Conversion of RNA Aptamer into Modified DNA Aptamers Provides for Prolonged Stability and Enhanced Antitumor Activity.
Aptamers, synthetic single-strand oligonucleotides that are similar in function to antibodies, are promising as therapeutics because of their minimal side effects. However, the stability and bioavailability of the aptamers pose a challenge. We developed aptamers converted from RNA aptamer to modified DNA aptamers that target phospho-AXL with improved stability and bioavailability. On the basis of the comparative analysis of a library of 17 converted modified DNA aptamers, we selected aptamer candidates, GLB-G25 and GLB-A04, that exhibited the highest bioavailability, stability, and robust antitumor effect in in vitro experiments. Backbone modifications such as thiophosphate or dithiophosphate and a covalent modification of the 5'-end of the aptamer with polyethylene glycol optimized the pharmacokinetic properties, improved the stability of the aptamers in vivo by reducing nuclease hydrolysis and renal clearance, and achieved high and sustained inhibition of AXL at a very low dose. Treatment with these modified aptamers in ovarian cancer orthotopic mouse models significantly reduced tumor growth and the number of metastases. This effective silencing of the phospho-AXL target thus demonstrated that aptamer specificity and bioavailability can be improved by the chemical modification of existing aptamers for phospho-AXL. These results lay the foundation for the translation of these aptamer candidates and companion biomarkers to the clinic.


Practical Approach to the Evaluation of Malignant Peritoneal Fluids in the Setting of Gynecologic Neoplasms.
Evaluation of peritoneal fluid cytology, either from ascitic fluids or as a result of peritoneal washings, is a fundamental aspect in the evaluation of women presenting with clinically concerning or histologically confirmed gynecologic neoplasms. Ascitic fluid samples are often the initial and only source of diagnostic material in women presenting with gynecologic malignancies, and important therapeutic decisions will result from the information provided in the cytology report. On the other hand, cytologic evaluation of peritoneal washing specimens obtained during surgical excision of a presumed gynecologic neoplasm provides crucial information to the clinical team regarding tumor staging, often with significant therapeutic implications. While recognition of high-grade tumors in either of these samples is generally straightforward, low-grade tumors and unusual neoplasms can prove to be more difficult to recognize, differentiate from benign mimics, and correctly diagnose, particularly in low-cellularity specimens. Even with high-grade tumors, a mere diagnosis of "positive for malignancy" in diagnostic ascitic fluid specimens might not suffice to guide clinical management, and the use of ancillary techniques to further and more definitively characterize the lesional cells is required. This review will focus on the clinically relevant issues surrounding interpretation of peritoneal fluid cytology specimens in the setting of gynecologic neoplasms, making emphasis on the salient cytomorphologic and immunocytochemical features of the various neoplastic processes, in an attempt to provide a practical yet effective guide on how to best evaluate, diagnose, and report these samples.


3-Nitrotyrosine: A biomarker of nitrogen free radical species modified proteins in systemic autoimmunogenic conditions.
The free radical-mediated damage to proteins results in the modification of amino acid residues, cross-linking of side chains and fragmentation. l-Tyrosine and protein bound tyrosine are prone to attack by various mediators and reactive nitrogen intermediates to form 3-nitrotyrosine (3-NT). Activated macrophages produce superoxide (O2(·-)) and NO, which are converted to peroxynitrite ONO2(-). 3-NT formation is also catalyzed by a class of peroxidases utilizing nitrite and hydrogen peroxide as substrates. Evidence supports the formation of 3-NT in vivo in diverse pathologic conditions and 3-NT is thought to be a relatively specific marker of oxidative damage mediated by peroxynitrite. Free/protein-bound tyrosines are attacked by various RNS, including peroxynitrite, to form free/protein-bound 3-NT, which may provide insight into the etiopathogenesis of autoimmune conditions. The formation of nitrotyrosine represents a specific peroxynitrite-mediated protein modification; thus, detection of nitrotyrosine in proteins is considered as a biomarker for endogenous peroxynitrite activity. The peroxynitrite-driven oxidation and nitration of biomolecules may lead to autoimmune diseases such as systemic lupus. The subsequent release of altered proteins may enable them to act as antigen-inducing antibodies against self-proteins. Hence, tyrosine nitrated proteins can act as neoantigens and lead to the generation of autoantibodies against self proteins in various autoimmune disorders.


The effects of latamoxef, cefotaxime, and cefoperazone on platelet function and coagulation in normal volunteers.
A bleeding diathesis characterized by in-vitro platelet dysfunction and prolongation of the template bleeding time (TBT) has been reported in patients receiving latamoxef ('moxalactam'), but not cefotaxime or cefoperazone. Hypoprothrombinaemia has been associated with the use of both latamoxef and cefoperazone in seriously ill and malnourished patients. We administered either latamoxef, cefotaxime or cefoperazone intravenously, at dosages within the range recommended by each manufacturer, to 14 normal volunteers. Latamoxef caused a dose and time dependent defect in platelet function characterized in vitro by abnormalities in aggregation to adenosine diphosphate and in vivo by prolongation of the template bleeding time. In two out of two subjects, a single 4 g dose of latamoxef caused neither prolongation of template bleeding times nor aggregation abnormalities. Two out of two subjects receiving latamoxef 6 g/day for six days had progressive prolongation of bleeding times to 12 and 15 min. Two additional subjects receiving latamoxef 12 g/day for four days had prolongation of template bleeding times to greater than 20 min. Of four subjects receiving cefotaxime 12 g/day for seven days, none had prolongation of template bleeding times or abnormalities in platelet aggregations. Of four subjects receiving cefoperazone 6 g/day, none had significant prolongation of template bleeding times and one had abnormalities in aggregation attributed to inadvertent salicylate ingestion. Prolongation of the prothrombin time or activated partial thromboplastin time did not occur in any of the 14 volunteers. Latamoxef is more likely to interfere with platelet function than either cefotaxime or cefoperazone.


Mitochondria of human Leydig cells as seen by high resolution scanning electron microscopy.
The three-dimensional ultrastructure of over 1000 mitochondria in human Leydig cells (from twelve sexually mature patients) was examined by high resolution scanning electron microscopy (HRSEM) of osmium-macerated specimens, as well as by transmission electron microscopy of conventional ultrathin sections. The stereo-pair imaging of the osmium-macerated specimens by HRSEM is also very useful for investigating the three-dimensional structure of cytoplasmic membranous organelles with great clarity. The mitochondria, which mainly are elongated (although some are ovate), possess cristae that are almost exclusively tubular and that occasionally display constrictions and terminal bulbules. Lamelliform cristae are quite rare. Occasionally, the tubular cristae are joined together to form a simple network. Classic crista junctions could not be identified with certainty, although the base of the tubular cristae might correspond functionally to such junctions. As a whole, in line with the identical and common embryological origin of adrenal cortex and gonads, mitochondria of human Leydig cell closely resemble those of steroidogenic cells of human suprarenal cortex treated by the same maceration method.


Craniofacial morphology in Treacher Collins syndrome.
Treacher Collins syndrome is a rare but well studied autosomal dominant craniofacial malformation syndrome. Most reports to date have focused on the facial morphology of this distinctive syndrome, but several more recent investigations have described abnormalities of the basicranium, mandible, and upper airway. The purpose of this report is to describe craniofacial morphology in a sample population of 12 patients with Treacher Collins syndrome. A special emphasis is placed on cranial base and mandibular structure because of the airway problems often encountered in this syndrome.


Ten years after entry into force of the Stockholm Convention: What do air monitoring data tell about its effectiveness?
More than a decade ago, the Stockholm Convention on Persistent Organic Pollutants (POPs), one of the multilateral environmental agreements administered by the United Nations Environment Programme (UNEP), entered into force. The objective of this Convention is to protect human health and the environment by controlling the releases of POPs. According to its Article 16, the effectiveness of the Stockholm Convention shall be evaluated using comparable monitoring data on the presence of POPs as well as their regional and global environmental transport. Here, we present a time series analysis on atmospheric POP concentrations from 15 monitoring stations in North America and Europe that provide long-term data and have started operations between 1990 and 2003. We systematically searched for temporal trends and significant structural changes in temporal trends that might result from the provisions of the Stockholm Convention. We find that such structural changes do occur, but they are related mostly to effects of national regulations enforced prior to the implementation of the Stockholm Convention, rather than to the enforcement of the provisions laid out in the Convention. One example is that concentrations of polychlorinated biphenyls, many of which started to decrease rapidly during the 1990s. Also effects of chemical transport and fate, for instance the re-volatilization of POPs from secondary sources, are thought to be a cause of some of the observed structural changes. We conclude that a decade of air monitoring data has not been sufficient for detecting general and statistically significant effects of the Stockholm Convention. Based on these lessons, we present recommendations for the future operation of existing monitoring programs and advocate for a stricter enforcement of the provisions of the Stockholm Convention, in the current absence of proof for its effectiveness.


Faculty performance evaluation in accredited U.S. public health graduate schools and programs: a national study.
To provide baseline data on evaluation of faculty performance in U.S. schools and programs of public health. The authors administered an anonymous Internet-based questionnaire using PHP Surveyor. The invited sample consisted of individuals listed in the Council on Education for Public Health (CEPH) Directory of Accredited Schools and Programs of Public Health. The authors explored performance measures in teaching, research, and service, and assessed how faculty performance measures are used. A total of 64 individuals (60.4%) responded to the survey, with 26 (40.6%) reporting accreditation/reaccreditation by CEPH within the preceding 24 months. Although all schools and programs employ faculty performance evaluations, a significant difference exists between schools and programs in the use of results for merit pay increases and mentoring purposes. Thirty-one (48.4%) of the organizations published minimum performance expectations. Fifty-nine (92.2%) of the respondents counted number of publications, but only 22 (34.4%) formally evaluated their quality. Sixty-two (96.9%) evaluated teaching through student course evaluations, and only 29 (45.3%) engaged in peer assessment. Although aggregate results of teaching evaluation are available to faculty and administrators, this information is often unavailable to students and the public. Most schools and programs documented faculty service activities qualitatively but neither assessed it quantitatively nor evaluated its impact. This study provides insight into how schools and programs of public health evaluate faculty performance. Results suggest that although schools and programs do evaluate faculty performance on a basic level, many do not devote substantial attention to this process.


