia in 54.5% and with reflux in 27.3%, while 18.2% were asymptomatic. The short-segment BE was more common than the long-segment BE (72.7% versus 27.3%). The predictive factors found after the bivariate and multivariate analysis (comparing cases versus population control group) were: hiatal hernia [OR = 12.1, CI 95% 2.25 64.75], consumption of AINES [OR = 6.72, CI 95% 1.6-29.1] and consumption of alcohol [OR = 4.22, CI 95% 1.1-16.91]. BE prevalence in the study was low. The presence of hiatal hernia, consumption of AINES, and consumption of alcohol were predictive risk factors to develop BE in the study.


EULAR recommendations for the treatment of systemic sclerosis: a report from the EULAR Scleroderma Trials and Research group (EUSTAR).
The optimal treatment of systemic sclerosis (SSc) is a challenge because the pathogenesis of SSc is unclear and it is an uncommon and clinically heterogeneous disease affecting multiple organ systems. The aim of the European League Against Rheumatism (EULAR) Scleroderma Trials and Research group (EUSTAR) was to develop evidence-based, consensus-derived recommendations for the treatment of SSc. To obtain and maintain a high level of intrinsic quality and comparability of this approach, EULAR standard operating procedures were followed. The task force comprised 18 SSc experts from Europe, the USA and Japan, two SSc patients and three fellows for literature research. The preliminary set of research questions concerning SSc treatment was provided by 74 EUSTAR centres. Based on discussion of the clinical research evidence from published literature, and combining this with current expert opinion and clinical experience, 14 recommendations for the treatment of SSc were formulated. The final set includes the following recommendations: three on SSc-related digital vasculopathy (Raynaud's phenomenon and ulcers); four on SSc-related pulmonary arterial hypertension; three on SSc-related gastrointestinal involvement; two on scleroderma renal crisis; one on SSc-related interstitial lung disease and one on skin involvement. Experts also formulated several questions for a future research agenda. Evidence-based, consensus-derived recommendations are useful for rheumatologists to help guide treatment for patients with SSc. These recommendations may also help to define directions for future clinical research in SSc.