Determination of Arabidopsis thaliana telomere length by PCR.
In humans, telomere length studies have acquired great relevance because the length of telomeres has been related to natural processes like disease, aging and cancer. However, very little is known about the influence of telomere length on the biology of wild type plants. The length of plant telomeres has been usually studied by Terminal Restriction Fragment (TRF) analyses. This technique requires high amounts of tissue, including multiple cell types, which might be the reason why very little is known about the influence of telomere length on plant natural processes. In contrast, many of the human telomere length studies have focused on homogenous cell populations. Most of these studies have been performed by PCR, using telomeric degenerated primers, which allow the determination of telomere length from small amounts of human cells. Here, we have adapted the human PCR procedure to analyze the length of Arabidopsis thaliana telomeres. This PCR approach will facilitate the analysis of telomere length from low amounts of tissue. We have used it to determine that CG and non CG DNA methylation positively regulates Arabidopsis telomere length.


CD4+ T Cell Activation During the Newborn Period: Barriers Against and Pathways Toward Th1 Immunity.
During the period of transition from intrauterine to extrauterine life, the neonatal immune system must learn to rapidly identify pathogens while balancing pro-inflammatory, antimicrobial responses with immune regulation that allows for resolution of inflammation and limits responses to commensal organisms and benign environmental antigens. However, the naive immune system of neonates is presented with several barriers that limit robust proinflammatory immune responses. Specifically, epigenetic modifications to neonatal naive CD4+ T cells, heightened neonatal regulatory T cell frequency and function, and limitations in the co-stimulatory potential of neonatal antigen presenting cells restrict development of CD4+ T cells with a T-helper 1 type functional profile. This restriction likely contributes to the increased risk of severe infection observed during early life. New research, however, suggests that neonates are capable of utilizing unique compensatory mechanisms to circumvent these restrictions and generate T-helper 1 type immunity under some circumstances. Understanding how to manipulate the immune responses of young infants to optimize development of T-helper 1 type immunity is key to the development of immune-based treatments and prevention strategies for severe infections in this vulnerable population.


Comparison of magnesium and methyldopa for the control of blood pressure in pregnancies complicated with hypertension.
Although magnesium is now the drug of choice for the prevention of eclamptic seizures only few studies have evaluated whether magnesium may reduce blood pressure in pregnancies complicated with hypertension. A total of 33 patients with pregnancy-induced hypertension were randomized to either magnesium or methyldopa treatment. Of these 16 received magnesium and 17 methyldopa. The treatment comprised a 48-hour magnesium infusion followed by oral magnesium tablets until 3 days after delivery or 250 mg methyldopa 4 times a day in a similar period. Patients treated with magnesium had 1 day after inclusion a statistically significantly lower systolic blood pressure compared to the level in the methyldopa group (138.1 +/- 11 vs. 147.6 +/- 11 mm Hg; p < 0.05), but no difference was observed in diastolic blood pressure (92.0 +/- 6.6 vs. 96.0 +/- 10.1 mm Hg; NS). From the 5th day of inclusion and until delivery both systolic and diastolic blood pressure were significantly lower in the magnesium group (p < 0.05). Including all blood pressure measurements in a single analysis showed that both systolic (138 +/- 13 vs. 148 +/- 15 mm Hg; p < 0.0001) and diastolic (92 +/- 10 vs. 94 +/- 10 mm Hg; p < 0.05) blood pressure were lower in the magnesium group compared to the methyldopa group. There was no difference between the two groups regarding gestational age at delivery, birth weight, Apgar scores and pH in umbilical cord blood. This preliminary study demonstrates that magnesium treatment lowers blood pressure in pregnancies complicated with hypertension. The effect is without any adverse effect on maternal and neonatal well-being.


Variants in myelin regulatory factor (MYRF) cause autosomal dominant and syndromic nanophthalmos in humans and retinal degeneration in mice.
Nanophthalmos is a rare, potentially devastating eye condition characterized by small eyes with relatively normal anatomy, a high hyperopic refractive error, and frequent association with angle closure glaucoma and vision loss. The condition constitutes the extreme of hyperopia or farsightedness, a common refractive error that is associated with strabismus and amblyopia in children. NNO1 was the first mapped nanophthalmos locus. We used combined pooled exome sequencing and strong linkage data in the large family used to map this locus to identify a canonical splice site alteration upstream of the last exon of the gene encoding myelin regulatory factor (MYRF c.3376-1G>A), a membrane bound transcription factor that undergoes autoproteolytic cleavage for nuclear localization. This variant produced a stable RNA transcript, leading to a frameshift mutation p.Gly1126Valfs*31 in the C-terminus of the protein. In addition, we identified an early truncating MYRF frameshift mutation, c.769dupC (p.S264QfsX74), in a patient with extreme axial hyperopia and syndromic features. Myrf conditional knockout mice (CKO) developed depigmentation of the retinal pigment epithelium (RPE) and retinal degeneration supporting a role of this gene in retinal and RPE development. Furthermore, we demonstrated the reduced expression of Tmem98, another known nanophthalmos gene, in Myrf CKO mice, and the physical interaction of MYRF with TMEM98. Our study establishes MYRF as a nanophthalmos gene and uncovers a new pathway for eye growth and development.


Membrane Binding of Parkinson's Protein α-Synuclein: Effect of Phosphorylation at Positions 87 and 129 by the S to D Mutation Approach.
Human α-synuclein, a protein relevant in the brain with so-far unknown function, plays an important role in Parkinson's disease. The phosphorylation state of αS was related to the disease, prompting interest in this process. The presumed physiological function and the disease action of αS involves membrane interaction. Here, we study the effect of phosphorylation at positions 87 and 129, mimicked by the mutations S87A, S129A (nonphosphorylated) and S87D, S129D (phosphorylated) on membrane binding. Local binding is detected by spin-label continuous-wave electron paramagnetic resonance. For S87A/D, six positions (27, 56, 63, 69, 76, and 90) are probed; and for S129A/D, three (27, 56, and 69). Binding to large unilamellar vesicles of 100 nm diameter of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine in a 1 : 1 composition is not affected by the phosphorylation state of S129. For phosphorylation at S87, local unbinding of αS from the membrane is observed. We speculate that modulating the local membrane affinity by phosphorylation could tune the way αS interacts with different membranes; for example, tuning its membrane fusion activity.


Rank-order inhibition by omega-conotoxins in human and animal autonomic nerve preparations.
The inhibitory effects of the omega-conotoxins GVIA, MVIIA and MVIIC on electrically-evoked, tetrodotoxin (10(-7) M)-sensitive, autonomic nerve activity were studied using human, rat or guinea-pig vas deferens and intestinal tissues. In each preparation from each species, nM concentrations of omega-conotoxins GVIA and MVIIA prevented the neuronally-mediated contractions, whereas omega-conotoxin MVIIC was either markedly less potent (IC(50)'s 1.4 or 2.9 log units more than for omega-conotoxin GVIA in guinea-pig ileum and rat vas deferens, respectively) or was without significant activity (human vas deferens, human Taenia coli) when tested at similar concentrations. In contrast the differences in potency between omega-conotoxins GVIA and MVIIC were considerably less when assayed directly on Ca(2+) channel currents evoked from rat superior cervical ganglion neurons in culture (approximately 0.1 log unit difference) and from a stable cell line expressing rat alpha(1B), alpha(2)delta, beta(1b) Ca(2+) channel subunits (approximately 0.9 log unit). These different rank-orders of inhibitory activity of the conotoxins support the suggestion that there are pharmacologically distinct N-type Ca(2+) channels in the peripheral nervous system, and that this tissue-dependent difference is seen in man.


Angle measurement based on the internal-reflection effect and the use of right-angle prisms.
A new development in angle measurement based on the internal-reflection effect (AMIRE) is described in which a pair of right-angle prisms is used to replace the previously used elongated critical-angle prisms, resulting in lower costs and a more compact size. Excellent linearity is achieved through careful alignment of the right-angle prisms. The measurement sensitivity and range can be selected through the use of light sources with different polarization states. Experiments with a prototype sensor demonstrated a measurement range of 1.6°, a resolution of 0.04 arcsec, and a nonlinearity error of ±0.1%. Both analytical and experimental results are presented.


Binary amplitude holograms made from dyed photoresist.
A method for fabricating binary amplitude holograms from a dyed photoresist is described. It is of particular interest for holograms that are used as null optics in the form metrology of aspheric surfaces and wavefronts. A pigment that strongly absorbs light near 633 nm was dissolved in a positive photoresist and the dyed resist was spun onto silica glass substrates. Stable resist layers were obtained that were essentially opaque at 633 nm with little effect on the transmittance of the resist in the UV. A Fresnel zone plate was fabricated from the dyed resist layer using contact lithography, and its performance was demonstrated at 633 nm.


Attenuation of cerebral vasospasm by systemic administration of an endothelin-A receptor antagonist, TBC 11251, in a rabbit model of subarachnoid hemorrhage.
Cerebral vasospasm is a major complication of subarachnoid hemorrhage (SAH) after the rupture of an intracranial aneurysm. Although the cause of cerebral vasospasm has not been fully established, several lines of evidence suggest that the vasoconstrictor peptide endothelin (ET) may play a crucial role. In the present study the potential of TBC 11251 (TBC), a newly developed ET(A) receptor antagonist, to prevent and/or reverse cerebral vasospasm was examined in a well-established rabbit model of SAH. Sixty-five New Zealand White rabbits were assigned to one of six groups. Experimental SAH was induced in rabbits comprising five of the groups by injecting autologous arterial blood into the cisterna magna. The treatment groups were as follows: 1) control (no SAH); 2) SAH only; 3) SAH + placebo at 24 and 36 hours (24/36); 4) SAH + TBC (24/36); 5) SAH + placebo twice daily (BID); and 6) SAH + TBC BID. All drug-treated animals received an intravenous dosage of 5 mg/kg TBC. After 48 hours, the animals were killed by intracardiac perfusion with fixative. The brainstems were removed and the basilar arteries (BAs) were prepared for histological examination. The cross-sectional area of each BA was measured using computer-assisted videomicroscopy by an investigator blind to the group from which it came. A one-way analysis of variance and paired group mean comparisons with the post-hoc Fisher least significant difference test were used for analysis of BA diameters and physiological parameters. The model provided reliable vasospasm, with the mean BA cross-sectional area constricting from 0.388 mm2 in the control group to 0.106 mm2 (27.4% of control) in the SAH only group. Treatment with TBC (24/36) after SAH (reversal protocol) produced a mean BA area of 0.175 mm2 (44.2% of control) which, although larger than the placebo group value of 0.135 mm2 (39.9% of control), was not statistically significant. However, treatment with TBC BID (prevention protocol) produced a mean BA area of 0.303 mm2 (78.1% of control) compared with the placebo BID value of 0.134 mm2 (34.6% of control); this effect was statistically significant (p < 0.01). There were no side effects noted and no differences in the mean arterial pressures between drug and placebo groups. These findings demonstrate that systemic administration of the ET(A) receptor antagonist TBC significantly attenuates cerebral vasospasm after SAH when given as a preventative therapy, and they provide additional support for the role of ET in the establishment of vasospasm.


