Photic and circadian expression of luciferase in mPeriod1-luc transgenic mice invivo.
A conserved transcription-translation negative feedback loop forms the molecular basis of the circadian oscillator in animals. Molecular interactions within this loop have been relatively well characterized in vitro and in cell culture; however, in vivo approaches are required to assess the functional significance of these interactions. Here, regulation of circadian gene expression was studied in vivo by using transgenic reporter mouse lines in which 6.75 kb of the mouse Period1 (mPer1) promoter drives luciferase (luc) expression. Six mPer1-luc transgenic lines were created, and all lines express a daily rhythm of luc mRNA in the suprachiasmatic nuclei (SCN). Each mPer1-luc line also sustains a long-term circadian rhythm of luminescence in SCN slice culture. A 6-h light pulse administered during the early subjective night rapidly induces luc mRNA expression in the SCN; however, high luc mRNA levels are sustained, whereas endogenous mPer1 mRNA levels return to baseline, suggesting that posttranscriptional events mediate the down-regulation of mPer1 after exposure to light. This approach demonstrates that the 6.75-kb mPer1 promoter fragment is sufficient to confer both circadian and photic regulation in vivo and reveals a potential posttranscriptional regulatory mechanism within the mammalian circadian oscillator.


Cross-linking immunoprecipitation-MS (xIP-MS): Topological Analysis of Chromatin-associated Protein Complexes Using Single Affinity Purification.
In recent years, cross-linking mass spectrometry has proven to be a robust and effective method of interrogating macromolecular protein complex topologies at peptide resolution. Traditionally, cross-linking mass spectrometry workflows have utilized homogenous complexes obtained through time-limiting reconstitution, tandem affinity purification, and conventional chromatography workflows. Here, we present cross-linking immunoprecipitation-MS (xIP-MS), a simple, rapid, and efficient method for structurally probing chromatin-associated protein complexes using small volumes of mammalian whole cell lysates, single affinity purification, and on-bead cross-linking followed by LC-MS/MS analysis. We first benchmarked xIP-MS using the structurally well-characterized phosphoribosyl pyrophosphate synthetase complex. We then applied xIP-MS to the chromatin-associated cohesin (SMC1A/3), XRCC5/6 (Ku70/86), and MCM complexes, and we provide novel structural and biological insights into their architectures and molecular function. Of note, we use xIP-MS to perform topological studies under cell cycle perturbations, showing that the xIP-MS protocol is sufficiently straightforward and efficient to allow comparative cross-linking experiments. This work, therefore, demonstrates that xIP-MS is a robust, flexible, and widely applicable methodology for interrogating chromatin-associated protein complex architectures.


Male-specific submaxillary gland protein, a lipocalin allergen of the golden hamster, differs from the lipocalin allergens of Siberian and Roborovski dwarf hamsters.
An increasing number of asthma cases upon exposure to hamsters and anaphylactic reactions following hamster bites are being reported, but the allergens responsible are still poorly characterized. In the Golden hamster, male-specific submaxillary gland protein (MSP), a lipocalin expressed in a sex- and tissue-specific manner in the submaxillary and lacrimal glands, is secreted in the saliva, tears and urine. The purpose of this study was to determine if MSP is an allergen, to identify IgE-reactive proteins of different hamster species and to analyse potential cross-reactivities. Fur extracts were prepared from four hamster species. Hamster-allergic patients were selected based on a history of positive IgE-test to hamster epithelium. The IgE-reactivity of patients' sera was investigated by means of immunoblot and ELISA. IgE-reactive proteins in fur extracts and the submaxillary gland were identified using anti-MSP antibodies, Edman sequencing or mass spectrometry. MSP was purified from Golden hamster and recombinant MSP was expressed in E. coli. Four patients had IgE-antibodies against 20.5-kDa and 24-kDa proteins of Golden hamster fur extract, which were identified as MSP. IgE-reactive MSP-like proteins were detected in European hamster fur extract. Three patient sera showed IgE-reactive bands at 17-21 kDa in Siberian and Roborovski hamster fur extracts. These proteins were identified as two closely related lipocalins. Immunoblot inhibition experiments showed that they are cross-reactive and are different from MSP. MSP lipocalin of the Golden hamster was identified as an allergen, and it is different from the cross-reactive lipocalin allergens of Siberian and Roborovski hamsters. Our findings highlight the need for specific tools for the in vitro and in vivo diagnosis of allergy to different hamster species.


High prevalence of lipoatrophy in pre-pubertal South African children on antiretroviral therapy: a cross-sectional study.
Despite changes in WHO guidelines, stavudine is still used extensively for treatment of pediatric HIV in the developing world. Lipoatrophy in sub-Saharan African children can be stigmatizing and have far-reaching consequences. The severity and extent of lipoatrophy in pre-pubertal children living in sub-Saharan Africa is unknown. In this cross-sectional study, children who were 3-12 years old, on antiretroviral therapy and pre-pubertal were recruited from a Family HIV Clinic in South Africa. Lipoatrophy was identified and graded by consensus between two HIV pediatricians using a standardized grading scale. A professional dietician performed formal dietary assessment and anthropometric measurements of trunk and limb fat. Previous antiretroviral exposures were recorded. In a Dual-Energy X-ray Absorbtiometry (DXA) substudy body composition was determined in 42 participants. Among 100 recruits, the prevalence of visually obvious lipoatrophy was 36% (95% CI: 27%-45%). Anthropometry and DXA measurements corroborated the clinical diagnosis of lipoatrophy: Both confirmed significant, substantial extremity fat loss in children with visually obvious lipoatrophy, when adjusted for age and sex. Adjusted odds ratio for developing lipoatrophy was 1.9 (95% CI: 1.3 - 2.9) for each additional year of accumulated exposure to standard dose stavudine. Cumulative time on standard dose stavudine was significantly associated with reductions in biceps and triceps skin-fold thickness (p=0.008). The prevalence of visually obvious lipoatrophy in pre-pubertal South African children on antiretroviral therapy is high. The amount of stavudine that children are exposed to needs review. Resources are needed to enable low-and-middle-income countries to provide suitable pediatric-formulated alternatives to stavudine-based pediatric regimens. The standard stavudine dose for children may need to be reduced. Diagnosis of lipoatrophy at an early stage is important to allow timeous antiretroviral switching to arrest progression and avoid stigmatization. Diagnosis using visual grading requires training and experience, and DXA and comprehensive anthropometry are not commonly available. A simple objective screening tool is needed to identify early lipoatrophy in resource-limited settings where specialized skills and equipment are not available.


Efficient assembly and annotation of the transcriptome of catfish by RNA-Seq analysis of a doubled haploid homozygote.
Upon the completion of whole genome sequencing, thorough genome annotation that associates genome sequences with biological meanings is essential. Genome annotation depends on the availability of transcript information as well as orthology information. In teleost fish, genome annotation is seriously hindered by genome duplication. Because of gene duplications, one cannot establish orthologies simply by homology comparisons. Rather intense phylogenetic analysis or structural analysis of orthologies is required for the identification of genes. To conduct phylogenetic analysis and orthology analysis, full-length transcripts are essential. Generation of large numbers of full-length transcripts using traditional transcript sequencing is very difficult and extremely costly. In this work, we took advantage of a doubled haploid catfish, which has two sets of identical chromosomes and in theory there should be no allelic variations. As such, transcript sequences generated from next-generation sequencing can be favorably assembled into full-length transcripts. Deep sequencing of the doubled haploid channel catfish transcriptome was performed using Illumina HiSeq 2000 platform, yielding over 300 million high-quality trimmed reads totaling 27 Gbp. Assembly of these reads generated 370,798 non-redundant transcript-derived contigs. Functional annotation of the assembly allowed identification of 25,144 unique protein-encoding genes. A total of 2,659 unique genes were identified as putative duplicated genes in the catfish genome because the assembly of the corresponding transcripts harbored PSVs or MSVs (in the form of pseudo-SNPs in the assembly). Of the 25,144 contigs with unique protein hits, around 20,000 contigs matched 50% length of reference proteins, and over 14,000 transcripts were identified as full-length with complete open reading frames. The characterization of consensus sequences surrounding start codon and the stop codon confirmed the correct assembly of the full-length transcripts. The large set of transcripts assembled in this study is the most comprehensive set of genome resources ever developed from catfish, which will provide the much needed resources for functional genome research in catfish, serving as a reference transcriptome for genome annotation, analysis of gene duplication, gene family structures, and digital gene expression analysis. The putative set of duplicated genes provide a starting point for genome scale analysis of gene duplication in the catfish genome, and should be a valuable resource for comparative genome analysis, genome evolution, and genome function studies.


A short history of MADS-box genes in plants.
Evolutionary developmental genetics (evodevotics) is a novel scientific endeavor which assumes that changes in developmental control genes are a major aspect of evolutionary changes in morphology. Understanding the phylogeny of developmental control genes may thus help us to understand the evolution of plant and animal form. The principles of evodevotics are exemplified by outlining the role of MADS-box genes in the evolution of plant reproductive structures. In extant eudicotyledonous flowering plants, MADS-box genes act as homeotic selector genes determining floral organ identity and as floral meristem identity genes. By reviewing current knowledge about MADS-box genes in ferns, gymnosperms and different types of angiosperms, we demonstrate that the phylogeny of MADS-box genes was strongly correlated with the origin and evolution of plant reproductive structures such as ovules and flowers. It seems likely, therefore, that changes in MADS-box gene structure, expression and function have been a major cause for innovations in reproductive development during land plant evolution, such as seed, flower and fruit formation.


Creatine supplementation does not affect human skeletal muscle glycogen content in the absence of prior exercise.
Due to the current lack of clarity, we examined whether 5 days of dietary creatine (Cr) supplementation per se can influence the glycogen content of human skeletal muscle. Six healthy male volunteers participated in the study, reporting to the laboratory on four occasions to exercise to the point of volitional exhaustion, each after 3 days of a controlled normal habitual dietary intake. After a familiarization visit, participants cycled to exhaustion in the absence of any supplementation (N), and then 2 wk later again they cycled to exhaustion after 5 days of supplementation with simple sugars (CHO). Finally, after a further 2 wk, they again cycled to exhaustion after 5 days of Cr supplementation. Muscle samples were taken at rest before exercise, at the time point of exhaustion in visit 1, and at subsequent visit time of exhaustion. There was a treatment effect on muscle total Cr content in Cr compared with N and CHO supplementation (P < 0.01). Resting muscle glycogen content was elevated above N following CHO (P < 0.05) but not after Cr. At exhaustion following N, glycogen content was no different from CHO and Cr measured at the same time point during exercise. Cr supplementation under conditions of controlled habitual dietary intake had no effect on muscle glycogen content at rest or after exhaustive exercise. We suggest that any Cr-associated increases in muscle glycogen storage are the result of an interaction between Cr supplementation and other mediators of muscle glycogen storage.


