The Solo Play of TERT Promoter Mutations.
The reactivation of telomerase reverse transcriptase (TERT) protein is the principal mechanism of telomere maintenance in cancer cells. Mutations in the TERT promoter (TERTp) are a common mechanism of TERT reactivation in many solid cancers, particularly those originating from slow-replicating tissues. They are associated with increased TERT levels, telomere stabilization, and cell immortalization and proliferation. Much effort has been invested in recent years in characterizing their prevalence in different cancers and their potential as biomarkers for tumor stratification, as well as assessing their molecular mechanism of action, but much remains to be understood. Notably, they appear late in cell transformation and are mutually exclusive with each other as well as with other telomere maintenance mechanisms, indicative of overlapping selective advantages and of a strict regulation of TERT expression levels. In this review, we summarized the latest literature on the role and prevalence of TERTp mutations across different cancer types, highlighting their biased distribution. We then discussed the need to maintain TERT levels at sufficient levels to immortalize cells and promote proliferation while remaining within cell sustainability levels. A better understanding of TERT regulation is crucial when considering its use as a possible target in antitumor strategies.


Catecholamine transporters and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine neurotoxicity: studies comparing the cloned human noradrenaline and human dopamine transporter.
The uptake and cytotoxicity of 1-methyl-4-phenylpyridinium (MPP+), the toxic metabolite of the parkinsonism inducing agent 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), were studied in COS-7 cells transiently transfected with the cloned human noradrenaline and dopamine transporters and in permanently transfected SK-N-MC neuroblastoma cells. MPP+ had a 10- to 20-fold lower K(m) value for the noradrenaline than for the dopamine transporter. In dopamine transporter expressing cells, the maximal transport rate (Vmax) of MPP+, dopamine and noradrenaline was the same, but in noradrenaline transporter expressing cells the Vmax of MPP+ and dopamine was only one-half of the Vmax of noradrenaline. The turnover numbers (Vmax of uptake/maximal binding sites of binding) were 5 times higher for the dopamine transporter (as measured with [3H]dopamine and [3H]-2 beta-carbomethoxy-3 beta-(4-fluorophenyl) tropane than for the noradrenaline transporter (as measured with [3H]noradrenaline and [3H]nisoxetine). In SK-N-MC cells with similar Vmax values for both catecholamines, noradrenaline transporter expressing cells were killed by lower concentrations of MPP+ in the medium than dopamine transporter expressing cells. Desipramine blocked the toxicity of MPP+ toward the noradrenaline transporter, but not the dopamine transporter expressing cells. We conclude that the toxic effect of MPTP at the striatal dopamine system in the MPTP primate model of Parkinson's disease is not correlated with the affinity profile of MPP+ for catecholamine transporters, but rather with the higher turnover number of MPP+ at the dopamine transporter. In contradistinction, the toxicity of MPTP at the noradrenaline neurons in the primate cerebral cortex (Pifl et al., 1991) may involve the higher affinity of MPP+ for the noradrenaline transporter.


Safety and efficacy of miltefosine alone and in combination with sodium stibogluconate and liposomal amphotericin B for the treatment of primary visceral leishmaniasis in East Africa: study protocol for a randomized controlled trial.
Treatment options for visceral leishmaniasis (VL) in East Africa are far from satisfactory due to cost, toxicity, prolonged treatment duration or emergence of parasite resistance. Hence there is a need to explore alternative treatment protocols such as miltefosine alone or in combinations including miltefosine, sodium stibogluconate (SSG) or liposomal amphotericin B. The aim of this trial is to identify regimen(s) which are sufficiently promising for future trials in East Africa. A phase II randomized, parallel arm, open-labelled trial is being conducted to assess the efficacy of each of the three regimens: liposomal amphotericin B with SSG, Liposomal amphotericin B with miltefosine and miltefosine alone. The primary endpoint is cure at day 28 with secondary endpoint at day 210 (6 months). Initial cure is a single composite measure based on parasitologic evaluation (bone marrow, spleen or lymph node aspirate) and clinical assessment. Repeated interim analyses have been planned after recruitment of 15 patients in each arm with a maximum sample size of 63 for each. These will follow group-sequential methods (the triangular test) to identify when a regimen is inadequate (<75% efficacy) or adequate (>90% efficacy). We describe a method to ensure consistency of the sequential analysis of day 28 cure with the non-sequential analysis of day 210 cure. A regimen with adequate efficacy would be a candidate for treatment of VL with reasonable costs. The design allows repeated testing throughout the trial recruitment period while maintaining good statistical properties (Type I & II error rates) and reducing the expected sample sizes. ClinicalTrials.gov: NCT01067443.


HIF-1α mediates tumor hypoxia to confer a perpetual mesenchymal phenotype for malignant progression.
Although tumor progression involves genetic and epigenetic alterations to normal cellular biology, the underlying mechanisms of these changes remain obscure. Numerous studies have shown that hypoxia-inducible factor 1α (HIF-1α) is overexpressed in many human cancers and up-regulates a host of hypoxia-responsive genes for cancer growth and survival. We recently identified an alternative mechanism of HIF-1α function that induces genetic alterations by suppressing DNA repair. Here, we show that long-term hypoxia, which mimics the tumor microenvironment, drives a perpetual epithelial-mesenchymal transition (EMT) through up-regulation of the zinc finger E-box binding homeobox protein ZEB2, whereas short-term hypoxia induces a reversible EMT that requires the transcription factor Twist1. Moreover, we show that the perpetual EMT driven by chronic hypoxia depends on HIF-1α induction of genetic alterations rather than its canonical transcriptional activator function. These mesenchymal tumor cells not only acquire tumorigenicity but also display characteristics of advanced cancers, including necrosis, aggressive invasion, and metastasis. Hence, these results reveal a mechanism by which HIF-1α promotes a perpetual mesenchymal phenotype, thereby advancing tumor progression.


An unusual copper(I) halide-based metal-organic framework with a cationic framework exhibiting the release/adsorption of iodine, ion-exchange and luminescent properties.
A copper(I) halide-based compound with a formula of [Cu4I3(DABCO)2]I3 (DABCO = N,N'-dimethyl-1,4-diazabicyclo[2.2.2]octane) has been prepared by solvothermal reactions. This compound has been characterized by single-crystal X-ray diffraction, elemental analysis, IR, TG, XPS and powder X-ray diffractions. Structure analyses reveal that this compound is constructed by unprecedented cationic cluster [Cu8I6](2+) and organic ligand DABCO and the channels of this compound are occupied by I2 and I(-). The guest I2 and I(-) can move freely in and out of the host-framework. UV/vis spectra confirm that the I2 molecules in the channels can release into some organic solvents and IR spectra confirm the I(-) was exchanged by SCN(-). In addition, the luminescent properties of this compound in the solid state have also been investigated.


Development and application of two novel monoclonal antibodies against overexpressed CD26 and integrin α3 in human pancreatic cancer.
Monoclonal antibody (mAb) technology is an excellent tool for the discovery of overexpressed cell surface tumour antigens and the development of targeting agents. Here, we report the development of two novel mAbs against CFPAC-1 human pancreatic cancer cells. Using ELISA, flow cytometry, immunoprecipitation, mass spectrometry, Western blot and immunohistochemistry, we found that the target antigens recognised by the two novel mAbs KU44.22B and KU44.13A, are integrin α3 and CD26 respectively, with high levels of expression in human pancreatic and other cancer cell lines and human pancreatic cancer tissue microarrays. Treatment with naked anti-CD26 mAb KU44.13A did not have any effect on the growth and migration of cancer cells nor did it induce receptor downregulation. In contrast, treatment with anti-integrin α3 mAb KU44.22B inhibited growth in vitro of Capan-2 cells, increased migration of BxPC-3 and CFPAC-1 cells and induced antibody internalisation. Both novel mAbs are capable of detecting their target antigens by immunohistochemistry but not by Western blot. These antibodies are excellent tools for studying the role of integrin α3 and CD26 in the complex biology of pancreatic cancer, their prognostic and predictive values and the therapeutic potential of their humanised and/or conjugated versions in patients whose tumours overexpress integrin α3 or CD26.


Hamstring injuries. Current trends in treatment and prevention.
Pre-exercise stretching and adequate warm-up are important in the prevention of hamstring injuries. A previous mild injury or fatigue may increase the risk of injury. Hamstring muscle tear is typically partial and takes place during eccentric exercise when the muscle develops tension while lengthening, but variation in injury mechanisms is possible. Diagnosis of typical hamstring muscle injury is usually based on typical injury mechanism and clinical findings of local pain and loss of function. Diagnosis of avulsion in the ischial tuberosity, with the need for longer immobilisation, and a complete rupture of the hamstring origin, in which immediate operative treatment is necessary, poses a challenge to the treating physician. X-rays, ultrasonography or magnetic resonance imaging (MRI) may be helpful in differential diagnostics. After first aid with rest, compression, cold and elevation, the treatment of hamstring muscle injury must be tailored to the grade of injury. Conservative treatment is based on a knowledge of the biological background of the healing process of the muscle. Experimental studies have shown that a short period of immobilisation is needed to accelerate formation of the granulation tissue matrix following injury. The length of the immobilisation is, however, dependent on the grade of injury and should be optimised so that the scar can bear the pulling forces operating on it without re-rupture. Mobilisation, on the other hand, is required in order to regain the original strength of the muscle and to achieve good final results in resorption of the connective tissue scar and re-capillarisation of the damaged area. Another important aim of mobilisation--especially in sports medical practice--is to avoid muscle atrophy and loss of strength and extensibility, which rapidly result from prolonged immobilisation. Complete ruptures with loss of function should be operated on, as should cases resistant to conservative therapy in which, in the late phase of repair, the scar and adhesions prevent the normal function of the hamstring muscle.


Changes in laminin during recovery from postischemic acute renal failure in rats.
Changes in the immunoreactivity of laminin have been demonstrated in a variety of tissues undergoing injury-regeneration process and during nephrogenesis. Studies in vitro revealed that laminin is important in many cellular functions such as growth, differentiation, and cell communication. This study examined the changes of laminin in the extracellular matrix (ECM) of proximal straight tubules (PST) in the outer stripe of the outer medulla after renal ischemia, an area where most morphological damage occurred. Anesthetized male Sprague-Dawley adult rats underwent a 30-min temporary occlusion of the left renal artery. The inulin clearance (Cin) of the postischemic left kidneys was significantly decreased at 30 min, 1 day, 2 days, and 5 days after the injury, and at 7 and 10 days after the injury it rose to a level not significantly different from the sham-operated controls. Using immunogold electron microscopy, the density of immunoreactive laminin (gold-conjugated laminin grains/micron2 ECM area) in PST where flattened PST cells were present was decreased at 1 day and 2 days, and at 5 days, a time when the renal function was still depressed, it returned to a level not significantly different from that in controls. These changes were not observed in those PST or proximal convoluted tubules in which morphology was intact. Further studies are needed to determine if these changes in laminin of damaged PST cells have any significant role during their recovery from the ischemic injury.