Age-related differences in white matter microstructure: region-specific patterns of diffusivity.
We collected MRI diffusion tensor imaging data from 80 younger (20-32 years) and 63 older (60-71 years) healthy adults. Tract-based spatial statistics (TBSS) analysis revealed that white matter integrity, as indicated by decreased fractional anisotropy (FA), was disrupted in numerous structures in older compared to younger adults. These regions displayed five distinct region-specific patterns of age-related differences in other diffusivity properties: (1) increases in both radial and mean diffusivity; (2) increases in radial diffusivity; (3) no differences in parameters other than FA; (4) a decrease in axial and an increase in radial diffusivity; and (5) a decrease in axial and mean diffusivity. These patterns suggest different biological underpinnings of age-related decline in FA, such as demyelination, Wallerian degeneration, gliosis, and severe fiber loss, and may represent stages in a cascade of age-related degeneration in white matter microstructure. This first simultaneous description of age-related differences in FA, mean, axial, and radial diffusivity requires histological and functional validation as well as analyses of intermediate age groups and longitudinal samples.


Development of Selective ADAMTS-5 Peptide Substrates to Monitor Proteinase Activity.
The dysregulation of proteinase activity is a hallmark of osteoarthritis (OA), a disease characterized by progressive degradation of articular cartilage by catabolic proteinases such as a disintegrin and metalloproteinase with thrombospondin type I motifs-5 (ADAMTS-5). The ability to detect such activity sensitively would aid disease diagnosis and the evaluation of targeted therapies. Förster resonance energy transfer (FRET) peptide substrates can detect and monitor disease-related proteinase activity. To date, FRET probes for detecting ADAMTS-5 activity are nonselective and relatively insensitive. We describe the development of rapidly cleaved and highly selective ADAMTS-5 FRET peptide substrates through in silico docking and combinatorial chemistry. The lead substrates 3 and 26 showed higher overall cleavage rates (∼3-4-fold) and catalytic efficiencies (∼1.5-2-fold) compared to the best current ADAMTS-5 substrate ortho-aminobenzoyl(Abz)-TESE↓SRGAIY-N-3-[2,4-dinitrophenyl]-l-2,3-diaminopropionyl(Dpa)-KK-NH2. They exhibited high selectivity for ADAMTS-5 over ADAMTS-4 (∼13-16-fold), MMP-2 (∼8-10-fold), and MMP-9 (∼548-2561-fold) and detected low nanomolar concentrations of ADAMTS-5.


Aluminum is a potential environmental factor for Crohn's disease induction: extended hypothesis.
Aluminum (Al) is a common environmental compound with immune-adjuvant activity and granulomatous inflammation inducer. Al exposure in food, additives, air, pharmaceuticals, and water pollution is ubiquitous in Western culture. Crohn's disease (CD) is a chronic relapsing intestinal inflammation in genetically susceptible individuals and is influenced by yet unidentified environmental factors. It is hypothesized, in the present review, that Al is a potential factor for induction or maintaining the inflammation in CD. Epidemiologically, CD incidence is higher in urban areas, where microparticle pollution is prevalent. Al immune activities share many characteristics with the immune pathology of CD: increased antigen presentation and APCs activation, many luminal bacterial or dietary compounds can be adsorbed to the metal and induce Th1 profile activity, promotion of humoral and cellular immune responses, proinflammatory, apoptotic, oxidative activity, and stress-related molecule expression enhancement, affecting intestinal bacterial composition and virulence, granuloma formation, colitis induction in an animal model of CD, and terminal ileum uptake. The Al-bacterial interaction, the microparticles homing the intestine together with the extensive immune activity, put Al as a potential environmental candidate for CD induction and maintenance.


An investigation of causes of false positive single nucleotide polymorphisms using simulated reads from a small eukaryote genome.
Single Nucleotide Polymorphisms (SNPs) are widely used molecular markers, and their use has increased massively since the inception of Next Generation Sequencing (NGS) technologies, which allow detection of large numbers of SNPs at low cost. However, both NGS data and their analysis are error-prone, which can lead to the generation of false positive (FP) SNPs. We explored the relationship between FP SNPs and seven factors involved in mapping-based variant calling - quality of the reference sequence, read length, choice of mapper and variant caller, mapping stringency and filtering of SNPs by read mapping quality and read depth. This resulted in 576 possible factor level combinations. We used error- and variant-free simulated reads to ensure that every SNP found was indeed a false positive. The variation in the number of FP SNPs generated ranged from 0 to 36,621 for the 120 million base pairs (Mbp) genome. All of the experimental factors tested had statistically significant effects on the number of FP SNPs generated and there was a considerable amount of interaction between the different factors. Using a fragmented reference sequence led to a dramatic increase in the number of FP SNPs generated, as did relaxed read mapping and a lack of SNP filtering. The choice of reference assembler, mapper and variant caller also significantly affected the outcome. The effect of read length was more complex and suggests a possible interaction between mapping specificity and the potential for contributing more false positives as read length increases. The choice of tools and parameters involved in variant calling can have a dramatic effect on the number of FP SNPs produced, with particularly poor combinations of software and/or parameter settings yielding tens of thousands in this experiment. Between-factor interactions make simple recommendations difficult for a SNP discovery pipeline but the quality of the reference sequence is clearly of paramount importance. Our findings are also a stark reminder that it can be unwise to use the relaxed mismatch settings provided as defaults by some read mappers when reads are being mapped to a relatively unfinished reference sequence from e.g. a non-model organism in its early stages of genomic exploration.


Juniors' hours: is the end in sight?
The question of how to reduce junior doctors' hours has taxed the government and hospital managers for nearly two decades. Now regional task forces are being asked to get them down to a maximum of 83 a week by 1 April next year. From a comparison of the way in which two different task forces (Northern and North West Thames) have responded to the challenge it emerges that most units will probably meet the 83 hour deadline by making simple rationalisations. Meeting the next deadline--of a 72 hour maximum by December 1996--will, however, require radical restructuring of working patterns.


[Karyomegalic interstitial nephritis: an atypical case].
Karyomegalic interstitial nephritis (KIN) is a rare disease entity that was first described by Burry in 1974. The prevalence of this disease is less than 1% and its pathogenesis is unclear. KIN is characterized by chronic tubulointerstitial nephritis associated with enlarged tubular epithelial cell nuclei, which leads to progressive decline of renal function. The disease has no known treatment. Here, we report on a 50-year-old female patient who presented with asymptomatic progressive decline of renal function. Renal biopsy demonstrated chronic tubulointerstitial nephritis with markedly enlarged and hyperchromic nuclei of tubule epithelial cells the hallmark of karyomegalic nephritis. Clinical and pathologic findings of this case are discussed in light of the available literature.


Extracellular 5-aminolevulinic acid production by Escherichia coli containing the Rhodopseudomonas palustris KUGB306 hemA gene.
The Rhodopseudomonas palustris KUGB306 hemA gene codes for 5-aminolevulinic acid (ALA) synthase. This enzyme catalyzes the condensation of glycine and succinyl-CoA to yield ALA in the presence of the cofactor pyridoxal 5'- phosphate. The R. palustris KUGB306 hemA gene in the pGEX-KG vector system was transformed into Escherichia coli BL21. The effects of physiological factors on the extracellular production of ALA by the recombinant E. coli were studied. Terrific Broth (TB) medium resulted in significantly higher cell growth and ALA production than did Luria-Bertani (LB) medium. ALA production was significantly enhanced by the addition of succinate together with glycine in the medium. Maximal ALA production (2.5 g/l) was observed upon the addition of D-glucose as an ALA dehydratase inhibitor in the late-log culture phase. Based on the results obtained from the shake-flask cultures, fermentation was carried out using the recombinant E. coli in TB medium, with the initial addition of 90 mM glycine and 120 mM succinate, and the addition of 45 mM D-glucose in the late-log phase. The extracellular production of ALA was also influenced by the pH of the culture broth. We maintained a pH of 6.5 in the fermenter throughout the culture process, achieving the maximal levels of extracellular ALA production (5.15 g/l, 39.3 mM).


PRIME: A Novel Low-Mass, High-Repetition Approach to Improve Function in Older Adults.
The ability to maintain functional independence in a rapidly aging population results in an increased life expectancy without corresponding increases in health care costs. The accelerated decline in V˙O2peak after the age of 65 yr is primarily due to peripheral tissue changes rather than centrally mediated factors. The purpose of this study was to determine whether the Peripheral Remodeling through Intermittent Muscular Exercise (PRIME) approach, consisting of a low-mass, high-repetition/duration skeletal muscle focused training regimen would provide superior functional benefits in participants older than 70 yr old and at risk for losing functional independence. In this clinical trial, 107 participants were randomized to 4 wk of either standard aerobic training (AT) or PRIME (phase 1). This was followed by 8 wk of a progressive whole-body aerobic and resistance training (AT + RT) for all participants (phase 2). The major outcome measures were cardiorespiratory fitness (peak oxygen consumption [V˙O2peak]), muscular fitness (1 repetition maximal strength [1RM]), and physical function (Senior Fitness Test [SFT] scores). Results were analyzed under a per-protocol criterion. Thirty-eight PRIME and 38 AT participants completed the 3-month protocols. V˙O2peak, 1RM, and SFT scores all increased significantly after 12 wk for both treatment groups (P < 0.05). However, relative to AT, participants randomized to PRIME demonstrated a greater increase in V˙O2peak (2.37 + 1.83 vs 1.50 + 1.82 mL·kg·min, P < 0.05), 1RM (48.52 + 27.03 vs 28.01 + 26.15 kg, P < 0.01) and SFT (22.50 + 9.98 vs 18.66 + 9.60 percentile, P < 0.05). Participants experienced greater increases in cardiorespiratory and muscular fitness and physical function when PRIME training was initiated before a combined AT + RT program. This novel exercise approach may be advantageous to individuals with other chronic disease conditions characterized by low functional capacity.


