Cell-specific, activation-dependent regulation of neutrophil CD32A ligand-binding function.
Neutrophils express 2 low-affinity FcgammaR, FcgammaRIIIB (CD16B), and FcgammaRIIA (CD32A). CD16B is a glycosyl-phosphatidyl inositol-anchored molecule, whereas CD32A is a polypeptide-anchored molecule. These 2 receptors also differ in their signaling. The biological significance of coexpression of 2 FcgammaRs with distinct membrane anchors and signaling capacities is not clearly understood. Using neutrophils from a CD16B-deficient donor and normal neutrophils treated with anti-CD16 monoclonal antibodies, the authors demonstrated that affinity modulation of CD32A is one of the mechanisms by which neutrophils regulate their FcgammaR-dependent functions. Neutrophils isolated from a CD16B(- )donor rosetted poorly with sheep erythrocytes opsonized with rabbit IgG (EA) (12% +/- 2% versus 80% +/- 6% for control) and were unable to mediate immunophagocytosis. However, activation of CD16B(-) neutrophils with fMLP, a bacterial chemotactic peptide, increased the CD32A-dependent EA rosetting to 58%. The CD32A-dependent rosetting of fMLP-activated normal neutrophils also increased nearly 5-fold, but there was no increase in CD32A expression. The CD32A-dependent immune complex (IC) binding was also increased in activated neutrophils. This affinity regulation was not observed with CD32A expressed on Chinese hamster ovary cells. These results suggest that in resting neutrophils CD32A is in a low-affinity state and that these cells primarily engage CD16B for IC binding. However, once the neutrophils are activated, the CD32A is converted to a high-affinity state that leads to CD32A-dependent ligand binding and signaling. These results suggest that neutrophils adopt a novel strategy to engage the 2 different FcgammaR selectively during physiologic and pathologic conditions to carry out their functions efficiently.


Cooperative and opposing effects of strategic and involuntary attention.
To assess whether working memory contents can effectively bias visual selection even when they do not represent the current target in the attention task, we recorded the ERP activity from participants performing both a memory task and, in the retention period, a visual search task. In this task, a distracter matching the memory content could be presented on the same side (congruent trials) or on the opposite side (incongruent trials) relative to the target location (Experiment 1 and Experiment 2). On some trials, only the matching distracter (but no target) was presented (catch trials, Experiment 2). Results showed that the N2pc component was modulated by the presence and location of a matching distracter. We interpret these results as evidence that the involuntary control exerted by the irrelevant memory contents coexists with the strategic mechanism related to the search target, influencing attention selection with roughly equal power. In Experiment 3, we found that the modulation of the N2pc is not strictly related to the active maintenance of the memory-target features but can also be elicited by repetition priming. Overall, these findings suggest that, together with the physical properties of the stimuli presented in the visual field, irrelevant memory contents represent a powerful class of factors that lead to involuntary attentional control.


CNTRICS imaging biomarkers final task selection: Long-term memory and reinforcement learning.
Functional imaging paradigms hold great promise as biomarkers for schizophrenia research as they can detect altered neural activity associated with the cognitive and emotional processing deficits that are so disabling to this patient population. In an attempt to identify the most promising functional imaging biomarkers for research on long-term memory (LTM), the Cognitive Neuroscience Treatment Research to Improve Cognition in Schizophrenia (CNTRICS) initiative selected "item encoding and retrieval," "relational encoding and retrieval," and "reinforcement learning" as key LTM constructs to guide the nomination process. This manuscript reports on the outcome of the third CNTRICS biomarkers meeting in which nominated paradigms in each of these domains were discussed by a review panel to arrive at a consensus on which of the nominated paradigms could be recommended for immediate translational development. After briefly describing this decision process, information is presented from the nominating authors describing the 4 functional imaging paradigms that were selected for immediate development. In addition to describing the tasks, information is provided on cognitive and neural construct validity, sensitivity to behavioral or pharmacological manipulations, availability of animal models, psychometric characteristics, effects of schizophrenia, and avenues for future development.


Genetic linkage of Wagner disease and erosive vitreoretinopathy to chromosome 5q13-14.
Wagner disease and erosive vitreoretinopathy are potentially blinding autosomal dominant diseases that share some similarities with Stickler syndrome. However, both disorders have associated retinal pigment epithelial changes, poor night vision, visual field defects, and abnormal electroretinographic findings, which are not found in families with COL2A1-associated Stickler syndrome. In addition, rhegmatogenous retinal detachments are uncommon in Wagner disease but occur in approximately 50% of patients with either Stickler syndrome or erosive vitreoretinopathy. To identify the chromosomal location of the genes involved in Wagner disease and erosive vitreoretinopathy and to distinguish these conditions genetically from Stickler syndrome. Fifteen affected members of a family affected with erosive vitreoretinopathy and 24 affected descendants of the pedigree described by Wagner were genotyped with a set of short tandem repeat polymorphisms distributed across the genome. Significant linkage was observed in each family between the disease phenotype and markers that map to chromosome 5q13-14. The highest lod score for the family affected with erosive vitreoretinopathy was 4.2 and was obtained with marker GATA3H06 (theta = 0). The highest lod score for the family affected with Wagner disease was 5.8 and was obtained with marker D5S815 (theta = 0). A candidate gene (cartilage link protein) that is known to lie near the linked interval was screened for mutations, but none was found in either family. These data suggest that erosive vitreoretinopathy and Wagner disease are allelic disorders and demonstrate that they are genetically distinct from COL2A1-associated Stickler syndrome.


An application of principal component analysis to the clavicle and clavicle fixation devices.
Principal component analysis (PCA) enables the building of statistical shape models of bones and joints. This has been used in conjunction with computer assisted surgery in the past. However, PCA of the clavicle has not been performed. Using PCA, we present a novel method that examines the major modes of size and three-dimensional shape variation in male and female clavicles and suggests a method of grouping the clavicle into size and shape categories. Twenty-one high-resolution computerized tomography scans of the clavicle were reconstructed and analyzed using a specifically developed statistical software package. After performing statistical shape analysis, PCA was applied to study the factors that account for anatomical variation. The first principal component representing size accounted for 70.5 percent of anatomical variation. The addition of a further three principal components accounted for almost 87 percent. Using statistical shape analysis, clavicles in males have a greater lateral depth and are longer, wider and thicker than in females. However, the sternal angle in females is larger than in males. PCA confirmed these differences between genders but also noted that men exhibit greater variance and classified clavicles into five morphological groups. This unique approach is the first that standardizes a clavicular orientation. It provides information that is useful to both, the biomedical engineer and clinician. Other applications include implant design with regard to modifying current or designing future clavicle fixation devices. Our findings support the need for further development of clavicle fixation devices and the questioning of whether gender-specific devices are necessary.


Recreating the tumor microenvironment in a bilayer, hyaluronic acid hydrogel construct for the growth of prostate cancer spheroids.
Cancer cells cultured in physiologically relevant, three-dimensional (3D) matrices can recapture many essential features of native tumor tissues. In this study, a hyaluronic acid (HA)-based bilayer hydrogel system that not only supports the tumoroid formation from LNCaP prostate cancer (PCa) cells, but also simulates their reciprocal interactions with the tumor-associated stroma was developed and characterized. HA hydrogels were prepared by mixing solutions of HA precursors functionalized with acrylate groups (HA-AC) and reactive thiols (HA-SH) under physiological conditions. The resultant viscoelastic gels have an average elastic modulus of 234 ± 30 Pa and can be degraded readily by hyaluronidase. The orthogonal and cytocompatible nature of the crosslinking chemistry permits facile incorporation of cytokine-releasing particles and PCa cells. In our bilayer hydrogel construct, the top layer contains heparin (HP)-decorated, HA-based hydrogel particles (HGPs) capable of releasing heparin-binding epidermal growth factor-like growth factor (HB-EGF) in a sustained manner at a rate of 2.5 wt%/day cumulatively. LNCaP cells embedded in the bottom layer receive the growth factor signals from the top, and in response form enlarging tumoroids with an average diameter of 85 μm by day 7. Cells in 3D hydrogels assemble into spherical tumoroids, form close cellular contacts through E-cadherin, and show cortical organization of F-actin, whereas those plated as 2D monolayers adopt a spread-out morphology. Compared to cells cultured on 2D, the engineered tumoroids significantly increased the expression of two pro-angiogenic factors, vascular endothelial growth factor-165 (VEGF(165)) and interleukin-8 (IL-8), both at mRNA and protein levels. Overall, the HA model system provides a useful platform for the study of tumor cell responses to growth factors and for screening of anticancer drugs targeting these pathways.


Closed cup vapor systems in percutaneous exposure studies: what is the dose?
Percutaneous vapor dosing studies have generally used saturated vapor concentration (SVC) measurements to estimate the exposure dose (Ct) of vapor produced from a volatile liquid within a closed system. The purpose of this study was to clarify whether the assumption was valid when translated to a biological system (pig skin) using sulfur mustard (SM) as a model skin penetrant. Three systems were evaluated, two containing skin and a control system (without skin). At set time points, samples from the headspace of each dosing system were extracted using a gas-tight syringe and analyzed by gas chromatography in conjunction with a flame-ionization detector. This demonstrated the rapid achievement of a constant vapor concentration within the biological and control systems and enabled a comparison with previously determined SVCs attained under ideal conditions. All three systems attained a constant vapor concentration within 2 min of exposure to SM. The control system reached an equilibrium vapor concentration of 1179 +/- 164 mg/m3, a value not significantly different from that derived from the SVC (1363 mg/m3). Because of absorption in the skin systems, SM vapor concentrations were significantly lower than that derived from the SVC and were dependent on the skin surface area within the dosing chamber (592 +/- 246 mg/m3 for a surface area of 10.15 cm2 and 740 +/- 224 mg/m3 for a surface area of 2.54 cm2). The assumption that SVC gives an acceptable measure of the Ct was shown to be valid by comparison with sulfur mustard recovered from the skin.