Safety and immunogenicity of multimeric-001--a novel universal influenza vaccine.
A new vaccine, Multimeric-001, containing conserved linear epitopes from the HA, NP, and M1 proteins of influenza type A and type B strains was designed to protect against seasonal and pandemic influenza virus strains, regardless of mutations. We assessed its safety and tolerability and characterized humoral and cellular immune responses elicited by its administration. Sixty healthy volunteers received either 250 or 500 μg injections, with or without adjuvant (Montanide ISA 51VG), or matching placebo. Two intramuscular injections were administered, 21 days apart. Treatment was well tolerated and no significant adverse events were noted. Forty-two days after first injection, there was a 50-fold and 37-fold increase in IgG titers against Multimeric-001 protein, following the adjuvanted 500 and 250 μg doses, respectively. Sera from immunized subjects lysed MDCK cells infected with strains of influenza representing the major strains that infect humans: H1N1, H3N2, and influenza B. Proliferation of peripheral blood mononuclear cells as well as increase in IL-2 and IFN-gamma secretion occurred following incubation with the vaccine. This vaccine model differs fundamentally from the current influenza virus vaccines, as it does not contain the variable regions of the virus hemagglutinin and hence does not induce hemagglutination inhibition antibodies that serve as surrogate markers for protection. In order to demonstrate the potential efficacy of the Multimeric-001, an alternative assay was employed, in which the lysis of MDCK cells infected with different virus strains was shown, with the involvement of the complement mechanism. The humoral and cellular responses suggest a cross-immunity of the vaccine toward influenza virus strains regardless of mutations. These results corroborate the protective effect of the vaccine, previously shown in animals. Larger-scale studies are under way to further substantiate the safety and efficacy of the vaccine candidate.


Clinical value of perilesional perfusion deficit measured by Technetium-99m-ECD single-photon emission computed tomography in hypertensive intracerebral hemorrhage.
Pathological and experimental studies indicate the existence of a "penumbra" of progressive tissue damage and edema in regions immediately surrounding a hematoma in patients of intracerebral hemorrhage (ICH). This zone of oligemia surrounding ICH has a potential for perfusion recovery. Improved understanding of the pathophysiology of perilesional blood flow changes and brain injury after ICH may result in improved treatment strategies. The aim was to study perilesional blood flow changes in ICH by perfusion deficit (PD) measured by single-photon emission computed tomography (SPECT) and to correlate it with the severity of ICH and outcome. Forty-four patients of computed tomography (CT) documented nonlobar deep ICH suggestive of hypertensive hematoma of <7 days duration were subjected to 99mTc-ethylene diacetate SPECT scans of the brain. Patients with significant midline shift (0.5 cm) or global blood flow reduction were excluded from the analysis. SPECT scan of the brain was analyzed by segmental analysis, a semi-quantitative method of cerebral blood flow. A difference of radiotracer uptake of >10% between the region of interest of ICH cases and the ratio between the two ROI below 0.9 was taken as a significant PD. A correlation of PD was analyzed with that of various parameters such as the severity of stroke, duration from onset of ictus, and imaging including CT scan of the brain and SPECT scan. A statistically significant difference in the percentage of radiotracer uptake on comparison of ipsilateral and contralateral to ICH (P < 0.001) was observed, suggesting a significant hypoperfusion in the perilesional area in patients with ICH. A statistically significant correlation was noted between the severity of stroke and PD indicated by various parameters such as the National Institutes of Health Stroke Scale (NIHSS) score at admission (r = 0.328, P = 0.016), Glasgow Coma Scale (GCS) score at admission (r = -0.388, P = 0.005), and ICH score at admission (r = 0.314, P = 0.020). This study demonstrated more severe hypoperfusion in clinically severe ICH which is a possible explanation of poor outcomes in severe ICH cases. We observed hypoperfusion on SPECT study in 25 of 34 (73.5%) patients with subacute ICH and 5 of 10 patients (50%) with acute ICH. The mean time from the onset of ictus to SPECT scan done was 5.04 ± 1.75 days with a range of 1-7 days, suggesting the persistence of hypoperfusion in subacute stages too. This finding may be of clinical importance for identifying the salvageable area surrounding ICH for any possible intervention in future to improve the outcome. This study demonstrates that perilesional PD occurs in acute and subacute cases of ICH. This hypoperfusion is possibly time related and appears to be more severe in patients having major ICH with poor clinical and imaging parameters. This area of hypoperfusion or ischemic penumbra is a potential site for perfusion recovery to improve clinical outcomes and to reduce long-term neurological deficits.


Assessment of temporary cerebral effects induced by focused ultrasound with optical coherence tomography angiography.
Focused ultrasound (FUS) in combination with microbubbles temporally and locally increases the permeability of the blood-brain barrier (BBB) for facilitating drug delivery. However, the temporary effects of FUS on the brain microstructure and microcirculation need to be addressed. We used label-free optical coherence tomography (OCT) and OCT angiography (OCTA) to investigate the morphological and microcirculation changes in mouse brains due to FUS exposure at different power levels. Additionally, the recovery progress of the induced effects was studied. The results show that FUS exposure causes cerebral vessel dilation and can be identified and quantitatively analyzed via OCT/OCTA. Micro-hemorrhages can be detected when an excessive FUS exposure power is applied, causing the degradation of OCTA signal owing to strong scattering by leaked red blood cells (RBCs) and weaker backscattered intensity from RBCs in vessels. The vessel dilation effect due to FUS exposure was found to abate in several hours. This study demonstrates that the FUS-induced cerebral transiently dilated effects can be in-vivo differentiated and monitored with OCTA, and shows the feasibility of using OCT/OCTA as a novel tool for long-time monitoring of cerebral vascular dynamics during FUS-BBB opening process.


Quantitative phosphoproteomics reveals SLP-76 dependent regulation of PAG and Src family kinases in T cells.
The SH2-domain-containing leukocyte protein of 76 kDa (SLP-76) plays a critical scaffolding role in T cell receptor (TCR) signaling. As an adaptor protein that contains multiple protein-binding domains, SLP-76 interacts with many signaling molecules and links proximal receptor stimulation to downstream effectors. The function of SLP-76 in TCR signaling has been widely studied using the Jurkat human leukaemic T cell line through protein disruption or site-directed mutagenesis. However, a wide-scale characterization of SLP-76-dependant phosphorylation events is still lacking. Quantitative profiling of over a hundred tyrosine phosphorylation sites revealed new modes of regulation of phosphorylation of PAG, PI3K, and WASP while reconfirming previously established regulation of Itk, PLCγ, and Erk phosphorylation by SLP-76. The absence of SLP-76 also perturbed the phosphorylation of Src family kinases (SFKs) Lck and Fyn, and subsequently a large number of SFK-regulated signaling molecules. Altogether our data suggests unique modes of regulation of positive and negative feedback pathways in T cells by SLP-76, reconfirming its central role in the pathway.


LDFF, the large molecular weight DNA fragmentation factor, is responsible for the large molecular weight DNA degradation during apoptosis in Xenopus egg extracts.
DNA degradation is a biochemical hallmark in apoptosis. It has been demonstrated in many cell types that there are two stages of DNA fragmentation during the apoptotic execution. In the early stage, chromatin DNA is cut into large molecular weight DNA fragments, although the responsible nuclease(s) has not been recognized. In the late stage, the chromatin DNA is cleaved further into short oligonucleosomal fragments by a well-characterized nuclease in apoptosis, the caspase-activated DNase (CAD/DFF40). In this study, we demonstrate that large molecular weight DNA fragmentation also occurs in Xenopus egg extracts in apoptosis. We show that the large molecular weight DNA fragmentation factor (LDFF) is not the Xenopus CAD homolog XCAD. LDFF is activated by caspase-3. The large molecular weight DNA fragmentation activity of LDFF is Mg2+-dependent and Ca2+-independent, can occur in both acidic and neutral pH conditions and can tolerate 45 degrees C treatment. These results indicate that LDFF in Xenopus egg extracts might be a new DNase (or DNases) responsible for the large DNA fragmentation.


Introducing a reward system in assessment in histology: a comment on the learning strategies it might engender.
Assessment, as an inextricable component of the curriculum, is an important factor influencing student approaches to learning. If assessment is to drive learning, then it must assess the desired outcomes. In an effort to alleviate some of the anxiety associated with a traditional discipline-based second year of medical studies, a bonus system was introduced into the Histology assessment. Students obtaining a year mark of 70% were rewarded with full marks for some tests, resulting in many requiring only a few percentage points in the final examination to pass Histology. In order to ascertain whether this bonus system might be impacting positively on student learning, thirty-two second year medical students (non-randomly selected, representing four academic groups based on their mid-year results) were interviewed in 1997 and, in 1999, the entire second year class completed a questionnaire (n = 189). Both groups were asked their opinions of the bonus system. Both groups overwhelming voted in favour of the bonus system, despite less than 45% of students failing to achieve it. Students commented that it relieved some of the stress of the year-end examinations, and was generally motivating with regard to their work commitment. Being satisfied with how and what we assess in Histology, we are of the opinion that this reward system may contribute to engendering appropriate learning approaches (i.e. for understanding) in students. As a result of its apparent positive influence on learning and attitudes towards learning, this bonus system will continue to operate until the traditional programme is phased out. It is hoped that other educators, believing that their assessment is a reflection of the intended outcomes, might recognise merit in rewarding students for consistent achievement.


A smart kitchen for ambient assisted living.
The kitchen environment is one of the scenarios in the home where users can benefit from Ambient Assisted Living (AAL) applications. Moreover, it is the place where old people suffer from most domestic injuries. This paper presents a novel design, implementation and assessment of a Smart Kitchen which provides Ambient Assisted Living services; a smart environment that increases elderly and disabled people's autonomy in their kitchen-related activities through context and user awareness, appropriate user interaction and artificial intelligence. It is based on a modular architecture which integrates a wide variety of home technology (household appliances, sensors, user interfaces, etc.) and associated communication standards and media (power line, radio frequency, infrared and cabled). Its software architecture is based on the Open Services Gateway initiative (OSGi), which allows building a complex system composed of small modules, each one providing the specific functionalities required, and can be easily scaled to meet our needs. The system has been evaluated by a large number of real users (63) and carers (31) in two living labs in Spain and UK. Results show a large potential of system functionalities combined with good usability and physical, sensory and cognitive accessibility.