PDP-1 links the TGF-β and IIS pathways to regulate longevity, development, and metabolism.
The insulin/IGF-1 signaling (IIS) pathway is a conserved regulator of longevity, development, and metabolism. In Caenorhabditis elegans IIS involves activation of DAF-2 (insulin/IGF-1 receptor tyrosine kinase), AGE-1 (PI 3-kinase), and additional downstream serine/threonine kinases that ultimately phosphorylate and negatively regulate the single FOXO transcription factor homolog DAF-16. Phosphatases help to maintain cellular signaling homeostasis by counterbalancing kinase activity. However, few phosphatases have been identified that negatively regulate the IIS pathway. Here we identify and characterize pdp-1 as a novel negative modulator of the IIS pathway. We show that PDP-1 regulates multiple outputs of IIS such as longevity, fat storage, and dauer diapause. In addition, PDP-1 promotes DAF-16 nuclear localization and transcriptional activity. Interestingly, genetic epistasis analyses place PDP-1 in the DAF-7/TGF-β signaling pathway, at the level of the R-SMAD proteins DAF-14 and DAF-8. Further investigation into how a component of TGF-β signaling affects multiple outputs of IIS/DAF-16, revealed extensive crosstalk between these two well-conserved signaling pathways. We find that PDP-1 modulates the expression of several insulin genes that are likely to feed into the IIS pathway to regulate DAF-16 activity. Importantly, dysregulation of IIS and TGF-β signaling has been implicated in diseases such as Type 2 Diabetes, obesity, and cancer. Our results may provide a new perspective in understanding of the regulation of these pathways under normal conditions and in the context of disease.


Neuraminidase treatment of isolated rat adipocytes and differential regulation of basal and insulin-stimulated glucose transport.
The role of membrane carbohydrate in the function of insulin receptors and glucose transporters was investigated with neuraminidase to release sialic acid from isolated rat adipocytes. Pretreatment of adipocytes with neuraminidase resulted in an increase in basal glucose transport. At the same time, insulin-stimulated glucose transport was reduced by an average of 30%. The enzyme action on glucose transport was not due to a nonspecific membrane perturbation because neuraminidase caused only a nonsignificant decrease in the uptake of the amino acid analog alpha-(methylamino)isobutyric acid and had no effect on basal or insulin-stimulated protein synthesis. Insulin binding was slightly increased in neuraminidase-treated cells, yet the shapes of the dose-response curves for insulin stimulation of glucose transport were similar (EC50 = 0.087 +/- 0.010 and 0.082 +/- 0.008 nM for control and treated cells, respectively). The neuraminidase-induced increase in basal transport was the result of an increase in the affinity of the glucose-transport system (Km = 7.3 +/- 1.4 and 3.6 +/- 0.7 mM before and after treatment, respectively) with no change in Vmax. Conversely, enzyme treatment decreased the Vmax of insulin-stimulated 3-O-methylglucose transport from 87.8 +/- 13.2 to 41.3 +/- 4.9 pmol.2 x 10(5) cells-1.s-1 while not altering the Km. These changes in glucose-transport activity resulting from enzyme treatment were not due to alterations in glucose-transporter concentration on the plasma membrane as measured by the D-glucose-inhibitable binding of [3H]cytochalasin B. Thus, sialic acid plays multiple roles in the control of glucose-transport activity.(ABSTRACT TRUNCATED AT 250 WORDS)


An Avocado Extract Enriched in Mannoheptulose Prevents the Negative Effects of a High-Fat Diet in Mice.
Beginning at 16 weeks of age and continuing for 44 weeks, male C57BL/6J were fed either a control (CON) diet; a high-fat (HF) diet (60% unsaturated); or the HF diet containing an extract of unripe avocados (AvX) enriched in the 7-carbon sugar mannoheptulose (MH), designed to act as a glycolytic inhibitor (HF + MH). Compared to the CON diet, mice on the HF diet exhibited higher body weights; body fat; blood lipids; and leptin with reduced adiponectin levels, insulin sensitivity, VO2max, and falls from a rotarod. Mice on the HF + MH diet were completely protected against these changes in the absence of significant diet effects on food intake. Compared to the CON diet, oxidative stress was also increased by the HF diet indicated by higher levels of total reactive oxygen species, superoxide, and peroxynitrite measured in liver samples by electron paramagnetic resonance spectroscopy, whereas the HF + MH diet attenuated these changes. Compared to the CON, the HF diet increased signaling in the mechanistic target of the rapamycin (mTOR) pathway, and the addition of the MH-enriched AvX to this diet attenuated these changes. Beyond generating further interest in the health benefits of avocados, these results draw further new attention to the effects of this rare sugar, MH, as a botanical intervention for preventing obesity.


Different effects of dominant negative mutants of desmocollin and desmoglein on the cell-cell adhesion of keratinocytes.
Desmosomes contain two types of cadherin: desmocollin (Dsc) and desmoglein (Dsg). In this study, we examined the different roles that Dsc and Dsg play in the formation of desmosomes, by using dominant-negative mutants. We constructed recombinant adenoviruses (Ad) containing truncated mutants of E-cadherin, desmocollin 3a, and desmoglein 3 lacking a large part of their extracellular domains (EcaddeltaEC, Dsc3adeltaEC, Dsg3deltaEC), using the Cre-loxP Ad system to circumvent the problem of the toxicity of the mutants to virus-producing cells. When Dsc3adeltaEC Ad-infected HaCaT cells were cultured with high levels of calcium, E-cadherin and beta-catenin, which are marker molecules for the adherens junction, disappeared from the cell-cell contact sites, and cell-cell adhesion was disrupted. This also occurred in the cells infected with EcaddeltaEC Ad. With Dsg3deltaEC Ad infection, keratin insertion at the cell-cell contact sites was inhibited and desmoplakin, a marker of desmosomes, was stained in perinuclear dots while the adherens junctions remained intact. Dsc3adeltaEC Ad inhibited the induction of adherens junctions and the subsequent formation of desmosomes with the calcium shift, while Dsg3deltaEC Ad only inhibited the formation of desmosomes. To further determine whether Dsc3adeltaEC directly affected adherens junctions, mouse fibroblast L cells transfected with E-cadherin (LEC5) were infected with these mutant Ads. Both Dsc3adeltaEC and EcaddeltaEC inhibited the cell-cell adhesion of LEC5 cells, as determined by the cell aggregation assay, while Dsg3deltaEC did not. These results indicate that the dominant negative effects of Dsg3deltaEC were restricted to desmosomes, while those of Dsc3adeltaEC were observed in both desmosomes and adherens junctions. Furthermore, the cytoplasmic domain of Dsc3adeltaEC coprecipitated both plakoglobin and beta-catenin in HaCaT cells. In addition, beta-catenin was found to bind the endogenous Dsc in HaCaT cells. These findings lead us to speculate that Dsc interacts with components of the adherens junctions through beta-catenin, and plays a role in nucleating desmosomes after the adherens junctions have been established.


ChatGPT in Drug Discovery: A Case Study on Anti-Cocaine Addiction Drug Development with Chatbots.
The birth of ChatGPT, a cutting-edge language model-based chatbot developed by OpenAI, ushered in a new era in AI. However, due to potential pitfalls, its role in rigorous scientific research is not clear yet. This paper vividly showcases its innovative application within the field of drug discovery. Focused specifically on developing anti-cocaine addiction drugs, the study employs GPT-4 as a virtual guide, offering strategic and methodological insights to researchers working on generative models for drug candidates. The primary objective is to generate optimal drug-like molecules with desired properties. By leveraging the capabilities of ChatGPT, the study introduces a novel approach to the drug discovery process. This symbiotic partnership between AI and researchers transforms how drug development is approached. Chatbots become facilitators, steering researchers towards innovative methodologies and productive paths for creating effective drug candidates. This research sheds light on the collaborative synergy between human expertise and AI assistance, wherein ChatGPT's cognitive abilities enhance the design and development of potential pharmaceutical solutions. This paper not only explores the integration of advanced AI in drug discovery but also reimagines the landscape by advocating for AI-powered chatbots as trailblazers in revolutionizing therapeutic innovation.


Germicidal ultraviolet irradiation in air conditioning systems: effect on office worker health and wellbeing: a pilot study.
The indoor environment of modern office buildings represents a new ecosystem that has been created totally by humans. Bacteria and fungi may contaminate this indoor environment, including the ventilation systems themselves, which in turn may result in adverse health effects. The objectives of this study were to test whether installation and operation of germicidal ultraviolet (GUV) lights in central ventilation systems would be feasible, without adverse effects, undetected by building occupants, and effective in eliminating microbial contamination. GUV lights were installed in the ventilation systems serving three floors of an office building, and were turned on and off during a total of four alternating 3 week blocks. Workers reported their environmental satisfaction, symptoms, as well as sickness absence, without knowledge of whether GUV lights were on or off. The indoor environment was measured in detail including airborne and surface bacteria and fungi. Airborne bacteria and fungi were not significantly different whether GUV lights were on or off, but were virtually eliminated from the surfaces of the ventilation system after 3 weeks of operation of GUV light. Of the other environmental variables measured, only total airborne particulates were significantly different under the two experimental conditions--higher with GUV lights on than off. Of 113 eligible workers, 104 (87%) participated; their environmental satisfaction ratings were not different whether GUV lights were on or off. Headache, difficulty concentrating, and eye irritation occurred less often with GUV lights on whereas skin rash or irritation was more common. Overall, the average number of work related symptoms reported was 1.1 with GUV lights off compared with 0.9 with GUV lights on. Installation and operation of GUV lights in central heating, ventilation and air conditioning systems of office buildings is feasible, cannot be detected by workers, and does not seem to result in any adverse effects.