Product state and speed distributions in photochemical triple fragmentations.
The clearest dynamical signature of a roaming reaction is a very cold distribution of energy into the rotational and translational degrees of freedom of the roaming donor fragment (e.g. CO) and an exceptionally hot vibrational distribution in the roaming acceptor fragment (e.g. H2, CH4). These signatures were initially identified in joint experimental/theoretical investigations of roaming in H2CO and CH3CHO and are now used to infer the presence of roaming mechanisms in other photodissociation reactions. In this paper we construct a phase space theory (PST) model of triple fragmentation (3F) and show that the dynamical signature of 3F is similar to that of the roaming donor fragment. The PST model starts with a calculation of two-body fragmentation (2F) of a generic molecule, ABC into AB + C. Every AB fragment with sufficient energy to undergo subsequence spontaneous dissociation is allowed to dissociate and the PST distribution of energy into A + B products is calculated for every initial AB state. Using CH3CHO --> HCO + CH3 --> H + CO + CH3 as an example, we calculate that the energy disposal into the rotational and translational degrees of freedom of the 3F products is very low, and is similar to the dynamical signature expected for production of CO via a roaming mechanism. We compare the 3F PST model with published experimental data for photodissociation of CH3CHO and CH3OCHO at energies above the 3F threshold.


Barratt Impulsivity in Healthy Adults Is Associated with Higher Gray Matter Concentration in the Parietal Occipital Cortex that Represents Peripheral Visual Field.
Impulsivity is a personality trait of clinical importance. Extant research focuses on fronto-striatal mechanisms of impulsivity and how executive functions are compromised in impulsive individuals. Imaging studies employing voxel based morphometry highlighted impulsivity-related changes in gray matter concentrations in a wide array of cerebral structures. In particular, whereas prefrontal cortical areas appear to show structural alterations in individuals with a neuropsychiatric condition, the findings are less than consistent in the healthy population. Here, in a sample (n = 113) of young adults assessed for Barratt impulsivity, we controlled for age, gender and alcohol use, and showed that higher impulsivity score is associated with increased gray matter volume (GMV) in bilateral medial parietal and occipital cortices known to represent the peripheral visual field. When impulsivity components were assessed, we observed that this increase in parieto-occipital cortical volume is correlated with inattention and non-planning but not motor subscore. In a separate behavioral experiment of 10 young adults, we demonstrated that impulsive individuals are more vulnerable to the influence of a distractor on target detection in an attention task. If replicated, these findings together suggest aberrant visual attention as a neural correlate of an impulsive personality trait in neurotypical individuals and need to be reconciled with the literature that focuses on frontal dysfunctions.


Tick and Tickborne Pathogen Surveillance as a Public Health Tool in the United States.
In recent decades, tickborne disease (TBD) cases and established populations of medically important ticks have been reported over expanding geographic areas, and an increasing number of tickborne bacteria, viruses, and protozoans have been recognized as human pathogens, collectively contributing to an increasing burden of TBDs in the United States. The prevention and diagnosis of TBDs depend greatly on an accurate understanding by the public and healthcare providers of when and where persons are at risk for exposure to human-biting ticks and to the pathogens these ticks transmit. However, national maps showing the distributions of medically important ticks and the presence or prevalence of tickborne pathogens are often incomplete, outdated, or lacking entirely. Similar deficiencies exist regarding geographic variability in host-seeking tick abundance. Efforts to accurately depict acarological risk are hampered by lack of systematic and routine surveillance for medically important ticks and their associated human pathogens. In this review, we: 1) outline the public health importance of tick surveillance; 2) identify gaps in knowledge regarding the distributions and abundance of medically important ticks in the United States and the presence and prevalence of their associated pathogens; 3) describe key objectives for tick surveillance and review methods appropriate for addressing those goals; and 4) assess current capacity and barriers to implementation and sustainability of tick surveillance programs.


Nomenclature of monocytes and dendritic cells in blood.
Monocytes and cells of the dendritic cell lineage circulate in blood and eventually migrate into tissue where they further mature and serve various functions, most notably in immune defense. Over recent years these cells have been characterized in detail with the use of cell surface markers and flow cytometry, and subpopulations have been described. The present document proposes a nomenclature for these cells and defines 3 types of monocytes (classical, intermediate, and nonclassical monocytes) and 3 types of dendritic cells (plasmacytoid and 2 types of myeloid dendritic cells) in human and in mouse blood. This classification has been approved by the Nomenclature Committee of the International Union of Immunological Societies, and we are convinced that it will facilitate communication among experts and in the wider scientific community.


Patients with heart failure as co-designers of an educational website: implications for medical education.
To identify the learning needs of patients with heart failure between outpatients follow-up visits from their perspective and to ascertain what they emphasize as being important in the design of an educational website for them. We conducted a two-step qualitative study at Aarhus University Hospital, Denmark. Twenty patients with heart failure participated either in focus group interviews, diary writing, or video-recorded design sessions. Data on learning needs were collected in step 1 and analyses, therefore, helped develop the preliminary prototypes of a website. In step 2, patients worked on the prototypes in video-recorded design sessions, employing a think-aloud method. The interviews were transcribed and a content analysis was performed on the text and video data. Patients' learning needs were multifaceted, driven by anxiety, arising from, and often influenced by, such daily situations and contexts as the medical condition, medication, challenges in daily life, and where to get support and how to manage their self-care. They emphasized different ways of adapting the design to the patient group to enable interaction with peers and professionals and specific interface issues. This study provided insights into the different learning needs of patients with heart failure, how managing daily situations is the starting point for these needs and how emotions play a part in patients' learning. Moreover, it showed how patient co-designers proved to be useful for understanding how to design a website that supports patients' learning: insights, which may become important in designing online learning tools for patients.


Assessment of the matrix degenerative effects of MMP-3, ADAMTS-4, and HTRA1, injected into a bovine intervertebral disc organ culture model.
In vitro study to develop an intervertebral disc degeneration organ culture model, using coccygeal bovine intervertebral discs (IVDs) and injection of proteolytic enzymes MMP-3, ADAMTS-4, and HTRA1. This study aimed to develop an in vitro model of enzyme-mediated intervertebral disc degeneration to mimic the clinical outcome in humans for investigation of therapeutic treatment options. Bovine IVDs are comparable with human IVDs in terms of cell composition and biomechanical behavior. Researchers injected papain and trypsin into them to create an intervertebral disc degeneration model with a degenerated nucleus pulposus (NP) area. They achieved macroscopic cavities as well as a loss of glycosaminoglycans (GAGs). However, none of these enzymes are clinically relevant. Bovine IVDs were harvested maintaining the endplates. Active forms of MMP-3, ADAMTS-4, and HTRA1 were injected at a dose of 10 μg/mL each. Phosphate-buffered saline was injected as a control. Discs were cultured for 8 days and loaded diurnally (days 1-4 with ≈0.4 MPa for 16 hr) and left under free swelling condition from days 4 to 8 to avoid expected artifacts because of dehydration of the NP. Outcome parameters included disc height, metabolic cell activity, DNA content, GAG content, total collagen content, relative gene expression, and histological investigation. The mean metabolic cell activity was significantly lower in the NP area of discs injected with ADAMTS-4 than the day 0 control discs. Disc height was decreased after injection with HTRA1 and was significantly correlated with changes in GAG/DNA of the NP tissue. Total collagen content tended to be lower in groups injected with ADAMTS4 and MMP-3. MMP-3, ADAMTS-4, and HTRA1 provoked neither visible matrix degradation nor major shifts in gene expression. However, cell activity was significantly reduced and HTRA1 induced loss of disc height that positively correlated with changes in GAG/DNA content. The use of higher doses of these enzymes or a combination thereof may, therefore, be necessary to induce disc degeneration.


United we stand: integrating the actin cytoskeleton and cell-matrix adhesions in cellular mechanotransduction.
Many essential cellular functions in health and disease are closely linked to the ability of cells to respond to mechanical forces. In the context of cell adhesion to the extracellular matrix, the forces that are generated within the actin cytoskeleton and transmitted through integrin-based focal adhesions are essential for the cellular response to environmental clues, such as the spatial distribution of adhesive ligands or matrix stiffness. Whereas substantial progress has been made in identifying mechanosensitive molecules that can transduce mechanical force into biochemical signals, much less is known about the nature of cytoskeletal force generation and transmission that regulates the magnitude, duration and spatial distribution of forces imposed on these mechanosensitive complexes. By focusing on cell-matrix adhesion to flat elastic substrates, on which traction forces can be measured with high temporal and spatial resolution, we discuss our current understanding of the physical mechanisms that integrate a large range of molecular mechanotransduction events on cellular scales. Physical limits of stability emerge as one important element of the cellular response that complements the structural changes affected by regulatory systems in response to mechanical processes.


Hemiballismus following closed head injury.
Movement disorders are relatively rare after closed head injury (CHI), but when present they can go unrecognized if clinicians are not aware of their occurrence. We are presenting a case of hemiballismus which was not recognized over 3 years and was labelled as malingering or as psychosomatic. The symptoms have responded significantly to pharmacological interventions. The SPECT scan of the brain showed the lesions in the subthalamic areas while MRI, CT scans of brain and EEGs were reported normal. It is concluded that one should be aware of the existence of movement disorders after mild to moderate CHI, and that SPECT scan of the brain should be considered if a patient is symptomatic and other neuroimaging studies prove 'normal'.


Impaired selection of invariant natural killer T cells in diverse mouse models of glycosphingolipid lysosomal storage diseases.
Glycolipid ligands for invariant natural killer T cells (iNKT cells) are loaded onto CD1d molecules in the late endosome/lysosome. Accumulation of glycosphingolipids (GSLs) in lysosomal storage diseases could potentially influence endogenous and exogenous lipid loading and/or presentation and, thus, affect iNKT cell selection or function. The percentages and frequency of iNKT cells were reduced in multiple mouse models of lysosomal GSL storage disease, irrespective of the specific genetic defect or lipid species stored. Reduced numbers of iNKT cells resulted in the absence of cytokine production in response to alpha-galactosylceramide (alpha-GalCer) and reduced iNKT cell-mediated lysis of wild-type targets loaded with alpha-GalCer. The reduction in iNKT cells did not result from defective expression of CD1d or a lack of antigen-presenting cells. Although H-2 restricted CD4(+) T cell responses were generally unaffected, processing of a lysosome-dependent analogue of alpha-GalCer was impaired in all the strains of mice tested. These data suggest that GSL storage may result in alterations in thymic selection of iNKT cells caused by impaired presentation of selecting ligands.