Perturbation of host ubiquitin systems by plant pathogen/pest effector proteins.
Microbial pathogens and pests of animals and plants secrete effector proteins into host cells, altering cellular physiology to the benefit of the invading parasite. Research in the past decade has delivered significant new insights into the molecular mechanisms of how these effector proteins function, with a particular focus on modulation of host immunity-related pathways. One host system that has emerged as a common target of effectors is the ubiquitination system in which substrate proteins are post-translationally modified by covalent conjugation with the small protein ubiquitin. This modification, typically via isopeptide bond formation through a lysine side chain of ubiquitin, can result in target degradation, relocalization, altered activity or affect protein-protein interactions. In this review, I focus primarily on how effector proteins from bacterial and filamentous pathogens of plants and pests perturb host ubiquitination pathways that ultimately include the 26S proteasome. The activities of these effectors, in how they affect ubiquitin pathways in plants, reveal how pathogens have evolved to identify and exploit weaknesses in this system that deliver increased pathogen fitness.


Tumor size-related DNA copy number changes occur in solitary fibrous tumors but not in hemangiopericytomas.
Solitary fibrous tumor (SFT) presenting in the pleura and other soft tissue sites and hemangiopericytoma (HPC) presenting at various soft tissue sites are mesenchymal tumors that share many histologic and immunohistochemical features. This raises the questions of whether these tumors are related and whether they belong within the spectrum of a single biologic entity. The behavior of both SFTs and HPCs is difficult to predict histologically. The genetic background of both SFTs and HPCs is poorly known, but it could be helpful in the evaluation of malignancy and could give clues to their possible relationship. In this study, we analyzed 15 SFTs and 11 HPCs by comparative genomic hybridization (CGH), a powerful molecular cytogenetic tool that can be applied to DNA extracted from formaldehyde-fixed and paraffin-embedded tissue. All of these tumors were immunohistochemically similar and showed reactivity for CD34-antigen but not for keratins, desmin, or muscle actins. Only 1 SFT smaller than 10 cm showed DNA copy number changes (a single loss in chromosome 13), but 7 of 8 SFTs larger than 10 cm (including all 4 tumors with more than 4 mitoses per 10 high power fields) showed changes, mostly chromosomal gains in 5q 7, 8, 12, and 18. Four cases showed losses, two of them in chromosome 13 and two others in 20q. These findings suggest that CGH might be useful in the evaluation of malignant transformation in SFT. The most common change, gain of the entire chromosome 8, seen in two cases as the only change, suggests trisomy 8 and parallels a similar finding previously described in other fibrous tumors, such as subsets of desmoid fibromatosis and infantile fibrosarcoma. In contrast, HPCs, including large and mitotically active tumors, showed no DNA copy number changes on CGH. This suggests that HPC is genetically different from SFT.


Transcriptional and posttranscriptional regulation of Bacillus sp. CDB3 arsenic-resistance operon ars1.
Bacillus sp. CDB3 possesses a novel eight-gene ars cluster (ars1, arsRYCDATorf7orf8) with some unusual features in regard to expression regulation. This study demonstrated that the cluster is a single operon but can also produce a short three-gene arsRYC transcript. A hairpin structure formed by internal inverted repeats between arsC and arsD was shown to diminish the expression of the full operon, thereby probably acting as a transcription attenuator. A degradation product of the arsRYC transcript was also identified. Electrophoretic mobility shift analysis demonstrated that ArsR interacts with the ars1 promoter forming a protein-DNA complex that could be impaired by arsenite. However, no interaction was detected between ArsD and the ars1 promoter, suggesting that the CDB3 ArsD protein may not play a regulatory role. Compared to other ars gene clusters, regulation of the Bacillus sp. CDB3 ars1 operon is more complex. It represents another example of specific mRNA degradation in the transporter gene region and possibly the first case of attenuator-mediated regulation of ars operons.


Tracking reduction of water lead levels in two homes during the Flint Federal Emergency.
A Federal Emergency was declared in Flint, MI, on January 16, 2016, 18-months after a switch to Flint River source water without phosphate corrosion control. Remedial actions to resolve the corresponding lead in water crisis included reconnection to the original Lake Huron source water with orthophosphate, implementing enhanced corrosion control by dosing extra orthophosphate, a "Flush for Flint" program to help clean out loose leaded sediment from service lines and premise plumbing, and eventually lead service line replacement. Independent sampling over a period of 37 months (January 2016-February 2019) was conducted by the United States Environmental Protection Agency and Virginia Tech to evaluate possible human exposure via normal flow (∼2-3 L/min) sampling at the cold kitchen tap, and to examine the status of loose deposits from the service line and the premise plumbing via high-velocity flushing (∼12-13 L/min) from the hose bib. The sampling results indicated that high lead in water persisted for more than a year in two Flint homes due to a large reservoir of lead deposits. The effects of a large reservoir of loose lead deposits persisted until the lead service line was completely removed in these two anomalous homes. As water conservation efforts are implemented in many areas of the country, problems with mobile lead reservoirs in service lines are likely to pose a human health risk.


Tissue microarrays analysis in chondrosarcomas: light microscopy, immunohistochemistry and xenograft study.
Chondrosarcoma (Chs) is the third most frequent primary malignant tumour of bone and can be primary or secondary, the latter results mainly from the malignant transformation of a benign pre-existing tumour. All the cases diagnosed as Chs (primary tumours, recurrences and/or metastasis and xenotransplanted Chs) from the files of our Department were collected. Only cases with paraffin blocks available were selected (Total 32 cases). Six Tissue Microarrays (TMAs) were performed and all the cases and biopsies were distributed into the following groups: a) only paraffin block available from primary and/or metastatic tumours (3 TMAs), b) paraffin block available from primary and/or metastatic tumours as well as from the corresponding Nude mice xenotransplant (2 TMAs), c) only paraffin block available from xenotransplanted Chs (1 TMA). A reclassification of all the cases was performed; in addition, conventional hematoxylin-eosin as well as immunohistochemistry staining (S100, SOX-9, Ki-67, BCL-2, p53, p16, CK, CD99, Survivin and Caveolin) was analyzed in all the TMA. The distribution of the cases according to the histopathological pattern and the location of tumours were as follows: fourteen Grade I Chs (all primaries), two primary Grade II Chs, ten Grade III Chs (all primaries), five dedifferentiated Chs (four primaries and one primary with metastasis), and two Chs from cell cultures (Ch grade III). One recurrent extraskeletal myxoid Chs was included as a control in the TMA. Although there was heterogeneity in immunohistochemistry results of the different material analyzed, S100, SOX-9, Caveolin and Survivin were more expressed. The number of passages in xenotransplants fluctuated between 1 and 13. Curiously, in Grade I Chs, these implanted tumours hardly grew, and the number of passages did not exceed one. The study of Chs by means of TMA techniques is very important because it will improve the assessment of different antibodies applied in the immunohistochemical assays. Xenotransplanted tumours in TMA improve knowledge concerning the variability in the morphological pattern shown by these tumours during the evolution in nudes.


Induction of anterior chamber associated immune deviation in rats receiving intracameral injections of retinal S antigen.
The injection of a variety of antigens, including transplantation antigens, haptens, and viral glycoproteins, into the anterior chamber of eyes of mice produces a characteristic spectrum of immune responses in which suppressed cell mediated immunity dominates. This phenomenon is called Anterior Chamber Associated Immune Deviation (ACAID). Having recently demonstrated that soluble antigens also can induce ACAID in mice, we examined the possibility that an important soluble ocular antigen, retinal S antigen (S Ag) may be capable of inducing ACAID in rats. We have found that injection of S Ag with adjuvant into the anterior chamber of eyes of adult Lewis rats evokes neither a humoral nor a cell mediated immune response that can be detected. However, animals that received S Ag intracamerally in this manner did respond to a subsequent immunogenic dose of S Ag in CFA injected into the hind foot pads, by producing serum anti S Ag antibodies. Importantly, these rats failed to display antigen-specific delayed hypersensitivity. Moreover, lymphoid cells from anterior chamber pre-treated animals, when adoptively transferred, inhibited the development of S Ag-specific delayed hypersensitivity in naive recipients indicating that an active suppression mechanism had been generated. Thus, a soluble antigen injected intracamerally in adult Lewis rats can generate ACAID. The possibility that ACAID may be relevant to the development of S Ag induced uveitis is discussed.


Quantifying recent ecological changes in remote lakes of North America and Greenland using sediment diatom assemblages.
Although arctic lakes have responded sensitively to 20(th)-century climate change, it remains uncertain how these ecological transformations compare with alpine and montane-boreal counterparts over the same interval. Furthermore, it is unclear to what degree other forcings, including atmospheric deposition of anthropogenic reactive nitrogen (Nr), have participated in recent regime shifts. Diatom-based paleolimnological syntheses offer an effective tool for retrospective assessments of past and ongoing changes in remote lake ecosystems. We synthesized 52 dated sediment diatom records from lakes in western North America and west Greenland, spanning broad latitudinal and altitudinal gradients, and representing alpine (n = 15), arctic (n = 20), and forested boreal-montane (n = 17) ecosystems. Diatom compositional turnover (beta-diversity) during the 20(th) century was estimated using Detrended Canonical Correspondence Analysis (DCCA) for each site and compared, for cores with sufficiently robust chronologies, to both the 19(th) century and the prior approximately 250 years (Little Ice Age). For both arctic and alpine lakes, beta-diversity during the 20(th) century is significantly greater than the previous 350 years, and increases with both latitude and altitude. Because no correlation is apparent between 20(th)-century diatom beta-diversity and any single physical or limnological parameter (including lake and catchment area, maximum depth, pH, conductivity, [NO(3)(-)], modeled Nr deposition, ambient summer and winter air temperatures, and modeled temperature trends 1948-2008), we used Principal Components Analysis (PCA) to summarize the amplitude of recent changes in relationship to lake pH, lake:catchment area ratio, modeled Nr deposition, and recent temperature trends. The ecological responses of remote lakes to post-industrial environmental changes are complex. However, two regions reveal concentrations of sites with elevated 20(th)-century diatom beta-diversity: the Arctic where temperatures are increasing most rapidly, and mid-latitude alpine lakes impacted by high Nr deposition rates. We predict that remote lakes will continue to shift towards new ecological states in the Anthropocene, particularly in regions where these two forcings begin to intersect geographically.


