Feasibility of (68)Ga-labeled Siglec-9 peptide for the imaging of acute lung inflammation: a pilot study in a porcine model of acute respiratory distress syndrome.
There is an unmet need for noninvasive, specific and quantitative imaging of inherent inflammatory activity. Vascular adhesion protein-1 (VAP-1) translocates to the luminal surface of endothelial cells upon inflammatory challenge. We hypothesized that in a porcine model of acute respiratory distress syndrome (ARDS), positron emission tomography (PET) with sialic acid-binding immunoglobulin-like lectin 9 (Siglec-9) based imaging agent targeting VAP-1 would allow quantification of regional pulmonary inflammation. ARDS was induced by lung lavages and injurious mechanical ventilation. Hemodynamics, respiratory system compliance (Crs) and blood gases were monitored. Dynamic examination using [(15)O]water PET-CT (10 min) was followed by dynamic (90 min) and whole-body examination using VAP-1 targeting (68)Ga-labeled 1,4,7,10-tetraaza cyclododecane-1,4,7-tris-acetic acid-10-ethylene glycol-conjugated Siglec-9 motif peptide ([(68)Ga]Ga-DOTA-Siglec-9). The animals received an anti-VAP-1 antibody for post-mortem immunohistochemistry assay of VAP-1 receptors. Tissue samples were collected post-mortem for the radioactivity uptake, histology and immunohistochemistry assessment. Marked reduction of oxygenation and Crs, and higher degree of inflammation were observed in ARDS animals. [(68)Ga]Ga-DOTA-Siglec-9 PET showed significant uptake in lungs, kidneys and urinary bladder. Normalization of the net uptake rate (Ki) for the tissue perfusion resulted in 4-fold higher uptake rate of [(68)Ga]Ga-DOTA-Siglec-9 in the ARDS lungs. Immunohistochemistry showed positive VAP-1 signal in the injured lungs. Detection of pulmonary inflammation associated with a porcine model of ARDS was possible with [(68)Ga]Ga-DOTA-Siglec-9 PET when using kinetic modeling and normalization for tissue perfusion.


Metabolome dynamic responses of Saccharomyces cerevisiae to simultaneous rapid perturbations in external electron acceptor and electron donor.
Rapid perturbation experiments are highly relevant to elaborate the in vivo kinetics for mathematical models of metabolism, which are needed for selecting gene targets for metabolic engineering. Perturbations were applied to chemostat-cultivated biomass (D=0.05 h(-1), aerobic glucose/ethanol-limited) using the BioScope of Saccharomyces cerevisiae CEN. PK 113-7D over time span of 90 and 180 s. The availability of the external electron acceptor oxygen was decreased from fully aerobic to anaerobic conditions. It was observed that the changes in metabolome response under these conditions were limited to the pyruvate node. Acetaldehyde supply was used as an extra external electron acceptor during glucose perturbation under fully aerobic conditions. This had a strong effect on the metabolome dynamics and resulted in a significantly higher initial glycolytic flux. Dynamic response of the adenine nucleotides indicated that their behavior is not dictated by the glycolytic flux but is much more coupled to the cytosolic NADH/NAD(+) ratio through the equilibrium pool of fructose 1,6-bisphosphate and 2/3-phosphoglycerate. Also, the electron donor availability (glucose) was decreased. This did not result in significant changes in the concentrations of the glycolytic and tricarboxylic acid cycle metabolites, whereas the adenine nucleotides, especially ADP and AMP, showed the opposite response to that observed in a glucose pulse experiment. Surprisingly, trehalose was not mobilized in the time frame of 180 s.


Starless bias and parameter-estimation bias in the likelihood-based phylogenetic method.
I analyzed various site pattern combinations in a 4-OTU case to identify sources of starless bias and parameter-estimation bias in likelihood-based phylogenetic methods, and reported three significant contributions. First, the likelihood method is counterintuitive in that it may not generate a star tree with sequences that are equidistant from each other. This behaviour, dubbed starless bias, happens in a 4-OTU tree when there is an excess (i.e., more than expected from a star tree and a substitution model) of conflicting phylogenetic signals supporting the three resolved topologies equally. Special site pattern combinations leading to rejection of a star tree, when sequences are equidistant from each other, were identified. Second, fitting gamma distribution to model rate heterogeneity over sites is strongly confounded with tree topology, especially in conjunction with the starless bias. I present examples to show dramatic differences in the estimated shape parameter α between a star tree and a resolved tree. There may be no rate heterogeneity over sites (with the estimated α > 10000) when a star tree is imposed, but α < 1 (suggesting strong rate heterogeneity over sites) when an (incorrect) resolved tree is imposed. Thus, the dependence of "rate heterogeneity" on tree topology implies that "rate heterogeneity" is not a sequence-specific feature, cautioning against interpreting a small α to mean that some sites are under strong purifying selection and others not. Thirdly, because there is no existing (and working) likelihood method for evaluating a star tree with continuous gamma-distributed rate, I have implemented the method for JC69 in a self-contained R script for a four-OTU tree (star or resolved), in addition to another R script assuming a constant rate over sites. These R scripts should be useful for teaching and exploring likelihood methods in phylogenetics.


The genome sequence of the common toad, Bufo bufo (Linnaeus, 1758).
We present a genome assembly from an individual male Bufo bufo (the common toad; Chordata; Amphibia; Anura; Bufonidae). The genome sequence is 5.04 gigabases in span. The majority of the assembly (99.1%) is scaffolded into 11 chromosomal pseudomolecules. Gene annotation of this assembly by the NCBI Eukaryotic Genome Annotation Pipeline has identified 21,517 protein coding genes.


Pemphigus erythematosus relapse associated with atorvastatin intake.
Statins, also known as 3-hydroxy-3-methylglutaril-CoA reductase inhibitors, are well-tolerated drugs used for prevention of atherosclerosis and cardiovascular events. Although they are generally considered safe, some serious adverse effects, such as myositis, myopathy, and rhabdomyolysis can rarely occur. Furthermore, recent data from long-term follow-up on patients who have been taking statins for a long period of time suggest that prolonged exposure to statins may trigger autoimmune reactions. The exact mechanism of statin-induced autoimmune reactions is unclear. Statins, as proapoptotic agents, release nuclear antigen into the circulation and may induce the production of pathogenic autoantibodies. Herein we report the case of a 70 year-old man who developed a relapse of pemphigus erythematosus, a syndrome with features of both lupus erythematosus and pemphigus, after atorvastatin intake.


Learning from a Feasibility Trial of a Simple Intervention: Is Research a Barrier to Service Delivery, or is Service Delivery a Barrier to Research?
(1) Background: Applied health services research (AHSR) relies upon coordination across multiple organizational boundaries. Our aim was to understand how competing organizational and professional goals enhance or impede the conduct of high quality AHSR. (2) Methods: A qualitative study was conducted in two local health care systems in the UK, linked to a feasibility trial of a clinic-based intervention in secondary care. Data collection involved 24 semi-structured interviews with research managers, clinical research staff, health professionals, and patients. (3) Results: This study required a dynamic network of interactions between heterogeneous health and social care stakeholders, each characterized by differing ways of organizing activities which constitute their core functions; cultures of collaboration and interaction and understanding of what research involves and how it contributes to patient care. These interrelated factors compounded the occupational and organizational boundaries that hindered communication and coordination. (4) Conclusions: Despite the strategic development of multiple organizations to foster inter-professional collaboration, the competing goals of research and clinical practice can impede the conduct of high quality AHSR. To remedy this requires the alignment and streamlining of organizational goals, so that all agencies involved in AHSR develop a shared understanding and mutual respect for the progress of evidence-based medicine and the complex and often nuanced environments in which it is created and practiced.


Global strategic partnerships in regenerative medicine.
The approach to research and development in biomedical science is changing. Increasingly, academia and industry seek to collaborate, and share resources and expertise, by establishing partnerships. Here, we explore the co-development partnership landscape in the field of regenerative medicine, focusing on agreements involving one or more private entities. A majority of the largest biopharmaceutical companies have announced strategic partnerships with a specific regenerative medicine focus, signifying the growth and widening appeal of this emerging sector.


Long-distance texting: Text messaging is linked with higher relationship satisfaction in long-distance relationships.
Due to the widespread use of smartphones, romantic couples can connect with their partners from virtually anywhere, at any time. Remote communication may be particularly important to long-distance relationships (LDRs), compared to geographically close relationships (GCRs). The goals of the current research were to examine differences between LDRs and GCRs in (1) the patterns of remote communication (video calls, voice calls, and texting), and (2) how frequency and responsiveness of remote communication are related to relationship satisfaction. Data were drawn from an online survey of emerging adults (n = 647) who were in a relationship or dating someone (36.5% were in an LDR). Participants in LDRs engaged in more frequent video calling, voice calling and texting, compared to those in GCRs. Long-distance relationship participants also perceived their partners to be more responsive during video and voice calls, compared to GCR participants. More frequent and responsive texting predicted significantly greater relationship satisfaction among participants in LDRs, but not GCRs. Meanwhile, frequency of voice calls was associated with greater relationship satisfaction in GCRs, but not in LDRs. The use of video calls was not significantly related to relationship satisfaction in either group. Overall, study findings add to a growing literature on remote communication in romantic couples and suggest a uniquely positive role of texting within LDRs. Further research is needed to examine the ways in which LDR and GCR couples can best capitalize on different forms of remote technology to maintain their relationships during periods of separation.


Continuous hydrogen peroxide production by organic buffers in phytoplankton culture media.
We investigated the production of hydrogen peroxide (HOOH) in illuminated seawater media containing a variety of zwitterionic buffers. Production rates varied extensively among buffers, with 4-(2-hydroxyethyl)1-piperazineethanesulfonic acid (HEPES) highest and N-Tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS) among the lowest. The rate of HOOH accumulation was remarkably consistent over many days, and increased linearly with buffer concentration, natural seawater concentration, and light level. Concentrations of HEPES commonly used in culture media (1-10 mM) generated enough HOOH to kill the axenic Prochlorococcus strain VOL1 during growth in enriched seawater media at lower, environmentally realistic cell concentrations and/or under high light exposure. We also demonstrated that HEPES can be used experimentally to study the biological effects of chronic exposure to sublethal levels of HOOH such as may be experienced by light-exposed microorganisms.