Report of phenol peel for Asians.
With the advancement of trichloroacetic acid peel technology and wound care, trichloroacetic acid peeling has become very successful in Korea. Its success has opened studies on the possibility of using phenol on Korean skin. Dr. Mee's phenol formula (molding mask technique) was chosen for experiments on Korean skin because of the presumed safety of use on non-Caucasian skin. Between January of 1996 and January of 1998, 30 cases of significant small pox scars were treated with phenol at the Korea University Anam Hospital. The age range was from 43 to 60 years, with a mean of 49 years. The average follow-up period was 13 months, ranging from 1 month to 2 years. All of the procedures were performed in the operating room while the patient was under deep i.v. sedation. After the entire face was peeled, it was covered with an occlusive tape mask. During the recovery period, the patients underwent a post-peel skin care program. All 30 patients showed significant improvement of the severe pox marks with a marked rejuvenation effect. There was no sign of hypopigmentation or hyperpigmentation. As a complication, two patients developed hypertrophic scars on the perioral area, which responded well to steroid injections. Another pair of patients had herpetic infection, which left minimal scarring. Five patients developed cardiac arrhythmia with this rapid technique, but this was safely managed by an anesthesiologist during the procedure. Korean skin belongs to Fitzpatrick types IV and V and occasionally to type III or VI. It is common knowledge that performing chemical peeling on the latter types of skin is dangerous, but in this report, excellent results were obtained from all 30 patients, even though the peeling itself was very deep. With more knowledge and experience, phenol peel can be safely conducted on Asian skin.


Metagenomic Exploration of the Marine Sponge Mycale hentscheli Uncovers Multiple Polyketide-Producing Bacterial Symbionts.
Marine sponges have been a prolific source of unique bioactive compounds that are presumed to act as a deterrent to predation. Many of these compounds have potential therapeutic applications; however, the lack of efficient and sustainable synthetic routes frequently limits clinical development. Here, we describe a metagenomic investigation of Mycale hentscheli, a chemically gifted marine sponge that possesses multiple distinct chemotypes. We applied shotgun metagenomic sequencing, hybrid assembly of short- and long-read data, and metagenomic binning to obtain a comprehensive picture of the microbiome of five specimens, spanning three chemotypes. Our data revealed multiple producing species, each having relatively modest secondary metabolomes, that contribute collectively to the chemical arsenal of the holobiont. We assembled complete genomes for multiple new genera, including two species that produce the cytotoxic polyketides pateamine and mycalamide, as well as a third high-abundance symbiont harboring a proteusin-type biosynthetic pathway that appears to encode a new polytheonamide-like compound. We also identified an additional 188 biosynthetic gene clusters, including a pathway for biosynthesis of peloruside. These results suggest that multiple species cooperatively contribute to defensive symbiosis in M. hentscheli and reveal that the taxonomic diversity of secondary-metabolite-producing sponge symbionts is larger and richer than previously recognized.IMPORTANCEMycale hentscheli is a marine sponge that is rich in bioactive small molecules. Here, we use direct metagenomic sequencing to elucidate highly complete and contiguous genomes for the major symbiotic bacteria of this sponge. We identify complete biosynthetic pathways for the three potent cytotoxic polyketides which have previously been isolated from M. hentscheli Remarkably, and in contrast to previous studies of marine sponges, we attribute each of these metabolites to a different producing microbe. We also find that the microbiome of M. hentscheli is stably maintained among individuals, even over long periods of time. Collectively, our data suggest a cooperative mode of defensive symbiosis in which multiple symbiotic bacterial species cooperatively contribute to the defensive chemical arsenal of the holobiont.


Proteomic profile, biological activities and antigenic analysis of the venom from Bothriopsis bilineata smaragdina ("loro machaco"), a pitviper snake from Peru.
In order to determine Bothriopsis bilineata smaragdina venom (BbsV) composition, proteomic approaches were performed. Venom components were analyzed by RP-HPLC, SDS- PAGE and nano LC on line with LTQ Orbitrap XL. Results showed a total of 189 identified proteins, grouped into 11 different subgroups, which include snake venom metalloproteinases (SVMPs, 54.67%), snake C-type lectins (Snaclecs, 15.78%), snake venom serine proteinases (SVSPs, 14.69%), cystein-rich secretory proteins (CRISP, 2.61%), phospholipases A2 (PLA2, 1.14%), phosphodiesterase (PDE, 1.17%), venom endothelial growth factor (VEGF, 1.06%) 5'nucleotidases (0.33%), L-amino acid oxidases (LAAOs, 0.28%) and other proteins. In vitro enzymatic activities (SVMP, SVSP, LAAO, Hyal and PLA2) of BbsV were also analyzed. BbsV showed high SVSP activity but low PLA2 activity, when compared to other Bothrops venoms. In vivo, BbsV induced hemorrhage and edema in mice and showed intraperitoneal median lethal dose (LD50) of 92.74 (± 0.15) μg/20 g of mice. Furthermore, BbsV reduced cell viability when incubated with VERO cells. Peruvian and Brazilian bothropic antivenoms recognize BbsV proteins, as detected by ELISA and Western Blotting. Both antivenoms were able to neutralize in vivo edema and hemorrhage. In Peru, snakebite is a public health problem, especially in the rain forest, as a result of progressive colonization of this geographical area. This country is the second in Latin America, after Brazil, to exhibit the largest variety of venomous snakes. B. atrox and B. b. smaragdina snakes are sympatric species in Peruvian Amazon region and are responsible for approximately 95% of the envenomings reported in this region. B. b. smaragdina may cause a smaller share (3 to 38%) of those accidents, due to its arboreal habits, that make human encounters with these snakes less likely to happen. Despite B. b. smaragdina recognized medical importance, its venom composition and biological activities have been poorly studied. Furthermore, BbsV is not a component of the antigenic pool used to produce the corresponding Peruvian bothropic antivenom (P-BAV). Our results not only provide new insights on BbsV composition and biological activity, but also demonstrate that both P-BAV and B-BAV polyvalent antivenoms have a considerable recognition of proteins from BbsV and, more importantly, neutralized hemorrhage and edema, the main local effects of bothropic envenomation.


Modulation of Human CXCL12 Binding Properties to Glycosaminoglycans To Enhance Chemotactic Gradients.
Controlled release of active biomolecules is an attractive approach to modulate chemotactic gradients and accordingly the recruitment of cells, e.g. endothelial progenitor cells to improve wound healing or stimulate angiogenesis after myocardial infarction. Here, we developed variants of hCXCL12, also named stromal cell-derived factor 1α, a chemokine that activates the CXCR4 and consequently recruits tissue specific stem and progenitor cells. hCXCL12 variants were designed to bind to glycosaminoglycans (GAGs) with different affinities in order to modulate its release. Sixteen analogs were recombinantly produced, characterized, and tested for their GAG-binding property. The most promising variants hCXCL12 K24/K27/R41/R47A and hCXCL12 Q48K were used for release studies from starPEG-heparin-hydrogels. The reduced GAG affinity led to a fast release of hCXCL12 K24/K27/R41/R47A, whereas hCXCL12 Q48K was slowly released over 2 weeks due to its increased binding strength compared to wild type hCXCL12. Migration of Jurkat cells and early endothelial progenitor cells was proven to demonstrate the applicability of the approach to endogenously CXCR4 expressing cell types. Thus, this work offers new options for enhancing chemotactic hCXCL12 gradients by a combination of native and modified hCXCL12 variants to improve and prolong the recruitment of CXCR4-positive stem and progenitor cells to injured sites.


A review of genetic methods and models for analysis of coronavirus-induced severe pneumonitis.
Coronaviruses (CoVs) have been studied for over 60 years, but have only recently gained notoriety as deadly human pathogens with the emergence of severe respiratory syndrome CoV and Middle East respiratory syndrome virus. The rapid emergence of these viruses has demonstrated the need for good models to study severe CoV respiratory infection and pathogenesis. There are, currently, different methods and models for the study of CoV disease. The available genetic methods for the study and evaluation of CoV genetics are reviewed here. There are several animal models, both mouse and alternative animals, for the study of severe CoV respiratory disease that have been examined, each with different pros and cons relative to the actual pathogenesis of the disease in humans. A current limitation of these models is that no animal model perfectly recapitulates the disease seen in humans. Through the review and analysis of the available disease models, investigators can employ the most appropriate available model to study various aspects of CoV pathogenesis and evaluate possible antiviral treatments that may potentially be successful in future treatment and prevention of severe CoV respiratory infections.


Soil nutrient additions increase invertebrate herbivore abundances, but not herbivory, across three grassland systems.
Resource availability may influence invertebrate communities, with important consequences for ecosystem function, such as biomass production. We assessed: (1) the effects of experimental soil nutrient additions on invertebrate abundances and feeding rates and (2) the resultant changes in the effects of invertebrates on aboveground plant biomass at three grassland sites spanning the North American Central Plains, across which plant tissue chemistry and biomass vary. Invertebrate communities and rates of herbivory were sampled within a long-term nutrient-addition experiment established at each site along the broad Central Plains precipitation gradient. Additionally, the effects of invertebrates on aboveground plant biomass were determined under ambient and elevated nutrient conditions. At the more mesic sites, invertebrate herbivore abundances increased and their per capita rate of herbivory decreased with nutrient additions. In contrast, at the semi-arid site where plant biomass is low and plant nutrient concentrations are high, invertebrate herbivore abundances did not vary and per capita rates of herbivory increased with nutrient additions. No change in the effect of invertebrate herbivores on aboveground plant biomass was observed at any of the sites. In sum, nutrient additions induced shifts in both plant biomass and leaf nutrient content, which altered invertebrate abundances and feeding rate. However, due to the inverse relationship between changes in herbivore abundance and per capita rates of herbivory, nutrient additions did not alter the effect of invertebrates on aboveground biomass. Overall, we suggest that this inverse response of herbivore abundance and per capita feeding rate may buffer ecosystems against changes in invertebrate damage in response to fluctuations in nutrient levels.