Alternative drug formulations of docetaxel: a review.
The anticancer drug docetaxel (Taxotere) is formulated in the nonionic surfactant polysorbate 80 (Tween 80). Early in the clinical development of docetaxel, it became clear that docetaxel administration is associated with the occurrence of unpredictable (acute) hypersensitivity reactions and cumulative fluid retention. These side-effects have been attributed, in part, to the presence of polysorbate 80 and have consequently initiated research focused on the development of a less-toxic, better-tolerated polysorbate 80-free formulation of docetaxel. More recently, there is an increasing interest in developing a (polysorbate 80-free) docetaxel formulation that selectively targets malignant tissue, thereby increasing efficacy while decreasing the occurrence of side-effects related to wide and nonspecific body distribution. This review aims to discuss the preclinical and clinical results of pharmaceutical strategies [PEGylated (immuno)liposomal docetaxel, docetaxel-fibrinogen-coated olive oil droplets, docetaxel encapsulated nanoparticle-aptamer bioconjugates, submicronic dispersion formulation] to develop an alternative, solvent-free, delivery form for docetaxel characterized by increased efficacy and decreased toxicity.


The Photobiology of Lutein and Zeaxanthin in the Eye.
Lutein and zeaxanthin are antioxidants found in the human retina and macula. Recent clinical trials have determined that age- and diet-related loss of lutein and zeaxanthin enhances phototoxic damage to the human eye and that supplementation of these carotenoids has a protective effect against photoinduced damage to the lens and the retina. Two of the major mechanisms of protection offered by lutein and zeaxanthin against age-related blue light damage are the quenching of singlet oxygen and other reactive oxygen species and the absorption of blue light. Determining the specific reactive intermediate(s) produced by a particular phototoxic ocular chromophore not only defines the mechanism of toxicity but can also later be used as a tool to prevent damage.


IGF2BP2 promotes glycolysis and hepatocellular carcinoma stemness by stabilizing CDC45 mRNA via m6A modification.
A growing number of studies have shown the prognostic importance of Cell division cycle protein 45 (CDC45) in hepatocellular carcinoma (HCC). This study aims to investigate the biological function and mechanism of CDC45 in HCC. The differential expression and prognostic significance of CDC45 in HCC and normal tissues were analyzed by bioinformatics. CDC45 was knocked down and the biological effects of CDC45 in HCC in vitro and in vivo were measured. Subsequently, using RNA m6A colorimetry and Methylated RNA Immunoprecipitation (MeRIP), the levels of m6A modification of total RNA and CDC45 were evaluated in cells. RIP was applied to establish that CDC45 and insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) interact. A test using actinomycin D was performed to gauge the stability of the CDC45 mRNA. Furthermore, the regulatory role of IGF2BP2 on CDC45 expression in HCC progression was explored by overexpressing IGF2BP2. High expression of CDC45 was correlated with poor prognosis in HCC patients. Knocking down CDC45 inhibited HCC cell proliferation, migration, invasion, EMT, stemness, and glycolysis, and promoted apoptosis, which was verified through in vitro experiments. Additionally, IGF2BP2 was highly expressed in HCC cells, and it was found to interact with CDC45. Knocking down IGF2BP2 resulted in reduced stability of CDC45 mRNA. Moreover, overexpression of IGF2BP2 promoted HCC cell proliferation, migration, invasion, EMT, stemness, and glycolysis, while inhibiting apoptosis, which was reversed by knocking down CDC45. In general, IGF2BP2 promoted HCC glycolysis and stemness by stabilizing CDC45 mRNA via m6A modification. [Figure: see text].


Climate change-induced vegetation shifts lead to more ecological droughts despite projected rainfall increases in many global temperate drylands.
Drylands occur worldwide and are particularly vulnerable to climate change because dryland ecosystems depend directly on soil water availability that may become increasingly limited as temperatures rise. Climate change will both directly impact soil water availability and change plant biomass, with resulting indirect feedbacks on soil moisture. Thus, the net impact of direct and indirect climate change effects on soil moisture requires better understanding. We used the ecohydrological simulation model SOILWAT at sites from temperate dryland ecosystems around the globe to disentangle the contributions of direct climate change effects and of additional indirect, climate change-induced changes in vegetation on soil water availability. We simulated current and future climate conditions projected by 16 GCMs under RCP 4.5 and RCP 8.5 for the end of the century. We determined shifts in water availability due to climate change alone and due to combined changes of climate and the growth form and biomass of vegetation. Vegetation change will mostly exacerbate low soil water availability in regions already expected to suffer from negative direct impacts of climate change (with the two RCP scenarios giving us qualitatively similar effects). By contrast, in regions that will likely experience increased water availability due to climate change alone, vegetation changes will counteract these increases due to increased water losses by interception. In only a small minority of locations, climate change-induced vegetation changes may lead to a net increase in water availability. These results suggest that changes in vegetation in response to climate change may exacerbate drought conditions and may dampen the effects of increased precipitation, that is, leading to more ecological droughts despite higher precipitation in some regions. Our results underscore the value of considering indirect effects of climate change on vegetation when assessing future soil moisture conditions in water-limited ecosystems.


2014 AMI Lifetime Achievement Award Recipient.
Our JBC Showcase section features collections of bioscientific photographs, medical illustrations, and other award-winning imagery from accomplished individuals in the field of biocommuniucation. The illustrators and photographers featured in our Showcase collections often are individials who have been awarded or honored by the Association of Medical Illustrators or the Biocommunications Association. The 41-1 JBC Showcase features the creative work of Gary Schnitz, the 2014 Recipient of AMI's Lifetime Acheivement Award.


The Interplay of Ethics, Animal Welfare, and IACUC Oversight on the Reproducibility of Animal Studies.
Reproducibility in animal studies has been defined as the ability of a result to be replicated through independent experiments within the same or among different laboratories. Over the past few years, much has been written and said about the lack of reproducibility of animal studies. Reasons that are commonly cited for this lack of reproducibility include inappropriate study design, errors in conducting the research, and potential fraud. In the quest to understand the basis for this lack of reproducibility, scientists have not fully considered the potential ramifications on ethical constructs for animal research, animal welfare considerations in animal research programs, the regulatory environment, and oversight by IACUCs. Here, we review how ethical theories behind animal research, policies, and practices meant to enhance animal welfare and the IACUC oversight process influence the reproducibility of animal studies, a previously undiscussed topic in the peer-reviewed literature.


Patient participation in medical consultations: why some patients are more involved than others.
Patients vary in their willingness and ability to actively participate in medical consultations. Because more active patient participation contributes to improved health outcomes and quality of care, it is important to understand factors affecting the way patients communicate with healthcare providers. The objectives of this study were to examine the extent to which patient participation in medical interactions is influenced by 1) the patient's personal characteristics (age, gender, education, ethnicity); 2) the physician's communication style (eg, use of partnership-building and supportive talk); and 3) the clinical setting (eg, the health condition, medical specialty). The authors conducted a post hoc cross-sectional analysis of 279 physician-patient interactions from 3 clinical sites: 1) primary care patients in Sacramento, California, 2) patients with systemic lupus erythematosus (SLE) from the San Francisco Bay area, and 3) patients with lung cancer from a VA hospital in Texas. The outcome measures included the degree to which patients asked questions, were assertive, and expressed concerns and the degree to which physicians used partnership-building and supportive talk (praise, reassurance, empathy) in their consultations. The majority of active participation behaviors were patient-initiated (84%) rather than prompted by physician partnership-building or supportive talk. Patients who were more active participants received more facilitative communication from physicians, were more educated, and were more likely to be white than of another ethnicity. Women more willingly expressed negative feelings and concerns. There was considerable variability in patient participation across the 3 clinical settings. Female physicians were more likely to use supportive talk than males, and physicians generally used less supportive talk with nonwhite compared with white patients. Patient participation in medical encounters depends on a complex interplay of personal, physician, and contextual factors. Although more educated and white patients tended to be more active participants than their counterparts, the strongest predictors of patient participation were situation-specific, namely the clinical setting and the physician's communicative style. Physicians could more effectively facilitate patient involvement by more frequently using partnership-building and supportive communication. Future research should investigate how the nuances of individual clinical settings (eg, the health condition, time allotted for the visit) impose constraints or opportunities for more effective patient involvement in care.


Stranger danger awareness in Williams syndrome.
The developmental disorder Williams syndrome (WS) is characterised by a distinctive cognitive profile and an intriguing social phenotype. Individuals with the disorder are often highly social engaging with familiar and unfamiliar people and once in an interaction they often show subtle abnormalities of social behaviour. Atypically increased approach to unfamiliar people is widely reported in the existing literature for both children and adults. Parents frequently report interactions with unfamiliar people as a major concern. In this study we aimed to evaluate 'stranger danger' awareness using a video vignette task with individuals who had WS. When linked to other components of the WS phenotype (e.g. reduced intellectual ability, increased social approach) an awareness of stranger danger is particularly important. Qualitative and quantitative data showed that young people with WS have difficulties making judgements about whether or not to trust and engage in conversation with unfamiliar people. Qualitative data showed that individuals with WS often suggested that they would engage in an interaction with an unfamiliar person. The findings have substantial implications for the safety of young people with the disorder and emphasise the need for intervention regarding this behaviour.


Emerging anthropogenic circularity science: principles, practices, and challenges.
Material depletion over reliance of linear economies and environmental pollution may be resolved by applying the principles and practices of anthropogenic circularity science. Here we systematically review the emergence of anthropogenic circularity science in the interdisciplinary development of green chemistry, supply chain, and industrial ecology at different scales. The first, second, and third laws of circularity chemistry are proposed as forming the basic principles of circularity science. To close the loop on critical materials, these three basic principles have been exemplified in the anthropogenic circularity practices. We highlight the spatial distribution of critical metal, waste generation, and recycling rate. Future opportunities and challenges for a circular economy and urban mining will predominate in anthropogenic circularity. Therefore, anthropogenic circularity science will play an increasing role in enabling a smooth transition to a circular society.