Non-linear analysis of heart rate variability for evaluating the acute effects of caffeinated beverages in young adults.
Caffeinated beverages are the most consumed substances in the world. High rate of uptake of these beverages leads to various cardiovascular disorders ranging from palpitations to coronary failure. The objective of the study is to ascertain how the complexity parameters of heart rate variability are affected by acute consumption of caffeinated beverages in young adults.Electrocardiogram measurements were performed before consuming drinks. After consuming the drinks, measurements were done again at 30 minutes and 60 minutes. Heart rate variability signals were acquired from electrocardiogram signals. Also, the signals were reconstructed in the phase space and largest Lyapunov exponent, correlation dimension, approximate entropy, and detrended fluctuation analysis values were calculated.Heart rate increased for energy drink and cola groups but not in coffee group. Non-linear parameter values of energy drink, coffee, and cola group are increased within 60 minutes after drink consumption. This change is statistically significant just for energy drink group.Energy drink consumption increases the complexity of the cardiovascular system in young adults significantly. Coffee and cola consumption have no significant effect on the non-linear parameters of heart rate variability.


Hot water epilepsy: Phenotype and single photon emission computed tomography observations.
We studied the anatomical correlates of reflex hot water epilepsy (HWE) using multimodality investigations viz. magnetic resonance imaging (MRI), electroencephalography (EEG), and single photon emission computed tomography (SPECT). Five men (mean age: 27.0 ΁ 5.8 years) with HWE were subjected to MRI of brain, video-EEG studies, and SPECT scan. These were correlated with phenotypic presentations. Seizures could be precipitated in three patients with pouring of hot water over the head and semiology of seizures was suggestive of temporal lobe epilepsy. Ictal SPECT showed hyperperfusion in: left medial temporal - one, left lateral temporal - one, and right parietal - one. Interictal SPECT was normal in all five patients and did not help in localization. MRI and interictal EEG was normal in all the patients. The clinical and SPECT studies suggested temporal lobe as the seizure onset zone in some of the patients with HWE.


Population Genomic Analysis of North American Eastern Wolves (Canis lycaon) Supports Their Conservation Priority Status.
The threatened eastern wolf is found predominantly in protected areas of central Ontario and has an evolutionary history obscured by interbreeding with coyotes and gray wolves, which challenges its conservation status and subsequent management. Here, we used a population genomics approach to uncover spatial patterns of variation in 281 canids in central Ontario and the Great Lakes region. This represents the first genome-wide single nucleotide polymorphism (SNP) dataset with substantial sample sizes of representative populations. Although they comprise their own genetic cluster, we found evidence of eastern wolf dispersal outside of the boundaries of protected areas, in that the frequency of eastern wolf genetic variation decreases with increasing distance from provincial parks. We detected eastern wolf alleles in admixed coyotes along the northeastern regions of Lake Huron and Lake Ontario. Our analyses confirm the unique genomic composition of eastern wolves, which are mostly restricted to small fragmented patches of protected habitat in central Ontario. We hope this work will encourage an innovative discussion regarding a plan for managed introgression, which could conserve eastern wolf genetic material in any genome regardless of their potential mosaic ancestry composition and the habitats that promote them.


HTLV-1-infected CD4+ T-cells display alternative exon usages that culminate in adult T-cell leukemia.
Reprogramming cellular gene transcription sustains HTLV-1 viral persistence that ultimately leads to the development of adult T-cell leukemia/lymphoma (ATLL). We hypothesized that besides these quantitative transcriptional effects, HTLV-1 qualitatively modifies the pattern of cellular gene expression. Exon expression analysis shows that patients' untransformed and malignant HTLV-1(+) CD4(+) T-cells exhibit multiple alternate exon usage (AEU) events. These affect either transcriptionally modified or unmodified genes, culminate in ATLL, and unveil new functional pathways involved in cancer and cell cycle. Unsupervised hierarchical clustering of array data permitted to isolate exon expression patterns of 3977 exons that discriminate uninfected, infected, and transformed CD4(+) T-cells. Furthermore, untransformed infected CD4+ clones and ATLL samples shared 486 exon modifications distributed in 320 genes, thereby indicating a role of AEUs in HTLV-1 leukemogenesis. Exposing cells to splicing modulators revealed that Sudemycin E reduces cell viability of HTLV-1 transformed cells without affecting primary control CD4+ cells and HTLV-1 negative cell lines, suggesting that the huge excess of AEU might provide news targets for treating ATLL. Taken together, these data reveal that HTLV-1 significantly modifies the structure of cellular transcripts and unmask new putative leukemogenic pathways and possible therapeutic targets.


Stabilization of local structures by pi-CH and aromatic-backbone amide interactions involving prolyl and aromatic residues.
Weakly polar interactions between the side-chain aromatic rings and hydrogens of backbone amides (Ar-HN) and CHn of aliphatic groups (pi-CH) are known to form local structures and to stabilize secondary structure in peptides and proteins. To investigate the existence of these interactions and to explore their possible role in constraining the structures of Pro-Xaa and Xaa-Pro fragments in proteins, a database search was performed in a non-redundant set of proteins from the Brookheaven Protein Data Bank for pi-CH and Ar-HN interactions in Pro-Xaa and Xaa-Pro fragments (where Xaa is either Phe, Tyr or Trp). In Xaa-Pro fragments, the percentage of pi-CH interactions and Ar-HN interactions, respectively, was 20.6 and 3.2%, in Pro-Xaa fragments 26.8, 8.6 and 4.0% of the Pro-Xaa fragments contained both interactions, while no Xaa-Pro fragments had both. The protein fragments containing Ar-HN and/or pi-CH interactions were clustered on the basis of similarity of selected torsion angles. The clustering resulted in well defined clusters. Thus, pi-CH and Ar(i)-HN(i) interactions were able to constrain individual conformations of the Pro-Xaa and Xaa-Pro fragments. These local structures were found to be independent of the secondary structure of the polypeptide chains in which the fragments were found.


Six widespread bacterial clones among Escherichia coli K1 isolates.
Variable properties among Escherichia coli isolates include serotype, electrophoretic migration of major outer membrane proteins, metabolic properties, production of hemolysin or colicin or both, and plasmid content. These characteristics were compared in E. coli strains of capsular types K1, K5, K92, and K100 and in non-encapsulated isolates. The 234 bacterial strains from the United States and Europe which we studied had been isolated from healthy or diseased individuals recently or as long ago as 1941. Regardless of source, most O7:K1, O16:K1, and O75:K100 isolates could be assigned to three unique, serotype-specific groups, which were interpreted as representing three bacterial clones. Two bacterial (sub)clones each were discerned among the O18:K1 and O18:K5 isolates, and two further, distinct clones were discerned among the O1:K1 isolates. The implications of these results for epidemiological analyses and for virulence are discussed.


Oxidant Signaling Mediated by Nox2 in Neutrophils Promotes Regenerative Myelopoiesis and Tissue Recovery following Ischemic Damage.
Ischemic tissue damage activates hematopoietic stem and progenitor cells (HSPCs) in the bone marrow (BM)-generating myeloid cells, and persistent HSPC activity may drive chronic inflammation and impair tissue recovery. Although increased reactive oxygen species in the BM regulate HSPC functions, their roles in myelopoiesis of activated HSPCs and subsequent tissue recovery during ischemic damage are not well understood. In this paper, we report that deletion of Nox2 NADPH oxidase in mice results in persistent elevations in BM HSPC activity and levels of inflammatory monocytes/macrophages in BM and ischemic tissue in a model of hindlimb ischemia. Ischemic tissue damage induces oxidants in BM such as elevations of hydrogen peroxide and oxidized phospholipids, which activate redox-sensitive Lyn kinase in a Nox2-dependent manner. Moreover, during tissue recovery after ischemic injury, this Nox2-ROS-Lyn kinase axis is induced by Nox2 in neutrophils that home to the BM, which inhibits HSPC activity and inflammatory monocyte generation and promotes tissue regeneration after ischemic damage. Thus, oxidant signaling in the BM mediated by Nox2 in neutrophils regulates myelopoiesis of HSPCs to promote regeneration of damaged tissue.


Reaction monitoring using hyperpolarized NMR with scaling of heteronuclear couplings by optimal tracking.
Off-resonance decoupling using the method of Scaling of Heteronuclear Couplings by Optimal Tracking (SHOT) enables determination of heteronuclear correlations of chemical shifts in single scan NMR spectra. Through modulation of J-coupling evolution by shaped radio frequency pulses, off resonance decoupling using SHOT pulses causes a user-defined dependence of the observed J-splitting, such as the splitting of 13C peaks, on the chemical shift offset of coupled nuclei, such as 1H. Because a decoupling experiment requires only a single scan, this method is suitable for characterizing on-going chemical reactions using hyperpolarization by dissolution dynamic nuclear polarization (D-DNP). We demonstrate the calculation of [13C, 1H] chemical shift correlations of the carbanionic active sites from hyperpolarized styrene polymerized using sodium naphthalene as an initiator. While off resonance decoupling by SHOT pulses does not enhance the resolution in the same way as a 2D NMR spectrum would, the ability to obtain the correlations in single scans makes this method ideal for determination of chemical shifts in on-going reactions on the second time scale. In addition, we present a novel SHOT pulse that allows to scale J-splittings 50% larger than the respective J-coupling constant. This feature can be used to enhance the resolution of the indirectly detected chemical shift and reduce peak overlap, as demonstrated in a model reaction between p-anisaldehyde and isobutylamine. For both pulses, the accuracy is evaluated under changing signal-to-noise ratios (SNR) of the peaks from reactants and reaction products, with an overall standard deviation of chemical shift differences compared to reference spectra of 0.02ppm when measured on a 400MHz NMR spectrometer. Notably, the appearance of decoupling side-bands, which scale with peak intensity, appears to be of secondary importance.