Network geometry with flavor: From complexity to quantum geometry.
Network geometry is attracting increasing attention because it has a wide range of applications, ranging from data mining to routing protocols in the Internet. At the same time advances in the understanding of the geometrical properties of networks are essential for further progress in quantum gravity. In network geometry, simplicial complexes describing the interaction between two or more nodes play a special role. In fact these structures can be used to discretize a geometrical d-dimensional space, and for this reason they have already been widely used in quantum gravity. Here we introduce the network geometry with flavor s=-1,0,1 (NGF) describing simplicial complexes defined in arbitrary dimension d and evolving by a nonequilibrium dynamics. The NGF can generate discrete geometries of different natures, ranging from chains and higher-dimensional manifolds to scale-free networks with small-world properties, scale-free degree distribution, and nontrivial community structure. The NGF admits as limiting cases both the Bianconi-Barabási models for complex networks, the stochastic Apollonian network, and the recently introduced model for complex quantum network manifolds. The thermodynamic properties of NGF reveal that NGF obeys a generalized area law opening a new scenario for formulating its coarse-grained limit. The structure of NGF is strongly dependent on the dimensionality d. In d=1 NGFs grow complex networks for which the preferential attachment mechanism is necessary in order to obtain a scale-free degree distribution. Instead, for NGF with dimension d>1 it is not necessary to have an explicit preferential attachment rule to generate scale-free topologies. We also show that NGF admits a quantum mechanical description in terms of associated quantum network states. Quantum network states evolve by a Markovian dynamics and a quantum network state at time t encodes all possible NGF evolutions up to time t. Interestingly the NGF remains fully classical but its statistical properties reveal the relation to its quantum mechanical description. In fact the δ-dimensional faces of the NGF have generalized degrees that follow either the Fermi-Dirac, Boltzmann, or Bose-Einstein statistics depending on the flavor s and the dimensions d and δ.


Comparison of the prognostic value of assessing tumor diameter versus tumor volume at diagnosis or in response to initial chemotherapy in rhabdomyosarcoma.
In this study on a series of 205 patients with rhabdomyosarcoma, we investigated whether the prognostic effect of tumor size, at diagnosis or in terms of tumor response after induction chemotherapy, differed when tumor diameter or tumor volume were considered. Tumor size was assessed radiologically at diagnosis and, for the 108 patients with measurable disease, after three courses of chemotherapy. The analysis was based on multivariable models (linear for association between size and patient/tumor characteristics, Cox for association with survival). The predictive performance of the Cox model (estimated by V measure) was compared for the tumor's diameter and volume. Initial tumor size was significantly larger in male or older patients and in T2 or alveolar tumors, but was not associated with the achievement of complete surgical resection. Initial tumor size significantly influenced overall survival. The risk of death was comparable for tumors 10 cm in maximum diameter and 194.0 cm(3) in volume. The predictive performance of the Cox model was much the same when the tumor's diameter or volume was considered. Tumor response was a significant predictor of survival, again irrespective of the type of tumor measurement considered. In our analysis, initial tumor size and tumor response were significant prognostic factors in rhabdomyosarcoma, regardless of whether tumor diameter or volume was considered. Three-dimensional tumor assessment was of no greater prognostic value than one-dimensional assessment, neither initially nor after induction treatment.


Watermarking in the space/spatial-frequency domain using two-dimensional Radon-Wigner distribution.
A two-dimensional (2-D) signal with a variable spatial frequency is proposed as a watermark in the spatial domain. This watermark is characterized by a linear frequency change. It can be efficiently detected by using 2-D space/spatial-frequency distributions. The projections of the 2-D Wigner distribution--the 2-D Radon-Wigner distribution, are used in order to emphasize the watermark detection process. The watermark robustness with respect to some very important image processing attacks, such as for example, the translation, rotation, cropping, JPEG compression, and filtering, is demonstrated and tested by using Stirmark 3.1.


A qualitative study on HIV risk behaviors and medical needs of sex workers in a China/Myanmar border town.
Ruili is a small border town between China and Myanmar where drugs and commercial sex are common, and rates of sexually transmitted diseases (STDs) including HIV, are high. A qualitative study was carried out on 89 commercial sex workers there in 2001 to understand more about their HIV awareness, medical-seeking behaviors and needs. We found that the sex workers were young and the turnover rates were high. Contrary to common belief, many came from nearby villages or cities, but were probably reluctant to participate in organized activities. Their medical knowledge was very limited, often acquired from peers and self-medication was common. The contraception they used was inappropriate and screening for cervical cancer was nonexistent. They were very stigma conscious. Condoms were purchased in small quantities when required and used only if the clients were agreeable. These findings have strong implications for the future planning of services and HIV/STD prevention.


Translation elongation factor-1 alpha interacts with the 3' stem-loop region of West Nile virus genomic RNA.
The conserved 3'-terminal stem-loop (3' SL) of the West Nile virus (WNV) genomic RNA was previously used to probe for cellular proteins that may be involved in flavivirus replication and three cellular proteins were detected that specifically interact with the WNV 3' SL RNA (J. L. Blackwell and M. A. Brinton, J. Virol. 69:5650-5658, 1995). In this study, one of these cellular proteins was purified to apparent homogeneity by ammonium sulfate precipitation and liquid chromatography. Amino acid sequence Western blotting, and supershift analyses identified the cellular protein as translation elongation factor-1 alpha (EF-1 alpha). Competition gel mobility shift assays demonstrated that the interaction between EF-1 alpha and WNV 3' SL RNA was specific. Dephosphorylation of EF-1 alpha by calf intestinal alkaline phosphatase inhibited its binding to WNV 3' SL RNA. The apparent equilibrium dissociation constant for the interaction between EF-1 alpha and WNV 3' SL RNA was calculated to be 1.1 x 10(-9) M. Calculation of the stoichiometry of the interaction indicated that one molecule of EF-1 alpha binds to each molecule of WNV 3' SL RNA. Using RNase footprinting and nitrocellulose filter binding assays, we detected a high-activity binding site on the main stem of the WNV 3' SL RNA. Interaction with EF-1 alpha at the high-activity binding site was sequence specific, since nucleotide substitution in this region reduced the binding activity of the WNV 3' SL RNA for EF-1 alpha by approximately 60%. Two low-activity binding sites were also detected, and each accounted for approximately 15 to 20% of the binding activity. Intracellular association between the host protein and the viral RNA was suggested by coimmunoprecipitation of WNV genomic RNA and EF-1 alpha, using an anti-EF-1 alpha antibody.


Monitoring large-scale changes in transcript abundance in drought- and salt-stressed barley.
Responses to drought and salinity in barley (Hordeum vulgare L. cv. Tokak) were monitored by microarray hybridization of 1463 DNA elements derived from cDNA libraries of 6 and 10 h drought-stressed plants. Functional identities indicated that many cDNAs in these libraries were associated with drought stress. About 38% of the transcripts were novel and functionally unknown. Hybridization experiments were analyzed for drought- and salinity-regulated sequences, with significant changes defined as a deviation from the control exceeding 2.5-fold. Responses of transcripts showed stress-dependent expression patterns and time courses. Nearly 15% of all transcripts were either up- or down-regulated under drought stress, while NaCl led to a change in 5% of the transcripts (24 h, 150 mM NaCl). Transcripts that showed significant up-regulation under drought stress are exemplified by jasmonate-responsive, metallothionein-like, late-embryogenesis-abundant (LEA) and ABA-responsive proteins. Most drastic down-regulation in a category was observed for photosynthesis-related functions. Up-regulation under both drought and salt stress was restricted to ESTs for metallothionein-like and LEA proteins, while increases in ubiquitin-related transcripts characterized salt stress. A number of functionally unknown transcripts from cDNA libraries of drought-stressed plants showed up-regulation by drought but down-regulation by salt stress, documenting how precisely transcript profiles report different growth conditions and environments.


Electroacupuncture Treats Myocardial Infarction by Influencing the Regulation of Substance P in the Neurovascular to Modulate PGI2/TXA2 Metabolic Homeostasis via PI3K/AKT Pathway: A Bioinformatics-Based Multiomics and Experimental Study.
Acute myocardial infarction (AMI) is the most severe form of coronary heart disease caused by ischemia and hypoxia. The study is aimed at investigating the role of neuropeptides and the mechanism of electroacupuncture (EA) in acute myocardial infarction (AMI) treatment. Compared with the normal population, a significant increase in substance P (SP) was observed in the serum of patients with AMI. PGI2 expression was increased in the SP-treated AMI mouse model, and TXA2 expression was decreased. And PI3K pathway-related genes, including Pik3ca, Akt, and Mtor, were upregulated in myocardial tissue of SP-treated AMI patients. Human cardiomyocyte cell lines (HCM) treated with SP increased mRNA and protein expression of PI3K pathway-related genes (Pik3ca, Pik3cb, Akt, and Mtor). Compared to MI control and EA-treated MI rat models, Myd88, MTOR, Akt1, Sp, and Irak1 were differentially expressed, consistent with in vivo and in vitro studies. EA treatment significantly enriched PI3K/AKT signaling pathway genes within MI-associated differentially expressed genes (DEGs) according to Kyoto Encyclopedia of Genes and Genomes (KEGG). Furthermore, it was confirmed by molecular docking analysis that PIK3CA, AKT1, and mTOR form stable dockings with neuropeptide SP. PI3K/AKT pathway activity may be affected directly or indirectly by EA via SP, which corrects the PGI2/TXA2 metabolic imbalance in AMI. MI treatment is now better understood as a result of this finding.


p53 modulates acquired resistance to EGFR inhibitors and radiation.
There is presently great interest in mechanisms of acquired resistance to epidermal growth factor receptor (EGFR) inhibitors that are now being used widely in the treatment of a variety of common human cancers. To investigate these mechanisms, we established EGFR inhibitor-resistant clones from non-small cell lung cancer cells. A comparative analysis revealed that acquired resistance to EGFR inhibitors was associated consistently with the loss of p53 and cross-resistance to radiation. To examine the role of p53, we first knocked down p53 in sensitive parental cells and found a reduction in sensitivity to both EGFR inhibitors and radiation. Conversely, restoration of functional p53 in EGFR inhibitor-resistant cells was sufficient to resensitize them to EGFR inhibitors or radiation in vitro and in vivo. Further studies indicate that p53 may enhance sensitivity to EGFR inhibitors and radiation via induction of cell-cycle arrest, apoptosis, and DNA damage repair. Taken together, these findings suggest a central role of p53 in the development of acquired resistance to EGFR inhibitors and prompt consideration to apply p53 restoration strategies in future clinical trials that combine EGFR inhibitors and radiation.