Vitamin A-sensitive tissues in transgenic mice expressing high levels of human cellular retinol-binding protein type I are not altered phenotypically.
The suggested function of cellular retinol-binding protein type I [CRBP(I)] is to carry retinol to esterifying or oxidizing enzymes. The retinyl esters are used in storage or transport, whereas oxidized forms such as all-trans or 9-cis retinoic acid are metabolites used in the mechanism of action of vitamin A. Thus, high expression of human CRBP(I) [hCRBP(I)] in transgenic mice might be expected to increase the production of retinoic acid in tissues, thereby inducing a phenotype resembling vitamin A toxicity. Alternatively, a vitamin A-deficient phenotype could also be envisioned as a result of an increased accumulation of vitamin A in storage cells induced by a high hCRBP(I) level. Signs of vitamin A toxicity or deficiency were therefore examined in tissues from transgenic mice with ectopic expression of hCRBP(I). Testis and intestine, the tissues with the highest expression of the transgene, showed normal gross morphology. Similarly, no abnormalities were observed in other tissues known to be sensitive to vitamin A status such as cornea and retina, and the epithelia in the cervix, trachea and skin. Furthermore, hematologic variables known to be influenced by vitamin A status such as the hemoglobin concentration, hematocrits and the number of red blood cells were within normal ranges in the transgenic mice. In conclusion, these transgenic mice have normal function of vitamin A despite high expression of hCRBP(I) in several tissues.


Innovative empirical approaches for inferring climate-warming impacts on plants in remote areas.
The prediction of the effects of climate warming on plant communities across the globe has become a major focus of ecology, evolution and biodiversity conservation. However, many of the frequently used empirical approaches for inferring how warming affects vegetation have been criticized for decades. In addition, methods that require no electricity may be preferred because of constraints of active warming, e.g. in remote areas. Efforts to overcome the limitations of earlier methods are currently under development, but these approaches have yet to be systematically evaluated side by side. Here, an overview of the benefits and limitations of a selection of innovative empirical techniques to study temperature effects on plants is presented, with a focus on practicality in relatively remote areas without an electric power supply. I focus on methods for: ecosystem aboveground and belowground warming; a fuller exploitation of spatial temperature variation; and long-term monitoring of plant ecological and microevolutionary changes in response to warming. An evaluation of the described methodological set-ups in a synthetic framework along six axes (associated with the consistency of temperature differences, disturbance, costs, confounding factors, spatial scale and versatility) highlights their potential usefulness and power. Hence, further developments of new approaches to empirically assess warming effects on plants can critically stimulate progress in climate-change biology.


Functionalization of hydrogenated silicene with alkali and alkaline earth metals for efficient hydrogen storage.
First principles density functional theory has been employed to investigate the electronic structure along with the stability, bonding mechanism, band gap and charge transfer of metal functionalized hydrogenated silicene (SiH), or silicane, in order to envisage the hydrogen storage capacity. Various metal adatoms including Li, Na, K, Be, Mg and Ca have been doped on the most stable chair like configuration of silicane. The corresponding binding energies and charge transfer mechanism have been discussed from the perspective of H2 storage ability. The Li and Na metal adatoms have been found to be ideally suitable not only for their strong metal to substrate binding and uniform distribution over the substrate but also for their high capacity for storage of hydrogen. The stability of both Li and Na functionalized SiH has also been confirmed by MD simulations. It was found that both Li(+) and Na(+) adsorbed four H2 molecules attaining reasonably high storage capacities of 6.30 wt% and 5.40 wt% respectively with average adsorption energies lying within the range suitable for practical H2 storage applications, in contrast with alkaline earth metals.


Reversibility of prion misfolding: insights from constant-pH molecular dynamics simulations.
The prion protein (PrP) is the cause of a group of diseases known as transmissible spongiform encephalopathies (TSEs). Creutzfeldt-Jakob disease and bovine spongiform encephalopathy are examples of TSEs. Although the normal form of PrP (PrP(C)) is monomeric and soluble, it can misfold into a pathogenic form (PrP(Sc)) that has a high content of β-structure and can aggregate forming amyloid fibrils. The mechanism of conversion of PrP(C) into PrP(Sc) is not known but different triggers have been proposed. It can be catalyzed by a PrP(Sc) sample, or it can be induced by an external factor, such as low pH. The pH effect on the structure of PrP was recently studied by computational methods [Campos et al. J. Phys. Chem. B 2010, 114, 12692-12700], and an evident trend of loss of helical structure was observed with pH decrease, together with a gain of β-structures. In particular, one simulation at pH 2 showed an evident misfolding transition. The main goal of the present work was to study the effects of a change in pH to 7 in several transient conformations of this simulation, in order to draw some conclusions about the reversibility of PrP misfolding. Although the most significant effect caused by the change of pH to 7 was a global stabilization of the protein structure, we could also observe that some conformational transitions induced by pH 2 were reversible in many of our simulations, namely those started from the early moments of the misfolding transition. This observation is in good agreement with experiments showing that, even at pH as low as 1.7, it is possible to revert the misfolding process [Bjorndahl et al. Biochemistry 2011, 50, 1162-1173].


Microvascular reconstruction of composite cranial defects.
Microvascular reconstruction of composite cranium defects is challenging, and operative planning differs in various anatomic regions of the cranium. The aim of this article is to detail surgical strategies for microvascular reconstruction in different areas of the cranium. Fifty-eight free tissue transfers were performed in 54 patients with composite cranial defects between 1995 and 2009. The cranium was divided into six zones to classify the defects. Data on patient demographics, location and cause of the defects, technical details of surgical management, and complications were analyzed. Defects were secondary to ablative procedures in 45 patients. Eight patients had posttraumatic defects, and one patient had congenital defect. Nine different flap types were used in the series. Choice of recipient vessels and the use of interpositional vein grafts varied according to the site of the cranial defects. Microsurgical reconstruction of cranial defects can be performed safely with predictable outcome by methodic assessment of the problem and by careful attention to the details of the reconstructive plan. In this article, valid conclusions on the planning of microvascular reconstruction in different areas of the cranium have been determined.


Impact of surgery and chemotherapy on von Willebrand factor and vascular endothelial growth factor levels in colorectal cancer.
von Willebrand factor (vWf) is thought to mediate binding of tumour cells to platelets and to favour their systemic spreading capacity. Platelets involved in tumour angiogenesis are capable of releasing vascular endothelial growth factor (VEGF). Hence, levels of vWf and VEGF may correlate with cancer stage. The objectives are determine the impact of surgery and chemotherapy on vWf and VEGF in colorectal cancer (CRC) patients. Twenty healthy volunteers (group 1), 14 patients with locally advanced CRC (group 2) and 12 patients with metastatic CRC (group 3) were enrolled. Blood samples were taken at recruitment in group 1, and before and after surgery and chemotherapy in groups 2 and 3, respectively. Blood levels of vWf, VEGF, platelet count, C-reactive protein (CRP), ceruloplasmin and carcinoembrionary antigen (CEA) were measured. At baseline, group 3 showed higher concentrations of vWf than the other groups (p<0.05). In group 2, vWf became elevated 40% post-surgery (p=0.016), independently of changes in CRP or ceruloplasmin. In group 3, chemotherapy caused a 42% reduction in VEGF (p=0.015). There was a strong correlation between higher vWf levels and more advanced CRC stage at diagnosis. These levels were elevated post-surgery in patients with locally advanced CRC. Chemotherapy significantly decreased VEGF in metastatic CRC patients before CEA showed any significant change.


Co-creation of services to maintain independence and optimise well-being: Learnings from Australia's Older Women Living Alone (OWLA) project.
For many populations at risk of social isolation, including Older Women Living Alone (OWLA), existing services to maintain independence and optimise well-being are difficult to access, unsuitable or unavailable. Co-creation is a strategy to develop 'person-centred' services that meet the needs of individuals. We adapted an existing framework for co-creation and used participatory action research methods, supported by an evidence base comprising a systematic review, analysis of routinely collected data and interviews, to develop person-centred services for OWLA. This approach achieved co-creation through an iterative process of consultation and review, involving a series of facilitated discussions with women living alone and stakeholders. A total of 13 women living alone, aged ≥55 years, and 11 stakeholders representing service providers and advocacy groups, were recruited to participate in these discussions. Sessions with between three and five OWLA, were held across Melbourne. The information was compiled and presented to service stakeholders in a single facilitated forum, held in central Melbourne. Smaller facilitated sessions with OWLA followed, to review and discuss the collated service stakeholder input. The information from these OWLA sessions were again compiled and directed back to the service stakeholders for consideration and further discussion. The two groups came together for a final forum to prioritise the co-created ten services that they believed would be feasible and would address unmet need to support OWLA maintain independence. The process of co-creation was time-consuming and required considerable preparation to facilitate input from the target population. Small groups, gathering at convenient local locations, with transport support were essential in removing barriers to participation. However, co-creation was a viable method of eliciting the women's preferences and developing services more likely to meet their needs.


Corneal Endothelial Cells Have an Absolute Requirement for Cysteine for Survival.
The aim of this study was to evaluate which amino acid(s) among the 20 standard protein amino acids is indispensable for the survival of cultured human corneal endothelial cells (HCECs). HCECs were cultured in amino acid screening media that were missing 1 specific amino acid, and cell growth was evaluated. After this first selection, we conducted a further evaluation of cell growth in response to the addition of 4 amino acids (cysteine, methionine, valine, and arginine) to amino acid-free culture media. We then evaluated the antioxidant effect of cysteine compared with other antioxidants in terms of apoptosis of HCECs, rabbit corneal endothelial cell (CECs), monkey CECs, and ex vivo human donor corneas. Culture in an amino acid-free Dulbecco Modified Eagle Medium (DMEM) decreased the cell numbers to 11.0% when compared with culture in normal DMEM. Removal of cysteine, methionine, valine, or arginine from DMEM significantly suppressed cell numbers (27.7%, 61.4%, 75.5%, and 60.6%, respectively) (P < 0.01), whereas removal of other amino acids did not significantly decrease cell numbers. A lack of cysteine induced apoptosis, but addition of antioxidants reversed this. Removal of cysteine induced in vitro apoptosis in HCECs, rabbit CECs, monkey CECs, and ex vivo human donor corneas, whereas the presence of cysteine almost completely suppressed this apoptosis. Cysteine seems to be an indispensable amino acid for HCEC growth and survival. Its necessity might reflect a high requirement for antioxidants to protect HCECs from oxidative stress, as HCECs have high aerobic metabolic activity.