Plantar shear stress distribution in patients with rheumatoid arthritis: relevance to foot pain.
Rheumatoid arthritis is an autoimmune disease that causes chronic, progressive joint inflammation; it commonly affects the joints of the feet. Biomechanical alterations and daily pain in the foot are the common outcomes of the disease. Earlier studies focusing on plantar pressure in such patients reported increased vertical loading along with peak pressure-pain associations. However, footwear designed according to the pressure profiles did not relieve symptoms effectively. We examined plantar shear and pressure distribution in patients with rheumatoid arthritis and compared the findings with those of controls, and we investigated a potential relationship between foot pain and local shear stresses. A custom-built platform was used to collect plantar pressure and shear stress data from nine patients with rheumatoid arthritis and 14 control participants. Seven patients reported the presence of pain under their feet. Pressure-time and shear-time integral values were also calculated. Peak pressure, pressure-time integral, resultant shear-time integral, and mediolateral shear stress magnitudes were higher in the complication group (P < .05). An association between peak shear-time integral and maximum pain locations was observed. Increased mediolateral shear stresses under the rheumatoid foot might be attributable to gait instability in such patients. A correlation between the locations of maximum shear-time integral and pain indicate the clinical significance of plantar shear in patients with rheumatoid arthritis.


The rational clinical examination. Is this patient hypovolemic?
To review, systematically, the physical diagnosis of hypovolemia in adults. We searched MEDLINE (January 1966-November 1997), personal files, and bibliographies of textbooks on physical diagnosis and identified 10 studies investigating postural vital signs or the capillary refill time of healthy volunteers, some of whom underwent phlebotomy of up to 1150 mL of blood, and 4 studies of patients presenting to emergency departments with suspected hypovolemia, usually due to vomiting, diarrhea, or decreased oral intake. When clinicians evaluate adults with suspected blood loss, the most helpful physical findings are either severe postural dizziness (preventing measurement of upright vital signs) or a postural pulse increment of 30 beats/min or more. The presence of either finding has a sensitivity for moderate blood loss of only 22% (95% confidence interval [CI], 6%-48%) but a much greater sensitivity for large blood loss of 97% (95% CI, 91%-100%); the corresponding specificity is 98% (95% CI, 97%-99%). Supine hypotension and tachycardia are frequently absent, even after up to 1150 mL of blood loss (sensitivity, 33%; 95% CI, 21%-47%, for supine hypotension). The finding of mild postural dizziness has no proven value. In patients with vomiting, diarrhea, or decreased oral intake, the presence of a dry axilla supports the diagnosis of hypovolemia (positive likelihood ratio, 2.8; 95% CI, 1.4-5.4), and moist mucous membranes and a tongue without furrows argue against it (negative likelihood ratio, 0.3; 95% CI, 0.1-0.6 for both findings). In adults, the capillary refill time and poor skin turgor have no proven diagnostic value. A large postural pulse change (> or =30 beats/min) or severe postural dizziness is required to clinically diagnose hypovolemia due to blood loss, although these findings are often absent after moderate amounts of blood loss. In patients with vomiting, diarrhea, or decreased oral intake, few findings have proven utility, and clinicians should measure serum electrolytes, serum blood urea nitrogen, and creatinine levels when diagnostic certainty is required.


Trajectories of Limitations in Instrumental Activities of Daily Living in Frail Older Adults With Vision, Hearing, or Dual Sensory Loss.
This study investigated the trajectories of decline in individual instrumental activities of daily living (IADL) with aging and the effect of hearing loss, vision loss, or dual sensory loss on these trajectories in community-living frail older persons. This longitudinal population-based study was conducted in 9,319 community-living frail Dutch persons aged 60 years and older. Self-reported hearing loss, vision loss, or dual sensory loss and nine IADL were assessed in 15 studies of the Dutch National Care for the Elderly Program (The Older Persons and Informal Caregivers Survey Minimum Dataset). Probabilities of limitations in IADL, odds ratios (per 5 years) for binary, and rate ratios (per 5 years) for score outcomes were calculated using mixed logistic and negative binomial models with age as the underlying timescale, stratified by sensory loss, and corrected for confounders. At baseline, the number of IADL limitations was higher in dual sensory loss (2.00 [interquartile range 1.00-4.00]) and vision loss (2.00 [interquartile range 1.00-4.00]) compared to no sensory loss (1.00 [interquartile range 0.00-2.00]) or hearing loss (1.00 [interquartile range 0.00-3.00]). Trajectories of individual IADL showed an increase in limitations in all IADL with age. Household tasks, traveling, shopping, preparing a meal, and walking showed the most rapid decline. Handling finances, traveling, and walking followed a different pattern of decline based on sensory loss status. The age effect on limitations in IADL appears to be similar across all types of sensory loss, with the exception of handling finances, traveling, and walking. At baseline, persons with self-reported sensory loss had higher levels of self-reported functional limitations. Trajectories depict a decline in IADL competence with age.


Probable photographic detection of the natural seventh-order rainbow.
We present a stacked and contrast-enhanced image comprised of 12 digital photographs that shows a series of color hues in the correct order and location to be part of the seventh-order rainbow. The observation was made on September 22, 2013, near Magdalena in New Mexico (USA). The seventh-order rainbow is located at 64° from the Sun in a region of the sky with little interference from the zero-order glow. The color hues in the image range from red to blue-violet, spanning about 12° in total extent; their locations generally agree with a numerical Debye-series simulation of the seventh-order rainbow. Despite the low color contrast of the seventh-order rainbow, the current observation indicates that it is feasible with current digital-imaging technology to detect this higher-order rainbow in near-ideal atmospheric conditions.


Minimal residual disease analysis in myeloma - when, why and where.
The primary hurdle in the path to curing multiple myeloma (MM) is defining a validated minimal residual disease (MRD) and its utility in the therapeutic decision making. A better definition of MRD will aid in tailoring MM therapy further to address the clonal heterogeneity and genomic instability and overcome patient's ineffective immune surveillance. MRD analysis can define the logical endpoint for maintenance therapy, in addition also aids in providing a better clinical end point for studies comparing novel agents in myeloma. MRD is a surrogate for the survival in MM. Guidelines for global incorporation of MRD in myeloma are fraught with lack of standardization, universal availability and abridged physicians' understanding of MRD modalities. We aimed at addressing some of the frequently asked questions in the MRD assessment and will also place in perspective some arguments in favor of MRD assessment in routine practice and clinical trial scenario.


Individual Differences in Hemodynamic Responses Measured on the Head Due to a Long-Term Stimulation Involving Colored Light Exposure and a Cognitive Task: A SPA-fNIRS Study.
When brain activity is measured by neuroimaging, the canonical hemodynamic response (increase in oxygenated hemoglobin ([O2Hb]) and decrease in deoxygenated hemoglobin ([HHb]) is not always seen in every subject. The reason for this intersubject-variability of the responses is still not completely understood. This study is performed with 32 healthy subjects, using the systemic physiology augmented functional near-infrared spectroscopy (SPA-fNIRS) approach. We investigate the intersubject variability of hemodynamic and systemic physiological responses, due to a verbal fluency task (VFT) under colored light exposure (CLE; blue and red). Five and seven different hemodynamic response patterns were detected in the subgroup analysis of the blue and red light exposure, respectively. We also found that arterial oxygen saturation and mean arterial pressure were positively correlated with [O2Hb] at the prefrontal cortex during the CLE-VFT independent of the color of light and classification of the subjects. Our study finds that there is substantial intersubject-variability of cerebral hemodynamic responses, which is partially explained by subject-specific systemic physiological changes induced by the CLE-VFT. This means that both subgroup analyses and the additional assessment of systemic physiology are of crucial importance to achieve a comprehensive understanding of the effects of a CLE-VFT on human subjects.


The neuropeptide neuromedin U stimulates innate lymphoid cells and type 2 inflammation.
The type 2 cytokines interleukin (IL)-4, IL-5, IL-9 and IL-13 have important roles in stimulating innate and adaptive immune responses that are required for resistance to helminth infection, promotion of allergic inflammation, metabolic homeostasis and tissue repair. Group 2 innate lymphoid cells (ILC2s) produce type 2 cytokines, and although advances have been made in understanding the cytokine milieu that promotes ILC2 responses, how ILC2 responses are regulated by other stimuli remains poorly understood. Here we demonstrate that ILC2s in the mouse gastrointestinal tract co-localize with cholinergic neurons that express the neuropeptide neuromedin U (NMU). In contrast to other haematopoietic cells, ILC2s selectively express the NMU receptor 1 (NMUR1). In vitro stimulation of ILC2s with NMU induced rapid cell activation, proliferation, and secretion of the type 2 cytokines IL-5, IL-9 and IL-13 that was dependent on cell-intrinsic expression of NMUR1 and Gαq protein. In vivo administration of NMU triggered potent type 2 cytokine responses characterized by ILC2 activation, proliferation and eosinophil recruitment that was associated with accelerated expulsion of the gastrointestinal nematode Nippostrongylus brasiliensis or induction of lung inflammation. Conversely, worm burden was higher in Nmur1-/- mice than in control mice. Furthermore, use of gene-deficient mice and adoptive cell transfer experiments revealed that ILC2s were necessary and sufficient to mount NMU-elicited type 2 cytokine responses. Together, these data indicate that the NMU-NMUR1 neuronal signalling circuit provides a selective mechanism through which the enteric nervous system and innate immune system integrate to promote rapid type 2 cytokine responses that can induce anti-microbial, inflammatory and tissue-protective type 2 responses at mucosal sites.


[Why do good drugs disappear?].
Old drugs, with proved efficacy and safety, are disappearing from the market. For instance, nitrofurantoin, an inexpensive effective agent for urinary tract infections with low incidence of bacterial resistance in comparison to other antibiotics, is becoming unavailable. Alpha-methyldopa and hydraLazine, drugs of choice for pregnancy hypertension, are no longer available. Chlorthalidone, a Long acting thiazide with best evidence on efficacy, is no Longer marketed in Israel. Switching to newer agents increases costs and is often associated with relative uncertainty about safety and efficacy. The reason for the disappearance of old drugs is a combination of market failures and failures in production and regulatory processes. System revisions are needed to allow continued availability of old, safe and effective drugs.