Coronary Artery Stenting Affects Wall Shear Stress Topological Skeleton.
Despite the important advancements in the stent technology for the treatment of diseased coronary arteries, major complications still affect the postoperative long-term outcome. The stent-induced flow disturbances, and especially the altered wall shear stress (WSS) profile at the strut level, play an important role in the pathophysiological mechanisms leading to stent thrombosis (ST) and in-stent restenosis (ISR). In this context, the analysis of the WSS topological skeleton is gaining more and more interest by extending the current understanding of the association between local hemodynamics and vascular diseases. This study aims to analyze the impact that a deployed coronary stent has on the WSS topological skeleton. Computational fluid dynamics (CFD) simulations were performed in three stented human coronary artery geometries reconstructed from clinical images. The selected cases presented stents with different designs (i.e., two contemporary drug-eluting stents and one bioresorbable scaffold) and included regions with stent malapposition or overlapping. A recently proposed Eulerian-based approach was applied to analyze the WSS topological skeleton features. The results highlighted that the presence of single or multiple stents within a coronary artery markedly impacts the WSS topological skeleton. In particular, repetitive patterns of WSS divergence were observed at the luminal surface, highlighting a WSS contraction action exerted proximal to the stent struts and a WSS expansion action distal to the stent struts. This WSS action pattern was independent from the stent design. In conclusion, these findings could contribute to a deeper understanding of the hemodynamics-driven processes underlying ST and ISR.


Unprecedented reactivity of polyamines with aldehydic DNA modifications: structural determinants of reactivity, characterization and enzymatic stability of adducts.
Apurinic/apyrimidinic (AP) sites, 5-formyluracil (fU) and 5-formylcytosine (fC) are abundant DNA modifications that share aldehyde-type reactivity. Here, we demonstrate that polyamines featuring at least one secondary 1,2-diamine fragment in combination with aromatic units form covalent DNA adducts upon reaction with AP sites (with concomitant cleavage of the AP strand), fU and, to a lesser extent, fC residues. Using small-molecule mimics of AP site and fU, we show that reaction of secondary 1,2-diamines with AP sites leads to the formation of unprecedented 3'-tetrahydrofuro[2,3,4-ef]-1,4-diazepane ('ribodiazepane') scaffold, whereas the reaction with fU produces cationic 2,3-dihydro-1,4-diazepinium adducts via uracil ring opening. The reactivity of polyamines towards AP sites versus fU and fC can be tuned by modulating their chemical structure and pH of the reaction medium, enabling up to 20-fold chemoselectivity for AP sites with respect to fU and fC. This reaction is efficient in near-physiological conditions at low-micromolar concentration of polyamines and tolerant to the presence of a large excess of unmodified DNA. Remarkably, 3'-ribodiazepane adducts are chemically stable and resistant to the action of apurinic/apyrimidinic endonuclease 1 (APE1) and tyrosyl-DNA phosphoesterase 1 (TDP1), two DNA repair enzymes known to cleanse a variety of 3' end-blocking DNA lesions.


PET (positron emission tomography) imaging of biomolecules using metal-DOTA complexes: a new collaborative challenge by chemists, biologists, and physicians for future diagnostics and exploration of in vivo dynamics.
Recently, PET has been paid a great deal of attention as a non-invasive imaging method. In this review, the recent advances of PET using biomolecules, such as peptides, monoclonal antibodies, proteins, oligonucleotides, and glycoproteins will be described. So far, PET of biomolecules has been mainly used for diagnosis of cancers. The biomolecules have been conjugated with the DOTA ligand, labeled with radiometals as the beta+ emitter, and targeted to specific tumors, where they have enabled visualization of even small metastatic lesions, due to the high sensitivity of the PET scanners. Some of the biomolecules have been used not only for PET diagnosis, but also for radiotherapeutic treatments by simply changing the radiometals to beta(-) emitters. Collaborative work between chemists, biologists, and physicians will be important for the future of biomolecule-based targeting and diagnosis.


[A case of initially unresectable pancreatic cancer with arterial invasion successfully resected after chemotherapy].
A 63-year-old man was diagnosed with pancreatic head cancer, and an operation was planned at his previous hospital. However, resection was not possible because of invasion of the superior mesenteric artery, and gemcitabine-based chemotherapy was administered for the tumor for 2 years. Imaging examinations after chemotherapy were unable to detect the tumor at the pancreatic head or the arterial invasion. The patient was subsequently referred to our hospital for surgery. Upon laparotomy, the tumor was judged to be resectable, and a pancreaticoduodenectomy was performed. Pathological examination indicated the presence of cancer cells in the resected specimens. The patient underwent adjuvant chemotherapy and is currently alive without recurrence. This case highlights the importance of ongoing follow-up in patients with locally advanced pancreatic cancer so that tumor resectability can be evaluated during chemotherapy.


Umbilical hernia in adults: day case local anaesthetic repair.
The waiting times for elective surgery of Umbilical hernia (UH) in adults are unacceptably long in some cases. During this period, irreducibility and strangulation are possible. We operate on adult patients under local anaesthesia (LA) as day cases to avoid this delay and describe our experience in this paper. The aims of our study were to look at the age and sex distribution, body weight, type and amount of local anaesthetic used, morbidity, admission and readmission rates, and waiting times of adult patients operated on for UH under LA. It was a retrospective study covering a 4 year period from July 1996 to June 2000 including all adult patients undergoing the above procedure under the care of a single consultant general surgeon. A standard Mayo repair using non absorbable material was used without a mesh or a drain. 32 patients with UH were operated on under LA, 23 males and 9 females with a median age of 51 years (range 20 to 86 years). The body weight ranged from 63 to 120 (median 87) kg. The average duration of the procedure was 30 (range 22-40) minutes. Sedation was needed in 4 patients. Two patients developed wound infections, one superficial and one deep. There was no mortality. The median period of follow-up was 24 (range 4-48) months and there was no recurrence. The median waiting time for the operation was 6 weeks. Day case local anaesthetic repair of UH in adults seems to be safe and feasible with an acceptable morbidity. Suture repair in the right patient has excellent results and the waiting times are acceptable.


Gemcitabine for the treatment of advanced nonsmall cell lung cancer.
Gemcitabine is a pyrimidine nucleoside antimetabolite agent which is active in several human malignancies, including nonsmall cell lung cancer (NSCLC). Because of its acceptable toxicity profile, with myelosuppression being the most common adverse event, gemcitabine can be safely combined with a number of cytotoxic agents, including platinum derivatives and new-generation anticancer compounds. In fact, the combination of gemcitabine and cisplatin is a first-line treatment for patients with advanced NSCLC, pharmacoeconomic data indicating that it represents the most cost-effective regimen among platinum-based combinations with third-generation cytotoxic drugs. The drug has been investigated in the context of nonplatinum-based regimens in a number of prospective clinical trials, and might provide a suitable alternative for patients with contraindications to platinum. Recently, gemcitabine-based doublets have been successfully tested in association with novel targeted agents with encouraging results, providing further evidence for the role of the drug in the treatment of NSCLC. In the last few years several attempts have been pursued in order to identify molecular predictors of gemcitabine activity, and recent data support the feasibility of genomic-based approaches to customize treatment with the ultimate goal of improving patient outcome.


Smoking and Alcohol Consumption Following a New Dementia Diagnosis.
Despite extensive research on lifestyle factors that influence the risk of developing dementia, limited evidence exists on whether older adults adopt healthier habits post-diagnosis in the hope of preserving their quality of life. Using panel data (1998-2014) from the Health and Retirement Study, this study investigated whether individuals who received a new dementia diagnosis were more likely to modify smoking and drinking behaviors than those without such a diagnosis. Propensity score weighting was used to adjust for observable differences between groups. Older adults with a new dementia diagnosis were 2.8 times more likely to reduce alcohol consumption than those without such a diagnosis. This result was mainly attributable to "light" drinkers at baseline and appeared to fade over time. We found no statistically significant effect of a dementia diagnosis on smoking cessation or on reduction in the number of cigarettes smoked. These results were robust to multiple sensitivity tests, including the use of cognition scores to indicate dementia onset instead of self-reported physician diagnosis. A new dementia diagnosis can serve as a window of opportunity that prompts some older adults to change habits related to alcohol consumption. This has important implications for clinical practice surrounding dementia diagnosis disclosure, the rates of which are currently much lower than other medical conditions.


[Quality of clinical studies published in the RBGO over one decade (1999-2009): methodological and ethical aspects and statistical procedures].
To evaluate the methodological and statistical design evolution of the publications in the Brazilian Journal of Gynecology and Obstetrics (RBGO) from resolution 196/96. A review of 133 articles published in 1999 (65) and 2009 (68) was performed by two independent reviewers with training in clinical epidemiology and methodology of scientific research. We included all original clinical articles, case and series reports and excluded editorials, letters to the editor, systematic reviews, experimental studies, opinion articles, besides abstracts of theses and dissertations. Characteristics related to the methodological quality of the studies were analyzed in each article using a checklist that evaluated two criteria: methodological aspects and statistical procedures. We used descriptive statistics and the χ2 test for comparison of the two years. There was a difference between 1999 and 2009 regarding the study and statistical design, with more accuracy in the procedures and the use of more robust tests between 1999 and 2009. In RBGO, we observed an evolution in the methods of published articles and a more in-depth use of the statistical analyses, with more sophisticated tests such as regression and multilevel analyses, which are essential techniques for the knowledge and planning of health interventions, leading to fewer interpretation errors.


Characterizing Neural Entrainment to Hierarchical Linguistic Units using Electroencephalography (EEG).
To understand speech, listeners have to combine the words they hear into phrases and sentences. Recent magnetoencephalography (MEG) and electrocorticography (ECoG) studies show that cortical activity is concurrently entrained/synchronized to the rhythms of multiple levels of linguistic units including words, phrases, and sentences. Here we investigate whether this phenomenon can be observed using electroencephalography (EEG), a technique that is more widely available than MEG and ECoG. We show that the EEG responses concurrently track the rhythms of hierarchical linguistic units such as syllables/words, phrases, and sentences. The strength of the sentential-rate response correlates with how well each subject can detect random words embedded in a sequence of sentences. In contrast, only a syllabic-rate response is observed for an unintelligible control stimulus. In sum, EEG provides a useful tool to characterize neural encoding of hierarchical linguistic units, potentially even in individual participants.