Erythrocyte microRNAs show biomarker potential and implicate multiple sclerosis susceptibility genes.
Multiple sclerosis is a demyelinating autoimmune disease, for which there is no blood-borne biomarker. Erythrocytes may provide a source of such biomarkers as they contain microRNAs. MicroRNAs regulate protein translation through complementary binding to messenger RNA. As erythrocytes are transcriptionally inactive, their microRNA profiles may be less susceptible to variation. The aim of this study was to assess the biomarker potential of erythrocyte microRNAs for multiple sclerosis and assess the potential contribution of erythrocyte-derived extracellular vesicle microRNAs to pathology. Erythrocytes were isolated from whole blood by density gradient centrifugation. Erythrocyte microRNAs of a discovery cohort (23 multiple sclerosis patients and 22 healthy controls) were sequenced. Increased expression of miR-183 cluster microRNAs (hsa-miR-96-5p, hsa-miR-182-5p and hsa-miR-183-5p) was validated in an independent cohort of 42 patients and 45 healthy and pathological (migraine) controls. Erythrocyte-derived extracellular vesicles were created ex vivo and their microRNAs were sequenced. Targets of microRNAs were predicted using miRDIP. Hsa-miR-182-5p and hsa-miR-183-5p were able to discriminate relapsing multiple sclerosis patients from migraine patients and/or healthy controls with 89-94% accuracy and around 90% specificity. Hsa-miR-182-5p and hsa-miR-183-5p expression correlated with measures of physical disability and hsa-miR-96-5p expression correlated with measures of cognitive disability in multiple sclerosis. Erythrocytes were found to selectively package microRNAs into extracellular vesicles and 34 microRNAs were found to be differentially packaged between healthy controls and multiple sclerosis patients. Several gene targets of differentially expressed and packaged erythrocyte microRNAs overlapped with multiple sclerosis susceptibility genes. Gene enrichment analysis indicated involvement in nervous system development and histone H3-K27 demethylation. Erythrocyte miR-183 cluster members may be developed into specific multiple sclerosis biomarkers that could assist with diagnosis and disability monitoring. Erythrocyte and their extracellular microRNAs were shown to target multiple sclerosis susceptibility genes and may be contributing to the pathophysiology via previously identified routes.


No evidence for directional evolution of body mass in herbivorous theropod dinosaurs.
The correlation between large body size and digestive efficiency has been hypothesized to have driven trends of increasing mass in herbivorous clades by means of directional selection. Yet, to date, few studies have investigated this relationship from a phylogenetic perspective, and none, to our knowledge, with regard to trophic shifts. Here, we reconstruct body mass in the three major subclades of non-avian theropod dinosaurs whose ecomorphology is correlated with extrinsic evidence of at least facultative herbivory in the fossil record--all of which also achieve relative gigantism (more than 3000 kg). Ordinary least-squares regressions on natural log-transformed mean mass recover significant correlations between increasing mass and geological time. However, tests for directional evolution in body mass find no support for a phylogenetic trend, instead favouring passive models of trait evolution. Cross-correlation of sympatric taxa from five localities in Asia reveals that environmental influences such as differential habitat sampling and/or taphonomic filtering affect the preserved record of dinosaurian body mass in the Cretaceous. Our results are congruent with studies documenting that behavioural and/or ecological factors may mitigate the benefit of increasing mass in extant taxa, and suggest that the hypothesis can be extrapolated to herbivorous lineages across geological time scales.


GABAergic neurons regulate lateral ventricular development via transcription factor Pax5.
Postmortem studies have revealed a downregulation of the transcription factor Pax5 in GABAergic neurons in bipolar disorder, a neurodevelopmental disorder, raising the question whether Pax5 in GABAergic neurons has a role in normal brain development. In a genetic approach to study functions of Pax5 in GABAergic neurons, Pax5 was specifically deleted in GABAergic neurons from Pax5 floxed mice using a novel Gad1-Cre transgenic mouse line expressing Cre recombinase in Gad1-positive, that is, GABAergic neurons. Surprisingly, these mice developed a marked enlargement of the lateral ventricles at approximately 7 weeks of age, which was lethal within 1-2 weeks of its appearance. This hydrocephalus phenotype was observed in mice homozygous or heterozygous for the Pax5 conditional knockout, with a gene dosage-dependent penetrance. By QTL (quantitative trait loci) mapping, a 3.5 Mb segment on mouse chromosome 4 flanked by markers D4Mit237 and D4Mit214 containing approximately 92 genes including Pax5 has previously been linked to differences in lateral ventricular size. Our findings are consistent with Pax5 being a relevant gene underlying this QTL phenotype and demonstrate that Pax5 in GABAergic neurons is essential for normal ventricular development.


A selective overview of feature screening methods with applications to neuroimaging data.
In neuroimaging studies, regression models are frequently used to identify the association of the imaging features and clinical outcome, where the number of imaging features (e.g., hundreds of thousands of voxel-level predictors) much outweighs the number of subjects in the studies. Classical best subset selection or penalized variable selection methods that perform well for low- or moderate-dimensional data do not scale to ultrahigh-dimensional neuroimaging data. To reduce the dimensionality, variable screening has emerged as a powerful tool for feature selection in neuroimaging studies. We present a selective review of the recent developments in ultrahigh-dimensional variable screening, with a focus on their practical performance on the analysis of neuroimaging data with complex spatial correlation structures and high-dimensionality. We conduct extensive simulation studies to compare the performance on selection accuracy and computational costs between the different methods. We present analyses of resting-state functional magnetic resonance imaging data in the Autism Brain Imaging Data Exchange study. This article is categorized under: Applications of Computational Statistics > Computational and Molecular BiologyStatistical Learning and Exploratory Methods of the Data Sciences > Image Data MiningStatistical and Graphical Methods of Data Analysis > Analysis of High Dimensional Data.


Similar qualitative and quantitative changes of mitochondrial respiration following strength and endurance training in normoxia and hypoxia in sedentary humans.
Endurance and strength training are established as distinct exercise modalities, increasing either mitochondrial density or myofibrillar units. Recent research, however, suggests that mitochondrial biogenesis is stimulated by both training modalities. To test the training "specificity" hypothesis, mitochondrial respiration was studied in permeabilized muscle fibers from 25 sedentary adults after endurance (ET) or strength training (ST) in normoxia or hypoxia [fraction of inspired oxygen (Fi(O(2))) = 21% or 13.5%]. Biopsies were taken from the musculus vastus lateralis, and cycle-ergometric incremental maximum oxygen uptake (VO(2max)) exercise tests were performed under normoxia, before and after the 10-wk training program. The main finding was a significant increase (P < 0.05) of fatty acid oxidation capacity per muscle mass, after endurance and strength training under normoxia [2.6- and 2.4-fold for endurance training normoxia group (ET(N)) and strength training normoxia group (ST(N)); n = 8 and 3] and hypoxia [2.0-fold for the endurance training hypoxia group (ET(H)) and strength training hypoxia group (ST(H)); n = 7 and 7], and higher coupling control of oxidative phosphorylation. The enhanced lipid oxidative phosphorylation (OXPHOS) capacity was mainly (87%) due to qualitative mitochondrial changes increasing the relative capacity for fatty acid oxidation (P < 0.01). Mitochondrial tissue-density contributed to a smaller extent (13%), reflected by the gain in muscle mass-specific respiratory capacity with a physiological substrate cocktail (glutamate, malate, succinate, and octanoylcarnitine). No significant increase was observed in mitochondrial DNA (mtDNA) content. Physiological OXPHOS capacity increased significantly in ET(N) (P < 0.01), with the same trend in ET(H) and ST(H) (P < 0.1). The limitation of flux by the phosphorylation system was diminished after training. Importantly, key mitochondrial adaptations were similar after endurance and strength training, regardless of normoxic or hypoxic exercise. The transition from a sedentary to an active lifestyle induced muscular changes of mitochondrial quality representative of mitochondrial health.


Molecular recognition in purinergic receptors. 2. Diastereoselectivity of the h-P2Y1-receptor.
In the companion paper, part 1, we described the construction of an improved molecular model for the h-P2Y1 receptor (h-P2Y1-R) and proposed a rational for the stereoelectronic selectivity of the receptor. Here, we extend our studies on the molecular recognition of the h-P2Y1-R to the exploration of the diastereoselectivity of this receptor. For this purpose, we implemented an integrative approach combining synthesis, spectral analysis, biochemical assays, and computational analysis. Specifically, we selected and synthesized novel ATP analogues bearing a chiral center on the phosphate chain. We analyzed the conformation of the chiral ATP analogues in solution by 1H/13C NMR and assigned the absolute configuration of the diastereoisomers. The coordination mode of these analogues with a Mg2+ ion was evaluated by 31P NMR. These chiral analogues were biochemically evaluated and found to be potent h-P2Y1-R ligands. An EC50 difference of ca. 20-fold was observed between the diastereoisomers. Their spectral absolute configuration assignment was confirmed by comparison of the biochemical results to those of ATP-alpha-S diastereoisomers whose chirality is known. Finally, a computational analysis was performed for the elucidation of molecular recognition employing molecular mechanics (docking) studies on the receptor:ligands complexes. On the basis of the current results, we hypothesize that h-P2Y1-R's chiral discrimination originates from the requirement that the nucleotide analogue interacts with a Mg2+ ion within the receptor binding site. This Mg2+ ion is possibly coordinated with both Asp204 and the ATP's alpha, beta, gamma-phosphates in a Lambda configuration.