Adjuvant pembrolizumab versus placebo in resected stage III melanoma (EORTC 1325-MG/KEYNOTE-054): distant metastasis-free survival results from a double-blind, randomised, controlled, phase 3 trial.
The European Organisation for Research and Treatment of Cancer (EORTC) 1325/KEYNOTE-054 trial assessed pembrolizumab versus placebo in patients with resected high-risk stage III melanoma. At 15-month median follow-up, pembrolizumab improved recurrence-free survival (hazard ratio [HR] 0·57 [98·4% CI 0·43-0·74], p<0·0001) compared with placebo, leading to its approval in the USA and Europe. This report provides the final results for the secondary efficacy endpoint, distant metastasis-free survival and an update of the recurrence-free survival results. This double-blind, randomised, controlled, phase 3 trial was done at 123 academic centres and community hospitals across 23 countries. Patients aged 18 years or older with complete resection of cutaneous melanoma metastatic to lymph node, classified as American Joint Committee on Cancer staging system, seventh edition (AJCC-7) stage IIIA (at least one lymph node metastasis >1 mm), IIIB, or IIIC (without in-transit metastasis), and with an Eastern Cooperative Oncology Group performance status of 0 or 1 were eligible. Patients were randomly assigned (1:1) via a central interactive voice response system to receive intravenous pembrolizumab 200 mg or placebo every 3 weeks for up to 18 doses or until disease recurrence or unacceptable toxicity. Randomisation was stratified according to disease stage and region, using a minimisation technique, and clinical investigators, patients, and those collecting or analysing the data were masked to treatment assignment. The two coprimary endpoints were recurrence-free survival in the intention-to-treat (ITT) population and in patients with PD-L1-positive tumours. The secondary endpoint reported here was distant metastasis-free survival in the ITT and PD-L1-positive populations. This study is registered with ClinicalTrials.gov, NCT02362594, and EudraCT, 2014-004944-37. Between Aug 26, 2015, and Nov 14, 2016, 1019 patients were assigned to receive either pembrolizumab (n=514) or placebo (n=505). At an overall median follow-up of 42·3 months (IQR 40·5-45·9), 3·5-year distant metastasis-free survival was higher in the pembrolizumab group than in the placebo group in the ITT population (65·3% [95% CI 60·9-69·5] in the pembrolizumab group vs 49·4% [44·8-53·8] in the placebo group; HR 0·60 [95% CI 0·49-0·73]; p<0·0001). In the 853 patients with PD-L1-positive tumours, 3·5-year distant metastasis-free survival was 66·7% (95% CI 61·8-71·2) in the pembrolizumab group and 51·6% (46·6-56·4) in the placebo group (HR 0·61 [95% CI 0·49-0·76]; p<0·0001). Recurrence-free survival remained longer in the pembrolizumab group 59·8% (95% CI 55·3-64·1) than the placebo group 41·4% (37·0-45·8) at this 3·5-year follow-up in the ITT population (HR 0·59 [95% CI 0·49-0·70]) and in those with PD-L1-positive tumours 61·4% (56·3-66·1) in the pembrolizumab group and 44·1% (39·2-48·8) in the placebo group (HR 0·59 [95% CI 0·49-0·73]). Pembrolizumab adjuvant therapy provided a significant and clinically meaningful improvement in distant metastasis-free survival at a 3·5-year median follow-up, which was consistent with the improvement in recurrence-free survival. Therefore, the results of this trial support the indication to use adjuvant pembrolizumab therapy in patients with resected high risk stage III cutaneous melanoma. Merck Sharp & Dohme.


A Time to Save.
Group B Streptococcus (GBS), characterized by Lancefield in 1933, was not recognized as a human pathogen until the early 1970s when it emerged and replaced Escherichia coli as the most common cause of sepsis and meningitis among neonates and young infants. This article briefly gives a personnel account of the discovery of clinical syndromes of GBS distinguished by age at onset, vertical mode of transmission for early-onset disease, meningeal tropism for GBS capsular (CPS) type III strains, and protective CPS epitopes. It also reviews the difficult evolution of the now routine program for antenatal GBS culture screening and intrapartum antibiotic prophylaxis, development of the first GBS candidate vaccines, clinical trials documenting the immunogenicity and safety of CPS tetanus toxoid conjugate vaccines, ongoing need to prevent morbidity and mortality in neonates and young infants, and critical need for commercial vaccines for routine use in pregnant women.


Fe3O4@GO magnetic nanocomposites protect mesenchymal stem cells and promote osteogenic differentiation of rat bone marrow mesenchymal stem cells.
Fe3O4 nanoparticles (Fe3O4 NPs) are typical magnetic materials for bone tissue regeneration. However, the accompanying oxidative stress during the reaction process of Fe3O4 NPs and H2O2 in bone remodeling and disease may hinder their application. In order to reduce this side effect, we selected graphene oxide (GO) to modify Fe3O4 NPs. We showed that Fe3O4@GO magnetic nanocomposites (Fe3O4@GO MNCs) eliminated 30% of H2O2 in 3 h, and reduced the amount of ˙OH, the intermediate product of the Fenton reaction. The cellular study demonstrated that Fe3O4@GO MNCs reduced the cell damage caused by reactive oxygen species (ROS) and improved the activity of mesenchymal stem cells (MSCs). Moreover, when the magnetic field and bone morphogenetic protein-2 (BMP2) delivered by Fe3O4@GO MNCs worked together, osteogenic differentiation of MSCs in vitro was well promoted.


Importance of luminal membrane mesothelin expression in intraductal papillary mucinous neoplasms.
The present study demonstrated that luminal membrane mesothelin expression is a reliable prognostic factor in gastric cancer. Intraductal papillary mucinous neoplasms (IPMNs) often exhibit a spectrum of dysplasia, ranging between adenoma and carcinoma. Therefore, an immunohistochemical analysis of mesothelin expression in IPMN was performed in the present study, focusing on the localization of mesothelin. IPMNs were classified into two groups, IPMNs associated with invasive carcinoma and low-high (L-H) grade dysplasias. The tumors were classified as mesothelin-positive or -negative and in the mesothelin-positive cases, the localization of mesothelin was evaluated as luminal membrane- or cytoplasmic-positive. Among the 37 IPMNs, mesothelin expression was observed in 21 samples (56.8%), including 46.2% (12 out of 26) of the L-H dysplasia and 81.8% (9 out of 11) of the invasive carcinoma samples (P=0.071). Luminal membrane localization was observed in 10 samples (27%), including 15.4% (4/26) of the L-H dysplasia samples and 54.5% (6 out of 11) of the invasive carcinoma samples (P=0.022). Six patients experienced post-operative recurrence, with five of the recurrent tumors exhibiting mesothelin expression and all six exhibiting luminal membrane localization. It was concluded that immunohistochemical examinations for mesothelin expression and localization are clinically useful for prognostic assessments and decision making regarding further treatment subsequent to surgical procedures in patients with IPMN.


[Co-incineration in cement kilns: health and environmental risks].
In this article we discuss the development of hazardous waste co-incineration in cement kilns in Brazil as well as its impacts on health and the environment. Information was gathered through an extensive review on social and environmental impacts of co-incineration, and case studies, chosen after discussion with social movement representatives concerned with the co-incineration issue and related to the Brazilian Network on Environmental Justice, are described to illustrate the reality of co-incineration in Brazil. Studies showed that workers and community health suffers negative impacts from such practices in high-, middle- and low-income countries. In the Brazilian context, the institutional and social vulnerability intensifies these problems. To conclude, we argue for the necessity of increasing the institutional capacity of health and environmental agencies in Brazil, through staff training and better infra-structure. Additionally, we also propose a return of the debate about political and ethical aspects of industrial waste trade.


Population and size distribution of solute-rich mesospecies within mesostructured aqueous amino acid solutions.
Aqueous solutions of highly soluble substances such as small amino acids are usually assumed to be essentially homogenous systems with some degree of short range local structuring due to specific interactions on the sub-nanometre scale (e.g. molecular clusters, hydration shells), usually not exceeding several solute molecules. However, recent theoretical and experimental studies have indicated the presence of much larger supramolecular assemblies or mesospecies in solutions of small organic and inorganic molecules as well as proteins. We investigated both supersaturated and undersaturated aqueous solutions of two simple amino acids (glycine and DL-alanine) using Dynamic Light Scattering (DLS), Brownian Microscopy/Nanoparticles Tracking Analysis (NTA) and Cryogenic Transmission Electron Microscopy (Cryo-TEM). Colloidal scale mesospecies (nanodroplets) were previously reported in supersaturated solutions of these amino acids and were implicated as intermediate species on non-classical crystallization pathways. Surprisingly, we have found that the mesospecies are also present in significant numbers in undersaturated solutions even when the solute concentration is well below the solid-liquid equilibrium concentration (saturation limit). Thus, mesopecies can be observed with mean diameters ranging from 100 to 300 nm and a size distribution that broadens towards larger size with increasing solute concentration. We note that the mesospecies are not a separate phase and the system is better described as a thermodynamically stable mesostructured liquid containing solute-rich domains dispersed within bulk solute solution. At a given temperature, solute molecules in such a mesostructured liquid phase are subject to equilibrium distribution between solute-rich mesospecies and the surrounding bulk solution.


Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix.
Orthotopic liver transplantation is the only available treatment for severe liver failure, but it is currently limited by organ shortage. One technical challenge that has thus far limited the development of a tissue-engineered liver graft is oxygen and nutrient transport. Here we demonstrate a novel approach to generate transplantable liver grafts using decellularized liver matrix. The decellularization process preserves the structural and functional characteristics of the native microvascular network, allowing efficient recellularization of the liver matrix with adult hepatocytes and subsequent perfusion for in vitro culture. The recellularized graft supports liver-specific function including albumin secretion, urea synthesis and cytochrome P450 expression at comparable levels to normal liver in vitro. The recellularized liver grafts can be transplanted into rats, supporting hepatocyte survival and function with minimal ischemic damage. These results provide a proof of principle for the generation of a transplantable liver graft as a potential treatment for liver disease.


Interventions for promoting physical activity.
Little is known about the effectiveness of strategies to enable people to achieve an increase in their physical activity. To assess the effects of interventions for promoting physical activity in adults aged 16 years and older, not living in an institution. We searched CENTRAL (Issue 4, 2001), MEDLINE, EMBASE, CINAHL, PsychLIT, BIDS ISI, SPORTDISCUS, SIGLE, SCISEARCH (from earliest date available to December 2001) and reference lists of articles. Randomised, controlled, trials comparing different interventions to encourage sedentary adults not living in an institution to become physically active. Studies required a minimum of six months follow up from the start of the intervention to the collection of final data and either used an intention to treat analysis or, failing that, had no more than 20% loss to follow up. At least two reviewers independently assessed each study quality and extracted data. Study authors were contacted for additional information where necessary. Standardised mean differences and 95% confidence intervals were calculated for continuous measures of self reported physical activity and cardio-respiratory fitness. For studies with dichotomous outcomes, odds ratios and 95% confidence intervals were calculated. The effect of interventions on self reported physical activity (11 studies; 3940 participants) was positive and moderate, with a pooled standardised mean difference of 0.31 (95% CI 0.12 to 0.50), as was the effect on cardio-respiratory fitness (7 studies; 1406 participants) pooled SMD 0.4 (95% CI 0.09 to 0.70). The effect of interventions in achieving a predetermined threshold of physical activity (6 studies; 2313 participants) was not significant with an odds ratio of 1.30 (95% CI 0.87 to 1.95). There was significant heterogeneity in the reported effects as well as heterogeneity in characteristics of the interventions. The heterogeneity in reported effects was reduced in higher quality studies, when physical activity was self-directed with some professional guidance and when there was on-going professional support. Our review suggests that physical activity interventions have a moderate effect on self reported physical activity and cardio-respiratory fitness, but not on achieving a predetermined level of physical activity. Due to the clinical and statistical heterogeneity of the studies, only limited conclusions can be drawn about the effectiveness of individual components of the interventions. Future studies should provide greater detail of the components of interventions.