Surgery and the information age.
Surgeons' basic competency relies upon their ability to combine their existing knowledge and practical skills in the care of their patients, and also upon keeping up to date and encompassing change. We now live in a society which has become accustomed to receive a constant flow of up-to-date information about the world around it. With recent developments, including the Internet, the World Wide Web, and advances in personal computer systems, the pace of change of information technology (IT) has been increasing. These changes have had a significant impact upon surgery, and it is now becoming necessary to be able to make use of these new resources to function effectively in modern practice. This article will outline some of the more important topics relating to IT in surgical practice as an introduction to a series on this subject which will be appearing in forthcoming issues of the Journal.


Immune Competence and Minimizing Susceptibility to COVID-19 and Other Immune System Threats.
Exposure to viruses, bacteria, and other pathogens is unavoidable. Yet, the mere presence of these threats is not enough to automatically predispose to illness. The susceptibility of an individual to viral or bacterial infections is dependent upon immune competence. Many factors can interfere with the functioning of the immune system. Epigenetic alterations in the form of lifestyle or environmental factors can lead to impaired immunity. For example, exposure to air pollution can increase the risk of complications and mortality from COVID-19. Obesity can also exacerbate the damaging effects of air pollution on the lungs and may enhance the association between air pollution and increased COVID-19 severity. Poor sleep is another factor leading to impaired immunity, likely due to the coinciding melatonin depletion. Melatonin has been found to have antiviral and immune-enhancing effects, and it has been proposed that this hormone may be beneficial in COVID-19 patients. Zinc and vitamins D and C have also been well studied for their ability to shorten the duration of upper respiratory infections, and vitamin D has been found to reduce mortality in COVID-19 patients. Cannabidiol can both directly and indirectly improve immunity by enhancing natural killer cell activity, reducing inflammation, and relieving stress. Other dietary supplements backed by solid scientific evidence to show they act as immune enhancers are astragalus, a yeast fermentate (EpiCor®), olive leaf extract, berberine, N-acetyl cysteine, and garlic.


Interaction and functional cooperation between the LIM protein FHL2, CBP/p300, and beta-catenin.
Transcriptional activation of gene expression by Wnt signaling is driven by the association of beta-catenin with TCF/LEF factors and the recruitment of transcriptional coactivators. It has been shown that the LIM protein FHL2 and the acetyltransferase CBP/p300 individually stimulate beta-catenin transactivating activity and that beta-catenin is acetylated by p300. Here, we report that FHL2 and CBP/p300 synergistically enhanced beta-catenin/TCF-mediated transcription from Wnt-responsive promoters and that the acetyltransferase activity of CBP/p300 was involved in the cooperation. CBP/p300 interacted directly with FHL2, predominantly through the CH3 domain but not the histone acetyltransferase domain, and different regions of CBP/p300 were involved in FHL2 and beta-catenin binding. We provided evidence for the formation of a ternary complex by FHL2, CBP/p300, and beta-catenin and for colocalization of the three proteins in the nucleus. In murine FHL2(-/-) embryo fibroblasts, the transactivation activity of beta-catenin/TCF was markedly reduced, and this defect could be restored by exogenous expression of FHL2. However, CBP/p300 were still able to coactivate the beta-catenin/TCF complex in FHL2(-/-) cells, suggesting that FHL2 is dispensable for the coactivator function of CBP/p300 on beta-catenin. Furthermore, we found that FHL2 significantly increased acetylation of beta-catenin by p300 in vivo. Finally, we showed that FHL2, CBP/p300, and beta-catenin could synergistically activate androgen receptor-mediated transcription, indicating that the synergistic coactivator function is not restricted to TCF/LEF.


NOD1 sensing of house dust mite-derived microbiota promotes allergic experimental asthma.
Asthma severity has been linked to exposure to gram-negative bacteria from the environment that are recognized by NOD1 receptor and are present in house dust mite (HDM) extracts. NOD1 polymorphism has been associated with asthma. We sought to evaluate whether either host or HDM-derived microbiota may contribute to NOD1-dependent disease severity. A model of HDM-induced experimental asthma was used and the effect of NOD1 deficiency was evaluated. Contribution of host microbiota was evaluated by fecal transplantation. Contribution of HDM-derived microbiota was assessed by 16S ribosomal RNA sequencing, mass spectrometry analysis, and peptidoglycan depletion of the extracts. In this model, loss of the bacterial sensor NOD1 and its adaptor RIPK2 improved asthma features. Such inhibitory effect was not related to dysbiosis caused by NOD1 deficiency, as shown by fecal transplantation of Nod1-deficient microbiota to wild-type germ-free mice. The 16S ribosomal RNA gene sequencing and mass spectrometry analysis of HDM allergen, revealed the presence of some muropeptides from gram-negative bacteria that belong to the Bartonellaceae family. While such HDM-associated muropeptides were found to activate NOD1 signaling in epithelial cells, peptidoglycan-depleted HDM had a decreased ability to instigate asthma in vivo. These data show that NOD1-dependent sensing of HDM-associated gram-negative bacteria aggravates the severity of experimental asthma, suggesting that inhibiting the NOD1 signaling pathway may be a therapeutic approach to treating asthma.


Does single, low-dose preoperative dexamethasone improve outcomes after colorectal surgery based on an enhanced recovery protocol? Double-blind, randomized clinical trial.
Preoperative single, high-dose methylprednisolone administration improves postoperative outcomes after colonic surgery. Several randomized studies, including major surgeries, assessed various high-dose steroid regimens; however, evidence about the effect of administration of lower doses on postoperative outcomes in colorectal surgery is not available. The aim of the present study is to determine whether the administration of a single, low dose of dexamethasone before surgery would confer an outcome advantage after colorectal surgery. Thirty patients undergoing colorectal surgery were included in this randomized, double-blind study. Patients received 8 mg dexamethasone or serum physiologic preoperatively. Levels of Interleukin-6 and C-reactive protein, pain scores, postoperative nausea and vomiting, mobilization, complications, hospital stay, and readmissions were compared. Age, sex, indications, and operations were similar in both groups (P > 0.05). C-reactive protein and Interleukin-6 levels increased significantly postoperatively in each group (P < 0.05), but there were no differences between groups when compared (P > 0.05). There were also no significant differences between pain scores, bowel functions, mobilization, hospital stay, complication rates, and readmission rates between the two groups (P > 0.05). Preoperative 8 mg dexamethasone administration has no significant effect on reducing postoperative inflammatory response and also does not improve outcomes of colorectal surgery.


Aging and Hearing Health: The Life-course Approach.
Sensory abilities decline with age. More than 5% of the world's population, approximately 360 million people, have disabling hearing loss. In adults, disabling hearing loss is defined by thresholds greater than 40 dBHL in the better hearing ear.Hearing disability is an important issue in geriatric medicine because it is associated with numerous health issues, including accelerated cognitive decline, depression, increased risk of dementia, poorer balance, falls, hospitalizations, and early mortality. There are also social implications, such as reduced communication function, social isolation, loss of autonomy, impaired driving ability, and financial decline. Furthermore, the onset of hearing loss is gradual and subtle, first affecting the detection of high-pitched sounds and with difficulty understanding speech in noisy but not in quiet environments. Consequently, delays in recognizing and seeking help for hearing difficulties are common. Age-related hearing loss has no known cure, and technologies (hearing aids, cochlear implants, and assistive devices) improve thresholds but do not restore hearing to normal. Therefore, health care for persons with hearing loss and people within their communication circles requires education and counseling (e.g., increasing knowledge, changing attitudes, and reducing stigma), behavior change (e.g., adapting communication strategies), and environmental modifications (e.g., reducing noise). In this article, we consider the causes, consequences, and magnitude of hearing loss from a life-course perspective. We examine the concept of "hearing health," how to achieve it, and implications for policy and practice.


Breathing Disturbances Without Hypoxia Are Associated With Objective Sleepiness in Sleep Apnea.
To determine whether defining two subtypes of sleep-disordered breathing (SDB) events-with or without hypoxia-results in measures that are more strongly associated with hypertension and sleepiness. A total of 1022 participants with 2112 nocturnal polysomnograms from the Wisconsin Sleep Cohort were analyzed with our automated algorithm, developed to detect breathing disturbances and desaturations. Breathing events were time-locked to desaturations, resulting in two indices-desaturating (hypoxia-breathing disturbance index [H-BDI]) and nondesaturating (nonhypoxia-breathing disturbance index [NH-BDI]) events-regardless of arousals. Measures of subjective (Epworth Sleepiness Scale) and objective (2981 multiple sleep latency tests from a subset of 865 participants) sleepiness were analyzed, in addition to clinically relevant clinicodemographic variables. Hypertension was defined as BP ≥ 140/90 or antihypertensive use. H-BDI, but not NH-BDI, correlated strongly with SDB severity indices that included hypoxia (r ≥ 0.89, p ≤ .001 with 3% oxygen-desaturation index [ODI] and apnea hypopnea index with 4% desaturations). A doubling of desaturation-associated events was associated with hypertension prevalence, which was significant for ODI but not H-BDI (3% ODI OR = 1.06, 95% CI = 1.00-1.12, p < .05; H-BDI OR 1.04, 95% CI = 0.98-1.10) and daytime sleepiness (β = 0.20 Epworth Sleepiness Scale [ESS] score, p < .0001; β = -0.20 minutes in MSL on multiple sleep latency test [MSLT], p < .01). Independently, nondesaturating event doubling was associated with more objective sleepiness (β = -0.52 minutes in MSL on MSLT, p < .001), but had less association with subjective sleepiness (β = 0.12 ESS score, p = .10). In longitudinal analyses, baseline nondesaturating events were associated with worsening of H-BDI over a 4-year follow-up, suggesting evolution in severity. In SDB, nondesaturating events are independently associated with objective daytime sleepiness, beyond the effect of desaturating events.


Mechanisms of Acquired Resistance to Tyrosine Kinase Inhibitors in Clear - Cell Renal Cell Carcinoma (ccRCC).
Clear - cell renal cell carcinoma (ccRCC) is a histological subtype of renal cell carcinoma - the most prevalent adult kidney cancer. Causes of ccRCC are not completely understood and therefore number of available therapies is limited. As a consequence of tumor chemo- and radioresistance as well as restrictions in offered targeted therapies, overall response rate is still unsatisfactory. Moreover, a significant group of patients (circa 1/4) does not respond to the targeted first-line treatment, while in other cases, after an initial period of stable improvement, disease progression occurs. Owing to this, more data on resistance mechanisms are needed, especially those concerning widely used, relatively lately approved and more successful than previous therapies - tyrosine kinase inhibitors (TKIs). Up to date, five TKIs have been licensed for ccRCC treatment: sunitinib (SUTENT®, Pfizer Inc.), sorafenib (Nexavar®, Bayer HealthCare/Onyx Pharmaceuticals), pazopanib (Votrient®, GlaxoSmithKline), axitinib (Inlyta®, Pfitzer Inc.) and tivozanib (AV-951®, AVEO Pharmaceuticals). Researchers have specified different subsets of tyrosine kinase inhibitors potential resistance mechanisms in clear-cell renal cell carcinoma. In most papers published until now, drug resistance is divided into intrinsic and acquired, and typically multi-drug resistance (MDR) protein is described. Herein, the authors focus on molecular analysis concerning acquired, non-genetic resistance to TKIs, with insight into specific biological processes.