Extranuclear estrogen receptors mediate the neuroprotective effects of estrogen in the rat hippocampus.
17beta-estradiol (E2) has been implicated to exert neuroprotective effects in the brain following cerebral ischemia. Classically, E2 is thought to exert its effects via genomic signaling mediated by interaction with nuclear estrogen receptors. However, the role and contribution of extranuclear estrogen receptors (ER) is unclear and was the subject of the current study. To accomplish this goal, we employed two E2 conjugates (E2 dendrimer, EDC, and E2-BSA) that can interact with extranuclear ER and exert rapid nongenomic signaling, but lack the ability to interact with nuclear ER due to their inability to enter the nucleus. EDC or E2-BSA (10 microM) was injected icv 60 min prior to global cerebral ischemia (GCI). FITC-tagged EDC or E2-BSA revealed high uptake in the hippocampal CA1 region after icv injection, with a membrane (extranuclear) localization pattern in cells. Both EDC and E2-BSA exerted robust neuroprotection in the CA1 against GCI, and the effect was blocked by the ER antagonist, ICI182,780. EDC and E2-BSA both rapidly enhanced activation of the prosurvival kinases, ERK and Akt, while attenuating activation of the proapoptotic kinase, JNK following GCI, effects that were blocked by ICI182,780. Administration of an MEK or PI3K inhibitor blocked the neuroprotective effects of EDC and E2-BSA. Further studies showed that EDC increased p-CREB and BDNF in the CA1 region in an ERK- and Akt-dependent manner, and that cognitive outcome after GCI was preserved by EDC in an ER-dependent manner. In conclusion, the current study demonstrates that activation of extranuclear ER results in induction of ERK-Akt-CREB-BDNF signaling in the hippocampal CA1 region, which significantly reduces ischemic neuronal injury and preserves cognitive function following GCI. The study adds to a growing literature that suggests that extranuclear ER can have important actions in the brain.


Improving parameter estimation for cell surface FRAP data.
Fluorescence Recovery After Photobleaching (FRAP) using the confocal laser scanning microscope has become a standard method used to determine the diffusion coefficient and mobile fraction of cell surface proteins. A common experimental approach is to bleach a stripe on the cell surface and fit the ensuing FRAP curve to a 1D diffusion model. This model is derived from the time course of recovery to an infinitely long stripe bleached on an infinite flat plane. This choice of model dictates the use of a long bleach stripe. We demonstrate that, in the case of a long bleach stripe, the finite extent of the cell leads to significant errors in parameter estimation. We further show that these errors are reduced when a relatively small stripe is bleached. Unfortunately, diffusion to such a region is fundamentally two dimensional and therefore applying the 1D model of diffusion leads to significant errors. We derive an equation suitable for fitting to FRAP data acquired from small bleach regions and analyze its accuracy using simulated data. We propose that the use of a small bleach region along with a two dimensional diffusion model is the ideal protocol for cell surface FRAP.


FGF ligands of the postnatal mammary stroma regulate distinct aspects of epithelial morphogenesis.
FGF signaling is essential for mammary gland development, yet the mechanisms by which different members of the FGF family control stem cell function and epithelial morphogenesis in this tissue are not well understood. Here, we have examined the requirement of Fgfr2 in mouse mammary gland morphogenesis using a postnatal organ regeneration model. We found that tissue regeneration from basal stem cells is a multistep event, including luminal differentiation and subsequent epithelial branching morphogenesis. Basal cells lacking Fgfr2 did not generate an epithelial network owing to a failure in luminal differentiation. Moreover, Fgfr2 null epithelium was unable to undergo ductal branch initiation and elongation due to a deficiency in directional migration. We identified FGF10 and FGF2 as stromal ligands that control distinct aspects of mammary ductal branching. FGF10 regulates branch initiation, which depends on directional epithelial migration. By contrast, FGF2 controls ductal elongation, requiring cell proliferation and epithelial expansion. Together, our data highlight a pleiotropic role of Fgfr2 in stem cell differentiation and branch initiation, and reveal that different FGF ligands regulate distinct aspects of epithelial behavior.


Polishing of pretreated dye wastewater using novel sequencing batch reactors.
Novel, aerobic sequencing batch reactor technology was tested as a polishing step for anaerobically treated textile wastewater containing dye. Operation cycle times of 6, 8 and 12 hours were studied using discrete phase periods. The SBRs were able to further remove influent dye concentration of as little as 5 mg/L, and remove highly variable loadings of COD and SS to effluent levels of 100 and 20 mg/L respectively.


RNA interference mutant induction in vivo demonstrates the essential nature of trypanosome flagellar function during mammalian infection.
We demonstrate that trypanosomes compromised in flagellar function are rapidly cleared from infected mice. Analysis of the PFR2 bloodstream RNA interference mutant revealed that defective cell motility occurred prior to cytokinesis failure. This validation provides a paradigm for the flagellum as a target for future assays and interventions against this human pathogen.


PAX6 suppression of glioma angiogenesis and the expression of vascular endothelial growth factor A.
We reported that PAX6 suppresses glioblastoma cell growth in vivo and anchorage-independent growth without significant alteration of cell proliferation in vitro, suggesting that PAX6 may alter the tumor microenvironment. Because we found that PAX6 downregulates expression of the gene encoding vascular endothelial growth factor A (VEGFA) in glioma cells, we used a subcutaneous xenograft model to verify PAX6 suppression of VEGFA-induced angiogenesis based on CD31-immunostaining of endothelial cells. The results showed a significant reduction of VEGFA at the transcription level in PAX6-transfected cells in xenografts and PAX6 has a suppressive effect on the microvascular amplification typically seen in glioblastoma. We showed that PAX6 suppression of VEGFA expression requires its DNA binding-domain. The C-terminal truncation mutant of PAX6, however, did not show the dominant negative function in regulating VEGFA expression that it showed previously in regulating MMP2 expression. In the glioma cell line U251HF, we further determined that blocking the PI3K/Akt signaling pathway with either adenoviral-mediated PTEN expression or LY294002 enhanced PAX6-mediated suppression of VEGFA in an additive manner; thus, PAX6-mediated suppression of VEGFA is not via the canonical pathway through HIF1A. These two VEGFA-regulatory pathways can also be similarly modulated in another malignant glioma cell line, U87, but not in LN229 where the basal VEGFA level is low and PTEN is wild-type. PAX6 suppression of VEGFA appears to be blocked in LN229. In conclusion, our data showed that PAX6 can initiate in glioma cells a new signaling pathway independent of PI3K/Akt-HIF1A signaling to suppress VEGFA expression.


Uncontrolled hypertension based on morning and evening home blood pressure measurements from the J-HOME study.
We evaluated the control condition of morning and evening home blood pressure (BP) and compared patients who had isolated uncontrolled morning hypertension and those who had sustained uncontrolled (morning and evening) hypertension using data from the Japan Home versus Office Blood Pressure Measurement Evaluation study. We evaluated 3303 treated hypertensive patients (mean age, 66.2+/-10.5 years; men, 44.7%) in Japan. We classified patients into controlled hypertension, isolated uncontrolled evening hypertension, isolated uncontrolled morning hypertension and sustained uncontrolled hypertension, based on the cutoff value of 135/85 mm Hg for both morning and evening home BP. Of the 3303 patients evaluated, 24.6% had isolated uncontrolled morning hypertension, and 42.0% had sustained uncontrolled hypertension. Factors associated with isolated uncontrolled morning hypertension included taking evening BP measurement after drinking alcohol or bathing. Factors associated with sustained uncontrolled hypertension were male gender, diabetes mellitus and renal disease. The regimen of antihypertensive medication was more complex in patients with uncontrolled morning hypertension than in controlled hypertension. Determinants of the difference between patients with isolated uncontrolled morning hypertension and those with sustained uncontrolled hypertension were diabetes mellitus, renal disease and lower prevalence in measuring evening BP after drinking alcohol or bathing. More than a half of the treated patients were classified into uncontrolled morning hypertension, which were associated with poorer prognosis. Classification of morning hypertensive patients into groups with or without evening hypertension may be useful for evaluating patients' total cardiovascular disease risk. Physicians should also consider evening BP measuring condition for adequate evaluation of evening BP values.


Identification of a gene cluster for telomestatin biosynthesis and heterologous expression using a specific promoter in a clean host.
Telomestatin, a strong telomerase inhibitor with G-quadruplex stabilizing activity, is a potential therapeutic agent for treating cancers. Difficulties in isolating telomestatin from microbial cultures and in chemical synthesis are bottlenecks impeding the wider use. Therefore, improvement in telomestatin production and structural diversification are required for further utilization and application. Here, we discovered the gene cluster responsible for telomestatin biosynthesis, and achieved production of telomestatin by heterologous expression of this cluster in the engineered Streptomyces avermitilis SUKA strain. Utilization of an optimal promoter was essential for successful production. Gene disruption studies revealed that the tlsB, tlsC, and tlsO-T genes play key roles in telomestatin biosynthesis. Moreover, exchanging TlsC core peptide sequences resulted in the production of novel telomestatin derivatives. This study sheds light on the expansion of chemical diversity of natural peptide products for drug development.


Environmental Adaptation of Robot Morphology and Control Through Real-World Evolution.
Robots operating in the real world will experience a range of different environments and tasks. It is essential for the robot to have the ability to adapt to its surroundings to work efficiently in changing conditions. Evolutionary robotics aims to solve this by optimizing both the control and body (morphology) of a robot, allowing adaptation to internal, as well as external factors. Most work in this field has been done in physics simulators, which are relatively simple and not able to replicate the richness of interactions found in the real world. Solutions that rely on the complex interplay among control, body, and environment are therefore rarely found. In this article, we rely solely on real-world evaluations and apply evolutionary search to yield combinations of morphology and control for our mechanically self-reconfiguring quadruped robot. We evolve solutions on two distinct physical surfaces and analyze the results in terms of both control and morphology. We then transition to two previously unseen surfaces to demonstrate the generality of our method. We find that the evolutionary search finds high-performing and diverse morphology-controller configurations by adapting both control and body to the different properties of the physical environments. We additionally find that morphology and control vary with statistical significance between the environments. Moreover, we observe that our method allows for morphology and control parameters to transfer to previously unseen terrains, demonstrating the generality of our approach.


Vaccination status of people living with HIV/AIDS in outpatient care in Fortaleza, Ceará, Brazil.
Antiretroviral therapy has increased the survival of patients with HIV/AIDS, thus necessitating health promotion practice with immunization. Vaccines are critical components for protecting people living with HIV/AIDS (PLWHA). The purpose of study was to analyze the vaccination status of PLWHA in outpatient care in Fortaleza, Ceará, Brazil. Cross-sectional study performed from June 2014 to June 2015. The screening was done with patients in antiretroviral therapy, 420 patients underwent screening, but only 99 met the inclusion criteria. Data were collected for interviews using forms to characterize sociodemographic, clinical and vaccination situations. Only 14 patients had complete vaccination schedules. The most used vaccines were hepatitis B, influenza vaccine and 23-valent pneumococcal. There was no difference between men and women regarding the proportion of PLWHA with full vaccination schedule or between sex, skin color, marital status, sexual orientation, religion or occupational status. There was no difference between having or not having a complete vaccination schedule and age, years of education, family income or number of hospitalizations. CD4+ T-cells count of patients with incomplete immunization was lower than patients with complete immunization. Health education strategies can be done individually or in groups to explain the importance of vaccination and to remind about doses to be administered. Most patients did not have proper adherence to vaccination schedules, especially due to lack of guidance. Results implied that education in health is important for vaccination adhesion, knowledge of adverse events and continuation of schemes.