Localization of complement component 3 on Streptococcus pneumoniae: anti-capsular antibody causes complement deposition on the pneumococcal capsule.
We have previously shown that complement component 3 (C3) deposited onto encapsulated Streptococcus pneumoniae by anti-capsular antibody (Ab) is a more efficient opsonin in vitro and in vivo than C3 deposited by anti-cell wall Ab (Brown et al., J. Clin. Invest. 69:85-98, 1982). In the present study, we explored the cellular location of C3b molecules that differ in opsonic efficiency by using avidin-ferritin to localize biotinylated Ab and C3 molecules on S. pneumoniae for electron microscopy. Anti-cell wall Ab and C3b molecules deposited by this Ab on unencapsulated S. pneumoniae were localized to S. pneumoniae cell walls. Anti-capsular Ab and C3b deposited by this Ab were seen in clusters on encapsulated S. pneumoniae at a distance from the cell wall. However, no avidin-ferritin staining of encapsulated S. pneumoniae was seen on incubation with biotinyl-anti-cell wall Ab, biotinylated C3 fixed by anti-cell wall Ab, or nonimmune serum containing biotinyl-C3. In each case, uptake of the biotinylated component was proven by radioactivity measurements, since biotinylated Ab and C3 were also radiolabeled with 125I. When avidin-ferritin did not bind to biotinylated components. Ouchterlony analysis indicated that C3 was bound to cell wall components on the encapsulated organisms. Thus, we conclude that, for encapsulated S. pneumoniae, opsonically efficient C3b molecules, deposited by anti-capsular Ab, are located on the S. pneumoniae capsule, whereas the opsonically inefficient C3b molecules deposited by anti-cell wall Ab or nonimmune serum are located on the cell wall. A major reason for the increased virulence of encapsulated compared to unencapsulated S. pneumoniae is that, in the absence of anti-capsular Ab, the S. pneumoniae capsule interferes with the recognition of cell wall-bound C3b molecules by phagocytic cell receptors.


Erythema nodosum and lung ailments at the clinic for dermatological and venereal diseases in Sarajevo from 1989 to 2005.
In 15 years period (1989 - 2004) we have treated at our Clinic a total of 101 patients diagnosed with erythema nodosum which makes 2.9% of the total number of patients treated in this period. In order to explore correlation between dermatological and lung diseases we applied statistical analysis with respect to number, sex, age and irregularities occurring on the skin surface. Female patients dominated with 96 of them (95.6%) in the group, while only five patients were male (4.3%). This makes 22 : 1 scale in favor of female patients, as opposed to usual 5:1 ratio. Similar predominance (86%) was found in the study conducted by Mert and Gurkan (1.2). Average age of the patients was 41.6 years, which is slightly higher than the average of 18 and 34. The youngest patient was 17 and oldest 77.


The effects of suspension particle size on the performance of air-jet, ultrasonic and vibrating-mesh nebulisers.
Using latex microspheres as model suspensions, the influence of suspension particle size (1, 4.5 and 10 μm) on the properties of aerosols produced using Pari LC Sprint (air-jet), Polygreen (ultrasonic), Aeroneb Pro (actively vibrating-mesh) and Omron MicroAir NE-U22 (passively vibrating-mesh) nebulisers was investigated. The performance of the Pari nebuliser was independent of latex spheres particle size. For both Polygreen and Aeroneb Pro nebulizers, total aerosol output increased when the size of latex spheres increased, with highest fine particle fraction (FPF) values being recorded. However, following nebulisation of 1 or 4.5 μm suspensions with the Polygreen device, no particles were detected in the aerosols deposited in a two-stage impinger, suggesting that the aerosols generated from this device consisted mainly of the continuous phase while the dispersed microspheres were excluded and remained in the nebuliser. The Omron nebuliser efficiently nebulised the 1 μm latex spheres, with high output rate and no particle aggregation. However, this device functioned inefficiently when delivering 4.5 or 10 μm suspensions, which was attributed to the mild vibrations of its mesh and/or the blockage of the mesh apertures by the microspheres. The Aeroneb Pro fragmented latex spheres into smaller particles, but uncontrolled aggregation occurred upon nebulisation. This study has shown that the design of the nebuliser influenced the aerosol properties using latex spheres as model suspensions. Moreover, for the recently marketed mesh nebulisers, the performance of the Aeroneb Pro device was less dependent on particle size of the suspension compared with the Omron MicroAir nebuliser.


Ruthenium tetroxide for fixing and staining cytoplasmic membranes.
Rat liver and kidney were fixed for electron microscopy with ruthenium tetroxide. When compared with tissues fixed with osmiun tetroxide, the membranes showed well without additional staining. The trilaminar structuire of the cytoplasmic membranes was clearly seen. In width the nuclear, mitochondrial, and cytoplasmic membranes investigated resembled the cell memnbrane.


Mortality and cause of death in a cohort of people who had ever injected drugs in Glasgow: 1982-2012.
To describe all-cause and cause-specific mortality in a cohort of people who had ever injected drugs (PWID) with a low prevalence of HIV over 20-30 years. Using a retrospective study design, identifying data from a cohort of PWID recruited between 1982 and 1993 through in-patient drug treatment services were linked to National Records for Scotland deaths data using probabilistic record linkage. We report all-cause and cause-specific mortality rates; standardized mortality ratios (SMR) across time, gender and age were estimated. Among 456 PWID, 139 (30.5%) died over 9024 person-years (PY) of follow-up. Mortality within the cohort was almost nine times higher than the general population, and remained elevated across all age groups. The greatest excess mortality rate was in the youngest age group, who were 15-24 years of age (SMR 31.6, 95% CI 21.2-47.1). Drug-related deaths declined over time and mortality was significantly higher among HIV positive participants. Although SMRs declined with follow-up, the SMR of the oldest age group (45-60) was 4.5 (95% CI 3.0-6.9). There were no significant differences in all-cause mortality rates between participants who were 25 years and older at cohort entry compared to younger participants. Mortality rates remained higher than the general population across all age groups. Screening services that identify a history of injecting drug use may be an opportunity to address risk factors faced by an ageing population of PWID and potentially have implications for future health care planning.


State disparities in time trends of adolescent body mass index percentile and weight-related behaviors in the United States.
Evidence is conflicting as to whether youth obesity prevalence has reached a plateau in the United States overall. Trends vary by state, and experts recommend exploring whether trends in weight-related behaviors are associated with changes in weight status trends. Thus, our objective was to estimate between-state variation in time trends of adolescent body mass index (BMI) percentile and weight-related behaviors from 2001 to 2007. A time series design combined cross-sectional Youth Risk Behavior Survey data from 272,044 adolescents in 29 states from 2001 to 2007. Self-reported height, weight, sports participation, physical education, television viewing, and daily consumption of 100% fruit juice, milk, and fruits and vegetables were collected. Linear mixed models estimated state variance in time trends of behaviors and BMI percentile. Across states, BMI percentile trends were consistent despite differences in behavioral trends. Boys experienced a modest linear increase in BMI percentile (ß = 0.18, 95% CI: 0.07, 0.30); girls experienced a non-linear increase, as the rate of increase declined over time from 1.02 units in 2001-2002 (95% CI: 0.68, 1.36) to 0.23 units in 2006-2007 (95% CI: -0.09, 0.56). States in which BMI percentile decreased experienced a greater decrease in TV viewing than states where BMI percentile increased. Otherwise, states with disparate BMI percentile trends did not differ with respect to behaviors. Future research should explore the role of other behaviors (e.g., soda consumption), measurement units (e.g., portion size), and societal trends (e.g., urban sprawl) on state and national adiposity trends.


Tuning of superconducting niobium nitride terahertz metamaterials.
Superconducting planar terahertz (THz) metamaterials (MMs), with unit cells of different sizes, are fabricated on 200 nm-thick niobium nitride (NbN) films deposited on MgO substrates. They are characterized using THz time domain spectroscopy over a temperature range from 8.1 K to 300 K, crossing the critical temperature of NbN films. As the gap frequency (f(g) = 2Δ0/h, where Δ0 is the energy gap at 0 K and h is the Plank constant) of NbN is 1.18 THz, the experimentally observed THz spectra span a frequency range from below f(g) to above it. We have found that, as the resonance frequency approaches f(g), the relative tuning range of MMs is quite wide (30%). We attribute this observation to the large change of kinetic inductance of superconducting film.


Relationship between retinopathy and asymptomatic atherosclerosis determined by measurement of carotid intima-media thickness in patients with type 2 diabetes mellitus.
Presence of diabetic retinopathy (DR) may be used as an early marker of atherosclerosis in type 2 diabetes mellitus (DM) patients. This study aimed to investigate the relationship between the presence of DR and carotid intima-media thickness (CIMT), which is an indicator of early atherosclerosis in patients with type 2 DM. Thirty DM patients with retinopathy (DR group), 28 DM patients without retinopathy (non-DR group), and 27 healthy controls (control group) were included in the study. CIMT was assessed using a high-resolution B-mode ultrasonography device. Mean CIMT was found to be 0.9±0.17 mm in the DR group, 0.8±0.16 mm in the non-DR group, and 0.7±0.13 mm in the control group. CIMT was found to be statistically significantly higher in the DR group compared to the other 2 groups (p<0.001). When multivariate analysis was performed, presence of DR still remained as an independent risk factor for increased CIMT values. Presence of DR in type 2 DM patients is an independent risk factor in terms of increased CIMT, which is considered to be a finding of subclinical atherosclerosis. Therefore, we believe that type 2 DM patients with retinopathy should be closely followed in terms of cardiovascular events.