Why Escherichia coli alpha-hemolysin induces calcium oscillations in mammalian cells--the pore is on its own.
Escherichia coli alpha-hemolysin (HlyA), archetype of a bacterial pore-forming toxin, has been reported to deregulate physiological Ca2+ channels, thus inducing periodic low-frequency Ca2+ oscillations that trigger transcriptional processes in mammalian cells. The present study was undertaken to delineate the mechanisms underlying the Ca2+ oscillations. Patch-clamp experiments were combined with single cell measurements of intracellular Ca2+ and with flowcytometric analyses. Application of HlyA at subcytocidal concentrations provoked Ca2+ oscillations in human renal and endothelial cells. However, contrary to the previous report, the phenomenon could not be inhibited by the Ca2+ channel blocker nifedipine and Ca2+ oscillations showed no constant periodicity at all. Ca2+ oscillations were dependent on the pore-forming activity of HlyA: application of a nonhemolytic but bindable toxin had no effect. Washout experiments revealed that Ca2+ oscillations could not be maintained in the absence of toxin in the medium. Analogously, propidium iodide flux into cells occurred in the presence of HlyA, but cells rapidly became impermeable toward the dye after toxin washout, indicating resealing or removal of the membrane lesions. Finally, patch-clamp experiments revealed temporal congruence between pore formation and Ca2+ influx. We conclude that the nonperiodic Ca2+ oscillations induced by HlyA are not due to deregulation of physiological Ca2+ channels but derive from pulsed influxes of Ca2+ as a consequence of formation and rapid closure of HlyA pores in mammalian cell membranes.


Accurate switching intensities and length scales in quasi-phase-matched materials.
We consider unseeded type I second-harmonic generation in quasi-phase-matched quadratic nonlinear materials and derive an accurate analytical expression for the evolution of the average intensity. The intensity-dependent nonlinear phase mismatch that is due to the cubic nonlinearity induced by quasi phase matching is found. The equivalent formula for the intensity of maximum conversion, the crossing of which changes the one-period nonlinear phase shift of the fundamental abruptly by pi , corrects earlier estimates [Opt. Lett. 23, 506 (1998)] by a factor of 5.3. We find the crystal lengths that are necessary to obtain an optimal flat phase versus intensity response on either side of this separatrix intensity.


Probing the hydrogen-bond network of water via time-resolved soft X-ray spectroscopy.
We report time-resolved studies of hydrogen bonding in liquid H(2)O, in response to direct excitation of the O-H stretch mode at 3 mum, probed via soft X-ray absorption spectroscopy at the oxygen K-edge. This approach employs a newly developed nanofluidic cell for transient soft X-ray spectroscopy in the liquid phase. Distinct changes in the near-edge spectral region (XANES) are observed, and are indicative of a transient temperature rise of 10 K following transient laser excitation and rapid thermalization of vibrational energy. The rapid heating occurs at constant volume and the associated increase in internal pressure, estimated to be 8 MPa, is manifested by distinct spectral changes that differ from those induced by temperature alone. We conclude that the near-edge spectral shape of the oxygen K-edge is a sensitive probe of internal pressure, opening new possibilities for testing the validity of water models and providing new insight into the nature of hydrogen bonding in water.


From the Cover: ROS-Induced Store-Operated Ca2+ Entry Coupled to PARP-1 Hyperactivation Is Independent of PARG Activity in Necrotic Cell Death.
2,3,5-tris(Glutathion-S-yl)hydroquinone, a potent nephrotoxic and nephrocarcinogenic metabolite of benzene and hydroquinone, generates reactive oxygen species (ROS) causing DNA strand breaks and the subsequent activation of DNA repair enzymes, including poly(ADP-ribose) polymerase (PARP)-1. Under robust oxidative DNA damage, PARP-1 is hyperactivated, resulting in the depletion of NAD+ and ATP with accompanying elevations in intracellular calcium concentrations (iCa2+), and ultimately necrotic cell death. The role of Ca2+ during PARP-dependent necrotic cell death remains unclear. We therefore sought to determine the relationship between Ca2+ and PARP-1 during ROS-induced necrotic cell death in human renal proximal tubule epithelial cells (HK-2). Our experiments suggest that store-operated Ca2+ channel (SOC) entry contributes to the coupling of PARP-1 activation to increases in iCa2+ during ROS-induced cell death. Poly(ADP-ribose)glycohydrolase (PARG), which catalyzes the degradation of PARs to yield free ADP-ribose (ADPR), is known to activate Ca2+ channels such as TRPM2. However, siRNA knockdown of PARG did not restore cell viability, indicating that free ADPR is not responsible for SOC activation in HK-2 cells. The data indicate that PARP-1 and iCa2+ are coupled through activation of SOC mediated Ca2+ entry in an apparently ADPR-independent fashion; alternative PAR-mediated signaling likely contributes to PARP-dependent necrotic cell death, perhaps via PAR-mediated signaling proteins that regulate iCa2+ homeostasis.


A comparison of the firing properties of putative excitatory and inhibitory neurons from CA1 and the entorhinal cortex.
The superficial layers of the entorhinal cortex (EC) provide the majority of the neocortical input to the hippocampus, and the deep layers of the EC receive the majority of neocortically bound hippocampal outputs. To characterize information transmission through the hippocampal and EC circuitry, we recorded simultaneously from neurons in the superficial EC, the CA1 region of hippocampus, and the deep EC while rodents ran for food reward in two environments. Spike waveform analysis allowed us to classify units as fast-spiking (FS) putative inhibitory cells or putative excitatory (PE) cells. PE and FS units' firing were often strongly correlated at short time scales, suggesting the presence a monosynaptic connection from the PE to FS units. EC PE units, unlike those found in CA1, showed little or no tendency to fire in bursts. We also found that the firing of FS and PE units from all regions was modulated by the approximately 8 Hz theta rhythm, although the firing of deep EC FS units tended to be less strongly modulated than that of the other types of units. When we examined the spatial specificity of FS units, we determined that FS units in all three regions showed low specificity. At the same time, retrospective coding, in which firing rates were related to past position, was present in FS units from all three regions and deep EC FS units often fired in a "path equivalent" manner in that they were active in physically different, but behaviorally related positions both within and across environments. Our results suggest that while the firing of FS units from CA1 and the EC show similarly low levels of position specificity, FS units from each region differ from one another in that they mirrored the associated PE units in terms of their tendency to show more complex positional firing properties like retrospective coding and path equivalence.


Insulin and glucagon regulate cytosolic phosphoenolpyruvate carboxykinase (GTP) mRNA in rat liver.
Insulin causes a 7-10-fold decrease of both the mRNA that codes for rat hepatic phosphoenolpyruvate carboxykinase (mRNAPEPCK) and of PEPCK synthesis, provided the animals are made diabetic and fed chow. mRNAPEPCK, measured either by in vitro translation or cDNA hybridization, decreases with a half-time of 30-60 min after insulin treatment. This coordinant decrease, which approximates the half-life of mRNAPEPCK measured in a variety of situations, suggests that insulin acts by decreasing mRNAPEPCK production, and that the hormone does not alter the activity of a fixed amount of this RNA, or enhance its degradation. Glucagon results in a ninefold induction of mRNAPEPCK. Half-maximal induction occurs with doses between 20-75 micrograms/100 g body wt and occurs within 30-45 min. Maximal induction requires 150 micrograms/100 g body wt and occurs about 80 min after a single glucagon injection. N6,O2'-dibutyryl cAMP and a cAMP analogue that is not metabolized, 8-(4-chlorophenyl-thio)cAMP, induce mRNAPEPCK as effectively as glucagon and with similar kinetics. Since sodium butyrate, adenosine, and dibutyryl cGMP are ineffective inducers, cAMP appears to be the active agent in the hepatocyte.


Dental visits to a North Carolina emergency department: a painful problem.
Emergency departments (EDs) act as the safety net and alternative care site for patients without insurance who have dental pain. We conducted a retrospective chart review of visits to an urban teaching hospital ED over a 12-month period, looking at patients who presented with a chief complaint or ICD code indicating dental pain, toothache, or dental abscess. The number of visits to this ED by patients with a dental complaint was 1,013, representing approximately 1.3% of all visits to this ED. Dental patients had a mean age of 32 (+/- 13) years, and 60% of all dental visits were made by African Americans. Dental patients were more likely to be self-pay than all other ED patients (61% versus 22%, P < 0.001). At the vast majority of dental ED visits (97%), the patient was treated and discharged; at most visits (90%) no dental procedure was performed. ED treatment typically consisted of pain control and antibiotics; at 81% of visits, the patient received an opiate prescription on discharge, and at 69% of visits, the patient received an antibiotic prescription on discharge. This retrospective chart review covered a limited period of time, included only patients at a large urban academic medical center, and did not incorporate follow-up analysis. Although they make up a small percentage of all ED visits, dental ED visits are more common among the uninsured, seldom result in definitive care or hospital admission, and often result in prescription of an opioid or antibiotic. These findings are cause for concern and have implications for public policy.


To the end! Distribution of attention along a tool in peri- and extrapersonal space.
This study investigated the distribution of visual attention along a handheld tool depending on functional tool practice and its position in peripersonal or extrapersonal space. We created a tool with two functional parts placed at the tool's middle and end. Participants held the tool over a display such that functional parts were aligned with stimuli for a 50/50, go/no-go, target detection task. In Experiment 1, two groups with no prior tool experience performed the task either in peripersonal (near the body; tool held horizontally) or extrapersonal space (beyond arms' reach; tool held straight). Faster response times (RTs) were found for targets at the tool's end and for the peripersonal space group. In Experiment 2, participants used the tool's middle part in a hockey-like game prior to the task to assess functional practice effects. Again, faster RTs were found for targets at the tool's end and in peripersonal space. However, a tool part × space interaction suggested that mid-tool practice reduced performance differences between tool parts but only in peripersonal space. Experiment 3 confirmed the interaction effect when mid-tool practice was constrained to only extrapersonal space. Results suggest that visual attention is naturally drawn to the tool's end but that functional tool use can redistribute attention when positioned in peripersonal space. In extrapersonal space, no change was found suggesting that the extension of peripersonal space is not uniform in regards to visual attention and that the visual attention component is perhaps dissociable from the remapping of spatial representation by tools.