Hamstrings stretch reflex in human spasticity.
In 16 patients with spastic paralysis the hamstrings stretch reflex was found to increase as the velocity of stretch increased, and generally to subside after movement ceased. These effects are attributable to the dynamic property of the primary spindle ending. The stretch reflex commonly appeared in only the last third of the stretching movement and was maximal as the knee became fully extended. This is consistent with the static properties of the primary and secondary spindle endings, and accounts for the absence of the clasp-knife phenomenon in the spastic hamstrings. The difference in the nature of the stretch reflex in spastic flexor and extensor muscles is best explained by the differential reflex effects of group II afferent fibres which facilitate flexor motoneurones and inhibit extensor motoneurones.


Assembly mechanism of a supramolecular MS-ring complex to initiate bacterial flagellar biogenesis in Vibrio species.
The bacterial flagellum is an organelle responsible for motility and has a rotary motor comprising the rotor and the stator. Flagellar biogenesis is initiated by the assembly of the MS-ring, a supramolecular complex embedded in the cytoplasmic membrane. The MS-ring consists of a few dozen copies of the transmembrane FliF protein, and is an essential core structure which is a part of the rotor. The number and location of the flagella are controlled by the FlhF and FlhG proteins in some species. However, there is no clarity on the factors initiating MS-ring assembly, and contribution of FlhF/FlhG to this process. Here, we show that FlhF and a C-ring component FliG facilitate Vibrio MS-ring formation. When Vibrio FliF alone was expressed in Escherichia coli cells, MS-ring formation rarely occurred, indicating the requirement of other factors for MS-ring assembly. Consequently, we investigated if FlhF aided FliF in MS-ring assembly. We found that FlhF allowed GFP-fused FliF to localize at the cell pole in a Vibrio cell, suggesting that it increases local concentration of FliF at the pole. When FliF was co-expressed with FlhF in E. coli cells, the MS-ring was effectively formed, indicating that FlhF somehow contributes to MS-ring formation. The isolated MS-ring structure was similar to the MS-ring formed by Salmonella FliF. Interestingly, FliG facilitates MS-ring formation, suggesting that FliF and FliG assist in each other's assembly into the MS-ring and C-ring. This study aids in understanding the mechanism behind MS-ring assembly using appropriate spatial/temporal regulations.Importance Flagellar formation is initiated by the assembly of the FliF protein into the MS-ring complex, embedded in the cytoplasmic membrane. The appropriate spatial/temporal control of MS-ring formation is important for the morphogenesis of the bacterial flagellum. Here, we focus on the assembly mechanism of Vibrio FliF into the MS-ring. FlhF, a positive regulator of the number and location of flagella, recruits the FliF molecules at the cell pole and facilitates MS-ring formation. FliG also facilitates MS-ring formation. Our study showed that these factors control flagellar biogenesis in Vibrio, by initiating the MS-ring assembly. Furthermore, it also implies that flagellar biogenesis is a sophisticated system linked with the expression of certain genes, protein localization and a supramolecular complex assembly.


Increased genetic stability of HeLa cells after connexin 43 gene transfection.
To test the hypothesis that intact gap-junctional intercellular communication (GJIC) is necessary for genomic stability, we compared the spontaneous and chemically induced mutation frequencies in GJIC-proficient and -deficient HeLa cells. Thus, we determined microsatellite instability and mutation frequency in the HPRT gene in parental HeLa cells, which have no GJIC ability, and in HeLa cells in which GJIC was restored by transfection with the connexin 43 (Cx43) gene. When HeLa cells with (Cx43+) or without Cx43 gene (Cx43-) were treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or methylnitrosourea, the Cx43+ cells survived better than Cx43- cells. The mutation frequency at CA repeats was measured with a shuttle vector; in the vector, the coding region of the beta-galactosidase gene was rendered out of frame by insertion of CA repeats, and the frame could be restored by insertion or deletion mutations of the CA repeats. The mutation frequency at CA repeats was 2-fold lower in Cx43+ cells than in Cx43-, both before and after exposure to MNNG or methylnitrosourea (P < 0.05). The frequency of spontaneous HPRT gene mutations, selected by their resistance to 6-thioguanine, was 3-fold lower in Cx43+ cells than Cx43- cells. Similarly, the frequency of MNNG-induced HPRT mutations was significantly higher in Cx43- cells (P < 0.001). Similar results were obtained even when the mutant selection process was carried out in the presence of alpha-glycyrrhetinic acid, a long-term inhibitor of GJIC, suggesting that the observed effect is not due to unwanted killing of cells by GJIC-mediated metabolic cooperation. Thus, our data demonstrate that HeLa cells transfected with the Cx43 gene become more resistant to spontaneous as well as chemically induced genetic changes.


Zebularine induces replication-dependent double-strand breaks which are preferentially repaired by homologous recombination.
Zebularine is a second-generation, highly stable hydrophilic inhibitor of DNA methylation with oral bioavailability that preferentially target cancer cells. It acts primarily as a trap for DNA methyl transferases (DNMTs) protein by forming covalent complexes between DNMT protein and zebularine-substrate DNA. It's well documented that replication-blocking DNA lesions can cause replication fork collapse and thereby to the formation of DNA double-strand breaks (DSB). DSB are dangerous lesions that can lead to potentially oncogenic genomic rearrangements or cell death. The two major pathways for repair of DSB are non-homologous end joining (NHEJ) and homologous recombination (HR). Recently, multiple functions for the HR machinery have been identified at arrested forks. Here we investigate in more detail the importance of the lesions induced by zebularine in terms of DNA damage and cytotoxicity as well as the role of HR in the repair of these lesions. When we examined the contribution of NHEJ and HR in the repair of DSB induced by zebularine we found that these breaks were preferentially repaired by HR. Also we show that the production of DSB is dependent on active replication. To test this, we determined chromosome damage by zebularine while transiently inhibiting DNA synthesis. Here we report that cells deficient in single-strand break (SSB) repair are hypersensitive to zebularine. We have observed more DSB induced by zebularine in XRCC1 deficient cells, likely to be the result of conversion of SSB into toxic DSB when encountered by a replication fork. Furthermore we demonstrate that HR is required for the repair of these breaks. Overall, our data suggest that zebularine induces replication-dependent DSB which are preferentially repaired by HR.


[Lymphocyte count in the blood of stomach cancer patients].
The content of T- and B-lymphocytes was studied in the blood of 72 patients with gastric ulcer and carcinoma. The amount of complete T- and B-lymphocytes and incomplete B-lymphocytes was found to dramatically decrease in patients with peptic ulcer and gastric carcinoma with distant metastases. The authors believe that the data obtained could be used in addition to others when choosing the specific treatment of patients with gastric carcinoma in the postoperative period.


Quantitation of influenza virus antigens on infected target cells and their recognition by cross-reactive cytotoxic T cells.
Monoclonal antibody to type-A influenza virus matrix (M)-protein was used to quantitate the appearance of M-protein on abortively infected P815 cells. After 16 h of infection with different type-A viruses, only a low amount of M-protein appears on the surface of infected cells (approximately 10(3) site/cell) in contrast to approximately 10(5) hemagglutinin molecules on each cell surface. However, virus replication is required for M-protein appearance. Analysis of solubilized membranes purified from 16-h-infected cells shows approximately 10(4) M-protein molecule/cell in the plasma membrane, a content that is consistent with the observed low surface expression, and that indicates that most of the M-protein is localized internally. We found no evidence that cross-reactive cytotoxic T cells could recognize M-protein; neither monoclonal antibody or hyperimmune anti-M-protein antiserum could inhibit T cell killing, either alone or in combination with monoclonal anti-H-2 antibody. Taken together, the low level of M-protein appearance and lack of T cell blocking by anti-M-protein antibody leaves doubt that M-protein is the antigen recognized by cross-reactive cytotoxic T cells.


Predictors for Mortality in Children with Scorpion Envenomation Admitted to Pediatric Intensive Care Unit, Qena Governorate, Egypt.
This study aimed to identify the clinical and laboratory manifestations that affect outcome of scorpion envenomation in children. It included 154 children admitted with scorpion sting envenomation over a period of 2 years. The epidemiological, clinical, and laboratory findings of patients were recorded, and grading of severity was performed based on local and systemic involvement. Organ failure was determined according to diagnostic criterion of multiple organ dysfunction syndrome, and severity of illness was assessed by the Pediatric Risk of Mortality (PRISM III) score. Of studied children, 58.4% were males and 41.6% were females. Children aged > 5 years suffered more scorpion stings (79.9%) than others did. The place of residence was rural more than urban, outdoor stings more than indoors, nocturnal more than diurnal, and most stings were on the exposed areas of the limbs. Based on clinical evaluation, 37.7% of patients were classified as class I severity followed by class II (48.7%) and class III (13.6%). Among studied cases, 21 deaths (13.6%) were registered; all of them belonged to class III severity. Mortality was significantly higher in children with agitation, coma, convulsions, arrhythmia, heart failure, pulmonary edema, and priapism. There were significantly higher values of leukocytes, platelets, creatinine, liver enzymes, glucose, and creatine phosphokinase in non-survivors than in survivors. The presence of organ failure was associated with mortality. In addition, the need for mechanical ventilation and inotropic support were at increased risk of mortality. Moreover, a significant association was found between PRISM score and the number of failed organs with fatal outcome.