On-chip constructive cell-network study (II): on-chip quasi-in vivo cardiac toxicity assay for ventricular tachycardia/fibrillation measurement using ring-shaped closed circuit microelectrode with lined-up cardiomyocyte cell network.
Conventional in vitro approach using human ether-a-go-go related gene (hERG) assay has been considered worldwide as the first screening assay for cardiac repolarization safety. However, it does not always oredict the potential QT prolongation risk or pro-arrhythmic risk correctly. For adaptable preclinical strategiesto evaluate global cardiac safety, an on-chip quasi-in vivo cardiac toxicity assay for lethal arrhythmia (ventricular tachyarrhythmia) measurement using ring-shaped closed circuit microelectrode chip has been developed. The ventricular electrocardiogram (ECG)-like field potential data, which includes both the repolarization and the conductance abnormality, was acquired from the self-convolutied extracellular field potentials (FPs) of a lined-up cardiomyocyte network on a circle-shaped microelectrode in an agarose microchamber. When Astemisol applied to the closed-loop cardiomyocyte network, self-convoluted FP profile of normal beating changed into an early afterdepolarization (EAD) like waveform, and then showed ventricular tachyarrhythmias and ventricular fibrilations (VT/Vf). QT-prolongation-like self-convoluted FP duration prolongation and its fluctuation increase was also observed according to the increase of Astemizole concentration. The results indicate that the convoluted FPs of the quasi-in vivo cell network assay includes both of the repolarization data and the conductance abnormality of cardiomyocyte networks has the strong potential to prediction lethal arrhythmia.


Inhibition of GNNQQNY prion peptide aggregation by trehalose: a mechanistic view.
Deposition of amyloid fibrils is the seminal event in the pathogenesis of numerous neurodegenerative diseases. The formation of this amyloid assembly is the manifestation of a cascade of structural transitions including toxic oligomer formation in the early stages of aggregation. Thus a viable therapeutic strategy involves the use of small molecular ligands to interfere with this assembly. In this perspective, we have explored the kinetics of aggregate formation of the fibril forming GNNQQNY peptide fragment from the yeast prion protein SUP35 using multiple all atom MD simulations with explicit solvent and provided mechanistic insights into the way trehalose, an experimentally known aggregation inhibitor, modulates the aggregation pathway. The results suggest that the assimilation process is impeded by different barriers at smaller and larger oligomeric sizes: the initial one being easily surpassed at higher temperatures and peptide concentrations. The kinetic profile demonstrates that trehalose delays the aggregation process by increasing both these activation barriers, specifically the latter one. It increases the sampling of small-sized aggregates that lack the beta sheet conformation. Analysis reveals that the barrier in the growth of larger stable oligomers causes the formation of multiple stable small oligomers which then fuse together bimolecularly. The PCA of 26 properties was carried out to deconvolute the events within the temporary lag phases, which suggested dynamism in lags involving an increase in interchain contacts and burial of SASA. The predominant growth route is monomer addition, which changes to condensation on account of a large number of depolymerisation events in the presence of trehalose. The favourable interaction of trehalose specifically with the sidechain of the peptide promotes crowding of trehalose molecules in its vicinity - the combination of both these factors imparts the observed behaviour. Furthermore, increasing trehalose concentration leads to faster expulsion of water molecules than interpeptide interactions. These expelled water molecules have larger translational movement, suggesting an entropy factor to favor the assembly process. Different conformations observed under this condition suggest the role of water molecules in guiding the morphology of the aggregates as well. A similar scenario exists on increasing peptide concentration.


Oscillating on borrowed time: diffusible signals from immortalized suprachiasmatic nucleus cells regulate circadian rhythmicity in cultured fibroblasts.
The capacity to generate circadian rhythms endogenously and to confer this rhythmicity to other cells was compared in immortalized cells derived from the suprachiasmatic nucleus (SCN) and a fibroblast line to differentiate SCN pacemaker properties from the oscillatory behavior of non-clock tissues. Only SCN2.2 cells were capable of endogenously generating circadian rhythms in 2-deoxyglucose uptake and Per gene expression. Similar to SCN function in vivo, SCN2.2 cells imposed rhythms of metabolic activity and Per gene expression on cocultured NIH/3T3 fibroblasts via a diffusible signal. The conferred rhythms in NIH/3T3 cells were phase delayed by 4-12 hr relative to SCN2.2 circadian patterns, thus resembling the phase relationship between SCN and peripheral tissue rhythms in vivo. Sustained metabolic rhythmicity in NIH/3T3 cells was dependent on continued exposure to SCN2.2-specific outputs. In response to a serum shock the NIH/3T3 fibroblasts exhibited recurrent oscillations in clock gene expression, but not in metabolic activity. These molecular rhythms in serum-shocked fibroblasts cycled in a phase relationship similar to that observed in the SCN in vivo; peak Per1 and Per2 mRNA expression preceded the rhythmic maxima in Cry1 and Cry2 mRNA levels by 4 hr. Despite these clock gene oscillations the serum-shocked NIH/3T3 cells failed to drive circadian rhythms of Per1 and Per2 expression in cocultures of untreated fibroblasts, suggesting that expression and circadian regulation of the Per and Cry genes are not sufficient to confer pacemaker function. Therefore, SCN-specific outputs are necessary to drive circadian rhythms of metabolic activity, and these output signals are not a direct product of clock gene oscillations.


Structural basis of the atypical activation mechanism of KRASV14I.
RAS regulation and signaling are largely accomplished by direct protein-protein interactions, making RAS protein dynamics a critical determinant of RAS function. Here, we report a crystal structure of GDP-bound KRASV14I, a mutated KRAS variant associated with the developmental RASopathy disorder Noonan syndrome (NS), at 1.5-1.6 Å resolution. The structure is notable for revealing a marked extension of switch 1 away from the G-domain and nucleotide-binding site of the KRAS protein. We found that this extension is associated with a loss of the magnesium ion and a tilt in the position of the guanine base because of the additional carbon introduced by the isoleucine substitution. Hydrogen-deuterium exchange MS analysis confirmed that this conformation occurs in solution, but also disclosed a difference in kinetics when compared with KRASA146T, another RAS mutant that displays a nearly identical conformation in previously reported crystal structures. This conformational change contributed to a high rate of guanine nucleotide-exchange factor (GEF)-dependent and -independent nucleotide exchange and to an increase in affinity for SOS Ras/Rac GEF 1 (SOS1), which appears to be the major mode of activation for this RAS variant. These results highlight a mechanistic connection between KRASA146T and KRASV14I that may have implications for the regulation of these variants and for the development of therapeutic strategies to manage KRAS variant-associated disorders.


Core-shell nanoparticles: synthesis and applications in catalysis and electrocatalysis.
Core-shell nanoparticles (CSNs) are a class of nanostructured materials that have recently received increased attention owing to their interesting properties and broad range of applications in catalysis, biology, materials chemistry and sensors. By rationally tuning the cores as well as the shells of such materials, a range of core-shell nanoparticles can be produced with tailorable properties that can play important roles in various catalytic processes and offer sustainable solutions to current energy problems. Various synthetic methods for preparing different classes of CSNs, including the Stöber method, solvothermal method, one-pot synthetic method involving surfactants, etc., are briefly mentioned here. The roles of various classes of CSNs are exemplified for both catalytic and electrocatalytic applications, including oxidation, reduction, coupling reactions, etc.


In vitro activity of Ro 23-6240, a new fluorinated 4-quinolone.
The in vitro activity of Ro 23-6240 (AM833), 6,8-difluoro-1-(2-fluoroethyl)-1,4-dihydro-4-oxo-7(4-methyl-1-piper azinyl) quinolone-3-carboxylic acid, was compared with those of norfloxacin, enoxacin, ofloxacin, and ciprofloxacin. Ro 23-6240 inhibited the majority of Enterobacteriaceae isolates at a concentration of less than or equal to 0.5 microgram/ml. It was especially active against Shigella sp., Salmonella sp., Escherichia coli, and Yersinia enterocolitica, with an MIC for 90% of the strains of less than or equal to 0.12 microgram/ml. The MIC for 90% of the strains was 1 microgram/ml for Serratia marcescens and 8 micrograms/ml for Pseudomonas aeruginosa. Staphylococcus aureus isolates, including methicillin-resistant strains, were inhibited by less than or equal to 1 microgram/ml. Streptococcal and anaerobic species were inhibited by 8 to 16 micrograms/ml. Ro 23-6240 inhibited beta-lactamase-producing bacteria resistant to broad-spectrum cephalosporins. The overall activity of Ro 23-6240 was similar to those of enoxacin, norfloxacin, and ofloxacin, but less than that of ciprofloxacin. The frequency of spontaneous resistance was low, although resistant bacteria could be isolated by repeated subculture. The activity of Ro 23-6240 was decreased in the presence of magnesium at concentrations similar to those present in urine.


Regulation of a truncated form of tropomyosin-related kinase B (TrkB) by Hsa-miR-185* in frontal cortex of suicide completers.
TrkB-T1 is a BDNF receptor lacking a tyrosine kinase domain that is highly expressed in astrocytes and regulates BDNF-evoked calcium transients. Previous studies indicate that downregulation of TrkB-T1 in frontal cortex may be involved in neurobiological processes underlying suicide. In a microarray screening study (N = 8), we interrogated all known microRNA in the frontal cortex of suicide completers with low expression of TrkB-T1 and normal controls. These findings were validated and followed up in a larger sample of cases and controls (N = 55). Functional analyses included microRNA silencing, microRNA overexpression and luciferase assays to investigate specificity and to validate interactions between differentially expressed microRNA and TrkB-T1. MicroRNAs Hsa-miR-185* and Hsa-miR-491-3p were upregulated in suicide completers with low expression of TrkB.T1 (P(nominal): 9.10(-5) and 1.8.10(-4) respectively; FDR-corrected p = 0.031). Bioinformatic analyses revealed five putative binding sites for the DiGeorge syndrome linked microRNA Hsa-miR-185*in the 3'UTR of TrkB-T1, but none for Hsa-miR-491-3P. The increase of Hsa-miR-185* in frontal cortex of suicide completers was validated then confirmed in a larger, randomly selected group of suicide completers, where an inverse correlation between Hsa-miR-185* and TrkB-T1 expression was observed (R = -0.439; p = 0.001). Silencing and overexpression studies performed in human cell lines confirmed the inverse relationship between hsa-mir-185* and trkB-T1 expression. Luciferase assays demonstrated that Hsa-miR-185* binds to sequences in the 3'UTR of TrkB-T1. These results suggest that an increase of Hsa-miR-185* expression levels regulates, at least in part, the TrkB-T1 decrease observed in the frontal cortex of suicide completers and further implicate the 22q11 region in psychopathology.