The embryonal carcinoma stem cell Ela-like activity involves a differentiation-regulated transcription factor.
Murine F9 embryonal carcinoma (EC) stem cells have an Ela-like transcription activity that is undetectable in F9 cells differentiated to parietal endoderm-like cells (F9-PE). The Ela-inducible adenovirus E2A promoter has been used to further define this activity and we show that in vitro the transcription of this promoter in F9 EC and F9-PE cell extracts reflects the regulation in vivo. In EC cell extracts several trans-acting protein factors bind to E2A promoter sequences. A distal domain containing a CRE binds proteins present in F9 EC, F9-PE and Hela cell extracts. Sequences between -71 and -50 define a multiplicity of binding activities, termed DRTF1, all of which are down regulated as EC stem cells differentiate. DRTF2, a low abundance, regulated binding activity requires DNA sequences that overlap those required by DRTF1. The CRE and the DRTF1 binding site compete for transcription in vitro, indicating that in EC cell extracts the respective proteins function as positively acting, binding site dependent transcription factors. Comparison of DRTF1 with the previously defined HeLa cell factor E2F, induced during adenovirus infection, indicates that although both factors recognise the same region of the promoter there are clear differences between them. These data indicate that multiple factors are necessary for efficient transcription of the E2A promoter in F9 EC cell extracts and suggest that DRTF1 is responsible, at least in part, for the developmental regulation of the cellular Ela-like activity.


AniPaint: interactive painterly animation from video.
This paper presents an interactive system for creating painterly animation from video sequences. Previous approaches to painterly animation typically emphasize either purely automatic stroke synthesis or purely manual stroke key framing. Our system supports a spectrum of interaction between these two approaches which allows the user more direct control over stroke synthesis. We introduce an approach for controlling the results of painterly animation: keyframed Control Strokes can affect automatic stroke's placement, orientation, movement, and color. Furthermore, we introduce a new automatic synthesis algorithm that traces strokes through a video sequence in a greedy manner, but, instead of a vector field, uses an objective function to guide placement. This allows the method to capture fine details, respect region boundaries, and achieve greater temporal coherence than previous methods. All editing is performed with a WYSIWYG interface where the user can directly refine the animation. We demonstrate a variety of examples using both automatic and user-guided results, with a variety of styles and source videos.


Developmental time course of acidic and basic fibroblast growth factors' expression in distinct cellular populations of the rat central nervous system.
Acidic and basic fibroblast growth factors (aFGF and bFGF, respectively) are expressed in high levels in adult central nervous system (CNS). We report the time course of developmental appearance and distribution of these factors and of two FGF receptors, FGFR-1 and FGFR-2, in the CNS of rats ranging in age from embryonic day 16 to adult. Immunohistochemical analysis showed that sensory neurons in the midbrain were the first cells to contain detectable aFGF immunoreactivity at embryonic day 18. The next cell group to contain aFGF were motor neurons, which were found to be aFGF-positive at the day of birth. A number of other subcortical neuronal populations were observed to contain aFGF immunoreactivity after postnatal day 7. Adult levels and distribution patterns of aFGF were reached in all CNS areas by postnatal day 28. Basic FGF immunoreactivity was observed at postnatal day 0 in neurons in the CA2 subfield of hippocampus. Astrocytes contained detectable bFGF immunoreactivity, starting at postnatal day 7. Adult levels and patterns of distribution of bFGF were reached in all CNS areas by postnatal day 28. These immunohistochemical observations were confirmed by using bioassay and Western blot techniques. FGFR-1 and FGFR-2 mRNA were expressed in significant levels in all CNS areas at all time points analyzed. The observation that aFGF and bFGF appear in specific and distinct cellular populations at relatively late developmental times suggests that these FGFs may be involved in specific mechanisms of CNS maturation, maintenance, and repair.


Human sarcomas contain RNA related to the RNA of a mouse leukemia virus.
Labeled DNA complementary to the RNA of the Rauscher leukemia virus was hybridized with RNA from the polysome fraction of human sarcomas. Eighteen out of 25 specimens contained RNA possessing homology to the RNA of the mouse leukemia virus but not to that of the unrelated viruses causing mammary tumors in mice or myeloblastosis in chickens. Further, no normal adult or fetal tissues showed significant amounts of RNA specific to mouse leukemia virus. It appears that human sarcomas contain RNA sequences homologous to those found in an agent related to a virus known to cause sarcomas in mice.


Treatment of Partial Rotator Cuff Tear with Ultrasound-guided Platelet-rich Plasma.
The treatment of symptomatic partial rotator cuff tear has presented substantial challenge to orthopaedic surgeons as it can vary from conservative to surgical repair. Researches have established the influence of platelet rich plasma in healing damaged tissue. Currently very few data are available regarding the evidence of clinical and radiological outcome of partial rotator cuff tear treated with ultrasound guided platelet rich plasma injection in English literature. 20 patients with symptomatic partial rotator cuff tears were treated with ultrasound guided platelet rich plasma injection. Before and after the injection of platelet rich plasma scoring was done with visual analogue score, Constant shoulder score, and UCLA shoulder score at 8 weeks and third month. A review ultrasound was performed 8 weeks after platelet rich plasma injection to assess the rotator cuff status. Our study showed statistically significant improvements in 17 patients in VAS pain score, constant shoulder score and UCLA shoulder score. No significant changes in ROM were noted when matched to the contra-lateral side (P < 0.001) at the 3 month follow-up. The study also showed good healing on radiological evaluation with ultrasonogram 8 weeks after platelet rich plasma injection. Ultrasound guided platelet rich plasma injection for partial rotator cuff tears is an effective procedure that leads to significant decrease in pain, improvement in shoulder functions, much cost-effective and less problematic compared to a surgical treatment.


Occupational heavy lifting and risk of ischemic heart disease and all-cause mortality.
Occupational heavy lifting is known to impose a high cardiovascular strain, but the risk of ischemic heart disease (IHD) from occupational heavy lifting is unknown. The objective was to investigate the association between occupational heavy lifting and risk of IHD and all-cause mortality, and the influence of occupational and leisure time physical activity on this association. Data were analyzed from 1987, 1994, and 2000 from the Danish National Health Interview Surveys providing a sample of 6,692 working men and 5,921 working women aged 16-85 years without cardiovascular disease at baseline. Conventional risk factors for the outcomes IHD and all-cause mortality were controlled for in Cox analyses. Among men, heavy lifting was associated with increased risk for IHD (hazard ratio (HR): 1.52, 95% Confidence interval (95% CI): 1.15, 2.02), while a decreased risk was associated with occupational (HR: 0.50, 95% CI: 0.37, 0.68) and leisure time (HR: 0.73, 95% CI: 0.56, 0.95) physical activity. Referencing men with high occupational physical activity and no heavy lifting, men with high occupational physical activity and heavy lifting did not have an increased risk (HR: 1.11, 95% CI:0.68, 1.82), while men with low occupational physical activity and heavy lifting had a substantial increased risk (HR: 2.56, 95% CI:1.52, 4.32). No significant associations were found for all-cause mortality or for females. These findings indicate an excessive risk for IHD from occupational heavy lifting among men, particularly among those with low occupational and leisure time physical activity.


Blood pressure and the brain: the neurology of hypertension.
Hypertension affects more than one in four adults. The brain is an early target of hypertension-induced organ damage, and may manifest as stroke, subclinical cerebrovascular abnormalities and dementia. Hypertension-related small vessel disease can cause vascular dementia and can potentiate Alzheimer's pathology, lowering the threshold at which signs and symptoms manifest. Many hypertensive emergencies may also have a neurological presentation, such as hypertensive encephalopathy, haemorrhagic stroke or pre-eclampsia. Here we highlight the importance of blood pressure in maintaining brain health and the brain's role in controlling blood pressure.


Difficulty in differentiating two cases of sigmoid stenosis by diverticulitis from cancer.
The incidence of colonic diverticulosis with or without diverticulitis has increased in the Japanese population due to the modernization of food and aging. The rate of diverticulitis in colon diverticulosis ranges from 8.1% to 9.6%. However, few cases of stenosis due to diverticulitis have been reported. These reports suggest that the differentiation between sigmoid diverticulitis and colon cancer is difficult. This report describes two cases of colon stenosis due to diverticulitis that were difficult to differentiate from colon cancer. Case 1 was a 70-year-old woman with narrowed stools for 1 month who underwent colonofiberscopy (CFS). CFS revealed a diverticulum and circumferential stenosis in the sigmoid colon. Barium enema revealed a marked, hourglass-shaped, 2-cm circumferential stenosis in the sigmoid colon. Fluorodeoxyglucose (FDG)-positron emission tomography computed tomography (CT) revealed an increased FDG uptake at the affected portion of the sigmoid colon. Sigmoid colon cancer was suspected, and laparoscopic sigmoidectomy was performed. Pathological examination demonstrated active inflammation with no evidence of malignancy. Case 2 was a 50-year-old man who presented to a nearby clinic with reduced stool output despite the urge to defecate. CFS detected severe stenosis in the sigmoid colon approximately 25 cm from the dentate line. Contrast-enhanced abdominal CT revealed multiple diverticula, wall thickening, and swelling of the lymph nodes around the peritoneal aorta and the inferior mesenteric artery. A partial sigmoidectomy was performed. Pathological examination of the resected specimen revealed no changes in the mucosal epithelial surface, but a marked infiltration of inflammatory cells was observed.


X-Ray diffraction and resonance shear measurement of nano-confined ionic liquids.
X-ray diffraction measurement at the SPring-8 synchrotron was employed to investigate the structures of two types of imidazolium-based ionic liquids (ILs), 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide ([C4mim][NTF2]) and 1-butyl-3-methylimidazolium tetrafluoroborate ([C4mim][BF4]), confined between silica surfaces by varying the surface separation distances of ca. 500 nm (bulk liquid), ca. 10 nm, and ca. 2 nm (hard wall thickness). The obtained diffraction profiles and intensities were discussed by considering the structures and properties of the nano-confined ILs between the silica surfaces investigated by resonance shear measurement (RSM) and molecular dynamics simulation (MD) in our previous reports. [C4mim][NTf2] showed two diffraction peaks at q = 8.8 nm-1 (spacing d = 0.71 nm) and at q = 14.0 nm-1 (spacing d = 0.45 nm) at the greatest distance (D = ca. 500 nm), which were assigned to the interval between the same ions (anion-anion or cation-cation) within the polar network of [C4mim][NTf2] and the interval between the neighboring anion-cation, respectively. The positions of these two peaks remained the same at D = ca. 10 nm and at the hard wall (D = ca. 2 nm) and their intensity factor increased, indicating that both the cation and anion existed in the same layer. This result was consistent with the checkerboard structure of [C4mim][NTf2] on the silica surface computer simulated in our previous studies. On the other hand, [C4mim][BF4] showed a peak at q = 15.4 nm-1 (spacing d = 0.41 nm) corresponding to the anion-cation interval at the greatest distance (D = ca. 500 nm). This peak became broader and weaker at D = ca. 12 nm and at D = ca. 2 nm.