Protective effect of catalpol on nicotine‑induced injury of alveolar bone and associated underlying mechanisms.
Smoking is an important factor that causes periodontitis, which manifests as alveolar bone injury and absorption, and has a high incidence and unfavorable treatment efficacy. Nicotine causes ischemia and inflammation of the periodontium and inhibits the mineralization of alveolar bones. Previous studies have revealed the anti‑tumor biological activities of catalpol, in addition to neuroprotection and anti‑inflammation. The present study therefore investigated the underlying protective mechanism of catalpol in alveolar bone injury. A total of 24 Wistar rats were infused with nicotine (0.7 mg/kg for 30 days), followed by subcutaneous injection of catalpol (2 µg/kg for 14 days). The loss of alveolar bone was examined, and bone alkaline phosphatase (AP) and osteocalcin levels were examined by ELISA. The expression of tumor necrosis factor (TNF)‑α and cyclooxygenase‑2 (COX‑2) was quantified by reverse transcription‑quantitative polymerase chain reaction analysis and western blotting. Treatment with nicotine decreased AP and osteocalcin levels, increased TNF‑α and COX‑2 expression levels, and led to alveolar bone loss compared with the control group. Catalpol decreased bone loss, increased AP and osteocalcin, and decreased TNF‑α and COX‑2 expression compared with the nicotine treatment group. Catalpol may alleviate nicotine‑induced injury and alveolar bone loss via inhibition of inflammatory factors, and facilitate the mineralization of alveolar bones.


Perspectives and experiences of elective surgery patients regarding pain management.
The purpose of this study was to explore the perspectives and experiences of elective surgery patients regarding pain management. A qualitative design, based on the content analysis approach, was used to collect and analyze the experience of 20 elective surgery patients who all had abdominal surgery in surgical wards in two teaching hospitals in Tehran, Iran. After employing purposeful sampling for the selection of the participants, semistructured interviews were held for data collection. During the data analysis, three main themes emerged: "perceptions of pain management goals", "patients' views of nurses' role in pain management", and "interaction in pain management". It was concluded that understanding the factors that influence pain management after surgery from the patients' viewpoint will contribute to the body of knowledge of nurses in order to promote the quality of nursing care.


The functional role of polyamines in eukaryotic cells.
Polyamines, consisting of putrescine, spermidine and spermine are essential for normal cell growth and viability in eukaryotic cells. Since polyamines are cations, they interact with DNA, ATP, phospholipids, specific kinds of proteins, and especially with RNA. Consequently, the functions of these acidic compounds and some proteins are modified by polyamines. In this review, the functional modifications of these molecules by polyamines are presented. Structural change of specific mRNAs by polyamines causes the stimulation of the synthesis of several different proteins, which are important for cell growth and viability. eIF5 A, the only known protein containing a spermidine derivative, i.e. hypusine, also functions at the level of translation. Experimental results thus far obtained strongly suggest that the most important function of polyamines is at the level of translation.


A prospective population-based study of pediatric trauma patients with mild alterations in consciousness (Glasgow Coma Scale score of 13-14).
Considerable controversy surrounds the appropriate evaluation of children with mild alterations in consciousness after closed head trauma (Glasgow Coma Scale [GCS] score of 13-14). The objective of the current study was to determine the incidence of intracranial lesions in pediatric patients with a field GCS score of 13 or 14 after closed head injuries. The current study is a population-based, multicenter prospective study of all patients to whom emergency medical services responded during a 12-month period. The setting was urban Los Angeles County, encompassing a patient population of 2.3 million children, 13 designated trauma centers, and 94 receiving hospitals. In the pediatric age group (<15 yr old), 8488 patients were transported by emergency medical services for injuries. Of these, 209 had a documented field GCS score of 13 or 14. One hundred fifty-seven patients were taken to trauma centers, and 135 (86%) underwent computed tomography. Forty-three patients (27.4%) had abnormal results on computed tomographic scans, 30 (19.1%) had an intracranial hemorrhage, and 5 required an operative neurosurgical procedure for hematoma evacuation. Positive and negative predictive values of deteriorating mental status (0.500 and 0.844, respectively), loss of consciousness (0.173 and 0.809), cranial fracture (0.483 and 0.875, and extracranial injuries (0.205 and 0.814) were poor predictors of intracranial hemorrhage. Pediatric patients who have mild alterations in consciousness in the field have a significant incidence of intracranial injury. The great majority of these patients will not require operative intervention, but the implications of missing these hemorrhages can be severe for this subgroup of head-injured patients. Because clinical criteria and cranial x-rays are poor predictors of intracranial hemorrhage, it is recommended that all children with a GCS score of 13 or 14 routinely undergo screening via non-contrast-enhanced computed tomography.


Mechanism of graphene-induced cytotoxicity: Role of endonucleases.
Graphene, a crystalline allotrope or carbon, presents numerous useful properties; however, its toxicity is yet to be determined. One of the most dramatic and irreversible toxic abilities of carbon nanomaterials is the induction of DNA fragmentation produced by endogenous cellular endonucleases. This study demonstrated that pristine graphene exposed to cultured kidney tubular epithelial cells is capable of inducing DNA fragmentation measured by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, which is usually associated with cell death. TUNEL (cell death) and endonuclease activity measured using a near infrared fluorescence probe was significantly higher in cells containing graphene aggregates detected by Raman spectroscopy. The elevation of TUNEL coincided with the increased abundance of heme oxygenase 1 (HO-1), heat shock protein 90 (HSP90), active caspase-3 and endonucleases (deoxyribonuclease I [DNase I] and endonuclease G [EndoG]), as measured by quantitative immunocytochemistry. Specific inhibitors for HO-1, HSP90, caspase-3, DNase I and EndoG almost completely blocked the DNA fragmentation induced by graphene exposure. Therefore, graphene induces cell death through oxidative injury, caspase-mediated and caspase-independent pathways; and endonucleases DNase I and EndoG are important for graphene toxicity. Inhibition of these pathways may ameliorate cell injury produced by graphene. Copyright © 2017 John Wiley & Sons, Ltd.


Osteohistology of Dromornis stirtoni (Aves: Dromornithidae) and the biological implications of the bone histology of the Australian mihirung birds.
The late Miocene Dromornis stirtoni is the largest of the giant flightless dromornithid birds. Here, we studied 22 long bones (femora, tibiotarsi, tarsometatarsi) of D. stirtoni to assess its osteohistology to deduce various aspects of its life history. Our results show that D. stirtoni took several years (likely, more than a decade), to reach adult body size, after which its growth rate slowed down, and skeletal maturity occurred. This growth strategy differs from that of its Pleistocene relative, Genyornis newtoni, which experienced faster rates of growth to reach adult body size. We propose that these mihirung birds, separated by millions of years, each responded to the prevailing environmental conditions of the time, by selecting for different growth strategies, with D. stirtoni having an extreme K-selected life history strategy. The presence of medullary bone permitted the identification of female D. stirtoni specimens, and its presence in some bones lacking an OCL layer showed sexual maturity preceded its formation. We postulate that while G. newtoni had a somewhat greater reproductive potential than D. stirtoni, it remained far less than that observed in the extant emu (Dromaius novaehollandiae). Genyornis newtoni survived into the late Pleistocene alongside extant emus and overlapped the arrival of the first humans in Australia, but the former species shortly thereafter became extinct while emus remain prolific.


Modeling and characterization of embedded electrode performance in transverse electrooptic modulators.
A mathematical model with experimental verification is presented to characterize the performance of surface and embedded electrodes in 2-D electrooptic modulators. From the solution of a discretized integral equation for the electrode surface charge, the electrode capacitance and the electric field penetration and uniformity are related to the switching voltage, speed, and uniformity of the electrooptic modulation. Fabricated surface and embedded electrodes in 9/65/35 PLZT are then evaluated with respect to the predictions of the model and the saturated quadratic response of the electrooptic material. These results provide important insight into the design trade-offs of switching speed, halfwave voltage, switching energy, and modulation uniformity of surface and embedded modulator geometries.


Trans catalysis in Tn5 transposition.
Synaptic complexes in prokaryotic transposons occur when transposase monomers bind to each of two specific end-binding sequences and then associate to bring the proteins and the two ends of the transposon together. It is within this complex of proteins and DNA that identical catalytic reactions are carried out by transposase on each of the ends of the transposon. In this study, we perform in vitro transposition reactions by combining the methylated inside end (IE(ME)) biased hyperactive Tn5 transposase, Tnp sC7 version 2.0, and the outside end (OE) biased hyperactive Tn5 transposase, Tnp EK/LP, with plasmid DNA containing a transposon defined by one IE(ME) and one OE. These two proteins cooperate to facilitate double end cleavage of the transposon from the plasmid and conversion into transposition products via strand transfer. When one of the hyperactive Tnps is replaced with a catalytically inactive version containing the mutation EA326 (DDE mutant), the predominant reaction product is a linearized plasmid resulting from single end cleavage. Restriction analysis of these linear products reveals that cleavage is occurring on the end distal to that which is bound by the transposase with an intact active site or in trans. Similar in vitro experiments performed with precut transposons and a supercoiled target plasmid demonstrated that the strand transfer reaction is also facilitated by a trans active DDE motif.


Pharmacokinetics of plasma-derived and recombinant factor IX - implications for prophylaxis and on-demand therapy.
Plasma-derived (pd) and recombinant (r) factor IX (FIX) differ in pharmacokinetic (PK) properties. These differences and their clinical implications have been debated since the introduction of rFIX. The aim of this review was to describe the comparative disposition of pdFIX and rFIX and will for this purpose begin with an overview of population PK modelling. In contrast to the model-independent method, a population PK model can analyse sparse data sets obtained in various settings, provide parameter values that can be used to predict coagulation factor levels with any kind of single or multiple dosing and include statistical analysis of variation between individuals. Population modelling has also clearly demonstrated the difference in PK between pdFIX and rFIX. Their distribution characteristics influence the FIX coagulant activity (FIX:C) level vs. time curve during the early hours after infusion. In vivo recovery and elimination half-life are consequently not adequate descriptors of the effective PK of FIX, and for new analogues with modified PK, differences in distribution might be clinically important. Calculated doses to maintain 1% trough levels during twice-weekly prophylactic treatment are considerably higher with rFIX than with pdFIX and roughly correspond to dosing in clinical studies. However, the putative relationship between FIX:C trough level and therapeutic outcome has never been confirmed in a clinical trial. Comparative studies on prophylaxis with different types of FIX are needed.