Antidiabetic and hypolipidemic activities of Dillenia indica extract in diabetic rats.
The present study was carried out to investigate the antidiabetic and hypolipidemic activities of bioactive fraction (ethyl acetate fraction) of Dillenia indica methanolic extract in experimental diabetic Wistar rats. Type-1 diabetes was induced by single intraperitoneal injection of streptozotocin (60 mg/kg body weight), and type-2 diabetes was induced by single intraperitoneal injection of streptozotocin (60 mg/kg body weight), 15 min after the intraperitoneal injection of 120 mg/kg nicotinamide. The rats were treated by administering graded oral doses of isolated ethyl acetate fraction of methanolic extract of D. indica (DIEE), 200 and 400 mg/kg body weight, respectively, for 21 d. The blood glucose level was estimated at weekly intervals by glucometer. Serum cholesterol, triglycerides and high-density lipoprotein cholesterol (HDL-C) were also evaluated in normal and diabetic rats by an autoanalyzer. In both experimental models, daily oral treatment with DIEE for 21 d resulted in a significant reduction in blood glucose (P<0.01), serum cholesterol (P<0.05) and triglycerides (P<0.05) levels whereas HDL-C level was found to be increased (P<0.05) as compared with the diabetic control group. DIEE at 400 mg/kg body weight has prominent antidiabetic effect in experimental type-1 and type-2 diabetes models in rats. It may therefore be used as an alternative remedy for the treatment of diabetes mellitus.


ON ANAPHYLATOXINS AND ENDOTOXINS OF THE TYPHOID BACILLUS.
The experiments recorded in this paper confirm the observations of Friedberger that acutely toxic bodies can be produced from typhoid bacilli by the action of sensitizer and complement and that, when small quantities of bacteria are used, an excess of sensitization either interferes with the formation of the poisons or leads to a cleavage of the bacterial proteid beyond the poisonous intermediate products spoken of as anaphylatoxins. Unlike the experience of other workers with poisons of this nature, however, our experiments have shown that the action of complement upon typhoid bacilli strongly sensitized or not at all sensitized may be carried on, at body temperature, for considerably longer than twelve hours without leading to a destruction of the poisons, and that this is true when the quantities of the bacteria used vary within the wide range of from one to twelve agar slants. It has been found, in fact, that in the case of this microorganism prolonged exposure at the higher temperature of considerable quantities of bacteria constitutes an unfailing method of regularly obtaining powerful poisons. The results obtained by the use of smaller quantities and the less vigorous complement action at low temperatures are far less regular or satisfactory. It would appear from this that complement action of considerable vigor is required to obtain from this bacillus any appreciable yield of anaphylatoxin, and that the poison, once formed, is not as unstable as that found in other microorganisms by Neufeld and Dold and others. In fact, although we have never observed complete lysis in vitro of the typhoid bacilli treated with antibody and complement, the sensitized bacteria exposed to the action of complement for as long as fifteen hours at 37.5 degrees C. showed, in our experiments, much disintegration, and yet powerful poisons were present. Were the influence of lysis or of the too vigorous action of the serum bodies as rapidly poison-destroying in the case of this bacillus as it has been shown to be in the case of some other bacteria, it would be hard to understand how anaphylatoxins could play any part in the toxemia of typhoid fever. This phase of our experiments, however, seems to indicate that the conditions prevailing in the infected body at the height of this disease would furnish ideal criteria for anaphylatoxin production, since, in such cases, vigorously sensitized bacilli, in large numbers, are under the prolonged influence of considerable quantities of complement, conditions exactly comparable to those prevailing in our experiments. Granted that this state of affairs is actually the case, then the subsidence of the disease might depend merely upon limitation of the supply of antigen, as the increasing bactericidal action of the blood constituents come into play, and upon the consequent diminution of the anaphylatoxin. For as the bacteria diminish and the sensitizer increases, a changed proportion between them is established which, finally, as experiment has shown, results in a failure of anaphylatoxin production. For although our experiments have shown that, within a wide latitude of relative proportions of bacteria and antibody, anaphylatoxin can be formed, beyond this range an excess of one or the other element eventually will prevent their formation. It is not, however, the purpose of this paper to discuss the mechanism of the subsidence of the disease since this phase of the work will necessitate further experimental study. In regard to the experiments with kaolin, we were unable to confirm the contention of Keysser and Wassermann, though it is more than likely that toxic bodies could be formed by the action of complement upon any foreign proteid rendered amenable to its action. We are not inclined to attribute too much importance to these negative results, recording them merely as they occurred. However, should it be found subsequently that anaphylatoxins can be formed in this way, it seems unlikely that they are formed from the sensitizer or amboceptor as matrix, since this was not specifically adsorbed out of concentrated serum by the kaolin in our experiments. On the basis of experiments with so called endotoxins, ,we feel that the existence of such preformed intracellular poisons as an element in typhoid toxemia has not been proved, and is not absolutely necessary for the explanation of the phenomena occurring in this disease. However, the diarrhea, the hemorrhagic lesions, and the protracted symptoms following the injection of extracts and filtrates of the bacillus, differing so strikingly from the acute illness with rapid death or equally rapid recovery resulting from anaphylatoxin poisoning, would justify the assumption that poisons of this nature may still play a part in the disease, adding an additional specific characteristic to the clinical picture. As stated before, however, it is not improbable that all these characteristics may represent merely a more protracted or subacute state of anaphylatoxin toxemia. The experiments with autolysates, although none of them were fatal in their results upon guinea pigs, have sufficiently indicated that poisons comparable to anaphylatoxins can be formed in this way. This would indicate that a reaction of proteolysis, which may take place slowly by autolysis, is hastened by the action of complement, and its velocity is still further augmented by the increase, within certain limits, of the sensitization,-a conception which would attribute to the combined action of complement and sensitizer a function not incomparable to that of the bodies spoken of as catalytic agents.


Brief overview of dye-sensitized solar cells.
Dye-sensitized solar cells (DSC) are based on molecular and nanometer-scale components. Record cell efficiencies of 12%, promising stability data and means of energy-efficient production methods have been accomplished. As selling points for the DSC technology the prospect of low-cost investments and fabrication are key features. DSCs offer the possibilities to design solar cells with a large flexibility in shape, color, and transparency. The basic principles of the operation of DSC, the state-of-the-art as well as the potentials for future development are described.


A new vibroplasty coupling technique as a treatment for conductive and mixed hearing losses: a report of 4 cases.
The primary objective was to report on experiences regarding the safety and efficacy of the Vibrant Soundbridge (VSB) using a floating mass transducer (FMT)-partial/total ossicular replacement prosthesis (PORP/TORP) assembly as a treatment for conductive and mixed hearing losses of mild-to-moderate/severe degree. The secondary objective was to gather information regarding device fitting, as well as to refine surgical procedures. Five German-speaking adults from 2 European study sites were implanted with a VSB using an FMT-PORP/TORP assembly and evaluated before and after surgery for air-and bone-conduction thresholds and speech recognition performance. Evaluating the safety and efficacy of the VSB in combination with a PORP or TORP to treat conductive and mixed hearing loss. Residual cochlear hearing was unaffected by implantation with the device. Functional gain (measured as the difference between preoperative unaided and postoperative VSB-aided thresholds) could be calculated in 2 of 5 subjects, demonstrating that the VSB is effective in treating bone-conduction hearing losses of moderate/severe degree. Word recognition tests in quiet and noise showed good improvement in 3 of the cases. One patient experienced several other medical problems, making her audiological outcomes limited. One patient was excluded from the study owing to insufficient benefit and subsequently underwent revision surgery with FMT placement at the round window. The use of the VSB, implanted using the FMT-PORP/TORP assembly, was safe in all and efficacious in 3 of the 5 cases in this study. These are patients who may have few, if any, other options to manage their hearing loss.


Assessment of eight HPV vaccination programs implemented in lowest income countries.
Cervix cancer, preventable, continues to be the third most common cancer in women worldwide, especially in lowest income countries. Prophylactic HPV vaccination should help to reduce the morbidity and mortality associated with cervical cancer. The purpose of the study was to describe the results of and key concerns in eight HPV vaccination programs conducted in seven lowest income countries through the Gardasil Access Program (GAP). The GAP provides free HPV vaccine to organizations and institutions in lowest income countries. The HPV vaccination programs were entirely developed, implemented and managed by local institutions. Institutions submitted application forms with institution characteristics, target population, communication delivery strategies. After completion of the vaccination campaign (3 doses), institutions provided a final project report with data on doses administered and vaccination models. Two indicators were calculated, the program vaccination coverage and adherence. Qualitative data were also collected in the following areas: government and community involvement; communication, and sensitization; training and logistics resources, and challenges. A total of eight programs were implemented in seven countries. The eight programs initially targeted a total of 87,580 girls, of which 76,983 received the full 3-dose vaccine course, with mean program vaccination coverage of 87.8%; the mean adherence between the first and third doses of vaccine was 90.9%. Three programs used school-based delivery models, 2 used health facility-based models, and 3 used mixed models that included schools and health facilities. Models that included school-based vaccination were most effective at reaching girls aged 9-13 years. Mixed models comprising school and health facility-based vaccination had better overall performance compared with models using just one of the methods. Increased rates of program coverage and adherence were positively correlated with the number of vaccination sites. Qualitative key insights from the school models showed a high level of coordination and logistics to facilitate vaccination administration, a lower risk of girls being lost to follow-up and vaccinations conducted within the academic year limit the number of girls lost to follow-up. Mixed models that incorporate both schools and health facilities appear to be the most effective at delivering HPV vaccine. This study provides lessons for development of public health programs and policies as countries go forward in national decision-making for HPV vaccination.


Microspheres for protein delivery prepared from amphiphilic multiblock copolymers. 1. Influence of preparation techniques on particle characteristics and protein delivery.
The entrapment of lysozyme in amphiphilic multiblock copolymer microspheres by emulsification and subsequent solvent removal processes was studied. The copolymers are composed of hydrophilic poly(ethylene glycol) (PEG) blocks and hydrophobic poly(butylene terephthalate) (PBT) blocks. Direct solvent extraction from a water-in-oil (w/o) emulsion in ethanol or methanol did not result in the formation of microspheres, due to massive polymer precipitation caused by rapid solvent extraction in these non-solvents. In a second process, microspheres were first prepared by a water-in-oil-in-water (w/o/w) emulsion system with 4% poly(vinyl alcohol) (PVA) as stabilizer in the external phase, followed by extraction of the remaining solvent. As non-solvents ethanol, methanol and mixtures of methanol and water were employed. However, the use of alcohols in the extraction medium resulted in microspheres which gave an incomplete lysozyme release at a non-constant rate. Complete lysozyme release was obtained from microspheres prepared by an emulsification-solvent evaporation method in PBS containing poly(vinyl pyrrolidone) (PVP) or PVA as stabilizer. PVA was most effective in stabilizing the w/o/w emulsion. Perfectly spherical microspheres were produced, with high protein entrapment efficiencies. These microspheres released lysozyme at an almost constant rate for approximately 28 days. The reproducibility of the w/o/w emulsion process was demonstrated by comparing particle characteristics and release profiles of three batches, prepared under similar conditions.