Social functioning and health-related quality of life trajectories in people with epilepsy after epilepsy surgery.
An improvement in quality of life (QoL) over time after epilepsy surgery has been demonstrated in people with epilepsy; however, social functioning has been less investigated. We conducted this study to examine whether postsurgical improvement is parallel between QoL and social functioning in patients with epilepsy. We retrospectively reviewed patients who underwent epilepsy surgery. All participants completed a comprehensive neuropsychological assessment, the Quality of Life in Epilepsy Inventory (QOLIE-89) questionnaire, and the Social and Occupational Functioning Scale for Epilepsy (SOFSE) before surgery and at 3 months, 6 months, and 1 year after surgery. Demographic and epilepsy-related information was also collected. Generalized estimating equations with identity links were used to model the QOLIE-89 and SOFSE over time and possible associated factors. A p < 0.05 was considered statistically significant. A total of 76 patients, including 36 males and 43 females aged 18 to 62 years were collected. Both total QOLIE-89 and overall SOFSE improved over time after epilepsy surgery (adjusted p value < 0.001 and 0.002, respectively, with Bonferroni's correction). Total QOLIE-89 improved 3 months after surgery, while overall SOFSE showed no significant improvement until 6 months after surgery. The presurgical Full-Scale Intelligence Quotient (FSIQ) of the Wechsler Adult Intelligence Scale-III and years of education were significantly associated with time-dependent improvement for both total QOLIE-89 and overall SOFSE (p value < 0.001). At one year after surgery, overall SOFSE and total QOLIE-89 scores were significantly higher in the seizure-free group than in the nonseizure-free group (p value = 0.040 and 0.032, respectively). Social functioning significantly improved in people with epilepsy after surgery as QoL, but it took more time to exhibit improvement. People with better FSIQ and more years of education had better improvement in social functioning over time. The early intervention of rehabilitation programs after epilepsy surgery might be necessary to facilitate the improvement in social functioning.


Cloud-Based Evaluation of Anatomical Structure Segmentation and Landmark Detection Algorithms: VISCERAL Anatomy Benchmarks.
Variations in the shape and appearance of anatomical structures in medical images are often relevant radiological signs of disease. Automatic tools can help automate parts of this manual process. A cloud-based evaluation framework is presented in this paper including results of benchmarking current state-of-the-art medical imaging algorithms for anatomical structure segmentation and landmark detection: the VISCERAL Anatomy benchmarks. The algorithms are implemented in virtual machines in the cloud where participants can only access the training data and can be run privately by the benchmark administrators to objectively compare their performance in an unseen common test set. Overall, 120 computed tomography and magnetic resonance patient volumes were manually annotated to create a standard Gold Corpus containing a total of 1295 structures and 1760 landmarks. Ten participants contributed with automatic algorithms for the organ segmentation task, and three for the landmark localization task. Different algorithms obtained the best scores in the four available imaging modalities and for subsets of anatomical structures. The annotation framework, resulting data set, evaluation setup, results and performance analysis from the three VISCERAL Anatomy benchmarks are presented in this article. Both the VISCERAL data set and Silver Corpus generated with the fusion of the participant algorithms on a larger set of non-manually-annotated medical images are available to the research community.


Integrated control and related technology of assistive devices.
Assistive devices are now available that allow persons with severe physical disabilities to complete tasks independently. When the user has severe physical limitations, it may be advantageous to have an integrated control system where a single control interface (e.g., joystick, head switches, voice recognition system, keypad) is used to operate two or more assistive devices (e.g., power wheelchairs, augmentative communication devices, computers, environmental control units, and other devices that are controlled electronically). The advantages of integrated control are that persons with limited motor control can access several devices with one access site without assistance, and the user does not need to learn a different operating mechanism for each device. The purpose of this review is to convey the depth and breadth of the research that has been conducted on integrated control systems, as well as to provide some insights into future directions. We reviewed research works pertaining to communication and environmental control, computer access, and wheelchair guidance systems. Information gathered in this study will help people become fully aware of the status of contemporary integrated control technology in order to increase the quality of life of people who use electronic assistive devices.


Cationic lipid-assisted nanoparticles for delivery of mRNA cancer vaccine.
Message RNA-based vaccines with prominent advantages such as facile production, no requirement for nuclear entry and high safety without the need for integration into host genome have been shown to be potent activators of the cytotoxic immune system. However, wider applications of mRNA-based therapeutics have been hindered because of their intrinsically high vulnerability to expressed nucleases and difficulty while entering antigen-presenting cells (APCs) directly. Here, we investigated the potential of cationic lipid-assisted nanoparticles (CLAN), which form a clinically translatable nucleic acid delivery system working as a carrier of an mRNA vaccine. We found that CLAN encapsulating mRNA encoding antigen could effectively stimulate the maturation of dendritic cells (DCs) and promote the activation and proliferation of antigen-specific T cells both in vitro and in vivo. Intravenous immunization of mice with CLAN containing mRNA encoding ovalbumin (OVA) provoked a strong OVA-specific T-cell response and slowed tumor growth in an aggressive E·G7-OVA lymphoma model. Collectively, CLAN proved to be a promising platform for mRNA vaccine delivery.


Anticonvulsant effect of cannabinoid receptor agonists in models of seizures in developing rats.
Although drugs targeting the cannabinoid system (e.g., CB1 receptor agonists) display anticonvulsant efficacy in adult animal models of seizures/epilepsy, they remain unexplored in developing animal models. However, cannabinoid system functions emerge early in development, providing a rationale for targeting this system in neonates. We examined the therapeutic potential of drugs targeting the cannabinoid system in three seizure models in developing rats. Postnatal day (P) 10, Sprague-Dawley rat pups were challenged with the chemoconvulsant methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) or pentylenetetrazole (PTZ), after treatment with either CB1/2 mixed agonist (WIN 55,212-2), CB1 agonist (arachidonyl-2'-chloroethylamide [ACEA]), CB2 agonist (HU-308), CB1 antagonist (AM-251), CB2 antagonist (AM-630), fatty acid amide hydrolase inhibitor (URB-597), or G protein-coupled receptor 55 agonist (O-1602). P20 Sprague-Dawley pups were challenged with DMCM after treatment with WIN, ACEA, or URB. Finally, after pretreatment with WIN, P10 Sprague-Dawley rats were challenged against acute hypoxia-induced seizures. The mixed CB1/2 agonist and the CB1-specific agonist, but no other drugs, displayed anticonvulsant effects against clonic seizures in the DMCM model. By contrast, both CB1 and CB2 antagonism increased seizure severity. Similarly, we found that the CB1/2 agonist displayed antiseizure efficacy against acute hypoxia-induced seizures (automatisms, clonic and tonic-clonic seizures) and tonic-clonic seizures evoked by PTZ. Anticonvulsant effects were seen in P10 animals but not P20 animals. Early life seizures represent a significant cause of morbidity, with 30-40% of infants and children with epilepsy failing to achieve seizure remission with current pharmacotherapy. Identification of new therapies for neonatal/infantile epilepsy syndromes is thus of high priority. These data indicate that the anticonvulsant action of the CB system is specific to CB1 receptor activation during early development and provide justification for further examination of CB1 receptor agonists as novel antiepileptic drugs targeting epilepsy in infants and children.


A Probabilistic Model for Estimating the Depth and Threshold Temperature of C-fiber Nociceptors.
The subjective experience of thermal pain follows the detection and encoding of noxious stimuli by primary afferent neurons called nociceptors. However, nociceptor morphology has been hard to access and the mechanisms of signal transduction remain unresolved. In order to understand how heat transducers in nociceptors are activated in vivo, it is important to estimate the temperatures that directly activate the skin-embedded nociceptor membrane. Hence, the nociceptor's temperature threshold must be estimated, which in turn will depend on the depth at which transduction happens in the skin. Since the temperature at the receptor cannot be accessed experimentally, such an estimation can currently only be achieved through modeling. However, the current state-of-the-art model to estimate temperature at the receptor suffers from the fact that it cannot account for the natural stochastic variability of neuronal responses. We improve this model using a probabilistic approach which accounts for uncertainties and potential noise in system. Using a data set of 24 C-fibers recorded in vitro, we show that, even without detailed knowledge of the bio-thermal properties of the system, the probabilistic model that we propose here is capable of providing estimates of threshold and depth in cases where the classical method fails.


Blood serum atherogenicity associated with coronary atherosclerosis. Evidence for nonlipid factor providing atherogenicity of low-density lipoproteins and an approach to its elimination.
To reveal the presence of atherogenic potential in the blood serum obtained from patients with angiographically assessed coronary atherosclerosis we used primary cultures of subendothelial cells isolated by collagenase from unaffected human aortic intima. Earlier, we have demonstrated that such cultures are made up mostly of typical and modified smooth muscle cells. Within 24 hours of cultivation with a 40% sera of patients suffering from coronary atherosclerosis, the total intracellular cholesterol level increased twofold to fivefold. Cultivation with the sera of healthy subjects had no effect on the intracellular cholesterol level. The sera of patients were separated by ultracentrifugation into two fractions: total lipoprotein fraction containing the main classes of lipoproteins and a lipoprotein-deficient fraction. The former, but not the lipoprotein-deficient fraction, was characterized by atherogenicity (i.e., the ability to induce the accumulation of intracellular cholesterol). Lipoproteins of the patients' serum were separated into main classes: low density lipoproteins (LDL), very low density lipoproteins (VLDL), and high density lipoproteins (HDL2 and HDL3). An atherogenic component of the serum capable of stimulating the deposition of intracellular cholesterol was represented by LDL and, in one case, by VLDL, but not by other classes of lipoproteins. LDL and other lipoproteins isolated from the blood serum of healthy subjects failed to raise the cholesterol content in cultured cells; that is, they were nonatherogenic.(ABSTRACT TRUNCATED AT 250 WORDS)


A dyslexia-associated variant in DCDC2 changes gene expression.
Reading disability (RD) or dyslexia is a common neurogenetic disorder. Two genes, KIAA0319 and DCDC2, have been identified by association studies of the DYX2 locus on 6p21.3. We previously identified a 2445 bp deletion, and a compound STR within the deleted region (BV677278), in intron 2 of DCDC2. The deletion and several alleles of the STR are strongly associated with RD (P = 0.00002). In this study we investigated whether BV677278 is a regulatory region for DCDC2 by electrophoretic mobility shift and luciferase reporter assays. We show that oligonucleotide probes from the STR bind nuclear protein from human brain, and that alleles of the STR have a range of DCDC2-specific enhancer activities. Five alleles displayed strong enhancer activity and increased gene expression, while allele 1 showed no enhancer activity. These studies suggest that the association of BV677278 with RD reflects a role as a modifier of DCDC2 expression.