Adaptation to high altitude in Sherpas: association with the insertion/deletion polymorphism in the Angiotensin-converting enzyme gene.
Sherpas are well-known for their physical strength at high altitudes. They adapt to high altitude so well that little acute or chronic mountain sickness has been documented in them. The possible genetic basis for this adaptation is, however, unclear. The objective of this study was to elucidate the genetic background underlying this characteristic among Sherpas with respect to the angiotension-converting enzyme (ACE) gene. We enrolled 105 Sherpa volunteers in Namche Bazaar (3440 meters) and 111 non-Sherpa Nepalese volunteers in Kathmandu Valley (1330 meters) in Nepal. Information about high-altitude exposure and physiological phenotypes was obtained via fieldwork investigation. The genotype of the insertion/deletion (I/D) polymorphism in the ACE gene was identified by polymerase chain reaction. Serum ACE activity was also measured. The distribution of the I dominant genotype (II & ID) and the I allelic frequency were significantly more prevalent in Sherpas (II & ID: 94.3%, I allele: 73.3%) than in non-Sherpas (II & ID: 85.6%, P = .035; I allele: 64.0%, P = .036). Moreover, despite residing at high altitude, the circulating ACE levels of Sherpas were statistically similar to those of non-Sherpas at low altitudes (Sherpas: 14.5 +/- 0.4 IU/L/37 degrees C; non-Sherpas: 14.7 +/- 0.4 IU/L/37 degrees C; P = .755). These findings suggest that the overrepresented I allele of the ACE gene in Sherpas might be one of the fundamental genetic factors responsible for maintaining physiological low-altitude ACE activity at high altitude, which may have an advantageous physiological role in adapting to a high-altitude environment.


Mitogenic growth factors in breast fluid obtained from healthy women: evaluation of biological and extraneous sources of variability.
Peptide growth factors (GFs), including epidermal GF (EGF) and transforming GF-alpha (TGF-alpha), are presumed to play an important role in the local regulation of breast cell proliferation. Breast fluid collected by nipple aspiration provides a potential means to assess the concentration of these factors in contact with the ductal epithelium. Although identification of immunoreactive EGF-like GFs in breast fluid has been reported previously, we performed this study to evaluate the sensitivity and reliability of newer RIA methods and to characterize the sources and amounts of both intra- and intersubject variability. We also evaluated the relationship of breast fluid EGF and TGF-alpha levels to each other and to plasma levels of estradiol and progesterone. Breast fluid and plasma samples were obtained two to four times at weekly intervals from 18 healthy, premenopausal women. EGF and TGF-alpha were measured by competitive binding RIA. Both GFs were detected with good precision in all breast fluid samples analyzed, using dilutions as low as 1:100 for EGF (1 microliter) and 1:25 for TGF-alpha (4 microliters). The correlations between the right and left breasts, sampled concurrently, were r = 0.78 (P = 0.003) for EGF and r = 0.89 (P = 0.0001) for TGF-alpha. For both GFs, the variation between women was substantially greater than the variation between breasts or over time in an individual woman, particularly for EGF, for which there were 100-fold differences between women in mean levels. When samples from multiple women were analyzed together, we found no apparent relationships between EGF and TGF-alpha levels or between either GF level and menstrual cycle phase or plasma hormone concentrations. However, in random effects analyses, EGF levels within an individual were significantly associated overall with both TGF-alpha (P = 0.02) and plasma estradiol levels (P = 0.01). These data, which are the first comprehensive results on the feasibility of measuring mitogenic GFs in breast fluid, support the conclusion that women secrete consistent and individually distinct levels of EGF and TGF-alpha and that, in at least some women, EGF secretion in vivo covaries with both TGF-alpha in breast fluid and circulating estradiol.


Modelling the association between patient characteristics and the change over time in a disease measure using observational cohort data.
In observational cohort studies we may wish to examine the associations between fixed patient characteristics and the longitudinal changes from baseline in a repeated outcome measure. Many biological and other outcome measures are known to be subject to measurement error and biological variation. In an initial analysis we may fit a regression model to all outcome measurements, accounting for all the identified sources of variability, and see how the characteristics are linked to the change for typical patients. However, the characteristics may also be linked to different distributions of the underlying outcome value at baseline, which itself may be correlated with the change over time. Therefore, if we wish to examine the change over time for patients of different characteristics but with the same underlying baseline value then the initial approach is confounded by the baseline values. Furthermore, if we attempt to remove this confounding by including the observed baseline measure as a covariate in a model for later measurements, then this may provide an approximate solution but is likely to introduce some bias. We propose a method based on first following the initial approach but then, applying a correction to the parameter estimates. This allows the predicted trajectories to be plotted and valid significance tests of association with characteristics. Our approach is compared with other methods and illustrated through a simulation study and an analysis of the association between HIV-1 subtype and immunological response after starting antiretroviral therapy.


The Montreal definition and classification of gastroesophageal reflux disease: a global evidence-based consensus.
A globally acceptable definition and classification of gastroesophageal reflux disease (GERD) is desirable for research and clinical practice. The aim of this initiative was to develop a consensus definition and classification that would be useful for patients, physicians, and regulatory agencies. A modified Delphi process was employed to reach consensus using repeated iterative voting. A series of statements was developed by a working group of five experts after a systematic review of the literature in three databases (Embase, Cochrane trials register, Medline). Over a period of 2 yr, the statements were developed, modified, and approved through four rounds of voting. The voting group consisted of 44 experts from 18 countries. The final vote was conducted on a 6-point scale and consensus was defined a priori as agreement by two-thirds of the participants. The level of agreement strengthened throughout the process with two-thirds of the participants agreeing with 86%, 88%, 94%, and 100% of statements at each vote, respectively. At the final vote, 94% of the final 51 statements were approved by 90% of the Consensus Group, and 90% of statements were accepted with strong agreement or minor reservation. GERD was defined as a condition that develops when the reflux of stomach contents causes troublesome symptoms and/or complications. The disease was subclassified into esophageal and extraesophageal syndromes. Novel aspects of the new definition include a patient-centered approach that is independent of endoscopic findings, subclassification of the disease into discrete syndromes, and the recognition of laryngitis, cough, asthma, and dental erosions as possible GERD syndromes. It also proposes a new definition for suspected and proven Barrett's esophagus. Evidence-based global consensus definitions are possible despite differences in terminology and language, prevalence, and manifestations of the disease in different countries. A global consensus definition for GERD may simplify disease management, allow collaborative research, and make studies more generalizable, assisting patients, physicians, and regulatory agencies.


New methods for objective angiogenesis evaluation of rat nerves using microcomputed tomography scanning and conventional photography.
Nerve regeneration involves multiple processes, which enhance blood supply that can be promoted by growth factors. Currently, tools are lacking to visualize the vascularization patterns in transplanted nerves in vivo. The purpose of this study was to describe three-dimensional visualization of the vascular system in the rat sciatic nerve and to quantify angiogenesis of nerve reconstruction. In 12 Lewis rats (weighing 250-300 g), 10 mm sciatic nerve gaps were repaired with ipsilateral reversed autologous nerve grafts. At 12 and 16 weeks of sacrifice, Microfil® contrast compound was injected in the aorta. Nerve autografts (N = 12) and contralateral untreated nerves (N = 12) were harvested and cleared while preserving the vasculature. The amount of vascularization was measured by quantifying the vascular surface area using conventional photography (two-dimensional) and the vascular volume was calculated with microcomputed tomography (three-dimensional). For each measurement, a vessel/nerve area ratio was calculated and expressed in percentages (vessel%). The vascular volume measured 3.53 ± 0.43% in autografts and 4.83 ± 0.45% vessels in controls at 12 weeks and 4.95 ± 0.44% and 6.19 ± 0.29% vessels at 16 weeks, respectively. The vascular surface area measured 25.04 ± 2.77% in autografts and 26.87 ± 2.13% vessels in controls at 12 weeks, and 28.11 ± 3.47% and 33.71 ± 2.60% vessels at 16 weeks, respectively. The correlation between both methods was statistically significant (p = .049). Both methods are considered to successfully reflect the degree of vascularization. Application of this technique could be used to visualize and objectively quantify angiogenesis of the transplanted nerve graft. Moreover, this simple method is easily reproducible and could be extrapolated to any other desired target organ ex vivo in small animals to investigate the vascular network.


Optical DNA Mapping Combined with Cas9-Targeted Resistance Gene Identification for Rapid Tracking of Resistance Plasmids in a Neonatal Intensive Care Unit Outbreak.
The global spread of antibiotic resistance among Enterobacteriaceae is largely due to multidrug resistance plasmids that can transfer between different bacterial strains and species. Horizontal gene transfer of resistance plasmids can complicate hospital outbreaks and cause problems in epidemiological tracing, since tracing is usually based on bacterial clonality. We have developed a method, based on optical DNA mapping combined with Cas9-assisted identification of resistance genes, which is used here to characterize plasmids during an extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae outbreak at a Swedish neonatal intensive care unit. The outbreak included 17 neonates initially colonized with ESBL-producing Klebsiella pneumoniae (ESBL-KP), some of which were found to carry additional ESBL-producing Escherichia coli (ESBL-EC) in follow-up samples. We demonstrate that all ESBL-KP isolates contained two plasmids with the blaCTX-M-15 gene located on the smaller one (~80 kbp). The same ESBL-KP clone was present in follow-up samples for up to 2 years in some patients, and the plasmid carrying the blaCTX-M-15 gene was stable throughout this time period. However, extensive genetic rearrangements within the second plasmid were observed in the optical DNA maps for several of the ESBL-KP isolates. Optical mapping also demonstrated that even though other bacterial clones and species carrying blaCTX-M group 1 genes were found in some neonates, no transfer of resistance plasmids had occurred. The data instead pointed toward unrelated acquisition of ESBL-producing Enterobacteriaceae (EPE). In addition to revealing important information about the specific outbreak, the method presented is a promising tool for surveillance and infection control in clinical settings.IMPORTANCE This study presents how a novel method, based on visualizing single plasmids using sequence-specific fluorescent labeling, could be used to analyze the genetic dynamics of an outbreak of resistant bacteria in a neonatal intensive care unit at a Swedish hospital. Plasmids are a central reason for the rapid global spread of bacterial resistance to antibiotics. In a single experimental procedure, this method replaces many traditional plasmid analysis techniques that together provide limited details and are slow to perform. The method is much faster than long-read whole-genome sequencing and offers direct genetic comparison of patient samples. We could conclude that no transfer of resistance plasmids had occurred between different bacteria during the outbreak and that secondary cases of ESBL-producing Enterobacteriaceae carriage were instead likely due to influx of new strains. We believe that the method offers potential in improving surveillance and infection control of resistant bacteria in hospitals.