DNA examination of ancient dental pulp incriminates typhoid fever as a probable cause of the Plague of Athens.
Until now, in the absence of direct microbiological evidence, the cause of the Plague of Athens has remained a matter of debate among scientists who have relied exclusively on Thucydides' narrations to introduce several possible diagnoses. A mass burial pit, unearthed in the Kerameikos ancient cemetery of Athens and dated back to the time of the plague outbreak (around 430 BC), has provided the required skeletal material for the investigation of ancient microbial DNA. To determine the probable cause of the Plague of Athens. Dental pulp was our material of choice, since it has been proved to be an ideal DNA source of ancient septicemic microorganisms through its good vascularization, durability and natural sterility. Six DNA amplifications targeted at genomic parts of the agents of plague (Yersinia pestis), typhus (Rickettsia prowazekii), anthrax (Bacillus anthracis), tuberculosis (Mycobacterium tuberculosis), cowpox (cowpox virus) and cat-scratch disease (Bartonella henselae) failed to yield any product in 'suicide' reactions of DNA samples isolated from three ancient teeth. On the seventh such attempt, DNA sequences of Salmonella enterica serovar Typhi were identified providing clear evidence for the presence of that microorganism in the dental pulp of teeth recovered from the Kerameikos mass grave. The results of this study clearly implicate typhoid fever as a probable cause of the Plague of Athens.


Electronic structural flexibility of heterobimetallic Mn/Fe cofactors: R2lox and R2c proteins.
The electronic structure of the Mn/Fe cofactor identified in a new class of oxidases (R2lox) described by Andersson and Högbom [Proc. Natl. Acad. Sci. U.S.A. 2009, 106, 5633] is reported. The R2lox protein is homologous to the small subunit of class Ic ribonucleotide reductase (R2c) but has a completely different in vivo function. Using multifrequency EPR and related pulse techniques, it is shown that the cofactor of R2lox represents an antiferromagnetically coupled Mn(III)/Fe(III) dimer linked by a μ-hydroxo/bis-μ-carboxylato bridging network. The Mn(III) ion is coordinated by a single water ligand. The R2lox cofactor is photoactive, converting into a second form (R2loxPhoto) upon visible illumination at cryogenic temperatures (77 K) that completely decays upon warming. This second, unstable form of the cofactor more closely resembles the Mn(III)/Fe(III) cofactor seen in R2c. It is shown that the two forms of the R2lox cofactor differ primarily in terms of the local site geometry and electronic state of the Mn(III) ion, as best evidenced by a reorientation of its unique (55)Mn hyperfine axis. Analysis of the metal hyperfine tensors in combination with density functional theory (DFT) calculations suggests that this change is triggered by deprotonation of the μ-hydroxo bridge. These results have important consequences for the mixed-metal R2c cofactor and the divergent chemistry R2lox and R2c perform.


Inhibition of stem cell factor-induced proliferation of primitive murine hematopoietic progenitor cells signaled through the 75-kilodalton tumor necrosis factor receptor. Regulation of c-kit and p53 expression.
TNF-alpha is a pleiotropic cytokine with stimulatory as well as inhibitory effects on hematopoiesis. We have previously demonstrated that TNF-alpha directly inhibits CSF-induced proliferation of primitive murine lineage-negative bone marrow progenitors (Lin-) and stem cell antigen-1 hematopoietic progenitors through the 75-kDa TNF receptor (TNF-R2), whereas TNF-alpha-induced inhibition of more committed Lin- progenitors is mediated through the 55-kDa TNF-R (TNF-R1), indicating a differential role of the two TNF-Rs in hematopoiesis. Numerous studies have demonstrated the ability of stem cell factor (SCF), a key regulator of hematopoiesis signaling through c-kit, to synergize with other hematopoietic growth factors, but little is known about cytokines capable of inhibiting hematopoiesis induced by SCF. While TNF-alpha has been demonstrated to enhance SCF-induced proliferation of myeloid leukemia blasts, the present report demonstrates that TNF-alpha, by signaling through TNF-R2, inhibits SCF-induced proliferation of normal murine Lin- and stem cell antigen-1 hematopoietic progenitors. SCF-stimulated proliferation of the hematopoietic cell line FDC-P1 was also potently inhibited by TNF-alpha and was accompanied by down-regulation of c-kit cell surface expression as well as c-kit mRNA levels. Finally, treatment of the FDC-P1 cell line with TNF-alpha resulted in increased levels of the tumor suppressor p53 mRNA, suggesting another mechanism by which hematopoietic effects of TNF-alpha may be mediated.


Three-dimensional bicontinuous nanoporous materials by vapor phase dealloying.
Three-dimensional bicontinuous open (3DBO) nanoporosity has been recognized as an important nanoarchitecture for catalysis, sensing, and energy storage. Dealloying, i.e., selectively removing a component from an alloy, is an efficient way to fabricate nanoporous materials. However, current electrochemical and liquid-metal dealloying methods can only be applied to a limited number of alloys and usually require an etching process with chemical waste. Here, we report a green and universal approach, vapor-phase dealloying, to fabricate nanoporous materials by utilizing the vapor pressure difference between constituent elements in an alloy to selectively remove a component with a high partial vapor pressure for 3DBO nanoporosity. We demonstrate that extensive elements, regardless of chemical activity, can be fabricated as nanoporous materials with tunable pore sizes. Importantly, the evaporated components can be fully recovered. This environmentally friendly dealloying method paves a way to fabricate 3DBO nanoporous materials for a wide range of structural and functional applications.


AMPA Receptor-mTOR Activation is Required for the Antidepressant-Like Effects of Sarcosine during the Forced Swim Test in Rats: Insertion of AMPA Receptor may Play a Role.
Sarcosine, an endogenous amino acid, is a competitive inhibitor of the type I glycine transporter and an N-methyl-d-aspartate receptor (NMDAR) coagonist. Recently, we found that sarcosine, an NMDAR enhancer, can improve depression-related behaviors in rodents and humans. This result differs from previous studies, which have reported antidepressant effects of NMDAR antagonists. The mechanisms underlying the therapeutic response of sarcosine remain unknown. This study examines the role of mammalian target of rapamycin (mTOR) signaling and α-amino-3-hydroxy-5-methylisoxazole-4-propionate receptor (AMPAR) activation, which are involved in the antidepressant-like effects of several glutamatergic system modulators. The effects of sarcosine in a forced swim test (FST) and the expression levels of phosphorylated mTOR signaling proteins were examined in the absence or presence of mTOR and AMPAR inhibitors. In addition, the influence of sarcosine on AMPAR trafficking was determined by analyzing the phosphorylation of AMPAR subunit GluR1 at the PKA site (often considered an indicator for GluR1 membrane insertion in neurons). A single injection of sarcosine exhibited antidepressant-like effects in rats in the FST and rapidly activated the mTOR signaling pathway, which were significantly blocked by mTOR inhibitor rapamycin or the AMPAR inhibitor 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(f)quinoxaline (NBQX) pretreatment. Moreover, NBQX pretreatment eliminated the ability of sarcosine to stimulate the phosphorylated mTOR signaling proteins. Furthermore, GluR1 phosphorylation at its PKA site was significantly increased after an acute in vivo sarcosine treatment. The results demonstrated that sarcosine exerts antidepressant-like effects by enhancing AMPAR-mTOR signaling pathway activity and facilitating AMPAR membrane insertion. Highlights-A single injection of sarcosine rapidly exerted antidepressant-like effects with a concomitant increase in the activation of the mammalian target of rapamycin mTOR signaling pathway.-The antidepressant-like effects of sarcosine occur through the activated AMPAR-mTOR signaling pathway.-Sarcosine could enhance AMPAR membrane insertion via an AMPAR throughput.


A comparative study of acquired amidase activity in Pseudomonas species.
Pseudomonas putida PP3 carrying dehalogenases I and II and Pseudomonas aeruginosa PAU3 carrying dehalogenase I coded for by plasmid pUU2 were able to grow on 2-monochloropropionic acid (2MCPA). Neither strain utilized 2-chloropropionamide (2CPA) as a carbon or nitrogen source for growth. Mutations in both strains to 2Cpa+ phenotypes (designated P. putida PPW3 and P. aeruginosa PAU5, respectively) involved the expression of an acquired 2CPA-amidase activity. The amidase followed by dehalogenase reactions in these strains constituted a novel metabolic pathway for growth on 2CPA. P. putida PPW3 synthesized a constitutive amidase of molecular mass 59 kDa consisting of two identical subunits of 29 kDa. For those amides tested this acquired enzyme was most active against chlorinated aliphatic amides, although substrate affinities (Km) and maximum rates of activity (Vmax) were poor. P. aeruginosa PAU5 acquired a 2Cpa+ phenotype by overproducing the A-amidase normally used by this species to hydrolyse aliphatic amides. The A-amidase had only slight activity towards 2CPA. However, with constitutive synthesis the mutant grew on the chlorinated substrates. Chloroacetamide (CAA) was a toxic substrate analogue for these Pseudomonas strains. A strain resistant to CAA was isolated from P. aeruginosa PAU5 when exposed to 1-10 mM-CAA. This mutant, P. aeruginosa PAU6, synthesized an inducible A-amidase. CAA-resistance depended upon the simultaneous expression of CAA-inducible amidase and dehalogenase activities.