Neonatal uterine and vaginal cell proliferation and adenogenesis are independent of estrogen receptor 1 (ESR1) in the mouse.
Neonatal uterus and vagina express estrogen receptor 1 (ESR1) and respond mitogenically to exogenous estrogens. However, neonatal ovariectomy does not inhibit preweaning uterine cell proliferation, indicating that this process is estrogen independent. Extensive literature suggests that ESR1 can be activated by growth factors in a ligand-independent manner and drive uterine cell proliferation. Alternatively, neonatal uterine cell proliferation could be ESR1 independent despite its obligatory role in adult luminal epithelial proliferation. To determine ESR1's role in uterine and vaginal development, we analyzed cell proliferation, apoptosis, and uterine gland development (adenogenesis) in wild-type (WT) and Esr1 knockout (Esr1KO) mice from Postnatal Day 2 to Postnatal Day 60. Uterine and vaginal cell proliferation, apoptosis, and uterine adenogenesis were comparable in WT and Esr1KO mice before weaning. By Days 29-60, glands had regressed, and uterine cell proliferation was reduced in Esr1KO mice in contrast to continued adenogenesis and proliferation in WT. Apoptosis in Esr1KO uterine epithelium was not increased compared to WT at any age, indicating that differences in cell proliferation, rather than apoptosis, cause divergence of uterine size in these two groups at puberty. Similarly, vaginal epithelial proliferation was reduced, and the epithelium became atrophic in Esr1KO mice by 29 days of age and later in Esr1KO mice. These results indicate that preweaning uterine and vaginal development is ESR1 independent but becomes dependent on ESR1 by Day 29 on. It is not yet clear what mechanisms drive preweaning vaginal and uterine development, but ligand-independent activation of ESR1 is not involved.


Dose response of a new phytase on dry matter, calcium, and phosphorus digestibility in weaned piglets.
The present study evaluated the dose response of Buttiauxella phytase on apparent total tract digestibility (ATTD) of DM, Ca, and P in weaned pigs at 2 locations. Experimental diets fed to weaned pigs were a positive control (PC), a negative control (NC), and NC supplemented with increasing levels of Buttiauxella phytase. In Trial A, ATTD of P was 57.2% for PC, 32.5% for NC, and 59.4, 62.0, 63.8, 66.0, and 67.3% for 250, 500, 750, 1000, and 2000 phytase units (FTU) added to NC, respectively. In Trial B, ATTD of P was 45.2% for PC, 28.4% for NC, and 58.7, 64.1, 67.9, and 70.9% for 250, 500, 1000, and 2000 FTU added to NC, respectively. In both studies, the reduction in P in the NC diets reduced (P < 0.01) ATTD of P when compared with the PC diets. Phytase supplementation linearly and quadratically increased (P < 0.01) ATTD of P at all inclusion levels to the NC diet. In conclusion, the average digestible P increase from Buttiauxella phytase (vs. the NC diet) was 1.3, 1.5, 1.6, and 1.7 g digestible P/kg feed for 250, 500, 1000, and 2000 FTU/kg, respectively.


Randomized Clinical Trial of Hybrid vs. Surgical vs. Percutaneous Multivessel Coronary Revascularization: 5‑year Follow-up of HREVS Trial.
Aim To evaluate 5-year results of the HREVS (Hybrid REvascularization Versus Standarts) study.Material and methods The study included 155 consecutive patients with multivessel coronary artery disease who were randomized into 3 groups: coronary artery bypass grafting (CABG) (n=50), hybrid coronary revascularization (HCR) (n=52) and percutaneous coronary intervention (PCI) (n=53) according to the consensus of the cardiology team on the technical and clinical feasibility of each of the three coronary revascularization strategies. The primary endpoint of the study was residual ischemia 12 months after revascularization according to data of single-photon emission computed tomography (SPECT). Secondary endpoints were major adverse cardiac and cerebrovascular events (MACCE) over 5 years of follow-up, which included all-cause death, myocardial infarction, stroke, and clinically determined repeat myocardial revascularization.Results Baseline characteristics of patients did not differ between study groups. Median residual ischemia determined by SPECT data after 12 months was not statistically significantly different in the CABG, HCR and PCI groups: 6.7 [4.6; 8.8]%, 6.4 [4.3; 8.5]% and 7.9 [5.9; 9.8]%, respectively (p=0.45). Mean follow-up period was 76.5 months (at least 60 months). There were no statistically significant differences in all-cause mortality between the CABG, HCR and PCI groups, 10.6, 12.8 and 8.2 %, respectively (p=0.23). Statistically significant differences between the groups of CABG, HCR and PCI in the incidence of myocardial infarction (12.8; 8.5 and 16.3 %; p=0.12), stroke (4.2; 6.4 and 10.2 % ; p=0.13), repeat revascularization for clinical indications (23.4; 23.4 and 34.7 %; p=0.11) were not observed either. However, the cumulative 5-year MACCE value was similar in the HCR group and the CABG group but significantly lower than in the PCI group (51.1, 51.1 and 69.4 %, respectively; p = 0.03).Conclusion HCR that combines advantages of PCI and CABG is a promising strategy for coronary revascularization in multivessel coronary artery disease. HCR demonstrates satisfactory long-term results comparable to those of CABG but superior to PCI. To confirm the safety and efficacy of HCR, a large multicenter study is required that would have a sufficient power to evaluate clinical endpoints.


Identification of the rhesus monkey HLA-G ortholog. Mamu-G is a pseudogene.
HLA-G is a nonclassical MHC class I molecule that is primarily expressed in the placenta. To investigate whether rhesus monkeys possess an HLA-G ortholog, we cloned and sequenced MHC class I cDNAs from the rhesus placenta. We identified two rhesus MHC class I cDNAs with sequence similarity to HLA-G. Each cDNA contained premature stop codons and frameshift mutations, suggesting that it was derived from an MHC class I pseudogene. Gene trees constructed using MHC class I alleles from human and nonhuman primates revealed that the rhesus placental pseudogene alleles clustered with HLA-G orthologs from the human, chimpanzee, and gorilla. These data suggested that this rhesus MHC class I pseudogene is an HLA-G ortholog. This locus was, therefore, designated Mamu (Macaca mulatta)-G. PCR amplification of a portion of Mamu-G from the genomic DNA of five rhesus monkeys resulted in the identification of five additional Mamu-G alleles and revealed the presence of four Mamu-G alleles in one rhesus monkey, suggesting that Mamu-G had been duplicated in this individual. Furthermore, the analysis of 81 MHC class I clones isolated from a rhesus placenta cDNA library did not result in the isolation of Mamu-G cDNAs, nor the isolation of any additional HLA-G homologs, suggesting that Mamu-G was transcribed at negligible levels. Given the similarity of rhesus monkey and human placenta structure and function, these data raise interesting questions regarding the role of HLA-G in pregnancy.


Structural investigation of the carbohydrate element of human pituitary follicle-stimulating hormone by methylation analysis.
A preparation of human follicle-stimulating hormone has been subjected to methylation analysis by using the methyl sulphinyl carbanion-dimethyl sulphoxide-methyl iodide method. The hydrolysis products of the methylated glycoprotein were reduced, acetylated, analysed by gas-phase chromatography and mass spectrometry and identified by comparison with standards. Methylation analysis demonstrated that (1) the d-galactose, mannose, fucose and 2-amino-2-deoxyglucose units exist in the pyranose forms, (2) the 2-amino-2-deoxyglucose units are N-acetylated, (3) the fucopyranose units occupy terminal non-reducing positions, (4) the d-galactopyranose units are linked in the 1- and 2-positions, (5) the mannopyranose units exist in three forms, some as terminal non-reducing residues, some as 1,6-linked residues and some as 1,3,4-linked branch points, and (6) the 2-acetamido deoxyglucopyranose units are 1,6-linked. These structural assignments are compared with other data previously obtained for the carbohydrate moieties of follicle-stimulating hormone.


Who mixes with whom? A method to determine the contact patterns of adults that may lead to the spread of airborne infections.
Although mixing patterns are thought to be important determinants of the spread of airborne infectious diseases, to our knowledge, there have been no attempts to directly quantify them for humans. We report on a preliminary study to identify such mixing patterns. A sample of 92 adults were asked to detail the individuals with whom they had conversed over the period of one, randomly assigned, day. Sixty-five (71%) completed the questionnaire, providing their age, the age of their contacts and the social context in which the contacts took place. The data were analysed using multilevel modelling. The study identified, and allowed the quantification of, contact patterns within this sample that may be of epidemiological significance. For example, the degree of assortativeness of mixing with respect to age was dependent not only on the age of participants but the number of contacts made. Estimates of the relative magnitude of contact rates between different social settings were made, with implications for outbreak potential. Simple questionnaire modifications are suggested which would yield information on the structure and dynamics of social networks and the intensity of contacts. Surveys of this nature may enable the quantification of who acquires infection from whom and from where.


Economic analysis of respiratory rehabilitation.
We report on the incremental costs associated with improvements in health-related quality of life (HRQL) following 6 months of respiratory rehabilitation compared with conventional community care. Prospective randomized controlled trial of rehabilitation. A respiratory rehabilitation unit. Eighty-four subjects who completed the rehabilitation trial. Two months of inpatient rehabilitation followed by 4 months of outpatient supervision. All costs (hospitalization, medical care, medications, home care, assistive devices, transportation) were included. Simultaneous allocation was used to determine capital and direct and indirect hospitalization costs. The incremental cost of achieving improvements beyond the minimal clinically important difference in dyspnea, emotional function, and mastery was $11,597 (Canadian). More than 90% of this cost was attributable to the inpatient phase of the program. Of the nonphysician health-care professionals, nursing was identified as the largest cost center, followed by physical therapy and occupational therapy. The number of subjects needed to be treated (NNT) to improve one subject was 4.1 for dyspnea, 4.4 for fatigue, 3.3 for emotion, and 2.5 for mastery. Cost estimates of various approaches to rehabilitation should be combined with valid, reliable, and responsive measures of outcome to enable cost-effectiveness measures to be reported. Comparison studies with the same method are necessary to determine whether the improvements in HRQL that follow inpatient rehabilitation are cheap or expensive. Such information will be important in identifying the extent to which alternative approaches to rehabilitation can influence resource allocation. A consideration of cost-effectiveness from the perspective of NNT may be useful in the evaluation of health-care programs.