Intraoperative optical intrinsic signal imaging: a clinical tool for functional brain mapping.
Optical imaging of intrinsic signals (OIS) is a well-established neuroimaging modality by which functional cortical activity is mapped by detecting activity-related changes in cortical light reflectance. Light reflectance changes are detected by a charged-coupled device camera that captures images of the exposed cortex both at rest and during activity. Although to date OIS has only been used for research purposes, intraoperative OIS (iOIS) holds promise as a clinical mapping tool. In general, iOIS demonstrates good spatial correlation with electrocortical stimulation mapping (ECSM) and other electrophysiological modalities. Additionally, iOIS offers high spatial resolution (in microns), does not make contact with the surface of the brain, and introduces no potentially harmful compounds. Moreover, mapping is relatively rapid. The authors review the potential contribution of iOIS to the intraoperative environment. Specifically, they review iOIS methodology, discuss signal origin, compare OIS with other functional mapping modalities, and explain its potential benefits and limitations. They propose that iOIS may, in the future, be used in conjunction with ECSM to improve the resolution and accuracy of intraoperative mapping, decrease total time of intraoperative mapping, and possibly improve neurological outcomes. Additional studies will be required to quantify the sensitivity and specificity of optical maps relative to ECSM before it can be implemented clinically.


Species-specific polymerase chain reactions for the detection of Mycoplasma buteonis, Mycoplasma falconis, Mycoplasma gypis, and Mycoplasma corogypsi in captive birds of prey.
Mycoplasmas are pathogens of different avian species, but the role of Mycoplasma in raptors is not yet completely determined. As Mycoplasma isolation and identification present several difficulties, species-specific polymerase chain reactions (PCRs) for the detection of mycoplasmas found in birds of prey (Mycoplasma buteonis, Mycoplasma corogypsi, Mycoplasma falconis, and Mycoplasma gypis) were established. The specificity of the PCR methods were investigated using known avian Mycoplasma reference strains and isolates as well as related bacteria and was found to be specific. Amplificons obtained with these PCRs from field samples showed no false-positive results in restriction enzyme analysis and sequencing. The sensitivities of the different PCR assays varied between 50 fg and 1 pg DNA. Twenty-five tracheal swabs from healthy captive birds of prey were investigated by culture and immunobinding assay as comparison to the PCRs. Mycoplasmal DNA was detected in 88% of the samples, with negative results only from vultures. Mycoplasma falconis and M. buteonis were regularly found in falcons, and M. gypis was found in a common buzzard. Mycoplasma corogypsi was not demonstrated. Several isolates could not be differentiated using an immunobinding assay as well as the described PCR methods.


Exploring Lung Cancer Screening Discussions on Twitter.
Lung cancer is the leading cause of cancer-related death in the United States. Low-dose computed tomography (LDCT) for lung cancer screening (LCS) can reduce lung cancer deaths by 20% compared to chest x-rays. Nevertheless, the uptake of LDCT is low for high-risk smokers. In this study we used Twitter data to understand laypeople's emotions towards LCS, find topics on LCS-related tweets, and assess the impact of promotional information on laypeople's discussions.


Polyamine oxidase-mediated intraerythrocytic killing of Plasmodium falciparum: evidence against the role of reactive oxygen metabolites.
The polyamines spermine and spermidine, in the presence of polyamine oxidase, were shown to be cytotoxic in vitro to various isolates of Plasmodium falciparum. Neither polyamines nor polyamine oxidase alone was cytotoxic. This cytotoxicity was manifested by the degeneration of the parasites into crisis forms and by the inhibition of methionine incorporation by the parasites. Only 2 to 2.5 h of exposure to the reaction mixture (polyamine oxidase, 100 micrograms/ml; spermine, 1 mM) resulted in parasite death. It was shown that ammonia, hydrogen peroxide, and associated reactive oxygen intermediates produced during the oxidation of polyamines were not the cause of the parasite death observed in this system. This suggested that aldehydes or further breakdown products of these, e.g., acrolein (or both), need to be considered as the effector substances of the polyamine oxidase-mediated killing of P. falciparum.


[Experimental research on animal model and treatment of Sjogren's syndrome by traditional chinese medicine (TCM)]
OBJECTIVE:To confirm the function of SS Syrup used in our department. METHODS: Under the basis of TCM and differentiation symptoms of SS,the animal model was established and applied to test the function of SS syrup.There are three indices to be observed,weight change,volume of drinking water daily,cAMP change. RESULTS: The three observed indices of dificiency symptom model had been rectified by SS syrup. CONCLUSION: SS syrup has two main functions:(1)reinforcing of the spleen and tonifying qi;(2)nourishing liver-yin and kidney-yin as well as removing pathogentic heat and fire.


Coptisine, the Characteristic Constituent from Coptis chinensis, Exhibits Significant Therapeutic Potential in Treating Cancers, Metabolic and Inflammatory Diseases.
Naturally derived alkaloids belong to a class of quite significant organic compounds. Coptisine, a benzyl tetrahydroisoquinoline alkaloid, is one of the major bioactive constituents in Coptis chinensis Franch., which is a famous traditional Chinese medicine. C. chinensis possesses many kinds of functions, including the ability to eliminate heat, expel dampness, purge fire, and remove noxious substances. In Asian countries, C. chinensis is traditionally employed to treat carbuncle and furuncle, diabetes, jaundice, stomach and intestinal disorders, red eyes, toothache, and skin disorders. Up to now, there has been plenty of research of coptisine with respect to its pharmacology. Nevertheless, a comprehensive review of coptisine-associated research is urgently needed. This paper was designed to summarize in detail the progress in the research of the pharmacology, pharmacokinetics, safety, and formulation of coptisine. The related studies included in this paper were retrieved from the following academic databases: The Web of Science, PubMed, Google scholar, Elsevier, and CNKI. The cutoff date was January 2023. Coptisine manifests various pharmacological actions, including anticancer, antimetabolic disease, anti-inflammatory disease, and antigastrointestinal disease effects, among others. Based on its pharmacokinetics, the primary metabolic site of coptisine is the liver. Coptisine is poorly absorbed in the gastrointestinal system, and most of it is expelled in the form of its prototype through feces. Regarding safety, coptisine displayed potential hepatotoxicity. Some novel formulations, including the [Formula: see text]-cyclodextrin-based inclusion complex and nanocarriers, could effectively enhance the bioavailability of coptisine. The traditional use of C. chinensis is closely connected with the pharmacological actions of coptisine. Although there are some disadvantages, including poor solubility, low bioavailability, and possible hepatotoxicity, coptisine is still a prospective naturally derived drug candidate, especially in the treatment of tumors as well as metabolic and inflammatory diseases. Further investigation of coptisine is necessary to facilitate the application of coptisine-based drugs in clinical practice.


Simulative and Experimental Characterization of a pH-Dependent Clamp-like DNA Triple-Helix Nanoswitch.
Here we couple experimental and simulative techniques to characterize the structural/dynamical behavior of a pH-triggered switching mechanism based on the formation of a parallel DNA triple helix. Fluorescent data demonstrate the ability of this structure to reversibly switch between two states upon pH changes. Two accelerated, half microsecond, MD simulations of the system having protonated or unprotonated cytosines, mimicking the pH 5.0 and 8.0 conditions, highlight the importance of the Hoogsteen interactions in stabilizing the system, finely depicting the time-dependent disruption of the hydrogen bond network. Urea-unfolding experiments and MM/GBSA calculations converge in indicating a stabilization energy at pH 5.0, 2-fold higher than that observed at pH 8.0. These results validate the pH-controlled behavior of the designed structure and suggest that simulative approaches can be successfully coupled with experimental data to characterize responsive DNA-based nanodevices.


Advantages of femtosecond laser in various corneal transplantation procedures: A narrative literature review.
The ability of the femtosecond laser (FSL) to create tissue incisions at desired depth and location with minimal collateral damage to surrounding tissue may offer potential advantages for corneal transplantation surgery. FSL may enable the precise and accurate preparation of both donor and recipient tissue, which can potentially result in enhanced wound integrity, facilitation of wound healing, and improved visual outcomes. Many studies have suggested that FSL may be a viable option in various corneal transplantation procedures, such as, penetrating keratoplasty, anterior lamellar keratoplasty, endothelial keratoplasty, and novel techniques, including Bowman layer transplantation and stromal lenticule addition keratoplasty. In this review, the author presents a brief overview on the application of FSL in various corneal transplantation procedures and discusses the results of studies regarding the safety and efficacy of FSL-assisted corneal transplantation procedures.


Spica Prunellae extract suppresses the growth of human colon carcinoma cells by targeting multiple oncogenes via activating miR-34a.
Spica Prunellae is the spike of the herb Prunella vulgaris L. in traditional Chinese medicine which is often used for the treatment of various cancers including colorectal cancer. In the present study, we found that a key tumor suppressor, microRNA-34a (miR-34a) is involved in the antitumor activity for Spica Prunellae. Human colon carcinoma HCT-8 cells treated with an ethanol extract of Spica Prunellae (EESP) had significantly decreased cell proliferation and viability, in a dose-dependent manner. Flow cytometry analysis with Annexin V/PI staining analysis revealed that EESP treatment could induce apoptosis of HCT-8 cells. The level of miR-34a was upregulated in HCT-8 cells following EESP treatment, whereas expression levels of its target genes Notch1, Notch2 and Bcl-2 were downregulated. Inhibition of miR-34a rescued the expression of these target genes. These results revealed that Spica Prunellae can suppress the growth of HCT-8 cells by targeting Notch1, Notch2 and Bcl-2 via activation of miR-34a.


Experimental evaluation of Bocopa monniera on rat gastric ulceration and secretion.
The anti-ulcerogenic effect of fresh juice from the whole plant of Bocapa monniera Wettst. (BMJ) commonly known as Brahmi in Hindi was examined using gastric ulcer models induced by ethanol, aspirin, 2 h cold restraint stress and 4 h pylorus ligation. Bocapa monniera juice (BMJ) at doses of 100 and 300 mg/kg and sucralfate at a dose of 250 mg/kg were given orally, twice daily for 5 days. BMJ 100-300 mg/kg produced significant antiulcer activity in all the experimental gastric ulcer models except in case of ethanol-induced ulcers where 100 mg/kg was not found to decrease it significantly. BMJ (100-300 mg/kg) was found to have little or no effect on the offensive acid-pepsin secretion, while cell shedding (microgram DNA/mg of protein) and mucin secretion in terms of total carbohydrates:protein ration (TC:P), the two important parameters of defensive factors were significantly decreased and increased respectively indicating enhancement of protective mucosal factors. Both BMJ (300 mg/kg) and SF showed tendency to increase the mucosal glycoproteins in terms of TC:P, though individual carbohydrates and total carbohydrates were either increased or showed a tendency to increase. Thus, ulcer protective effect of BMJ may be due to its effect on mucosal defensive factors like enhanced mucin secretion, mucosal glycoprotein and decreased cell shedding rather than on offensive factors such as acid and pepsin.