Outcome of untreated infection with Entamoeba histolytica in homosexual men with and without HIV antibody.
Among homosexual men the prevalence of infection with Entamoeba histolytica is high. To determine the clinical importance of this infection 55 homosexual men carrying the parasite were investigated in detail. No clinical, serological, or histological evidence of invasive amoebiasis was found in any of them. The patients were not treated and were followed up for 12 to 29 months (mean 21.6 months), during which period none developed symptoms that could be attributed to E histolytica. Spontaneous loss of the parasite occurred in 17 patients, some of whom later became reinfected. Sixteen patients had antibody to human immunodeficiency virus, and infection with E histolytica showed the same benign course in them as in the patients who did not have antibody. Throughout the study classification of the isolates of E histolytica consistently showed that they belonged only to non-pathogenic zymodemes. The findings provide further evidence that E histolytica in homosexual men is a commensal organism.


A randomized comparison of misoprostol 6 to 8 hours versus 24 hours after mifepristone for abortion.
To demonstrate equivalence between mifepristone 200 mg followed 6 to 8 hours later and 24 hours later by misoprostol 800 microg vaginally for abortion in women up to 63 days of gestation. Mifepristone 200 mg was swallowed by 1,080 women after which they were randomly assigned to self-administer misoprostol intravaginally 6 to 8 hours later (group 1) or 23 to 25 hours later (group 2) at home. Participants returned for an evaluation, including transvaginal ultrasonography, 7 +/- 1 days after initiating treatment. Subjects who had not aborted were offered a second dose of misoprostol. All participants returned approximately 2 weeks after receiving mifepristone. Telephone contact was also attempted approximately 5 weeks after treatment. Treatment was considered a failure if a suction aspiration was performed for any indication. Complete abortion rates for groups 1 and 2 were 503 of 525 (95.8%, 95% confidence interval 93.7%, 97.3%) and 521 of 531 (98.1%, 95% confidence interval 96.6%, 99.1%), respectively, which were statistically equivalent. Side effects were significantly more common after mifepristone administration for women in group 2. Nausea, vomiting, and heavy bleeding were also significantly greater for women in group 2 after misoprostol treatment. Pain and subject acceptability were similar between groups. Mifepristone 200 mg followed 6 to 8 hours later by misoprostol 800 microg vaginally is as effective for abortion and has significantly fewer side effects as compared with regimens using a 24-hour dosing interval. Women receiving mifepristone and vaginal misoprostol for abortion can have the flexibility to administer the misoprostol as soon as 6 hours after using the mifepristone.


Reproducibility and Repeatability of Semiquantitative 18F-Fluorodihydrotestosterone Uptake Metrics in Castration-Resistant Prostate Cancer Metastases: A Prospective Multicenter Study.
18F-fluorodihydrotestosterone (18F-FDHT) is a radiolabeled analog of the androgen receptor's primary ligand that is currently being credentialed as a biomarker for prognosis, response, and pharmacodynamic effects of new therapeutics. As part of the biomarker qualification process, we prospectively assessed its reproducibility and repeatability in men with metastatic castration-resistant prostate cancer. Methods: We conducted a prospective multiinstitutional study of metastatic castration-resistant prostate cancer patients undergoing 2 (test/retest) 18F-FDHT PET/CT scans on 2 consecutive days. Two independent readers evaluated all examinations and recorded SUVs, androgen receptor-positive tumor volumes, and total lesion uptake for the most avid lesion detected in each of 32 predefined anatomic regions. The relative absolute difference and reproducibility coefficient (RC) of each metric were calculated between the test and retest scans. Linear regression analyses, intraclass correlation coefficients (ICCs), and Bland-Altman plots were used to evaluate repeatability of 18F-FDHT metrics. The coefficient of variation and ICC were used to assess interobserver reproducibility. Results: Twenty-seven patients with 140 18F-FDHT-avid regions were included. The best repeatability among 18F-FDHT uptake metrics was found for SUV metrics (SUVmax, SUVmean, and SUVpeak), with no significant differences in repeatability among them. Correlations between the test and retest scans were strong for all SUV metrics (R2 ≥ 0.92; ICC ≥ 0.97). The RCs of the SUV metrics ranged from 21.3% (SUVpeak) to 24.6% (SUVmax). The test and retest androgen receptor-positive tumor volumes and TLU, respectively, were highly correlated (R2 and ICC ≥ 0.97), although variability was significantly higher than that for SUV (RCs > 46.4%). The prostate-specific antigen levels, Gleason score, weight, and age did not affect repeatability, nor did total injected activity, uptake measurement time, or differences in uptake time between the 2 scans. Including the most avid lesion per patient, the 5 most avid lesions per patient, only lesions 4.2 mL or more, only lesions with an SUV of 4 g/mL or more, or normalizing of SUV to area under the parent plasma activity concentration-time curve did not significantly affect repeatability. All metrics showed high interobserver reproducibility (ICC > 0.98; coefficient of variation < 0.2%-10.8%). Conclusion: Uptake metrics derived from 18F-FDHT PET/CT show high repeatability and interobserver reproducibility.


When health technology assessment is confidential and experts have no power: the case of Hungary.
Health technology assessment (HTA) is not simply a mechanistic technical exercise as it takes place within a specific institutional context. Yet, we know little about how this context influences the operation of HTA and its ability to influence policy and practice. We seek to demonstrate the importance of considering institutional context, using a case study of Hungary, a country that has pioneered HTA in Central and Eastern Europe. We conducted 26 in-depth, semi-structured interviews with public- and private-sector stakeholders. We found that while the HTA Department, the Hungarian HTA organisation, fulfilled its formal role envisaged in the legislation, its potential for supporting evidence-based decision-making was not fully realised given the low levels of transparency and stakeholder engagement. Further, the Department's practical influence throughout the reimbursement process was perceived as being constrained by the payer and policy-makers, as well as its own limited organisational capacity. There was also scepticism as to whether the current operational form of the HTA process delivered 'good value for money'. Nevertheless, it still had a positive impact on the development of a broader institutional HTA infrastructure in Hungary. Our findings highlight the importance of considering institutional context in analysing the HTA function within health systems.


Task-dependent extraction of information from videos of iridescent and glossy samples.
We present an exploratory study on iridescence that revealed systematic differences in the perceptual clustering of glossy and iridescent samples that was driven by instructions to focus on either the material or the color properties of the samples. Participants' similarity ratings of pairs of video stimuli, showing the samples from multiple views, were analyzed using multidimensional scaling (MDS), and differences between the MDS solutions for the two tasks were consistent with flexible weighting of information from different views of the samples. These findings point to ecological implications for how viewers perceive and interact with the color-changing properties of iridescent objects.


Characterization of the Photochemical Properties of 5-Benzyluracil via Time-Dependent Density Functional Theory.
We present a detailed study of the excited state properties of 5-benzyluracil (5BU) in the gas phase and in implicit solvent using different electronic structure approaches ranging from time-dependent density functional theory in the linear response regime (LR-TDDFT) to a set of different wave-function-based methods for excited states, namely perturbed coupled cluster (CC2), algebraic diagrammatic construction method to second order (ADC(2)), and perturbed configuration interaction (CIS(D)). 5BU has been used to investigate DNA base-amino acid interactions. In particular, it served as a model of protein-DNA photoinduced cross-linking. While LR-TDDFT is computationally the most efficient first-principles approach for static and dynamic simulations of this bichromophoric system, its accuracy is difficult to assess due to the presence of excited states with charge transfer character. In this work, the performance of different exchange correlation functionals is compared against accurate benchmarks obtained either from high level wave-function-based methods or directly from experimental absorption spectra. Our investigation shows that accurate results for the excitation energies can be obtained using the hybrid meta-GGA functional M06. In view of dynamical studies of the relaxation of 5BU after photoexcitation, we also show that the PBE functional, while failing in the Franck-Condon region, provides qualitatively good results for the characterisation of a possible photocyclization path.


EGF signalling activates the ubiquitin proteasome system to modulate C. elegans lifespan.
Epidermal growth factor (EGF) signalling regulates growth and differentiation. Here, we examine the function of EGF signalling in Caenorhabditis elegans lifespan. We find that EGF signalling regulates lifespan via the Ras-MAPK pathway and the PLZF transcription factors EOR-1 and EOR-2. As animals enter adulthood, EGF signalling upregulates the expression of genes involved in the ubiquitin proteasome system (UPS), including the Skp1-like protein SKR-5, while downregulating the expression of HSP16-type chaperones. Using reporters for global UPS activity, protein aggregation, and oxidative stress, we find that EGF signalling alters protein homoeostasis in adults by increasing UPS activity and polyubiquitination, while decreasing protein aggregation. We show that SKR-5 and the E3/E4 ligases that comprise the ubiquitin fusion degradation (UFD) complex are required for the increase in UPS activity observed in adults, and that animals that lack SKR-5 or the UFD have reduced lifespans and indications of oxidative stress. We propose that as animals enter fertile adulthood, EGF signalling switches the mechanism for maintaining protein homoeostasis from a chaperone-based approach to an approach involving protein elimination via augmented UPS activity.


Antibody-Antigen Interaction Dynamics Revealed by Analysis of Single-Molecule Equilibrium Fluctuations on Individual Plasmonic Nanoparticle Biosensors.
Antibody-antigen interactions are complex events central to immune response, in vivo and in vitro diagnostics, and development of therapeutic substances. We developed an ultrastable single-molecule localized surface plasmon resonance (LSPR) sensing platform optimized for studying antibody-antigen interaction kinetics over very long time scales. The setup allowed us to perform equilibrium fluctuations analysis of the PEG/anti-PEG interaction. By time and frequency domain analysis, we demonstrate that reversible adsorption of monovalently bound anti-PEG antibodies is the dominant factor affecting the LSPR fluctuations. The results suggest that equilibrium fluctuation analysis can be an alternative to established methods for determination of interaction rates. In particular, the methodology is suited to analyze molecular systems whose properties change during the initial interaction phases, for example, due to mass transport limitations or, as demonstrated here, because the effective association rate constant varies with surface concentration of adsorbed molecules.


Urban Farming with Enhanced Rock Weathering As a Prospective Climate Stabilization Wedge.
With no single carbon capture and sequestration solution able to limit the global temperature rise to 1.5-2.0 °C by 2100, additional climate stabilization measures are needed to complement current mitigation approaches. Urban farming presents an easy-to-adopt pathway toward carbon neutrality, unlocking extensive urban surface areas that can be leveraged to grow food while sequestering CO2. Urban farming involves extensive surface areas, such as roofs, balconies, and vertical spaces, allowing for soil presence and atmospheric carbon sequestration through air-to-soil contact. In this viewpoint we also advocate the incorporation of enhanced rock weathering (ERW) into urban farming, providing a further opportunity for this recognized negative emissions technology that is gaining momentum worldwide to gain greater utilization.