Patient time costs associated with cancer care.
Although costs of medical care for cancer have been investigated extensively, patient time costs associated with cancer care have rarely been estimated systematically. In this study, we estimated patient time costs associated with cancer care in patients aged 65 years and older in the United States. We identified 763,527 patients with breast, colorectal, corpus uteri, gastric, head and neck, lung, melanoma of the skin, ovary, prostate, renal, and urinary bladder cancers from linked Surveillance, Epidemiology, and End Results-Medicare files and 1,145,159 noncancer control subjects among Medicare enrollees who were matched by sex, age-group, and geographic location. Frequency of service use was calculated by category for patients and control subjects using Medicare claims data from 1995 to 2001. For each service category, time estimates were combined with service frequency and an hourly value of patient time. Net patient time costs were summed in the initial, continuing, and last-year-of-life phases of care for each tumor site. Net time cost estimates for the initial phase of care were applied to national estimates of numbers of new cancers in 2005 to obtain national time costs for the initial phase of care. Net patient time costs during the initial phase of care ranged from 271 dollars (95% confidence interval [CI] = 213 dollars to 329 dollars) and 842 dollars (95% CI = 806 dollars to 878 dollars) for melanoma of the skin and prostate cancer, respectively, to 5348 dollars (95% CI = 4978 dollars to 5718 dollars) and 5605 dollars (95% CI = 5273 dollars to 5937 dollars) for gastric and ovarian cancers, respectively. Net patient time costs for care during the last year of life ranged from 1509 dollars (95% CI = 1343 dollars to 1675 dollars) for melanoma of the skin to 7799 dollars (95% CI = 7433 dollars to 8165 dollars), 7435 dollars (95% CI = 7207 dollars to 7663 dollars), and 7388 dollars (95% CI = 7018 dollars to 7758 dollars) for gastric, lung, and ovarian cancers, respectively. In 2005, patient time costs for the initial phase of care were 2.3 billion dollars. Patient time costs for cancer care in the United States are substantial and vary by tumor site and phase of care, likely reflecting differences in stage at diagnosis and availability and intensity of treatment.


Comparison of two methodologies to measure agricultural occupational fatalities.
Agricultural occupational fatalities in Pennsylvania for the years 1985-87 were followed up. Supplemental data concerning the occupation of the deceased and circumstances of the fatal accident were obtained from a family member. The number of fatalities designated as agriculturally and occupationally related by the National Traumatic Occupational Fatality (NTOF) system was compared with the number of fatalities identified by using alternative criteria for classification of agriculturally and occupationally related fatalities. There may be nearly a 30 percent error in the NTOF method resulting in a 20 percent undercount.


Sex differences in antinociceptive tolerance to delta-9-tetrahydrocannabinol in the rat.
Sex differences in cannabinoid effects have been reported in rodents, with adult females typically being more sensitive than adult males. The present study compared the development of antinociceptive tolerance to delta-9-tetrahydrocannabinol (THC) in adult, gonadally intact female vs. male rats. Cumulative dose-effect curves were obtained for THC (1.0-18 mg/kg i.p.) on warm water tail withdrawal and paw pressure tests. Vehicle or the sex-specific ED80 dose for THC was administered twice daily for 9 days; THC dose-effect curves were then re-determined. On the pre-chronic test day, THC was significantly more potent in females than males in producing antinociception on the tail withdrawal and paw pressure tests. After 9 days of twice-daily THC treatment (5.4 mg/kg/injection in females and 7.6 mg/kg/injection in males), THC potency on both tests decreased more in females than males. On the tail withdrawal test, chronic THC produced 4.2- vs. 2.8-fold increases in ED50 values in females vs. males, respectively. On the paw pressure test, chronic THC produced 4.4- vs. 2.9-fold increases in ED50 values in females vs. males, respectively. Chronic THC treatment did not significantly disrupt estrous cycling in females. These results demonstrate that--even when sex differences in acute THC potency are controlled--females develop more antinociceptive tolerance to THC than males. Given the importance of drug tolerance in the development of drug dependence, these results suggest that females may be more vulnerable than males to developing dependence after chronic cannabinoid exposure.


Benthic Carbon Mineralization and Nutrient Turnover in a Scottish Sea Loch: An Integrative In Situ Study.
Based on in situ microprofiles, chamber incubations and eddy covariance measurements, we investigated the benthic carbon mineralization and nutrient regeneration in a ~65-m-deep sedimentation basin of Loch Etive, UK. The sediment hosted a considerable amount of infauna that was dominated by the brittle star A. filiformis. The numerous burrows were intensively irrigated enhancing the benthic in situ O2 uptake by ~50 %, and inducing highly variable redox conditions and O2 distribution in the surface sediment as also documented by complementary laboratory-based planar optode measurements. The average benthic O2 exchange as derived by chamber incubations and the eddy covariance approach were similar (14.9 ± 2.5 and 13.1 ± 9.0 mmol m-2 day-1) providing confidence in the two measuring approaches. Moreover, the non-invasive eddy approach revealed a flow-dependent benthic O2 flux that was partly ascribed to enhanced ventilation of infauna burrows during periods of elevated flow rates. The ratio in exchange rates of ΣCO2 and O2 was close to unity, confirming that the O2 uptake was a good proxy for the benthic carbon mineralization in this setting. The infauna activity resulted in highly dynamic redox conditions that presumably facilitated an efficient degradation of both terrestrial and marine-derived organic material. The complex O2 dynamics of the burrow environment also concurrently stimulated nitrification and coupled denitrification rates making the sediment an efficient sink for bioavailable nitrogen. Furthermore, bioturbation mediated a high efflux of dissolved phosphorus and silicate. The study documents a high spatial and temporal variation in benthic solute exchange with important implications for benthic turnover of organic carbon and nutrients. However, more long-term in situ investigations with like approaches are required to fully understand how environmental events and spatio-temporal variations interrelate to the overall biogeochemical functioning of coastal sediments.


[Validation of the French version of the Body Shape Questionnaire].
In today's societies, pressures from the idea of thinness are omnipresent and lead to a corporal dissatisfaction with an excessive preoccupation of the body's image. It seems important to have, in France, a device that can evaluate the corporal dissatisfaction degree, for the reason that the troubles of the body's image is a common diagnostical category to the anorexia and bulimia (DSM IV, 1994). Cooper et al. (1987) have developed one-dimensional questionnaire of 34 items in order to measure the worries towards the weight and the shape of the body, called the "Body Shape Questionnaire" (BSQ). Its concurrent validity has been shown with the using of the corporal dissatisfaction under-scale of the Eating Disorders Inventory (EDI; Garner et Olmsted, 1984) and the using of the diagnostical questionnaire: Eating Attitude Test (EAT; Garner et Garfinkel, 1979). The BSQ gives us a way to explore the role of extreme worries towards the body's apparence in the development, the keeping and the treatment of eating disorders. From this point the BSQ is a tool widely used on an international level in researches on the eating disorders. It has been validited in Spain and in Germany whereas it has not been yet subject to a validation in France. For this reason, we proposed to use the BSQ on the french population. The sample is made of 242 university girl students. The average age is 20.7 years old with an standard deviation of 2.26. The size and the weight helped us to calculate the Body Mass Index (BMI) which is in fact the weight divided by the size squared. The average BMI is 21.06 kg/m2 +/- 2.87. Regarding the evaluating devices, the BSQ is presented as a scale of 34 items marked by 6 different points: 1 never, 2 barely, 3 sometimes, 4 often, 5 very often, 6 always. After getting the authorization form the authors to respect during these kinds of procedures, we have started validation. The corporal dissatisfaction under-scale of Eating Disorder Inventory (EDI, Garner et al., 1991) is made of 9 items and has a scale of 6 points going from "never" to "always". In order to evaluate the accuracy of the Body Shape Questionnaire (BSQ) in France, we made a test/retest. The concurrent validity of the BSQ has been evaluated by the EDI. The 242 subjects have been asked to fill in both questionnaires during the test and the re-test (4 weeks after the test). We have evaluated the accuracy of the BSQ with the calculation of the constancy test/retest (Pearson's r) and the measure of the internal consistency (Cronbach's alpha). Then, we tested the validity regarding an external criteria. The validation procedure is based here on an examination of the correlations between the tests results (BSQ) and another measure taken as criteria (EDI). The solutions proposed by the factorial exploratory analysis have been tested by confirmatory analysis using the 2 index of adjustment: the GFI ("Goodness of Fit Index") and the CFI (Comparative Index of Bentler) which, greater than 0.85, shows a satisfying adjustment and the RMC (redidue) which has to be less than 0.10 for a satisfying adjustment. Finally, inferential statistical analysis have studied the relations between the weight's category and the total result at the BSQ and the results of the factors sprung from factorial analysis. Correlations between the items of test/re-test BSQ are greater than 0.93. The alpha coefficient is high for the test (0.95) and also for the re-test (0.94). This shows a very good internal consistency between the 34 items of the BSQ. Concerning the EDI, the correlation between the test/re-test are greater than 0.98. The alpha coefficient of Cronbach is high: 0.85 for the test and 0.84 for the re-test. The Pearson's r of the BSQ/EDI test and the BSQ/EDI re-test greater than 0.7 point out the concomitant validity of the BSQ with the EDI. We made a factorial analysis of the BSQ (test) on the 242 subjects. We used the extraction of the main components method with the extraction's rule of the curve for the real values. The transformation method used here is the normalised Varimax. The factorial analysis shows four factors with their own value greater or equal to 1 (15.1; 1.77; 1.48; 1.08). This solution concerning these four factors explains 55.2% of the total variance. The oblical rotations analysis of the four factors seems to point out a high correlation between these latters (from 0.54 to 0.77), this suggests a hierarchical pattern with a single factor which is confirmed with an Cronbach's alpha of 0.95. Then, we chose to gather around each selected factors the items presenting a substantial saturation, greater than or equal to 0.5 and which are saturated by only one factor. The first factor obtained by the factorial analysis of the BSQ has been called "social avoidance and shame of the exposure of the body". The internal coherence of this factor is satisfactory (Cronbach's alpha = 0.87). The second factor obtained has been called "body dissatisfaction compared to the lower parts ot the body (Cronbach's alpha = 0.90). The third factor has been called "using laxatives and vomiting in order to reduce body dissatisfaction" The fourth factor has been called "unsuited cognitions and behaviours in order to control the weight (Cronbach's alpha = 0.76). The solution with 1 and 4 factors has been tested by a confirmatory analysis. The adjustment parameters of the unifactorial pattern were not satisfactory (GFI = 0.76, CFI = 0.83, RMC = 0.5). The 4 factors pattern adjusted better to the data (GFI = 0.86, CFI = 0.90, RMC = 0.5). In order to determinate the differences between the weight/score categories to the BSQ) score in comparison to the 4 factors (coming from the factorial analysis), several Anova have been released as well as post-hoc tests (test of Sheffé). There is a significant effect of the weight category on the corporal dissatisfaction (BSQ result), F (2.230) = 11.34; p < 0.0001. Scheffe's test placed in a prominent position the fact that the subjects having an overweight (99.85 +/- 31.34) have a corporal dissatisfaction significantly greater than the subjects having a normal weight (74.08 +/- 27.94) and presenting a thinness (63.19 +/- 23.61). We pointed out a significant effect of the weight category on factor one "social avoidance and shame of the exposure of the body" [F (2.232) = 20.18; p < 0.001], on factor 2 "body dissatisfaction compared to the lower parts of the body" [F (2.232) = 6.38; p < 0.005] and factor 4 "reduce body dissatisfaction and unsuited cognitions and behaviours in order to control the weight" [F (2.232) = 5.67; p < 0.005]. However, there is no effect in the weight category for factor 3 "using laxatives and cominting in order to reduce the corporal dissatisfaction" [F (2.232) = 5.67; p > 0.05]. Sheffe's test highlighted the fact that the subjects having overweight have a result more important than the ones having a normal weight and presenting a thinness for factor 1.2 and 4. The BSQ "French version" seems in fact to present the same metrical qualities than its original Anglo-Saxon and Spanish versions. The concluding results invite us to continue our study of the BSQ including in our patients, the bulimic subjects and the ones who are worried about corporal aspect. Its implications on the clinical field could be really interesting especially on people suffering from the eating disorders. Then, the BSQ could provide us a way to explore the role of an extreme worry of the body's appearance in the development, the maintenance and the treatment of the eating disorders.