Effects of caffeine on skin and core temperatures, alertness, and recovery sleep during circadian misalignment.
Caffeine promotes wakefulness during night shift work, although it also disturbs subsequent daytime sleep. Increased alertness by caffeine is associated with a higher core body temperature (CBT). A lower CBT and a narrow distal-to-proximal skin temperature gradient (DPG) have been reported to be associated with improved sleep, yet whether caffeine influences the DPG is unknown. We tested the hypothesis that the use caffeine during nighttime total sleep deprivation would reduce the DPG, increase CBT and alertness, and disturb subsequent daytime recovery sleep. We also expected that a greater widening of the DPG prior to sleep would be associated with a greater degree of sleep disturbance. Thirty healthy adults (9 females) aged 21.6 ± 3.5 years participated in a double-blind, 28-h modified constant routine protocol. At 23 h of wakefulness, participants in the treatment condition (n = 10) were given 2.9 mg/kg caffeine, equivalent to ~200 mg (or 2 espressos) for a 70-kg adult, 5 h before a daytime recovery sleep episode. Throughout the protocol, core and skin body temperatures, DPG, sleep architecture, and subjective alertness and mood were measured. Prior to sleep, caffeine significantly widened the DPG and increased CBT, alertness, and clear-headedness (p < 0.05). Caffeine also disturbed daytime recovery sleep (p < 0.05). Increased CBT and a wider DPG prior to sleep were associated with a longer latency to sleep, and a wider DPG was associated with disturbed recovery sleep (i.e., increased wakefulness after sleep onset, increased stage 1 sleep, decreased sleep efficiency, and decreased slow wave sleep) (p < 0.05). A widening of the DPG following nighttime caffeine may represent a component of the integrated physiological response by which caffeine improves alertness and disturbs subsequent daytime recovery sleep. Furthermore, our findings highlight that sleep disturbances associated with caffeine consumed near the circadian trough of alertness are still present when daytime recovery sleep occurs 5 h or approximately 1 half-life later.


Controllable double tunneling induced transparency and solitons formation in a quantum dot molecule.
We consider the coupling effect between interdot tunneling coupling and external optical control field to study the linear optical property and the formation of temporal optical solitons in a quantum dot molecules system, analytically. The results show that the double tunneling induced transparency (TIT) windows are appeared in the absorption curve of probe field because of the formation of dynamic Stark splitting and quantum destructive interference effect from the two upper levels. Interestingly, the width of the TIT window becomes wider with the increasing intensity of the optical control field. We also find that the Kerr nonlinear effect of the probe field can be modulated effectively through coherent control both the control field and the interdot tunneling coupling in this system. Meanwhile, we demonstrate that the formation of dark or bright solitons can be practical regulated by varying the intensity of the optical control field.


Drosophila Amphiphysin is a post-synaptic protein required for normal locomotion but not endocytosis.
Clathrin-mediated endocytosis is required to recycle synaptic vesicles for fast and efficient neurotransmission. Amphiphysins are thought to be multiprotein adaptors that may contribute to this process by bringing together many of the proteins required for endocytosis. Their in vivo function, however, has yet to be determined. Here, we show that the Drosophila genome encodes a single amphiphysin gene that is broadly expressed during development. We also show that, unlike its vertebrate counterparts, Drosophila Amphiphysin is enriched postsynaptically at the larval neuromuscular junction. To determine the role of Drosophila Amphiphysin, we also generated null mutants which are viable but give rise to larvae and adults with pronounced locomotory defects. Surprisingly, the locomotory defects cannot be accounted for by alterations in the morphology or physiology of the neuromuscular junction. Moreover, using stimulus protocols designed to test endocytosis under moderate and extreme vesicle cycling, we could not detect any defect in the neuromuscular junction of the amphiphysin mutant. Taken together, our findings suggest that Amphiphysin is not required for viability, nor is it absolutely required for clathrin-mediated endocytosis. However, Drosophila Amphiphysin function is required in both larvae and adults for normal locomotion.


Effects of early dexamethasone therapy on pulmonary mechanics and chronic lung disease in very low birth weight infants: a randomized, controlled trial.
To determine the changes in pulmonary mechanics before and during early dexamethasone therapy, and to evaluate the effect of dexamethasone on the duration of mechanical ventilation in very low birth weight (VLBW) ventilator-dependent infants at risk for chronic lung disease (CLD). A prospective randomized trial was conducted. Forty-three patients (birth weight 600 to 1500 g, gestational age 24 to 32 weeks) who failed to be weaned from the respirator at 7 to 14 days of age were enrolled; 23 infants received a 7-day course of dexamethasone (0.5 mg/kg/day intravenously for 3 days, 0.25 mg/kg/day for 3 days, and 0.1 mg/kg/day for 1 day), and 20 patients were in the control group. At similar mean airway pressure (MAP) and fractional inspired oxygen concentration (FiO2), respiratory system mechanics were measured before and on days 2, 5, and 7 of the study. Airway pressure, flow and tidal volume (VT) were recorded and only mechanical breaths were analyzed. Respiratory compliance (Crs) and respiratory resistance (Rrs) were calculated by two factor least mean square analysis. Eighty-three percent of infants in the dexamethasone group and 90% in the control group received surfactant in the first 24 hours of life. There was a significant increase in Crs and VT in the dexamethasone group as compared with the control group (P < .001). No major changes in Rrs were observed. Dexamethasone therapy significantly decreased FiO2 and MAP P < .001) and facilitated successful weaning from mechanical ventilation. In addition to a shorter duration of mechanical ventilation (P < .01), the occurrence of CLD (FiO2 > 0.21 at 36 weeks of corrected gestational age, chest radiograph changes) was significantly decreased in the dexamethasone group (P < .01). Except for a transient increase in blood pressure and serum glucose, there were no significant differences in infection rates, intraventricular hemorrhage, or retinopathy of prematurity. Thirteen patients in the control group received dexamethasone at a later age. Our findings indicate that: 1) early dexamethasone therapy in VLBW infants markedly improves respiratory compliance and tidal volume, reduces FiO2 and MAP requirements, and facilitates extubation in these infants; 2) early dexamethasone therapy reduces the duration of mechanical ventilation and decreases CLD (at 28 days and 36 weeks) in a population of VLBW infants largely treated with surfactant.


Genetics and smoking-associated cancers. A study of 485 families.
Cancer risk was evaluated in relatives of 254 consecutively ascertained probands with histologically verified lung cancer, and 231 probands with other smoking-related cancers. Findings disclosed a lack of any strong evidence for increased risk in lung cancer per se when only lung cancer in relatives was considered. Confounding factors, most prominent of which was the effect of cigarette smoking, variation of secular trends, and the heritability of the smoking phenotype itself, tended to obfuscate identification of an inherited effect presenting itself exclusively as lung cancer liability. On the other hand, a significant increase was observed in cancers of all anatomic sites among the relatives of lung cancer probands (P less than 0.001). Most of these neoplastic lesions were not associated with smoking and were not greatly influenced by secular trends. Furthermore, no significant excesses of cancer at all anatomic sites in relatives of probands with other smoking-associated carcinomas were observed. Thus, it may be concluded that the observation of increased risk for cancer at all anatomic sites in relatives of lung cancer probands may be a reflection of an underlying susceptibility to malignancy in these families.


Hypoxia control to normalize pathologic angiogenesis: potential role for endothelial precursor cells and miRNAs regulation.
Tumor microenvironment is a complex and highly dynamic milieu that provides very important clues on tumor development and progression mechanisms. Tumor-associated endothelial cells play a key role in stroma organization. They achieve tumor angiogenesis, a formation of tumor-associated (angiogenic) vessels mainly through sprouting from locally preexisting vessels and/or recruitment of bone marrow-derived endothelial progenitor cells. This process participates to supply nutritional support and oxygen to the growing tumor. Endothelial cells constitute the interface between circulating blood cells, tumor cells and the extracellular matrix, thereby controlling leukocyte recruitment, tumor cell behavior and metastasis formation. Hypoxia, a critical parameter of the tumor microenvironment, controls endothelial/tumor cell interactions and is the key to tumor angiogenesis development. Under hypoxic stress, tumor cells produce factors that promote angiogenesis, vasculogenesis, tumor cell motility, metastasis and cancer stem cell selection. Targeting tumor vessels is a therapeutic strategy that has lately been fast evolving from antiangiogenesis to vessel normalization as discussed in this review. We shall focus on the pivotal role of endothelial cells within the tumor microenvironment, the specific features and the part played by circulating endothelial precursors cells. Attention is stressed on their recruitment to the tumor site and their role in tumor angiogenesis where they are submitted to miRNAs-mediated de/regulation. Here the compensation of the tumor deregulated angiogenic miRNAs - angiomiRs - is emphasized as a potential therapeutic approach. The strategy is to over express anti-angiomiRs in the tumor angiogenesis site upon selective delivery by precursor endothelial cells as miRs carriers.


Assessing the belief bias effect with ROCs: reply to Dube, Rotello, and Heit (2010).
Dube, Rotello, and Heit (2010) argued (a) that the so-called receiver operating characteristic is nonlinear for data on belief bias in syllogistic reasoning; (b) that their data are inconsistent with Klauer, Musch, and Naumer's (see record 2000-02818-008) model of belief bias; (c) that their data are inconsistent with any of the existing accounts of belief bias and only consistent with a theory provided by signal detection theory; and (d) that in fact, belief bias is a response bias effect. In this reply, we present reanalyses of Dube et al.'s data and of old data suggesting (a) that the receiver operating characteristic is linear for binary "valid" versus "invalid" responses, as employed by the bulk of research in this field; (b) that Klauer et al.'s model describes the old data significantly better than does Dube et al.'s model and that it describes Dube et al.'s data somewhat better than does Dube et al.'s model; (c) that Dube et al.'s data are consistent with the account of belief bias by misinterpreted necessity, whereas Dube et al.'s signal detection model does not fit their data; and (d) that belief bias is more than a response bias effect.