Structure and Distribution of an Unrecognized Interstitium in Human Tissues.
Confocal laser endomicroscopy (pCLE) provides real-time histologic imaging of human tissues at a depth of 60-70 μm during endoscopy. pCLE of the extrahepatic bile duct after fluorescein injection demonstrated a reticular pattern within fluorescein-filled sinuses that had no known anatomical correlate. Freezing biopsy tissue before fixation preserved the anatomy of this structure, demonstrating that it is part of the submucosa and a previously unappreciated fluid-filled interstitial space, draining to lymph nodes and supported by a complex network of thick collagen bundles. These bundles are intermittently lined on one side by fibroblast-like cells that stain with endothelial markers and vimentin, although there is a highly unusual and extensive unlined interface between the matrix proteins of the bundles and the surrounding fluid. We observed similar structures in numerous tissues that are subject to intermittent or rhythmic compression, including the submucosae of the entire gastrointestinal tract and urinary bladder, the dermis, the peri-bronchial and peri-arterial soft tissues, and fascia. These anatomic structures may be important in cancer metastasis, edema, fibrosis, and mechanical functioning of many or all tissues and organs. In sum, we describe the anatomy and histology of a previously unrecognized, though widespread, macroscopic, fluid-filled space within and between tissues, a novel expansion and specification of the concept of the human interstitium.


Cholesterol-lowering effects of calcium carbonate in patients with mild to moderate hypercholesterolemia.
In recent years, several authors have noted that oral calcium treatment was associated with a reduction in serum cholesterol level. Calcium carbonate was examined for its ability to lower serum cholesterol levels in hypercholesterolemic patients. Fifty-six patients with mild to moderate hypercholesterolemia were examined in this randomized, double-blind, placebo-controlled crossover study. Patients were treated with a low-fat, low-cholesterol diet targeted at the American Heart Association Step-1 diet for 8 weeks before and while receiving placebo or calcium carbonate (9.98 mmol [400 mg] of elemental calcium) three times daily with meals for 6 weeks. Patients were then crossed over to the alternate treatment for an additional 6-week period. Compared with placebo, calcium carbonate achieved a 4.4% reduction in the low-density lipoprotein cholesterol level, and a 4.1% increase in the high-density lipoprotein cholesterol level. The ratio of low-density lipoprotein cholesterol to high-density lipoprotein cholesterol significantly decreased by 6.5% with calcium carbonate treatment. Calcium carbonate treatment did not significantly affect blood pressure or serum levels of triglycerides, lipoprotein Apo B, or calcium. Relative urinary saturation ratios of calcium oxalate levels were unchanged during calcium carbonate therapy. Compliance with diet and treatment was excellent and no significant adverse effects were noted. Thus, calcium carbonate was a modestly effective and well-tolerated adjunct to diet in the management of mild to moderate hypercholesterolemia in this clinical study.


Quarantine protects Falkland Islands (Malvinas) cats from feline coronavirus infection.
Feline coronavirus (FCoV) causes feline infectious peritonitis (FIP). Since 2002, when 20 cats on the Falkland Islands were found to be FCoV seronegative, only seronegative cats could be imported. Between 2005-2007, 95 pet and 10 feral cats tested negative by indirect immunofluorescence antibody (IFA) analysis using two strains of type II FCoV, two transmissible gastroenteritis virus assays, an enzyme-linked immunosorbent assay and rapid immunomigration test. Twenty-four samples (23%) showed non-specific fluorescence, mostly attributable to anti-nuclear antibodies (ANA). The reason for ANA was unclear: reactive samples were negative for Erhlichia canis antibodies; seven were feline immunodeficiency virus positive, but 15 were negative. It was not possible to determine retrospectively whether the cats had autoimmune disease, hyperthyroidism treatment, or recent vaccination which may also cause ANA. The FCoV/ FIP-free status of the Falkland Islands cats should be maintained by FCoV testing incoming cats. However, ANA can complicate interpretation of IFA tests.


Induction of bradykinesia with pallidal deep brain stimulation in patients with cranial-cervical dystonia.
Deep brain stimulation (DBS) of the globus pallidus internus (GPi) is an effective and well-tolerated treatment for idiopathic generalized dystonia. More recently, it has been applied as a treatment for focal and segmental dystonias. This patient population offers an opportunity to study the effects of alteration of pallidal outflow on previously normal limb function. We sought to retrospectively characterize the extent of novel GPi DBS-induced adverse motor effects in patients with adult-onset cervical and cranial-cervical dystonia using a questionnaire, and compared the findings to dystonia improvement as measured by standard scales. Despite significant improvement in dystonia (65% in mean Burke-Fahn-MarsdenDystonia Rating Scale motor score, p < 0.005, and 59% in mean Toronto Western Spasmodic Torticollis Rating Scale score, p < 0.008), slowing and difficulty with normal motor function was reported in previously nondystonic extremities in 10 of 11 patients. Symptoms were common in both upper and lower extremities and included new difficulties with handwriting (82%), getting up from a chair or in/out of a car (73%), and walking (45%), and were not associated with aberrant lead placement near the internal capsule. Although GPi DBS was shown to be effective in these patients, the influence of GPi DBS on nondystonic body regions deserves further investigation.


Resveratrol alleviates the cytotoxicity induced by the radiocontrast agent, ioxitalamate, by reducing the production of reactive oxygen species in HK-2 human renal proximal tubule epithelial cells in vitro.
Radiocontrast-induced nephropathy (RIN) is one of the leading causes of hospital-acquired acute kidney injury (AKI). The clinical strategies currently available for the prevention of RIN are insufficient. In this study, we aimed to determine whether resveratrol, a polyphenol phytoalexin, can be used to prevent RIN. For this purpose, in vitro experiments were performed using a human renal proximal tubule epithelial cell line (HK-2 cells). Following treatment for 48 h, the highly toxic radiocontrast agent, ioxitalamate, exerted cytotoxic effects on the HK-2 cells in a concentration-dependent manner, as shown by MTT assay. The half maximal inhibitory concentration (IC50) was found to be approximately 30 mg/ml. Flow cytometry also revealed a marked increase in the number of apoptotic cells following exposure to ioxitalamate. In addition, the number of necrotic, but not necroptotic cells was increased. However, treatment with resveratrol (12.5 µM) for 48 h significantly alleviated ioxitalamate (30 mg/ml)-induced cytotoxicity, by reducing cytosolic DNA fragmentation, increasing the expression of the anti-apoptotic protein, Bcl-2 (B-cell lymphoma 2), and survivin, activating caspase-3, preventing autophagic death and suppressing the production of reactive oxygen species (ROS). Resveratrol also suppressed the ioxitalamate-induced formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of oxidative DNA damage. N-acetylcysteine (NAC), a ROS scavenger commonly used to prevent RIN, also reduced ioxitalamate-induced cytotoxicity, but at a high concentration of 1 mM. Sirtuin (SIRT)1 and SIRT3 were not found to play a role in these effects. Overall, our findings suggest that resveratrol may prove to be an effective adjuvant therapy for the prevention of RIN.


Gene expression changes with age in skin, adipose tissue, blood and brain.
Previous studies have demonstrated that gene expression levels change with age. These changes are hypothesized to influence the aging rate of an individual. We analyzed gene expression changes with age in abdominal skin, subcutaneous adipose tissue and lymphoblastoid cell lines in 856 female twins in the age range of 39-85 years. Additionally, we investigated genotypic variants involved in genotype-by-age interactions to understand how the genomic regulation of gene expression alters with age. Using a linear mixed model, differential expression with age was identified in 1,672 genes in skin and 188 genes in adipose tissue. Only two genes expressed in lymphoblastoid cell lines showed significant changes with age. Genes significantly regulated by age were compared with expression profiles in 10 brain regions from 100 postmortem brains aged 16 to 83 years. We identified only one age-related gene common to the three tissues. There were 12 genes that showed differential expression with age in both skin and brain tissue and three common to adipose and brain tissues. Skin showed the most age-related gene expression changes of all the tissues investigated, with many of the genes being previously implicated in fatty acid metabolism, mitochondrial activity, cancer and splicing. A significant proportion of age-related changes in gene expression appear to be tissue-specific with only a few genes sharing an age effect in expression across tissues. More research is needed to improve our understanding of the genetic influences on aging and the relationship with age-related diseases.


Early bilingualism enhances mechanisms of false-belief reasoning.
In their first years, children's understanding of mental states seems to improve dramatically, but the mechanisms underlying these changes are still unclear. Such 'theory of mind' (ToM) abilities may arise during development, or have an innate basis, developmental changes reflecting limitations of other abilities involved in ToM tasks (e.g. inhibition). Special circumstances such as early bilingualism may enhance ToM development or other capacities required by ToM tasks. Here we compare 3-year-old bilinguals and monolinguals on a standard ToM task, a modified ToM task and a control task involving physical reasoning. The modified ToM task mimicked a language-switch situation that bilinguals often encounter and that could influence their ToM abilities. If such experience contributes to an early consolidation of ToM in bilinguals, they should be selectively enhanced in the modified task. In contrast, if bilinguals have an advantage due to better executive inhibitory abilities involved in ToM tasks, they should outperform monolinguals on both ToM tasks, inhibitory demands being similar. Bilingual children showed an advantage on the two ToM tasks but not on the control task. The precocious success of bilinguals may be associated with their well-developed control functions formed during monitoring and selecting languages.