Prevention of cardiac surgery-associated acute kidney injury.
Numerous strategies have been evaluated to prevent early CSA-AKI. Although correction of hemodynamic problems is paramount, there are no clinical studies that compare different hemodynamic management or monitoring strategies with regard to their effect on kidney function. Pharmacologic strategies including diuretics, different classes of vasodilators and drugs with anti-inflammatory effects such as N-acetyl-cysteine, do not appear to be effective. Most of the studies are underpowered and use physiological rather than clinical endpoints. Further trials are warranted with fenoldopam and nesiritide (rhBNP). Observational and underpowered randomized studies show beneficial renal effects of off-pump technique and avoidance of aortic manipulation. There is very limited evidence for preoperative fluid loading and preemptive RRT. Potentially nephrotoxic agents should be used with caution in patients at risk of CSA-AKI. Tranexamic acid or aminocaproic acid should be preferred over aprotinin. No pharmacologic intervention has been adequately tested in the prevention of late CSA-AKI. A singlecenter study, including a predominance of patients after cardiac surgery, showed a decrease of kidney injury with tight glycemic control.


Hypnosis and anesthesia: back to the future.
Hypnosis is a physiological mind activity characterized by focused attention, absorption, dissociation and plastic imagination. In the early 19th century, several hundred surgical interventions were described with hypnosis as the sole anesthetic, in an epoch when no anesthetic drugs were available; then hypnosis was prejudicially abandoned and forgotten after its introduction. In the past two decades, an increasing number of studies on hypnosis has shown its capacity to modify the activity of the prefrontal cortex, default mode network and pain neuromatrix (including the anterior cingulate cortex, amygdala, thalamus, insula and somatosensory cortex) and increase pain threshold up to the level of surgical anesthesia. Hypnotic analgesia also prevents pain-related cardiovascular response: therefore, it may stand comparison with pharmacological anesthesia, yielding true protection from stress for the patient. The wealth of data available in the literature provides clear evidence of its meaningful effects on perioperative emotional distress, pain, medication consumption, physiological parameters, duration of surgery and outcome. Hypnosis may be used as follows: 1) as sole anesthetic, in minor surgery and invasive maneuvers and/or selected patients; 2) as adjuvant of pharmacological anesthesia (local anesthesia and/or sedation); 3) as an adjuvant technique in both pre- and postoperative phases in patients submitted to general anesthesia. Hypnosis, unlike any other therapeutic tools, does not call for drugs or equipment and is an attractive technique: it is free of charge, not burdened with proved adverse events and promises to help improving cost/benefits ratio.


Possible ecological risks of transgenic organism release when transgenes affect mating success: sexual selection and the Trojan gene hypothesis.
Widespread interest in producing transgenic organisms is balanced by concern over ecological hazards, such as species extinction if such organisms were to be released into nature. An ecological risk associated with the introduction of a transgenic organism is that the transgene, though rare, can spread in a natural population. An increase in transgene frequency is often assumed to be unlikely because transgenic organisms typically have some viability disadvantage. Reduced viability is assumed to be common because transgenic individuals are best viewed as macromutants that lack any history of selection that could reduce negative fitness effects. However, these arguments ignore the potential advantageous effects of transgenes on some aspect of fitness such as mating success. Here, we examine the risk to a natural population after release of a few transgenic individuals when the transgene trait simultaneously increases transgenic male mating success and lowers the viability of transgenic offspring. We obtained relevant life history data by using the small cyprinodont fish, Japanese medaka (Oryzias latipes) as a model. Our deterministic equations predict that a transgene introduced into a natural population by a small number of transgenic fish will spread as a result of enhanced mating advantage, but the reduced viability of offspring will cause eventual local extinction of both populations. Such risks should be evaluated with each new transgenic animal before release.


Upper airway viruses and bacteria in urban Aboriginal and Torres Strait Islander children in Brisbane, Australia: a cross-sectional study.
Respiratory morbidity in Australian Indigenous children is higher than their non-Indigenous counterparts, irrespective of urban or remote residence. There are limited studies addressing acute respiratory illness (ARI) in urban Indigenous children, particularly those that address the upper airway microbiome and its relationship to disease. We aimed to describe the prevalence of upper airway viruses and bacteria in symptomatic and asymptomatic urban-based Australian Indigenous children aged less than 5 years. A cross-sectional analysis of data collected at baseline in an ongoing prospective cohort study of urban Aboriginal and Torres Strait Islander children registered with a primary health care service in the northern suburbs of Brisbane, Australia. Clinical, demographic and epidemiological data and bilateral anterior nasal swabs were collected on enrolment. Polymerase chain reaction was performed on nasal swabs to detect 17 respiratory viruses and 7 bacteria. The primary outcome was the prevalence of these microbes at enrolment. Logistic regression was performed to investigate differences in microbe prevalence between children with and without acute respiratory illness with cough as a symptom (ARIwC) at time of specimen collection. Between February 2013 and October 2015, 164 children were enrolled. The median age at enrolment was 18.0 months (IQR 7.2-34.3), 49.4% were boys and 56 children (34.2%) had ARIwC. Overall, 133/164 (81%) nasal swabs were positive for at least one organism; 131 (79.9%) for any bacteria, 59 (36.2%) for any virus and 57 (34.8%) for both viruses and bacteria. Co-detection of viruses and bacteria was more common in females than males (61.4% vs 38.6%, p = 0.044). No microbes, alone or in combination, were significantly associated with the presence of ARIwC. The prevalence of upper airways microbes in asymptomatic children is similar to non-Indigenous children with ARIwC from the same region. Determining the aetiology of ARIwC in this community is complicated by the high prevalence of multiple respiratory pathogens in the upper airways. Australia New Zealand Clinical Trial Registry Registration Number: 12,614,001,214,628. Retrospectively registered.


Multiscale corner detection by using wavelet transform.
A multiscale corner detection algorithm based on the wavelet transform of contour orientation is proposed. It can utilize both the information of the local extrema and modulus of transform results to detect corners and arcs effectively. The ramp-width of contour orientation profile, which can be computed using the transformed modulus of two scales, reveals the difference between corner and arc and is utilized in the determination of corner points. The experimental results have shown that the detector is more effective than both the single- and multiple-scale detectors. They also demonstrate that the detector is insensitive to boundary noise. In addition, the proposed method is more efficient than the other multiscale corner detector because it operates on fewer number of scales, which can be implemented by a fast transform algorithm.


The Keap1-Nrf2 system as an in vivo sensor for electrophiles.
The Keap1-Nrf2 regulatory system plays a central role in cytoprotection from electrophilic and oxidative stress. In unstressed conditions, Nrf2 is constantly ubiquitinated by the Cul3-Keap1 ubiquitin E3 ligase complex and is degraded in the proteasome. Upon the exposure to electrophilic and oxidative stress, reactive cysteine residues in Keap1 are covalently modified, which abrogates the E3 ligase activity of the Cul3-Keap1 complex. Consequently Nrf2 is stabilized and induces the transcription of various cytoprotective genes. Structural analyses have revealed the overall structure of the Keap1 homodimer as well as structural features of the association between Keap1 and Nrf2, which has greatly enhanced our understanding of the molecular mechanisms involved in the regulation of the Keap1-Nrf2 system. Recently nitric oxide signaling has been shown to activate Nrf2, suggesting that Nrf2 is a mediator of the cytoprotective effect of nitric oxide. Analyses of Nrf2-null mice have revealed a critical contribution of Nrf2 to the protection from various diseases caused by electrophilic and oxidative stress. In contrast, constitutive activation of Nrf2 has been found in many cancers, resulting in resistance against chemotherapy and radiotherapy in cancer cells. Thus, Nrf2 is a promising target for drug development. The development of Nrf2 inducers and inhibitors is an important challenge for enhancing therapies for stress-induced diseases and cancers, respectively.


Characterization of the Lactococcus lactis transcription factor FlpA and demonstration of an in vitro switch.
The commercially important bacterium Lactococcus lactis contains two FNR-like proteins (FlpA and FlpB) which have a high degree of identity to each other and to the FLP of Lactobacillus casei. FlpA was isolated from a GST-FlpA fusion protein produced in Escherichia coli. Like FLP, isolated FlpA is a homodimeric protein containing both Zn and Cu. However, the properties of FlpA were more like those of the E. coli oxygen-responsive transcription factor FNR than the FLP of L. casei. As prepared FlpA recognized an FNR site (TTGAT-N4-ATCAA) but not an FLP site (CCTGA-N4-TCAGG) in band-shift assays. In contrast to FLP, DNA binding by FlpA did not require the formation of an intramolecular disulphide bond. However, despite containing only two cysteine residues per monomer, FlpA was able to acquire an FNR-like, oxygen-labile [4Fe 4S] cluster. But, whereas the incorporation of a [4Fe 4S] cluster into FNR enhances interaction with target DNA, it abolished DNA binding by FlpA. An FlpA variant (FlpA') with an N-terminal region designed to be more FLP-like failed to incorporate an iron-sulphur cluster but could now form an intramolecular disulphide. This simple example of protein engineering, converting an oxygen-labile [4Fe 4S] containing FNR-like protein into a dithiol-disulphide FLP-like redox sensor demonstrates the versatility of the basic CRP structure. Attempts to demonstrate an FlpA-based aerobic-anaerobic switch in the heterologous host E. coli were unsuccessful. However, studies with a series of FNR-dependent lac reporter fusions in strains of E. coli expressing flpA or flpB revealed that both homologues were able to activate expression of FNR-dependent promoters in vivo but only when positioned 61 base pairs upstream of the transcription start.