Small-molecule BCL6 inhibitor effectively treats mice with nonsclerodermatous chronic graft-versus-host disease.
Patient outcomes for steroid-dependent or -refractory chronic graft-versus-host diesease (cGVHD) are poor, and only ibrutinib has been US Food and Drug Administration (FDA) approved for this indication. cGVHD is often driven by the germinal center (GC) reaction, in which T follicular helper cells interact with GC B cells to produce antibodies that are associated with disease pathogenesis. The transcriptional corepressor B-cell lymphoma 6 (BCL6) is a member of the Broad-complex, Tramtrack, and Bric-abrac/poxvirus and zinc finger (BTB/POZ) transcription factor family and master regulator of the immune cells in the GC reaction. We demonstrate that BCL6 expression in both donor T cells and B cells is necessary for cGVHD development, pointing to BCL6 as a therapeutic cGVHD target. A small-molecule BCL6 inhibitor reversed active cGVHD in a mouse model of multiorgan system injury with bronchiolitis obliterans associated with a robust GC reaction, but not in cGVHD mice with scleroderma as the prominent manifestation. For cGVHD patients with antibody-driven cGVHD, targeting of BCL6 represents a new approach with specificity for a master GC regulator that would extend the currently available second-line agents.


Structural Insights into Plasticity and Discovery of Remdesivir Metabolite GS-441524 Binding in SARS-CoV-2 Macrodomain.
The nsP3 macrodomain is a conserved protein interaction module that plays essential regulatory roles in the host immune response by recognizing and removing posttranslational ADP-ribosylation sites during SARS-CoV-2 infection. Thus targeting this protein domain may offer a therapeutic strategy to combat current and future virus pandemics. To assist inhibitor development efforts, we report here a comprehensive set of macrodomain crystal structures complexed with diverse naturally occurring nucleotides, small molecules, and nucleotide analogues including GS-441524 and its phosphorylated analogue, active metabolites of remdesivir. The presented data strengthen our understanding of the SARS-CoV-2 macrodomain structural plasticity and provide chemical starting points for future inhibitor development.


Reduced surround suppression in monocular motion perception.
Motion discrimination of large stimuli is impaired at high contrast and short durations. This psychophysical result has been linked with the center-surround suppression found in neurons of area MT. Recent physiology results have shown that most frontoparallel MT cells respond more strongly to binocular than to monocular stimulation. Here we measured the surround suppression strength under binocular and monocular viewing. Thirty-nine participants took part in two experiments: (a) where the nonstimulated eye viewed a blank field of the same luminance (n = 8) and (b) where it was occluded with a patch (n = 31). In both experiments, we measured duration thresholds for small (1 deg diameter) and large (7 deg) drifting gratings of 1 cpd with 85% contrast. For each subject, a Motion Suppression Index (MSI) was computed by subtracting the duration thresholds in logarithmic units of the large minus the small stimulus. Results were similar in both experiments. Combining the MSI of both experiments, we found that the strength of suppression for binocular condition (MSIbinocular = 0.249 ± 0.126 log10 (ms)) is 1.79 times higher than under monocular viewing (MSImonocular = 0.139 ± 0.137 log10 (ms)). This increase is too high to be explained by the higher perceived contrast of binocular stimuli and offers a new way of testing whether MT neurons account for surround suppression. Potentially, differences in surround suppression reported in clinical populations may reflect altered binocular processing.


Awareness and uptake of recommended vaccines among immunosuppressed patients.
The Green Book recommended in 2009 that immunosuppressed patients should be receiving the yearly seasonal flu vaccine, the pandemic swine influenza A H1N1/09 vaccine and should have had the single pneumococcal vaccination. A retrospective audit in 2010 involving 60 immunosuppressed patients revealed that 83.3% of participants were aware of their entitlement to the vaccines. The majority were informed by their GP practice rather than the prescribing specialist. In 2009, 70% of participants received the seasonal flu vaccine, 40% received the H1N1 vaccine and 21.6% had received the pneumococcal vaccine. Reasons given for not receiving the recommended vaccines were lack of awareness, reported by 37.5%, followed by worries regarding side-effects reported by 25%. The data suggest that uptake rates, particularly for pneumococcal vaccination, could be improved with targeted information and promotion at the point of commencing immunosuppressants and approaching the influenza season. Prescribing physicians should take a more active role in routinely promoting and planning vaccination for at-risk groups and should provide information on how to receive the recommended vaccines and their side-effect profiles.


Roles of porphyrins and host iron transport proteins in regulation of growth of Porphyromonas gingivalis W50.
Porphyromonas gingivalis (Bacteroides gingivalis) requires iron in the form of hemin for growth and virulence in vitro, but the contributions of the porphyrin ring structure, porphyrin-associated iron, host hemin-sequestering molecules, and host iron-withholding proteins to its survival are unknown. Therefore, the effects of various porphyrins, host iron transport proteins, and inorganic iron sources on the growth of P. gingivalis W50 were examined to delineate the various types of iron molecules used for cellular metabolism. Cell envelope-associated hemin and iron stores contributed to the growth of P. gingivalis in hemin-free culture, and depletion of these endogenous reserves required eight serial transfers into hemin-free medium for total suppression of growth. Comparable growth of P. gingivalis was observed with 7.7 microM equivalents of hemin as hemoglobin (HGB), methemoglobin, myoglobin, hemin-saturated serum albumin, lactoperoxidase, cytochrome c, and catalase. Unrestricted growth was recorded in the presence of haptoglobin-HGB and hemopexin-hemin complexes, indicating that these host defense proteins do not sequester HGB and hemin from P. gingivalis. The iron chelator 2,2'-bipyridyl functionally chelated hemin-associated iron, resulting in dose-dependent inhibition of growth in hemin-restricted cultures at 1 to 25 microM 2,2'-bipyridyl concentrations. In the absence of an exogenous iron source, protoporphyrin IX did not support P. gingivalis growth. These findings suggest that the iron atom in the hemin molecule is the critical constituent for growth and that the tetrapyrrole porphyrin ring structure may represent an important vehicle for delivery of iron into the P. gingivalis cell. P. gingivalis does not have a strict requirement for porphyrins, since growth occurred with nonhemin iron sources, including high concentrations (200 muM) of ferric, ferrous, and nitrogenous inorganic iron, and P. gingivalis exhibited unrestricted growth in the presence of host transferrin, lactoferrin, and serum albumin. The diversity of iron substrates utilized by P. gingivalis and the observation that growth was not affected by the bacteriostatic effects of host iron-withholding proteins, which it may encounter in the periodontal pocket, may explain why P. gingivalis is such a formidable pathogen in the periodontal disease process.


Preoperative and perioperative use of levosimendan in cardiac surgery: European expert opinion.
In cardiac surgery, postoperative low cardiac output has been shown to correlate with increased rates of organ failure and mortality. Catecholamines have been the standard therapy for many years, although they carry substantial risk for adverse cardiac and systemic effects, and have been reported to be associated with increased mortality. On the other hand, the calcium sensitiser and potassium channel opener levosimendan has been shown to improve cardiac function with no imbalance in oxygen consumption, and to have protective effects in other organs. Numerous clinical trials have indicated favourable cardiac and non-cardiac effects of preoperative and perioperative administration of levosimendan. A panel of 27 experts from 18 countries has now reviewed the literature on the use of levosimendan in on-pump and off-pump coronary artery bypass grafting and in heart valve surgery. This panel discussed the published evidence in these various settings, and agreed to vote on a set of questions related to the cardioprotective effects of levosimendan when administered preoperatively, with the purpose of reaching a consensus on which patients could benefit from the preoperative use of levosimendan and in which kind of procedures, and at which doses and timing should levosimendan be administered. Here, we present a systematic review of the literature to report on the completed and ongoing studies on levosimendan, including the newly commenced LEVO-CTS phase III study (NCT02025621), and on the consensus reached on the recommendations proposed for the use of preoperative levosimendan.


Layer-specific potentiation of network GABAergic inhibition in the CA1 area of the hippocampus.
One of the most important functions of GABAergic inhibition in cortical regions is the tight control of spatiotemporal activity of principal neuronal ensembles. However, electrophysiological recordings do not provide sufficient spatial information to determine the spatiotemporal properties of inhibitory plasticity. Using Voltage Sensitive Dye Imaging (VSDI) in mouse hippocampal slices, we demonstrate that GABAA-mediated field inhibitory postsynaptic potentials undergo layer-specific potentiation upon activation of metabotropic glutamate receptors (mGlu). VSDI recordings allowed detection of pharmacologically isolated GABAA-dependent hyperpolarization signals. Bath-application of the selective group-I mGlu receptor agonist, (S)-3,5-Dihydroxyphenylglycine (DHPG), induces an enhancement of the GABAergic VSDI-recorded signal, which is more or less pronounced in different hippocampal layers. This potentiation is mediated by mGlu5 and downstream activation of IP3 receptors. Our results depict network GABAergic activity in the hippocampal CA1 region and its sub-layers, showing also a novel form of inhibitory synaptic plasticity tightly coupled to glutamatergic activity.


Trabecular bone volume and osteoprotegerin expression in uremic rats given high calcium.
Calcium (Ca)-containing phosphate binders have been recommended for the treatment of hyperphosphatemia in children with chronic kidney disease. To study the effects of high Ca levels on trabecular bone volume (BV) and osteoprotegerin (OPG) expression in uremic young rats, a model of marked overcorrection of secondary hyperparathyroidism was created by providing a diet of high Ca to 5/6 nephrectomized young rats (Nx-Ca) for 4 weeks. The results of chondrocyte proliferation and apoptosis, osteoclastic activity, OPG expression and BV were compared among intact rats given the control diet, intact rats given a high Ca diet and 5/6 nephrectomized rats given the control diet (Nx-Control) and the high Ca diet (Nx-Ca). Ionized Ca levels were higher and parathyroid hormone levels were lower in Nx-Ca rats than in the other groups. Final weight, final length and final tibial length of Nx-Ca rats were significantly less than those of the other groups, although the length gain did not differ among the groups. The hypertrophic zone width was markedly enlarged in Nx-Ca rats. Chondrocyte proliferation rates did not differ among the groups, whereas osteoclastic activity was decreased in Nx-Ca rats compared with the Nx-Control animals. The OPG expression and BV were increased in Nx-Ca rats compared with the Nx-Control rats. Increased BV should improve bone strength, whereas disturbance of osteoclastogenesis interferes with bone remodeling. Bone quality has yet to be determined in high Ca-fed uremic young rats.