Tomographic evaluation of TMJ loading affected by occlusal pivots.
The purpose of this study was to clarify the direction and magnitude of condyle displacement during clenching with occlusal pivots. Temporomandibular joint (TMJ) loading and flexibility and the clinical significance of orthopedic appliance therapy are also discussed. Participants were 16 volunteers with a mean age of 25.8 years. Maxillary and mandibular flat occlusal devices were fabricated. TMJ tomogram series were taken first with tripod support and then with maxillary clenched anterior pivot support and/or unilateral posterior pivot support. Condyle positions on the TMJ tomograms were analyzed using the digital subtraction technique. When clenching with the anterior pivot, the anterior reference point (A) on the mandibular condyle was translated 0.1 mm posteriorly (P > 0.05) and 0.2 mm superiorly (P < 0.01) from the tripod-support reference point. In the case of the ipsilateral posterior pivot, point A was rotated 1.2 degrees (P < 0.01) and was translated 0.4 mm (P < 0.01) posteriorly and 0.18 mm superiorly (P > 0.05). Condyle displacement during clenching with the anterior and posterior unilateral pivot was found, which supports previous findings. However, the direction and magnitude of the displacement differed from those of previous studies using jaw-tracking devices, and individual values differed greatly among subjects.


A comparison of recovery times and characteristics with sevoflurane and isoflurane anaesthesia in horses undergoing magnetic resonance imaging.
To compare recovery times and quality following maintenance of anaesthesia with sevoflurane or isoflurane after a standard intravenous induction technique in horses undergoing magnetic resonance imaging (MRI). Prospective, randomised, blinded clinical study. Animals One hundred ASA I/II horses undergoing MRI. Pre-anaesthetic medication with intravenous acepromazine and romifidine was followed by induction of anaesthesia with diazepam and ketamine. The animals were randomised into two groups to receive either sevoflurane or isoflurane in oxygen. Horses were subjectively scored (0-5) for temperament before sedation, for quality of sedation, induction and maintenance and anaesthetic depth on entering the recovery area. Recoveries were videotaped and scored by an observer, unaware of the treatment, using two scoring systems. Times to the first movement, head lift, sternal recumbency and standing were recorded along with the number of attempts to achieve sternal and standing positions. Variables were compared using a Student t-test or Mann-Whitney U-test (p < 0.05), while the correlation between subjective recovery score and other relevant variables was tested calculating the Spearman Rank correlation coefficient and linear regression modelling performed when significant. Seventy-seven horses entered the final analysis, 38 received isoflurane and 39 sevoflurane. Body mass, age and duration of anaesthesia were similar for both groups. There were no differences in recovery times, scoring or number of attempts to achieve sternal recumbency and standing between groups. Weak, but significant, correlations were found between the subjective recovery score for the pooled data from both groups and both temperament and time in sternal recumbency. No differences in recovery times or quality were detected following isoflurane or sevoflurane anaesthesia after intravenous induction. Clinical relevance Sevoflurane affords no obvious advantage in recovery over isoflurane following a standard intravenous induction technique in horses not undergoing surgery.


Cardiac myocyte-protective effect of microRNA-22 during ischemia and reperfusion through disrupting the caveolin-3/eNOS signaling.
MicroRNA-22 (miR-22) was previously reported to elicit cardiac myocyte hypertrophy and had an anti-apoptotic effect on neurons. However, its effects on cardiac myocyte apoptosis and cardiac function during ischemia and reperfusion (I/R) are not clear. In the present study, we demonstrate that pre-administration of miR-22 mimic reduced I/R-induced cardiac dysfunction significantly in a rat model. We found that miR-22 overexpression inhibited cardiac myocyte apoptosis, and reduced cardiac remodeling during I/R. Significant cardiac myocyte apoptosis was also observed in a cardiac myocyte model after hypoxia/reoxygenation (H/R), a representative process of I/R. Further experiments showed that eNOS activity and the following NO production were significantly decreased during I/R and H/R, while such decrease was inhibited by overexpression of miR-22. Mechanistically, overexpression of miR-22 had little effect on the total protein level of eNOS, but restored the level of p-eNOS (Ser1177) which was down-regulated during H/R. Further RT-PCR results demonstrated that Caveolin 3 (Cav3), an upstream negative regulator of eNOS, was upregulated during H/R, resulting in a decrease of p-eNOS. However, such upregulation of Cav3 transcript level was inhibited directly by miR-22 during H/R, leading to a restored p-eNOS level and followed NO production in cardiac myocytes. Together, the present study revealed that miR-22 down-regulated Cav3, leading to restored eNOS activity and NO production, which further inhibited cardiac myocyte apoptosis and promoted cardiac function after I/R. Of clinical interest, the present study may highlight miR-22 as a potential therapeutic agent for reducing I/R induced cardiac injury.


A Pilot Randomised Controlled Trial of a School-Based Resilience Intervention to Prevent Depressive Symptoms for Young Adolescents with Autism Spectrum Disorder: A Mixed Methods Analysis.
Despite increased depression in adolescents with Autism Spectrum Disorder (ASD), effective prevention approaches for this population are limited. A mixed methods pilot randomised controlled trial (N = 29) of the evidence-based Resourceful Adolescent Program-Autism Spectrum Disorder (RAP-A-ASD) designed to prevent depression was conducted in schools with adolescents with ASD in years 6 and 7. Quantitative results showed significant intervention effects on parent reports of adolescent coping self-efficacy (maintained at 6 month follow-up) but no effect on depressive symptoms or mental health. Qualitative outcomes reflected perceived improvements from the intervention for adolescents' coping self-efficacy, self-confidence, social skills, and affect regulation. Converging results remain encouraging given this population's difficulties coping with adversity, managing emotions and interacting socially which strongly influence developmental outcomes.


FANCJ/BACH1 acetylation at lysine 1249 regulates the DNA damage response.
BRCA1 promotes DNA repair through interactions with multiple proteins, including CtIP and FANCJ (also known as BRIP1/BACH1). While CtIP facilitates DNA end resection when de-acetylated, the function of FANCJ in repair processing is less well defined. Here, we report that FANCJ is also acetylated. Preventing FANCJ acetylation at lysine 1249 does not interfere with the ability of cells to survive DNA interstrand crosslinks (ICLs). However, resistance is achieved with reduced reliance on recombination. Mechanistically, FANCJ acetylation facilitates DNA end processing required for repair and checkpoint signaling. This conclusion was based on the finding that FANCJ and its acetylation were required for robust RPA foci formation, RPA phosphorylation, and Rad51 foci formation in response to camptothecin (CPT). Furthermore, both preventing and mimicking FANCJ acetylation at lysine 1249 disrupts FANCJ function in checkpoint maintenance. Thus, we propose that the dynamic regulation of FANCJ acetylation is critical for robust DNA damage response, recombination-based processing, and ultimately checkpoint maintenance.


Naphthyl Thio- and Carba-xylopyranosides for Exploration of the Active Site of β-1,4-Galactosyltransferase 7 (β4GalT7).
Xyloside analogues with substitution of the endocyclic oxygen atom by sulfur or carbon were investigated as substrates for β-1,4-galactosyltransferase 7 (β4GalT7), a key enzyme in the biosynthesis of glycosaminoglycan chains. The analogues with an endocyclic sulfur atom proved to be excellent substrates for β4GalT7, and were galactosylated approximately fifteen times more efficiently than the corresponding xyloside. The 5a-carba-β-xylopyranoside in the d-configuration proved to be a good substrate for β4GalT7, whereas the enantiomer in the l-configuration showed no activity. Further investigations by X-ray crystallography, NMR spectroscopy, and molecular modeling provided a rationale for the pronounced activity of the sulfur analogues. Favorable π-π interactions between the 2-naphthyl moiety and a tyrosine side chain of the enzyme were observed for the thio analogues, which open up for the design of efficient GAG primers and inhibitors.


Luminal lymphoid cells in the rabbit intestine.
In normal rabbits, lymphocytes have been identified in large numbers on the luminal epithelial aspects of both Peyer's patches and the appendix by scanning and transmission electron microscopy. Luminal cells shared an intimate relationship with intestinal microorganisms. Irrigation of the appendiceal lumen has proved a useful technique for collecting 8.5 x 10(6) (mean) viable lymphocytes (73%) and macrophages (22%). Functional characteristics (mitogen responsiveness, T-cell numbers and cells with positive cytoplasmic immunoglobulin A fluorescence) were similar to isolated intestinal mucosal cell preparations. In vivo cytokinetic studies indicated that 50% of luminal appendiceal lymphocytes were recently divided (within 5 h). The passage of lymphoid cells into the the appendix lumen was antigen responsive, as was the size of the appendiceal lymphoid mass. This evidence suggests that migration of lymphoid cells into the lumen of the intestine in healthy animals occurs normally probably from the gut-associated lymphoid tissue--a phenomenon similar to that observed in the bronchial tract. Both mucosal surfaces may, therefore, be important sites of lymphocyte and macrophage traffic.


Protein cage nanoparticles bearing the LyP-1 peptide for enhanced imaging of macrophage-rich vascular lesions.
Cage-like protein nanoparticles are promising platforms for cell- and tissue-specific targeted delivery of imaging and therapeutic agents. Here, we have successfully modified the 12 nm small heat shock protein from Methanococcus jannaschii (MjHsp) to detect atherosclerotic plaque lesions in a mouse model system. As macrophages are centrally involved in the initiation and progression of atherosclerosis, targeted imaging of macrophages is valuable to assess the biologic status of the blood vessel wall. LyP-1, a nine residue peptide, has been shown to target tumor-associated macrophages. Thus, LyP-1 was genetically incorporated onto the exterior surface of MjHsp, while a fluorescent molecule (Cy5.5) was conjugated on the interior cavity. This bioengineered protein cage, LyP-Hsp, exhibited enhanced affinity to macrophage in vitro. Furthermore, in vivo injection of LyP-Hsp allowed visualization of macrophage-rich murine carotid lesions by in situ and ex vivo fluorescence imaging. These results demonstrate the potential of LyP-1-conjugated protein cages as nanoscale platforms for delivery of imaging agents for the diagnosis of atherosclerosis.