Comparative effectiveness of tenofovir in HIV-infected treatment-experienced patients: systematic review and meta-analysis.
Antiretroviral therapy (ART) regimens for HIV infection are frequently changed. We conducted a systematic review of randomized trials (RCTs) on the benefits and harms of switching to tenofovir disoproxil fumarate (TDF)-based regimens in ART-experienced patients. We included RCTs in HIV-infected adults comparing switching to a TDF-containing regimen with maintaining or switching to another regimen. We searched MEDLINE, EMBASE, CENTRAL, LILACS, SCI, and the WHO Global Health Library. We assessed bias with the Cochrane tool and synthesized data using random-effects meta-analyses and Peto's approach. For further analyses, we added data from a previous systematic review in treatment-naïve patients. 17 RCTs with 2210 patients were included. All but one study had a high risk of bias. There was no significant association of switching to TDF-based regimens with mortality, fractures, CD4-cell count, body fat, virological failure, LDL-, and HDL-cholesterol. TDF-based regimens decreased total cholesterol (mean difference -12.05 mg/dL; 95% CI -20.76 to -3.34), trigylcerides (-14.33 mg/dL; -23.73 to -4.93), and bone mineral density (BMD; hip: -2.46%; -3.9 to -1.03; lumbar spine -1.52%; -2.69 to -0.34). Effects on estimated glomerular filtration (eGFR) were inconsistent and depended on the measurement. Adding 22 RCTs from 8297 treatment-naïve patients gave consistent results with then significant reductions of LDL (-7.57 mg/dL; -10.37 to -4.78), HDL (-2.38 mg/dL; -3.83 to -0.93), and eGFR (-3.49 ml/min; -5.56 to -1.43). Switching to TDF-based regimens is associated with reductions of BMD and lipid levels and possibly lowered kidney function. The evidence is limited by the high risk of bias.


Human prostasomes express glycolytic enzymes with capacity for ATP production.
Prostasomes are prostate-derived, exosome-like microvesicles that transmit signaling complexes between the acinar epithelial cells of the prostate and sperm cells. The vast majority of prostasomes have a diameter of 30-200 nm, and they are generally surrounded by a classical membrane bilayer. Using a selected proteomic approach, it became increasingly clear that prostasomes harbor distinct subsets of proteins that may be linked to adenosine triphosphate (ATP) metabolic turnover that in turn might be of importance in the role of prostasomes as auxiliary instruments in the fertilization process. Among the 21 proteins identified, most of the enzymes of anaerobic glycolysis were represented, and three of the glycolytic enzymes present are among the top 10 proteins found in most exosomes, once again linking prostasomes to the exosome family. Other prostasomal enzymes involved in ATP turnover were adenylate kinase, ATPase, 5'-nucleotidase, and hexose transporters. The identified enzymes in their prostasomal context were operational for ATP formation when supplied with substrates. The net ATP production was low due to a high prostasomal ATPase activity that could be partially inhibited by vanadate that was utilized to profile the ATP-forming ability of prostasomes. Glucose and fructose were equivalent as glycolytic substrates for prostasomal ATP formation, and the enzymes involved were apparently surface located on prostasomes, since an alternative substrate not being membrane permeable (glyceraldehyde 3-phosphate) was operative, too. There is no clear-cut function linked to this subset of prostasomal proteins, but some possible roles are discussed.


Predictive accuracy in the neuroprediction of rearrest.
A recently published study by the present authors reported evidence that functional changes in the anterior cingulate cortex within a sample of 96 criminal offenders who were engaged in a Go/No-Go impulse control task significantly predicted their rearrest following release from prison. In an extended analysis, we use discrimination and calibration techniques to test the accuracy of these predictions relative to more traditional models and their ability to generalize to new observations in both full and reduced models. Modest to strong discrimination and calibration accuracy were found, providing additional support for the utility of neurobiological measures in predicting rearrest.


Multistimuli-Responsive Enaminitrile Molecular Switches Displaying H+-Induced Aggregate Emission, Metal Ion-Induced Turn-On Fluorescence, and Organogelation Properties.
Multistimuli-responsive enaminitrile-based configurational switches displaying aggregation-induced emission (AIE), fluorescence turn-on effects, and supergelation properties are presented. The E-isomers dominated (>97%) in neutral/basic solution, and the structures underwent precisely controlled switching around the enamine C═C bond upon addition of acid/base. Specific fluorescence output was observed in response to different external input in the solution and solid states. In response to H+, configurational switching resulted in complete formation of the nonemissive Z-H+-isomers in solution, however displaying deep-blue to blue fluorescence (ΦF up to 0.41) in the solid state. In response to CuII in the solution state, the E-isomers exhibited intense, turn-on, blue-green fluorescence, which could be turned off by addition of competitive coordination. The acid/base-activated switching, together with the induced AIE-effects, further enabled the accomplishment of a responsive superorganogelator. In nonpolar solvents, a blue-fluorescent supramolecular gel was formed upon addition of acid to the E-isomer suspension. The gelation could be reversed by addition of base, and the overall, reversible process could be repeated at least five cycles.


RNAsnoop: efficient target prediction for H/ACA snoRNAs.
Small nucleolar RNAs are an abundant class of non-coding RNAs that guide chemical modifications of rRNAs, snRNAs and some mRNAs. In the case of many 'orphan' snoRNAs, the targeted nucleotides remain unknown, however. The box H/ACA subclass determines uridine residues that are to be converted into pseudouridines via specific complementary binding in a well-defined secondary structure configuration that is outside the scope of common RNA (co-)folding algorithms. RNAsnoop implements a dynamic programming algorithm that computes thermodynamically optimal H/ACA-RNA interactions in an efficient scanning variant. Complemented by an support vector machine (SVM)-based machine learning approach to distinguish true binding sites from spurious solutions and a system to evaluate comparative information, it presents an efficient and reliable tool for the prediction of H/ACA snoRNA target sites. We apply RNAsnoop to identify the snoRNAs that are responsible for several of the remaining 'orphan' pseudouridine modifications in human rRNAs, and we assign a target to one of the five orphan H/ACA snoRNAs in Drosophila. The C source code of RNAsnoop is freely available at http://www.tbi.univie.ac.at/ -htafer/RNAsnoop


Clinical implications of nervus intermedius variants in patients with geniculate neuralgia: Let anatomy be the guide.
Geniculate neuralgia (GN) is an uncommon, but severe, condition that is characterized by excruciating paroxysmal pain in the seventh cranial nerve's cutaneous distribution of general somatic afferent fibers carried through the nervus intermedius (NI). GN becomes a surgical disease in refractory cases of pain after exhaustive medical management. Surgical intervention in the form of microvascular decompression and nerve sectioning has been investigated with good patient outcomes. Despite this, there are limited guidelines on either technique's appropriateness in specific operative scenarios. In our 30-year experience in GNs surgical management, we have found that a detailed knowledge of the NIs anatomy, variants, and intraoperative surgical anatomic findings are the key to choosing the most appropriate intervention, and may provide the answer to why some patients fail to experience pain relief after surgery. These anatomic variants also may explain why many patients commonly do not experience side effects related to the visceral efferent and special afferent fibers after nerve sectioning.


Hyperelastic "bone": A highly versatile, growth factor-free, osteoregenerative, scalable, and surgically friendly biomaterial.
Despite substantial attention given to the development of osteoregenerative biomaterials, severe deficiencies remain in current products. These limitations include an inability to adequately, rapidly, and reproducibly regenerate new bone; high costs and limited manufacturing capacity; and lack of surgical ease of handling. To address these shortcomings, we generated a new, synthetic osteoregenerative biomaterial, hyperelastic "bone" (HB). HB, which is composed of 90 weight % (wt %) hydroxyapatite and 10 wt % polycaprolactone or poly(lactic-co-glycolic acid), could be rapidly three-dimensionally (3D) printed (up to 275 cm(3)/hour) from room temperature extruded liquid inks. The resulting 3D-printed HB exhibited elastic mechanical properties (~32 to 67% strain to failure, ~4 to 11 MPa elastic modulus), was highly absorbent (50% material porosity), supported cell viability and proliferation, and induced osteogenic differentiation of bone marrow-derived human mesenchymal stem cells cultured in vitro over 4 weeks without any osteo-inducing factors in the medium. We evaluated HB in vivo in a mouse subcutaneous implant model for material biocompatibility (7 and 35 days), in a rat posterolateral spinal fusion model for new bone formation (8 weeks), and in a large, non-human primate calvarial defect case study (4 weeks). HB did not elicit a negative immune response, became vascularized, quickly integrated with surrounding tissues, and rapidly ossified and supported new bone growth without the need for added biological factors.


A real-time proximity querying algorithm for haptic-based molecular docking.
Intermolecular binding underlies every metabolic and regulatory processes of the cell, and the therapeutic and pharmacological properties of drugs. Molecular docking systems model and simulate these interactions in silico and allow us to study the binding process. Haptic-based docking provides an immersive virtual docking environment where the user can interact with and guide the molecules to their binding pose. Moreover, it allows human perception, intuition and knowledge to assist and accelerate the docking process, and reduces incorrect binding poses. Crucial for interactive docking is the real-time calculation of interaction forces. For smooth and accurate haptic exploration and manipulation, force-feedback cues have to be updated at a rate of 1 kHz. Hence, force calculations must be performed within 1 ms. To achieve this, modern haptic-based docking approaches often utilize pre-computed force grids and linear interpolation. However, such grids are time-consuming to pre-compute (especially for large molecules), memory hungry, can induce rough force transitions at cell boundaries and cannot be applied to flexible docking. Here we propose an efficient proximity querying method for computing intermolecular forces in real time. Our motivation is the eventual development of a haptic-based docking solution that can model molecular flexibility. Uniquely in a haptics application we use octrees to decompose the 3D search space in order to identify the set of interacting atoms within a cut-off distance. Force calculations are then performed on this set in real time. The implementation constructs the trees dynamically, and computes the interaction forces of large molecular structures (i.e. consisting of thousands of atoms) within haptic refresh rates. We have implemented this method in an immersive, haptic-based, rigid-body, molecular docking application called Haptimol_RD. The user can use the haptic device to orientate the molecules in space, sense the interaction forces on the device, and guide the molecules to their binding pose. Haptimol_RD is designed to run on consumer level hardware, i.e. there is no need for specialized/proprietary hardware.


Intraarticular distal humerus fracture complicated by osteogenesis imperfecta treated with primary total elbow arthroplasty: a case report.
Intraarticular fractures of the distal humerus are challenging problems for the treating surgeon. When these fractures are complicated by severe comminution and poor bone quality, open reduction and internal fixation may lead to poor clinical outcomes when compared with those treated with primary total elbow arthroplasty. The population in which this clinical scenario most often arises is the elderly. An unusual case is presented in which a 38-year-old individual was successfully treated with primary total elbow arthroplasty for a highly comminuted, intraarticular distal humerus fracture with severe osteopenia due to osteogenesis imperfecta in which standard plate osteosynthesis was unlikely to provide sufficient stable fixation.