Associations between renal hyperfiltration and serum alkaline phosphatase.
Renal hyperfiltration, which is associated with renal injury, occurs in diabetic or obese individuals. Serum alkaline phosphatase (ALP) level is also elevated in patients with diabetes (DM) or metabolic syndrome (MS), and increased urinary excretion of ALP has been demonstrated in patients who have hyperfiltration and tubular damage. However, little was investigated about the association between hyperfiltration and serum ALP level. A retrospective observational study of the 21,308 adults in the Korea National Health and Nutrition Examination Survey IV-V databases (2008-2011) was performed. Renal hyperfiltration was defined as exceeding the age- and sex-specific 97.5th percentile. We divided participants into 4 groups according to their estimated glomerular filtration rate (eGFR): >120, 90-119, 60-89, and <60 mL/min/1.73 m2. The participants with eGFR >120 mL/min/1.73 m2 showed the highest risk for MS, in the highest ALP quartiles (3.848, 95% CI, 1.876-7.892), compared to the lowest quartile. Similarly, the highest risk for DM, in the highest ALP quartiles, was observed in participants with eGFR >120 ml/min/1.73 m2 (2.166, 95% CI, 1.084-4.329). ALP quartiles were significantly associated with albuminuria in participants with eGFR ≥ 60 ml/min/1.73m2. The highest ALP quartile had a 1.631-fold risk elevation for albuminuria with adjustment of age and sex. (95% CI, 1.158-2.297, P = 0.005). After adjustment, the highest ALP quartile had a 1.624-fold risk elevation, for renal hyperfiltration (95% CI, 1.204-2.192, P = 0.002). In addition, hyperfiltration was significantly associated with hemoglobin, triglyceride, white blood cell count, DM, smoking, and alcohol consumption (P<0.05). The relationship between serum ALP and metabolic disorders is stronger in participants with an upper-normal range of eGFR. Higher ALP levels are significantly associated with renal hyperfiltration in Korean general population.


Post-resuscitation care for neonates receiving positive pressure ventilation at birth.
To investigate if postresuscitation care (PRC) is indicated for all infants ≥35 weeks' gestation who receive positive pressure ventilation (PPV) at birth, explore the aspects of this care and the factors most predictive of it. Our hospital admits any infant who requires PPV at birth to special (intermediate/intensive) neonatal care unit (SNCU) for observation for at least 6 hours. All infants ≥35 weeks' gestation born between 1994 and 2013, who received PPV at birth, were reviewed. We examined perinatal factors that could predict the need for PRC after short (<1 minute) and prolonged (≥1 minute) PPV, admission course, neonatal morbidities, and the aspects of care given. Among 87 464 infants born, 3658 (4.2%) had PPV at birth with 3305 (90%) admitted for PRC. Of those, 1558 (42.6%) were in the short PPV group and 2100 (57.4%) in the prolonged PPV group. Approximately 59% of infants who received short PPV stayed in the SNCU for ≥1 day. Infants who received prolonged PPV were more likely to have morbidities and require special neonatal care. Multiple logistic regression analysis revealed the risk factors of placental abruption, assisted delivery, small-for-dates, gestational age <37 weeks, low 5-minute Apgar score, and need for intubation at birth to be independent predictors for SNCU stay ≥1 day and need for assisted ventilation, central lines, and parenteral nutrition. Our data support the need for PRC even for infants receiving short PPV at birth.


Mechanism of transdominant inhibition of CCR5-mediated HIV-1 infection by ccr5delta32.
Human chemokine receptor 5 (CCR5) functions as a co-receptor for Human immunodeficiency virus (HIV-1) infection. CCR5 is a seven-transmembrane cell surface receptor. Recently, a naturally occurring mutation of CCR5, ccr5Delta32, has been described. A small number of Caucasians are homozygously ccr5Delta32/ccr5Delta32, while a larger number of individuals are heterozygously CCR5/ccr5Delta32. The ccr5Delta32/ccr5Delta32 genotype has been linked to a phenotype that is "highly" protected from HIV-1 infection. On the other hand, several studies have shown that the CCR5/ccr5Delta32 genotype confers "relative" protection from AIDS with onset of disease being delayed by 2-4 years. Although it is known that peripheral blood lymphocytes from heterozygous individuals (CCR5/ccr5Delta32) support ex vivo HIV-1 replication at a reduced level compared with CCR5/CCR5 cells, the molecular basis for this observation is unknown. Here we report on events that post-translationally modify CCR5. We show that CCR5 progresses through the endoplasmic reticulum prior to appearing on the cell surface. Mature CCR5 can be post-translationally modified by phosphorylation and/or co-translationally by multimerization. By contrast, mutant ccr5Delta32, although retaining the capacity for multimerization, was incapable of being phosphorylated. ccr5Delta32 heterocomplexes with CCR5, and this interaction retains CCR5 in the endoplasmic reticulum resulting in reduced cell surface expression. Thus, co-expression in cells of ccr5Delta32 with CCR5 produces a trans-inhibition by the former of ability by the latter to support HIV-1 infection. Taken together, our findings suggest CCR5/ccr5Delta32 heterodimerization as a molecular explanation for the delayed onset of AIDS in CCR5/ccr5Delta32 individuals.


Causal interpretations of family GWAS in the presence of heterogeneous effects.
Family-based genome-wide association studies (GWAS) have emerged as a gold standard for assessing causal effects of alleles and polygenic scores. Notably, family studies are often claimed to provide an unbiased estimate of the average causal effect (or average treatment effect; ATE) of an allele, on the basis of an analogy between the random transmission of alleles from parents to children and a randomized controlled trial. Here, we show that this interpretation does not hold in general. Because Mendelian segregation only randomizes alleles among children of heterozygotes, the effects of alleles in the children of homozygotes are not observable. Consequently, if an allele has different average effects in the children of homozygotes and heterozygotes, as can arise in the presence of gene-by-environment interactions, gene-by-gene interactions, or differences in LD patterns, family studies provide a biased estimate of the average effect in the sample. At a single locus, family-based association studies can be thought of as providing an unbiased estimate of the average effect in the children of heterozygotes (i.e., a local average treatment effect; LATE). This interpretation does not extend to polygenic scores, however, because different sets of SNPs are heterozygous in each family. Therefore, other than under specific conditions, the within-family regression slope of a PGS cannot be assumed to provide an un-biased estimate for any subset or weighted average of families. Instead, family-based studies can be reinterpreted as enabling an unbiased estimate of the extent to which Mendelian segregation at loci in the PGS contributes to the population-level variance in the trait. Because this estimate does not include the between-family variance, however, this interpretation applies to only (roughly) half of the sample PGS variance. In practice, the potential biases of a family-based GWAS are likely smaller than those arising from confounding in a standard, population-based GWAS, and so family studies remain important for the dissection of genetic contributions to phenotypic variation. Nonetheless, the causal interpretation of family-based GWAS estimates is less straightforward than has been widely appreciated.


[Experience in the use of biostator in the treatment of diabetes mellitus in children of the pubertal age].
The authors discuss the possibilities of the use of the apparatus "artificial pancreas biostator" in multiple modality treatment of patients suffering from type I diabetes. The own research data are provided. They are based on the use of the apparatus "biostator" in 62 children aged 9-15 years with diabetes mellitus in order to improve insulin therapy. After treatment sessions with the aid of the apparatus all the patients noted improvement of the well-being and abatement of the signs of diabetes mellitus lability. In some cases, the treatment using the apparatus promoted a more rapid attainment of disease compensation as compared to the application of conventional treatment methods. It has been shown that high insulin requirement of pubertal children with diabetes decompensation is determined in many respects by activation of the hypothalamus-pituitary-adrenal system.


The small GTPase RSG1 controls a final step in primary cilia initiation.
Primary cilia, which are essential for normal development and tissue homeostasis, are extensions of the mother centriole, but the mechanisms that remodel the centriole to promote cilia initiation are poorly understood. Here we show that mouse embryos that lack the small guanosine triphosphatase RSG1 die at embryonic day 12.5, with developmental abnormalities characteristic of decreased cilia-dependent Hedgehog signaling. Rsg1 mutant embryos have fewer primary cilia than wild-type embryos, but the cilia that form are of normal length and traffic Hedgehog pathway proteins within the cilium correctly. Rsg1 mother centrioles recruit proteins required for cilia initiation and dock onto ciliary vesicles, but axonemal microtubules fail to elongate normally. RSG1 localizes to the mother centriole in a process that depends on tau tubulin kinase 2 (TTBK2), the CPLANE complex protein Inturned (INTU), and its own GTPase activity. The data suggest a specific role for RSG1 in the final maturation of the mother centriole and ciliary vesicle that allows extension of the ciliary axoneme.


Classification of Cryptosporidium species from patients with sporadic cryptosporidiosis by use of sequence-based multilocus analysis following mutation scanning.
In the present study, we analyzed genetic variation in Cryptosporidium species from humans (n = 62) with clinical cryptosporidiosis in South Australia. Sequence variation was assessed in regions within the small subunit of nuclear rRNA (p-SSU), the 70-kDa heat shock protein (p-hsp70), and the 60-kDa glycoprotein (p-gp60) genes by employing single-strand conformation polymorphism analysis and sequencing. Based on the analyses of p-SSU and p-hsp70, Cryptosporidium hominis (n = 38) and Cryptosporidium parvum (n = 24) were identified. The analysis of p-gp60 revealed eight distinct subgenotypes, classified as C. hominis IaA17R1 (n = 3), IbA9G3R2 (n = 14), IbA10G2R2 (n = 20), and IfA12G1R1 (n = 1), as well as C. parvum IIaA18G3R1 (n = 15), IIaA20G3R1 (n = 6), IIaA22G4R1 (n = 2), and IIcA5G3R2 (n = 1). Subgenotypes IaA17R1 and IIaA22G4R1 are new. Of the six other subgenotypes, IbA10G2R2, IIaA18G3R1, IIaA20G3R1, and IIcA5G3R2 were reported previously from the state of Victoria. This is the fourth record in Australia of C. parvum subgenotype IIaA18G3R1 from humans, which, to date, has been isolated only from cattle in other countries. This subgenotype might be a significant contributor to sporadic human cryptosporidiosis and may indicate a greater zoonotic contribution to the infection of humans in the area of study. Comparative analyses revealed, for the first time, the differences in the genetic makeup of Cryptosporidium populations between two relatively close, major metropolitan cities.