20-HETE contributes to ischemia-induced angiogenesis.
Angiogenesis is an important adaptation for recovery from peripheral ischemia. Here, we determined whether 20-hydroxyeicosatetraenoic acid (20-HETE) contributes to ischemia-induced angiogenesis and assessed its underlying molecular and cellular mechanisms using a mouse hindlimb-ischemia angiogenesis model. Hindlimb blood flow was measured by Laser Doppler Perfusion Imaging and microvessel density was determined by CD31 and tomato lectin staining. We found that systemic and local administration of a 20-HETE synthesis inhibitor, DDMS, or a 20-HETE antagonist, 6,15-20-HEDGE significantly reduced blood flow recovery and microvessel formation in response to ischemia. 20-HETE production, measured by LC/MS/MS, was markedly increased in ischemic muscles (91±11 vs. 8±2pg/mg in controls), which was associated with prominent upregulation of the 20-HETE synthase, CYP4A12. Immunofluorescence co-localized increased CYP4A12 expression in response to ischemia to CD31-positive EC in the ischemic hindlimb microvessels. We further showed that ischemia increased HIF-1α, VEGF, and VEGFR2 expression in gracilis muscles and that these increases were negated by DDMS and 6,15-20-HEDGE. Lastly, we showed that ERK1/2 of MAPK is a component of 20-HETE regulated ischemic angiogenesis. Taken together, these data indicate that 20-HETE is a critical contributor of ischemia-induced angiogenesis in vivo.


[Developmental dysphasia and epileptiform EEG activity in children].
Objective. To study the electrical activity of the brain in children with developmental dysphasia (alalia). Material and methods. We analyzed the EEGs of 65 children with developmental dysphasia, including 48 boys and 17 girls, aged from 3 to 4 years 11 months. General speech underdevelopment (GSU) of the 1st level (with active vocabulary less than 15-20 words) was found in 31 children and GSU of the 2nd level (with active vocabulary of 20-50 words) - in 34 children. To specify the changes in the brain electrical activity, we conducted video-EEG-monitoring during sleep and waking states in 27 patients. Results. Focal epileptiform EEG changes with no concomitant paroxysmal symptoms were recorded in 12,3% of children with dysphasia. The epileptiform activity was more frequent in GSU of the 1st level (5 (16.1%) patients) than in GSU of the 2nd level (3 (8.8%) patients). Benign epileptiform discharges of childhood with low index were identified in 2 (6,5%) children with GSU of the 1st level and in1 (2,9%) child with GSU of the 2nd level; low index spike-waves were recorded in 3 (9,7%) children with GSU of the 1st level and in 2 (5,9%) with GSU of the 2nd level. Conclusion. The data allow to clarify the frequency of epileptiform EEG activity in those children with developmental dysphasia, who do not have autism or history of seizures. The differential diagnosis with rare epileptic encephalopathies is needed, such as epilepsy with electrical status epilepticus during slow sleep (ESES) and Landau-Kleffner syndrome.


Patient-reported outcomes at discontinuation of anti-angiogenesis therapy in the randomized trial of chemotherapy with bevacizumab for advanced cervical cancer: an NRG Oncology Group study.
To describe patient-reported outcomes and toxicities at time of treatment discontinuation secondary to progression or toxicities in advanced/recurrent cervical cancer patients receiving chemotherapy with bevacizumab. Summarize toxicity, grade, and health-related quality of life within 1 month of treatment discontinuation for women receiving chemotherapy with bevacizumab in GOG240. Of the 227 patients who received chemotherapy with bevacizumab, 148 discontinued study protocol treatment (90 for disease progression and 58 for toxicity). The median survival time from treatment discontinuation to death was 7.9 months (95% CI 5.0 to 9.0) for those who progressed versus 12.1 months (95% CI 8.9 to 23.2) for those who discontinued therapy due to toxicities. The most common grade 3 or higher toxicities included hematologic, gastrointestinal, and pain. Some 57% (84/148) of patients completed quality of life assessment within 1 month of treatment discontinuation. Those patients who discontinued treatment due to progression had a mean decline in the FACT-Cx TOI of 3.2 points versus 2.2 in patients who discontinued therapy due to toxicity. This was a 9.9 point greater decline in the FACT-Cx TOI scores than those who discontinued treatment due to progression (95% CI 2.8 to 17.0, p=0.007). The decline in quality of life was due to worsening physical and functional well-being. Those who discontinued treatment due to toxicities had worse neurotoxicity and pain. Patients who discontinued chemotherapy with bevacizumab for toxicity experienced longer post-protocol survival but significantly greater declination in quality of life than those with progression. Future trial design should include supportive care interventions that optimize physiologic function and performance status for salvage therapies.


Phenotypic regionalization of the vertebral column in the thorny skate Amblyraja radiata: Stability and variation.
Regionalization of the vertebral column occurred early during vertebrate evolution and has been extensively investigated in mammals. However, less data are available on vertebral regions of crown gnathostomes. This is particularly true for batoids (skates, sawfishes, guitarfishes, and rays) whose vertebral column has long been considered to be composed of the same two regions as in teleost fishes despite the presence of a synarcual. However, the numerous vertebral units in chondrichthyans may display a more complex regionalization pattern than previously assumed and the intraspecific variation of such pattern deserves a thorough investigation. In this study, we use micro-computed tomography (µCT) scans of vertebral columns of a growth series of thorny skates Amblyraja radiata to provide the first fine-scale morphological description of vertebral units in a batoids species. We further investigate axial regionalization using a replicable clustering analysis on presence/absence of vertebral elements to decipher the regionalization of the vertebral column of A. radiata. We identify four vertebral regions in this species. The two anteriormost regions, named synarcual and thoracic, may undergo strong developmental or functional constraints because they display stable patterns of shapes and numbers of vertebral units across all growth stages. The third region, named hemal transitional, is characterized by high inter-individual morphological variation and displays a transition between the monospondylous (one centrum per somite) to diplospondylous (two centra per somite) conditions. The posteriormost region, named caudal, is subdivided into three sub-regions with shapes changing gradually along the anteroposterior axis. These regionalized patterns are discussed in light of ecological habits of skates.


Investigating the separate and interactive associations of trauma and depression on neurocognition in urban dwelling adults.
Trauma and depression have each been associated with neurocognitive alterations, but their combined effect on neurocognition is unclear. We investigated the separate and interactive associations of trauma and depression on neurocognition in a sample of ethnically diverse urban dwellers, and explored the impact of age on these effects. 284 adults aged 30-89 were divided into groups based on their current depression and trauma history. Individuals meeting DSM-IV criteria for depression were considered Depressed (D+) and individuals rated through diagnostic interview as having trauma history were considered positive for Trauma (T+). Resulting Ns were 73 D+T+, 56 D+T-, 68 D-T+, and 87 D-T-. A principal component analysis of neuropsychological scores resulted in a 3-factor solution representing verbal learning/memory/recognition (VERBAL-LMR), visual learning/memory/recognition, and speeded attention/cognitive flexibility accounting for 70.21% of the variance. Multivariable linear regressions adjusting for age revealed that Trauma, regardless of Depression, is associated with worse VERBAL-LMR performance. This Trauma association was driven by verbal list and prose passages learning and memory, but not recognition memory. Age-stratified (<60 versus ≥60 years) regressions revealed the Trauma association was only significant for older adults. No main or interactive effects for Depression were observed. Trauma, regardless of Depression, is associated with worse verbal learning and memory, but not recognition performance. These results suggest that trauma exposure may negatively impact neurocognition. Clinicians working with adults in urban settings should query for trauma in addition to depression when considering subjective and objective measures of neurocognitive functioning, particularly in older adults.


Total pancreatectomy with islet autotransplantation: recent updates and outcomes.
Total pancreatectomy with islet autotransplantation (TPIAT) is a reliable therapy to retain endocrine function, to alleviate pain and improve quality of life in adult and pediatric patients suffering from refractory chronic pancreatitis and recurrent acute pancreatitis. Recently, an expansion of indications to include benign and malignant pancreatic disease has been suggested. Improved methods for evaluating the functional quality of islets and predicting transplant outcome have been discussed. Furthermore, this review will discuss the recent advances in the procedure, anti-inflammatory strategies and outcomes of TPIAT. New assays to measure posttransplant islet damage and improved methods to assess islet quality by monitoring the oxygen consumption rate have shown great promise. Anti-inflammatory strategies such as an interleukin-1 receptor antagonist, α-1 antitrypsin, tumor necrosis factor α inhibitor and an inhibitor of CXCR1/2 have been widely investigated under clinical trials. The primary objective of TPIAT, which is to relieve pain and reduce narcotic usage, has been shown to have long-term success. However, achieving long-term insulin independence is still a challenge that needs to be addressed. The mechanism that leads to chronic graft failure in TPIAT needs to be delineated.


Domain organization and function in GluK2 subtype kainate receptors.
Glutamate receptor ion channels (iGluRs) are excitatory neurotransmitter receptors with a unique molecular architecture in which the extracellular domains assemble as a dimer of dimers. The structure of individual dimer assemblies has been established previously for both the isolated ligand-binding domain (LBD) and more recently for the larger amino terminal domain (ATD). How these dimers pack to form tetrameric assemblies in intact iGluRs has remained controversial. Using recently solved crystal structures for the GluK2 kainate receptor ATD as a guide, we performed cysteine mutant cross-linking experiments in full-length tetrameric GluK2 to establish how the ATD packs in a dimer of dimers assembly. A similar approach, using a full-length AMPA receptor GluA2 crystal structure as a guide, was used to design cysteine mutant cross-links for the GluK2 LBD dimer of dimers assembly. The formation of cross-linked tetramers in full-length GluK2 by combinations of ATD and LBD mutants which individually produce only cross-linked dimers suggests that subunits in the ATD and LBD layers swap dimer partners. Functional studies reveal that cross-linking either the ATD or the LBD inhibits activation of GluK2 and that, in the LBD, cross-links within and between dimers have different effects. These results establish that kainate and AMPA receptors have a conserved extracellular architecture and provide insight into the role of individual dimer assemblies in activation of ion channel gating.


Organization and evolution of repeated DNA sequences in closely related plant genomes.
In common with many other eukaryotic species, the genomes of species in the genus Allium contain a high proportion of repeated DNA sequences, which may be implicated in the considerable differences in genome size that are seen between even very closely related species. The gross organization of repetitive sequences within the genome of Allium sativum and of some other related species has been investigated using DNA/DNA hybridization studies. Such studies show that there has been much modulation in the amounts of different repeated DNA families during the evolution of the genus Allium; these repetitive elements are interspersed in all species with sequences of low repetition. The organization and distribution of one particular repetitive family within the genus has been examined using a cloned hybridization probe. Hybridization of this probe to DNA from related genomes reveals that this element is present in all Allium species examined, but with large-scale modulation of its abundance, and some considerable changes in its sequence environment. The evolution of such genome-specific arrangements of common repetitive elements and the possible mechanisms by which they might be maintained are discussed.