Study protocol for a 9-month randomised controlled trial assessing the effects of almonds versus carbohydrate-rich snack foods on weight loss and weight maintenance.
Epidemiological studies indicate an inverse association between nut consumption and body mass index (BMI). However, clinical trials evaluating the effects of nut consumption compared with a nut-free diet on adiposity have reported mixed findings with some studies reporting greater weight loss and others reporting no weight change. This paper describes the rationale and detailed protocol for a randomised controlled trial assessing whether the inclusion of almonds or carbohydrate-rich snacks in an otherwise nut-free energy-restricted diet will promote weight loss during 3 months of energy restriction and limit weight regain during 6 months of weight maintenance. One hundred and thirty-four adults aged 25-65 years with a BMI of 27.5-34.9 kg/m2 will be recruited and randomly allocated to either the almond-enriched diet (AED) (15% energy from almonds) or a nut-free control diet (NFD) (15% energy from carbohydrate-rich snack foods). Study snack foods will be provided. Weight loss will be achieved through a 30% energy restriction over 3 months, and weight maintenance will be encouraged for 6 months by increasing overall energy intake by ~120-180 kcal/day (~500-750kJ/day) as required. Food will be self-selected, based on recommendations from the study dietitian. Body composition, resting energy expenditure, total daily energy expenditure (via doubly labelled water), physical activity, appetite regulation, cardiometabolic health, gut microbiome, liver health, inflammatory factors, eating behaviours, mood and personality, functional mobility and pain, quality of life and sleep patterns will be measured throughout the 9-month trial. The effects of intervention on the outcome measures over time will be analysed using random effects mixed models, with treatment (AED or NFD) and time (baseline, 3 months and 9 months) being the between and within factors, respectively in the analysis. Ethics approval was obtained from the University of South Australia Human Research Ethics Committee (201436). Results from this trial will be disseminated through publication in peer-reviewed journals, national and international presentations. Australian New Zealand Clinical Trials Registry (ACTRN12618001861246).


Bidirectional Relations of Impulsive Personality and Alcohol Use Across Three Waves of Data Collection.
To explore the bidirectional relations between alcohol use and three impulsive personality traits, to advance understanding of risk processes. 525 college students (mean age = 18.95 years) recruited in August 2008 and 2009 and followed up annually for three years. Personality and past/current substance use were assessed. T2 sensation seeking mediated the predictive relationship between T1 and T3 alcohol use, and T2 alcohol use mediated the predictive relationship between T1 and T3 sensation seeking. In addition, T2 alcohol problems mediated the predictive relationship between T1 alcohol use and T3 negative urgency. Findings support a bidirectional relationship between sensation seeking and alcohol use, and drinking anticipates drinking problems, which predict increases in negative urgency. For some individuals, there appears to be an ongoing process of increased risk in the form of increases in both drinking and high-risk personality traits.


Highly Amino Acid Selective Hydrolysis of Myoglobin at Aspartate Residues as Promoted by Zirconium(IV)-Substituted Polyoxometalates.
SDS-PAGE/Edman degradation and HPLC MS/MS showed that zirconium(IV)-substituted Lindqvist-, Keggin-, and Wells-Dawson-type polyoxometalates (POMs) selectively hydrolyze the protein myoglobin at Asp-X peptide bonds under mildly acidic and neutral conditions. This transformation is the first example of highly sequence selective protein hydrolysis by POMs, a novel class of protein-hydrolyzing agents. The selectivity is directed by Asp residues located on the surface of the protein and is further assisted by electrostatic interactions between the negatively charged POMs and positively charged surface patches in the vicinity of the cleavage site.


Benzene oxidation coupled to sulfate reduction.
Highly reduced sediments from San Diego Bay, Calif., that were incubated under strictly anaerobic conditions metabolized benzene within 55 days when they were exposed initially to 1 (mu)M benzene. The rate of benzene metabolism increased as benzene was added back to the benzene-adapted sediments. When a [(sup14)C]benzene tracer was included with the benzene added to benzene-adapted sediments, 92% of the added radioactivity was recovered as (sup14)CO(inf2). Molybdate, an inhibitor of sulfate reduction, inhibited benzene uptake and production of (sup14)CO(inf2) from [(sup14)C]benzene. Benzene metabolism stopped when the sediments became sulfate depleted, and benzene uptake resumed when sulfate was added again. The stoichiometry of benzene uptake and sulfate reduction was consistent with the hypothesis that sulfate was the principal electron acceptor for benzene oxidation. Isotope trapping experiments performed with [(sup14)C]benzene revealed that there was no production of such potential extracellular intermediates of benzene oxidation as phenol, benzoate, p-hydroxybenzoate, cyclohexane, catechol, and acetate. The results demonstrate that benzene can be oxidized in the absence of O(inf2), with sulfate serving as the electron acceptor, and suggest that some sulfate reducers are capable of completely oxidizing benzene to carbon dioxide without the production of extracellular intermediates. Although anaerobic benzene oxidation coupled to chelated Fe(III) has been documented previously, the study reported here provides the first example of a natural sediment compound that can serve as an electron acceptor for anaerobic benzene oxidation.


Exploring the meaning of everyday life, for those suffering from dementia.
Living with dementia means struggling to preserve a sense of self, to retain (and reevaluate) one's values in a new situation, and to search for a new way of life. This study explored the meaning of everyday life as expressed by 11 people suffering from dementia. Data were collected through interviews with people in the early stage of a diagnosed dementia disease. The findings indicate that many people with dementia feel shame, sorrow, and sadness when their life with the disease begins but also indicate prospects of a more manageable life. Since the reaction and adjustment to dementia differs from person to person, professional caregivers need to use a case-specific approach in the provision of care.


Recent developments in the use of immunotherapy in non-small cell lung cancer.
Targeted therapies have significantly improved the prognosis of subsets of patients with advanced non-small-cell lung cancer (NSCLC) harboring somatically activated oncogenes, such as mutant EGFR and rearranged ALK. However, the efficacy of these agents is limited by the development of acquired resistance which occurs after variable periods of time. Therefore, there is an urgent need for novel therapeutic strategies to achieve long lasting disease control, as well as new therapies for those patients without targetable driver mutations. A deeper understanding of interactions between the immune system and tumor cells has led to the development of a number of immunotherapeutic agents. We review current data on immunotherapy for lung cancer treatment, with a focus on checkpoint inhibitors and therapeutic vaccines. References for this review were identified through searches of PubMed, congress proceedings and reference lists from key original and review papers. Expert commentary: While most vaccines have been unsuccessful, inhibitors of specific immune checkpoints, including CTLA-4 and PD-1/PD-L1 pathway, have shown significant clinical activity and manageable toxicities and recently, two anti-PD-1 monoclonal antibodies have been approved by the FDA for treatment of patients with advanced NSCLC. Identification of reliable predictive biomarkers for patient selection and novel rational combinations are currently active areas of research to further improve the efficacy of immunotherapy.


Reconstitution of glutamate receptor proteins purified from Xenopus central nervous system into artificial bilayers.
Excitatory amino acid (EAA) receptor (EAAR) proteins purified from Xenopus central nervous system using a domoate affinity column and then separated into fractions using sucrose density gradient centrifugation were reconstituted, first into liposomes and then into planar lipid bilayers, using pipette-dipping and black lipid membrane techniques. Although the protein was eluted from the column with either alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) or kainate and could not be eluted with N-methyl-D-aspartate (NMDA), channel openings were obtained after exposure of the bilayers to kainate, AMPA, or NMDA (usually only in the presence of glycine). In bilayers exhibiting a single open channel conductance level this was approximately 6 pS with AMPA, approximately 9 pS with kainate, and approximately 50 pS with NMDA. However, with a few batches of protein unitary channel openings of up to 400 pS were observed, suggesting that reconstituted EAAR may sometimes form functional aggregates. The protein eluted from the domoate column was divided into two fractions on a sucrose density gradient. After reconstitution, one fraction responded to all three EAAs, whereas the other responded only to the non-NMDA receptor agonists. An explanation for these results is that some of the EAAR eluted from the column contain NMDA receptor subunits in addition to non-NMDA receptor subunits.


Analysis of American College of Surgeons trauma consultation program.
To identify the criteria deficiencies found during peer consultation of hospitals and the relationship to subsequent verification. Between September 1987 and December 1992, 52 hospitals had consultation visits using American College of Surgeons criteria. Each report was studied for deficiencies, frequency of deficiencies, and relationship to verification. There are 108 American College of Surgeons criteria. Thirty-five different criteria deficiencies were found. The number of deficiencies per hospital ranged from zero to 12. The more frequent deficiencies included a lack of the following: quality improvement, 35 (67%); trauma service, 20 (38%); trauma surgeon in emergency department, 20 (38%); 24-hour operating room availability, 17 (33%); trauma registry, 17 (33%); trauma continuing medical education, 16 (31%); trauma director, 15 (29%); computed tomography technician in hospital, 15 (29%); research, 14 (27%); trauma coordinator, 14 (27%); and neurosurgeon availability, 13 (25%). No hospital that lacked commitment of surgeons (n = 12) or hospital (n = 3) requested a verification visit. Twenty-four hospitals (46%) achieved verification by February 1994. Twenty-eight hospitals had six or fewer deficiencies, with 19 (68%) verified. Twenty-four hospitals had seven or more deficiencies, with only five (21%) subsequently verified. Verification visits followed consultation by 3 to 52 months. Two hospitals with nine deficiencies were verified after 30 and 48 months, although one failed its first verification visit. American College of Surgeons consultation assists hospitals to identify their trauma center capability and appears to improve their ability to pass subsequent trauma center verification. Most criteria deficiencies are correctable. Lack of commitment by the surgeons or hospital is difficult to correct. There is an inverse relationship between the number of deficiencies and subsequent verification.


Interactions in lipid stabilised foam films.
The interaction between lipid bilayers in water has been intensively studied over the last decades. Osmotic stress was applied to evaluate the forces between two approaching lipid bilayers in aqueous solution. The force-distance relation between lipid mono- or bilayers deposited on mica sheets using a surface force apparatus (SFA) was also measured. Lipid stabilised foam films offer another possibility to study the interactions between lipid monolayers. These films can be prepared comparatively easy with very good reproducibility. Foam films consist usually of two adsorbed surfactant monolayers separated by a layer of the aqueous solution from which the film is created. Their thickness can be conveniently measured using microinterferometric techniques. Studies with foam films deliver valuable information on the interactions between lipid membranes and especially their stability and permeability. Presenting inverse black lipid membrane (BLM) foam films supply information about the properties of the lipid self-organisation in bilayers. The present paper summarises results on microscopic lipid stabilised foam films by measuring their thickness and contact angle. Most of the presented results concern foam films prepared from dispersions of the zwitterionic lipid 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC) and some of its mixtures with the anionic lipid -- 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DMPG). The strength of the long range and short range forces between the lipid layers is discussed. The van der Waals attractive force is calculated. The electrostatic repulsive force is estimated from experiments at different electrolyte concentrations (NaCl, CaCl₂) or by modification of the electrostatic double layer surface potential by incorporating charged lipids in the lipid monolayers. The short range interactions are studied and modified by using small carbohydrates (fructose and sucrose), ethanol (EtOH) or dimethylsulfoxide (DMSO). Some results are compared with the structure of lipid monolayers deposited at the liquid/air interface (monolayers spread in Langmuir trough), which are one of most studied biomembrane model system. The comparison between the film thickness and the free energy of film formation is used to estimate the contribution of the different components of the disjoining pressure to the total interaction in the film and their dependence on the composition of the film forming solution.