Fast Identification of Possible Drug Treatment of Coronavirus Disease -19 (COVID-19) Through Computational Drug Repurposing Study.
The recent outbreak of novel coronavirus disease -19 (COVID-19) calls for and welcomes possible treatment strategies using drugs on the market. It is very efficient to apply computer-aided drug design techniques to quickly identify promising drug repurposing candidates, especially after the detailed 3D-structures of key virous proteins are resolved. Taking the advantage of a recently released crystal structure of COVID-19 protease in complex with a covalently-bonded inhibitor, N3,1 I conducted virtual docking screening of approved drugs and drug candidates in clinical trials. For the top docking hits, I then performed molecular dynamics simulations followed by binding free energy calculations using an endpoint method called MM-PBSA-WSAS.2-4 Several promising known drugs stand out as potential inhibitors of COVID-19 protease, including Carfilzomib, Eravacycline, Valrubicin, Lopinavir and Elbasvir. Carfilzomib, an approved anti-cancer drug acting as a proteasome inhibitor, has the best MM-PBSA-WSAS binding free energy, -13.82 kcal/mol. Streptomycin, an antibiotic and a charged molecule, also demonstrates some inhibitory effect, even though the predicted binding free energy of the charged form (-3.82 kcal/mol) is not nearly as low as that of the neutral form (-7.92 kcal/mol). One bioactive, PubChem 23727975, has a binding free energy of -12.86 kcal/mol. Detailed receptor-ligand interactions were analyzed and hot spots for the receptor-ligand binding were identified. I found that one hotspot residue HIS41, is a conserved residue across many viruses including COVID-19, SARS, MERS, and HCV. The findings of this study can facilitate rational drug design targeting the COVID-19 protease.


Poor nutritional status, risk of sarcopenia and nutrition related complaints are prevalent in COVID-19 patients during and after hospital admission.
Patients with COVID-19 infection presents with a broad clinical spectrum of symptoms and complications. As a consequence nutritional requirements are not met, resulting in weight- and muscle loss, and malnutrition. The aim of the present study is to delineate nutritional complaints, the (course of the) nutritional status and risk of sarcopenia of COVID-19 patients, during hospitalisation and after discharge. In this prospective observational study in 407 hospital admitted COVID-19 patients in four university and peripheral hospitals, data were collected during dietetic consultations. Presence of nutrition related complaints (decreased appetite, loss of smell, changed taste, loss of taste, chewing and swallowing problems, nausea, vomiting, feeling of being full, stool frequency and consistency, gastric retention, need for help with food intake due to weakness and shortness of breath and nutritional status (weight loss, BMI, risk of sarcopenia with SARC-F ≥4 points) before, during hospital stay and after discharge were, where possible, collected. Included patients were most men (69%), median age of 64.8 ± 12.4 years, 60% were admitted to ICU at any time point during hospitalisation with a median LOS of 15 days and an in-hospital mortality rate of 21%. The most commonly reported complaints were: decreased appetite (58%), feeling of being full (49%) and shortness of breath (43%). One in three patients experienced changed taste, loss of taste and/or loss of smell. Prior to hospital admission, 67% of the patients was overweight (BMI >25 kg/m2), 35% of the patients was characterised as malnourished, mainly caused by considerable weight loss. Serious acute weight loss (>5 kg) was showed in 22% of the patents during the hospital stay; most of these patients (85%) were admitted to the ICU at any point in time. A high risk of sarcopenia (SARC-F ≥ 4 points) was scored in 73% of the patients during hospital admission. In conclusion, one in five hospital admitted COVID-19 patients suffered from serious acute weight loss and 73% had a high risk of sarcopenia. Moreover, almost all patients had one or more nutritional complaints. Of these complaints, decreased appetite, feeling of being full, shortness of breath and changed taste and loss of taste were the most predominant nutrition related complaints. These symptoms have serious repercussions on nutritional status. Although nutritional complaints persisted a long time after discharge, only a small group of patients received dietetic treatment after hospital discharge in recovery phase. Clinicians should consider the risks of acute malnutrition and sarcopenia in COVID-19 patients and investigate multidisciplinary treatment including dietetics during hospital stay and after discharge.


How molecular interactions affect crystal morphology: the case of haloperidol.
The tableting behaviour of drugs can be dramatically affected by changes in the crystal habit of the drug molecule. Pharmaceutical companies are therefore interested in the morphology prediction as a possible tool to optimise the industrial process. Molecular mechanics calculations embedded in dedicated software together with X-ray diffraction analysis were used to enlighten the structural properties of 4-[4-(4-chlorophenyl)-4-hydroxypiperidin-1-yl]-1-(4-fluorophenyl)butan-1-one-whose commercial name is haloperidol--an antipsychotic drug that contributed to the progress and revolution of psychiatric care. We defined, by means of X-ray powder diffraction, which--or how much--of the two crystallographic structures present in the Cambridge Crystallographic Database represents the commercial crystalline powder. Once the correct structure was selected, the whole structural analysis was carried out as a comparison with the already deposited structures. The available single crystal structure was used to model the X-ray powder diffraction pattern. The "real" structure was then optimised by means of molecular mechanics and the crystal morphology of the compounds was predicted with different computational methods. Analogies and differences among the different morphologies, together with the potential role of several solvents were used to try to bridge the gap between the molecular structure--that is, the atomic point of view--and the crystal habit.


Ageing in Werner syndrome.
Oxidative stress markers including pentosidine and homocysteine were examined comparing them with inflammation markers including highly sensitive C-reactive protein (hsCRP) and matrix metalloproteinase-9 (MMP-9) in serum from patients with Werner syndrome (WS) and healthy individuals. Elevation of serum pentosidine correlated significantly with normal aging in healthy individuals (p < 0.0004). Serum pentosidine in WS increased significantly compared with age-matched healthy individuals (p < 0.05). Serum homocysteine levels increased insignificantly with normal aging in healthy individuals and in WS compared with age-matched healthy individuals. As both pentosidine and homocysteine levels did not correlate with hsCRP or MMP-9, both oxidative stress markers may be differentially regulated by inflammation.


Loss of wild-type ATRX expression in somatic cell hybrids segregates with activation of Alternative Lengthening of Telomeres.
Alternative Lengthening of Telomeres (ALT) is a non-telomerase mechanism of telomere lengthening that occurs in about 10% of cancers overall and is particularly common in astrocytic brain tumors and specific types of sarcomas. Somatic cell hybridization analyses have previously shown that normal telomerase-negative fibroblasts and telomerase-positive immortalized cell lines contain repressors of ALT activity, indicating that activation of ALT results from loss of one or more unidentified repressors. More recently, ATRX or DAXX was shown to be mutated both in tumors with telomere lengths suggestive of ALT activity and in ALT cell lines. Here, an ALT cell line was separately fused to each of four telomerase-positive cell lines, and four or five independent hybrid lines from each fusion were examined for expression of ATRX and DAXX and for telomere lengthening mechanism. The hybrid lines expressed either telomerase or ALT, with the other mechanism being repressed. DAXX was expressed normally in all parental cell lines and in all of the hybrids. ATRX was expressed normally in each of the four telomerase-positive parental cell lines and in every telomerase-positive hybrid line, and was abnormal in the ALT parental cells and in all but one of the ALT hybrids. This correlation between ALT activity and loss of ATRX expression is consistent with ATRX being a repressor of ALT.


Human CD4 binds immunoglobulins.
T cell glycoprotein CD4 binds to class II major histocompatibility molecules and to the human immunodeficiency virus (HIV) envelope protein gp120. Recombinant CD4 (rCD4) bound to polyclonal immunoglobulin (Ig) and 39 of 50 (78%) human myeloma proteins. This binding depended on the Fab and not the Fc portion of Ig and was independent of the light chain. Soluble rCD4, HIV gp120, and sulfated dextrans inhibited the CD4-Ig interaction. With the use of a panel of synthetic peptides, the region critical for binding to Ig was localized to amino acids 21 to 38 of the first extracellular domain of CD4. CD4-bound antibody (Ab) complexed with antigen approximately 100 times better than Ab alone. This activity may contribute to the Ab-mediated enhancement of cellular HIV interaction that appears to depend on a trimolecular complex of HIV, antibodies to gp120, and CD4.