3D scintigraphic imaging and navigation in radioguided surgery: freehand SPECT technology and its clinical applications.
Freehand SPECT (fhSPECT) is a technology platform for providing 3-dimensional (3D) navigation for radioguided surgical procedures, such as sentinel lymph node (SLN) biopsy (SLNB). In addition to the information provided by conventional handheld gamma detection probes, fhSPECT allows for direct visualization of the distribution of radioactivity in any given region of interest, allowing for improved navigation to radioactive target lesions and providing accurate lesion depth measurements. Herein, we will review the currently available clinical data on the use of fhSPECT: (i) for SLNB of various malignancies, including difficult-to-detect SLNs, and (ii) for radioguided localization of solid tumors. Moreover, the combination of fhSPECT with other technologies (e.g., small field-of-view gamma cameras, and diagnostic ultrasound) is discussed. These technical advances have the potential to greatly expand the clinical application of radioguided surgery in the future.


The effect of temperature on cardiovascular disease hospital admissions among elderly people in Thai Nguyen Province, Vietnam.
Projected increases in weather variability due to climate change will have severe consequences on human health, increasing mortality, and disease rates. Among these, cardiovascular diseases (CVD), highly prevalent among the elderly, have been shown to be sensitive to extreme temperatures and heat waves. This study aimed to find out the relationship between daily temperature (and other weather parameters) and daily CVD hospital admissions among the elderly population in Thai Nguyen province, a northern province of Vietnam. Retrospective data of CVD cases were obtained from a data base of four hospitals in Thai Nguyen province for a period of 5 years from 2008 to 2012. CVD hospital admissions were aggregated by day and merged with daily weather data from this period. Distributed lag non-linear model (DLNM) was used to derive specific estimates of the effect of weather parameters on CVD hospital admissions of up to 30 days, adjusted for time trends using b-splines, day of the week, and public holidays. This study shows that the average point of minimum CVD admissions was at 26°C. Above and below this threshold, the cumulative CVD admission risk over 30 lag days tended to increase with both lower and higher temperatures. The cold effect was found to occur 4-15 days following exposure, peaking at a week's delay. The cumulative effect of cold exposure on CVD admissions was statistically significant with a relative risk of 1.12 (95% confidence interval: 1.01-1.25) for 1°C decrease below the threshold. The cumulative effect of hot temperature on CVD admissions was found to be non-significant and was estimated to be at a relative risk of 1.17 (95% confidence interval: 0.90-1.52) for 1°C increase in the temperature. No significant association was found between CVD admissions and the other weather variables. Exposure to cold temperature is associated with increasing CVD admission risk among the elderly population.


The effect of phenylephrine on the onset time of rocuronium.
Several studies have demonstrated that ephedrine shortens the onset time of muscle relaxants, and it does so probably by increasing the cardiac output. However, elevation of the systemic blood pressure through α adrenergic stimulation via ephedrine may affect the onset of muscle relaxants during the induction of anesthesia. We investigated the effect of phenylephrine, which is a selective α-1 agonist, on the onset time of rocuronium and the intubating conditions in adults after the administration of propofol. Sixty-four patients were randomly assigned to two groups. Phenylephrine (0.9 µg/kg) (P group) or the same volume of saline (S group) was injected before rocuronium (0.6 mg/kg) administration. Anesthesia was induced with fentanyl 2 µg/kg and propofol 2 mg/kg. The onset time was defined as the time from the end of rocuronium injection to the time when a single twitch height gets to 0% or the minimum level. A well-trained anesthesiologist who was 'blinded' to the treatment groups evaluated the intubating conditions. The mean arterial pressure and heart rate were recorded before induction, before intubation, immediately after intubation and 1 minute and 2 minutes after intubation. The onset time was 84 ± 18 sec in the P-group and 72 ± 14 sec in the S-group. There was no difference of the intubating conditions, the mean arterial pressure and the heart rate between the two groups. A small dose of phenylephrine, which has a limited effect on blood pressure, delayed the onset time of rocuronium after the administration of propofol, and the vasoconstriction effect of phenylephrine may affect the prolongation of the rocuronium onset time at the induction of anesthesia with using propofol.


Homeostatic bioenergetic network regulation - a novel concept to avoid pharmacoresistance in epilepsy.
INTRODUCTION: Despite epilepsy being one of the most prevalent neurological disorders, one third of all patients with epilepsy cannot adequately be treated with available antiepileptic drugs. One of the significant causes for the failure of conventional pharmacotherapeutic treatment is the development of pharmacoresistance in many forms of epilepsy. The problem of pharmacoresistance has called for the development of new conceptual strategies that improve future drug development efforts. AREAS COVERED: A thorough review of the recent literature on pharmacoresistance in epilepsy was completed and select examples were chosen to highlight the mechanisms of pharmacoresistance in epilepsy and to demonstrate how those mechanistic findings might lead to improved treatment of pharmacoresistant epilepsy. The reader will gain a thorough understanding of pharmacoresistance in epilepsy and an appreciation of the limitations of conventional drug development strategies. EXPERT OPINION: Conventional drug development efforts aim to achieve specificity of symptom control by enhancing the selectivity of drugs acting on specific downstream targets; this conceptual strategy bears the undue risk of development of pharmacoresistance. Modulation of homeostatic bioenergetic network regulation is a novel conceptual strategy to affect whole neuronal networks synergistically by mobilizing multiple endogenous biochemical and receptor-dependent molecular pathways. In our expert opinion we conclude that homeostatic bioenergetic network regulation might thus be used as an innovative strategy for the control of pharmacoresistant seizures. Recent focal adenosine augmentation strategies support the feasibility of this strategy.


Assessing the risk of ignition in the Russian far east within a modeling framework of fire threat.
The forests of high biological importance in the Russian Far East (RFE) have been experiencing increasing pressure from growing demands for natural resources under the changing economy of post-Soviet Russia. This pressure is further amplified by the rising threat of large and catastrophic fire occurrence, which threatens both the resources and the economic potential of the region. In this paper we introduce a conceptual Fire Threat Model (FTM) and use it to provide quantitative assessment of the risk of ignition in the Russian Far East. The remotely sensed data driven FTM is aimed at evaluating potential wildland fire occurrence and its impact and recovery potential for a given resource. This model is intended for use by resource managers to assist in assessing current levels of fire threat to a given resource, projecting the changes in fire threat under changing climate and land use, and evaluating the efficiency of various management approaches aimed at minimizing the fire impact. Risk of ignition (one of the major uncertainties within fire threat modeling) was analyzed using the MODIS active fire product. The risk of ignition in the RFE is shown to be highly variable in spatial and temporal domains. However, the number of ignition points is not directly proportional to the amount of fire occurrence in the area. Fire ignitions in the RFE are strongly linked to anthropogenic activity (transportation routes, settlements, and land use). An increase in the number of fire ignitions during summer months could be attributed to (1) disruption of the summer monsoons and subsequent changes in fire weather and (2) an increase in natural sources of fire ignitions.


Procalcitonin versus C-reactive protein for guiding antibiotic therapy in sepsis: a randomized trial.
We sought to evaluate whether procalcitonin was superior to C-reactive protein in guiding antibiotic therapy in intensive care patients with sepsis. Randomized open clinical trial. Two university hospitals in Brazil. Patients with severe sepsis or septic shock. Patients were randomized in two groups: the procalcitonin group and the C-reactive protein group. Antibiotic therapy was discontinued following a protocol based on serum levels of these markers, according to the allocation group. The procalcitonin group was considered superior if the duration of antibiotic therapy was at least 25% shorter than in the C-reactive protein group. For both groups, at least seven full-days of antibiotic therapy were ensured in patients with Sequential Organ Failure Assessment greater than 10 and/or bacteremia at inclusion, and patients with evident resolution of the infectious process had antibiotics stopped after 7 days, despite biomarkers levels. Ninety-four patients were randomized: 49 patients to the procalcitonin group and 45 patients to the C-reactive protein group. The mean age was 59.8 (SD, 16.8) years. The median duration of antibiotic therapy for the first episode of infection was 7.0 (Q1-Q3, 6.0-8.5) days in the procalcitonin group and 6.0 (Q1-Q3, 5.0-7.0) days in the C-reactive protein group (p=0.13), with a hazard ratio of 1.206 (95% CI, 0.774-1.3; p=0.13). Overall, protocol overruling occurred in only 13 (13.8%) patients. Twenty-one patients died in each group (p=0.836). C-reactive protein was as useful as procalcitonin in reducing antibiotic use in a predominantly medical population of septic patients, causing no apparent harm.


Structures and functions linked to genome-wide adaptation of human influenza A viruses.
Human influenza A viruses elicit short-term respiratory infections with considerable mortality and morbidity. While H3N2 viruses circulate for more than 50 years, the recent introduction of pH1N1 viruses presents an excellent opportunity for a comparative analysis of the genome-wide evolutionary forces acting on both subtypes. Here, we inferred patches of sites relevant for adaptation, i.e. being under positive selection, on eleven viral protein structures, from all available data since 1968 and correlated these with known functional properties. Overall, pH1N1 have more patches than H3N2 viruses, especially in the viral polymerase complex, while antigenic evolution is more apparent for H3N2 viruses. In both subtypes, NS1 has the highest patch and patch site frequency, indicating that NS1-mediated viral attenuation of host inflammatory responses is a continuously intensifying process, elevated even in the longtime-circulating subtype H3N2. We confirmed the resistance-causing effects of two pH1N1 changes against oseltamivir in NA activity assays, demonstrating the value of the resource for discovering functionally relevant changes. Our results represent an atlas of protein regions and sites with links to host adaptation, antiviral drug resistance and immune evasion for both subtypes for further study.