Neuronal activity regulates the regional vulnerability to amyloid-β deposition.
Amyloid-β (Aβ) plaque deposition in specific brain regions is a pathological hallmark of Alzheimer's disease. However, the mechanism underlying the regional vulnerability to Aβ deposition in Alzheimer's disease is unknown. Herein, we provide evidence that endogenous neuronal activity regulates the regional concentration of interstitial fluid (ISF) Aβ, which drives local Aβ aggregation. Using in vivo microdialysis, we show that ISF Aβ concentrations in several brain regions of APP transgenic mice before plaque deposition were commensurate with the degree of subsequent plaque deposition and with the concentration of lactate, a marker of neuronal activity. Furthermore, unilateral vibrissal stimulation increased ISF Aβ, and unilateral vibrissal deprivation decreased ISF Aβ and lactate, in contralateral barrel cortex. Long-term unilateral vibrissal deprivation decreased amyloid plaque formation and growth. Our results suggest a mechanism to account for the vulnerability of specific brain regions to Aβ deposition in Alzheimer's disease.


Zinc-alpha2-glycoprotein: a new biomarker of breast cancer?
Obesity increases the risk of breast cancer. It is established that adipocyte secretions, i.e. adipokines, may play a role in mammary carcinogenesis. We have shown that two major adipokines, leptin and adiponectin, were expressed in mammary adenocarcinoma. Here, we evaluated zinc-alpha2-glycoprotein (ZAG) expression in tumor (n=55) and healthy (n=6) breast tissue by immunohistochemistry and examined whether it was correlated with that of major adipokines, usual tumor biomarkers (sex steroids receptors, i.e. estrogen (ER) and progesterone; Ki-67; cErb2), or apoptosis markers (Bcl2 and Bax). ZAG expression was detected in ductal carcinoma and normal epithelial adjacent tissue but not in normal tissue of healthy women. In cancer tissue, its expression was correlated positively to leptin receptor and negatively to adiponectin receptor and ER. These preliminary results suggest both a relationship between ZAG expression and pathways involving adipokines or estrogen and that ZAG may be a potential breast cancer biomarker.


A systematic review and meta-analysis of the psychosis continuum: evidence for a psychosis proneness-persistence-impairment model of psychotic disorder.
A systematic review of all reported incidence and prevalence studies of population rates of subclinical psychotic experiences reveals a median prevalence rate of around 5% and a median incidence rate of around 3%. A meta-analysis of risk factors reveals associations with developmental stage, child and adult social adversity, psychoactive drug use, and also male sex and migrant status. The small difference between prevalence and incidence rates, together with data from follow-up studies, indicates that approximately 75-90% of developmental psychotic experiences are transitory and disappear over time. There is evidence, however, that transitory developmental expression of psychosis (psychosis proneness) may become abnormally persistent (persistence) and subsequently clinically relevant (impairment), depending on the degree of environmental risk the person is additionally exposed to. The psychosis proneness-persistence-impairment model considers genetic background factors impacting on a broadly distributed and transitory population expression of psychosis during development, poor prognosis of which, in terms of persistence and clinical need, is predicted by environmental exposure interacting with genetic risk.


[Surgical anatomy of the perforating veins of Hunter's canal].
The vein perforating Hunter's Canal is often mentioned in the books on anatomy but is never described. After injecting coloured Neoprene Latex, ten long saphenous veins were dissected in the thigh. It was therefore possible to specify the origin, the course, the relation and the terminus of the perforating vein. The origin appeared variable in the long saphena, sometimes because of variations in the saphena itself; the course of the perforating vein is essentially along the posterior edge of the Couturier, then along the deep side. The terminus is found in the femoral vein after crossing the upper part of Hunter's fascia. The course of Hunter's perforating vein can therefore be divided into two parts: one variable, superficial, the other deep and constant.


Temporal and palaeoclimatic context of the evolution of insular woodiness in the Canary Islands.
Insular woodiness (IW), referring to the evolutionary transition from herbaceousness toward woodiness on islands, has arisen more than 30 times on the Canary Islands (Atlantic Ocean). One of the IW hypotheses suggests that drought has been a major driver of wood formation, but we do not know in which palaeoclimatic conditions the insular woody lineages originated. Therefore, we provided an updated review on the presence of IW on the Canaries, reviewed the palaeoclimate, and estimated the timing of origin of woodiness of 24 insular woody lineages that represent a large majority of the insular woody species diversity on the Canaries. Our single, broad-scale dating analysis shows that woodiness in 60%-65% of the insular woody lineages studied originated within the last 3.2 Myr, during which Mediterranean seasonality (yearly summer droughts) became established on the Canaries. Consequently, our results are consistent with palaeoclimatic aridification as a potential driver of woodiness in a considerable proportion of the insular woody Canary Island lineages. However, the observed pattern between insular woodiness and palaeodrought during the last couple of million years could potentially have emerged as a result of the typically young age of the native insular flora, characterized by a high turnover.


Statins for the treatment of dementia.
The use of statin therapy in established Alzheimer's disease (AD) or vascular dementia (VaD) is a relatively unexplored area. In AD ss-amyloid protein (Ass) is deposited in the form of extracellular plaques and previous studies have determined Ass generation is cholesterol dependent. Hypercholesterolaemia has also been implicated in the pathogenesis of VaD. Due to the role of statins in cholesterol reduction it is biologically plausible they may be efficacious in the treatment of AD and dementia. To assess the clinical efficacy and tolerability of statins in the treatment of dementia. We searched the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group, The Cochrane Library, MEDLINE, EMBASE, PsycINFO, CINAHL and LILACS, as well as many trials registries and grey literature sources (27 October 2008). Double-blind, randomized controlled trials of statins given for at least six months in people with a diagnosis of dementia. Two independent authors extracted and assessed data independently against the inclusion criteria. Data were pooled where appropriate and entered into a meta-analysis. Three studies were identified (748 participants, age range 50-90 years). All patients had a diagnosis of probable or possible AD according to standard criteria and most patients were established on a cholinesterase inhibitor. Treatment in ADCLT 2005 consisted of 80mg atorvastatin compared to placebo for 52 weeks, serum low density lipoprotein (LDL) cholesterol was reduced by 54% in the atorvastatin group. Treatment in Simons 2002 consisted of 40mg simvastatin compared to placebo for 26 weeks, serum LDL cholesterol was reduced by 52% in the simvastatin group. Treatment in LEADe 2010 consisted of 80mg atorvastatin compared to placebo for 72 weeks, LDL cholesterol was reduced by 50.2% by month 3 and remained constant through month 18. Change in Alzheimer's Disease Assessment Scale- cognitive subscale (ADAS-Cog) from baseline was a primary outcome in 3 studies; when data were pooled there was considerable heterogeneity so the random effects model was used, statins did not provide any beneficial effect in this cognitive measure [mean difference -1.12, 95% CI -3.99, 1.75, p = 0.44]. All studies provided change in Mini Mental State Examination (MMSE) from baseline; again random effects model was used due to considerable heterogeneity: there was no significant benefit from statins in this cognitive measure when the data were pooled [mean difference -1.53, 95% CI -3.28, 0.21, p = 0.08]. There was some evidence that patients on statins in ADCLT 2005 maintained better cognitive function if serum cholesterol was high at baseline, MMSE was higher at baseline or if they had an apolipoprotein E4 allele present. This would need to be confirmed in larger studies however. Treatment related adverse effects were available from two studies, LEADe 2010 and Simons 2002; when data were pooled there was no significant difference between statins and placebo [odds ratio 2.45, 95% CI 0.69, 8.62, p = 0.16]. There was no significant difference in global function, behaviour or activities of daily living in the statin and placebo groups. One large randomised controlled trial (RCT) ( CLASP 2008) has not yet published its results. There were no studies identified assessing role of statins in treatment of VaD. There was no evidence that statins were detrimental to cognition. There is insufficient evidence to recommend statins for the treatment of dementia. Analysis from the studies available, including one large RCT, indicate statins have no benefit on the outcome measures ADAS-Cog or MMSE. We need to await full results from CLASP 2008 before we can be certain. This Cochrane review will be updated as these results become available.


Characteristics of cisternal cerebrospinal fluid associated with primary brain tumors in the dog: a retrospective study.
Results of cisternal CSF analyses of 77 dogs with primary brain tumors (1970 to 1984) were examined retrospectively. The types of primary brain tumors were astrocytomas, choroid plexus papillomas, ependymomas, meningiomas, and oligodendrogliomas. Fifty-three dogs had complete CSF analyses performed (total WBC count, total protein content, and pressure). Of these 53 CSF, 39.6% had a pattern of change consistent with current descriptions of CSF associated with brain neoplasia. Results of the remaining 60.4% of the 53 CSF were normal (9.4%) or had an atypical pattern of abnormality (50.9%). The CSF associated with meningiomas was unique in having CSF with a WBC count greater than 50 cells/microliter and a WBC differential count greater than 50% polymorphonuclear (PMN) cells. Within the meningioma group, a predominance of PMN cells in the CSF correlated with necrosis or PMN cell infiltration of the tumors. Additional correlations between specific CSF characteristics and histologic features of the tumors could not be made. Statistical analysis of the 77 CSF revealed that the mean CSF total WBC count of the oligodendroglioma group was significantly less than the mean WBC count of the choroid plexus papilloma group and the meningioma group. The mean CSF total protein contents of the astrocytoma group and the meningioma group were significantly less than the mean protein of the choroid plexus papilloma group. The mean CSF pressure of the tumor groups was not significantly different. For all 77 tumors, the most common abnormality was an increased total protein content (69.4%); the least common abnormality was an increased total WBC count (41.3%).(ABSTRACT TRUNCATED AT 250 WORDS)


Papulopustular and Ocular Rosacea with an Alleged Coincidence of Cutaneous Lupus Erythematosus: A Case Report.
Rosacea is a chronic inflammatory skin disease characterized by central facial erythema with or without ocular involvement. It is often difficult to distinguish rosacea from other malar rashes, one of which is acute cutaneous lupus erythematosus (CLE), particularly when there is an increase in antinuclear antibody (ANA) level. We report the case of a 16-year old woman with facial erythematous plaque accompanied by papules and pustules, reddened eyes, and swollen eyelids since the last one year. Dermoscopic examination revealed telangiectasia, and skin scraping examination with 20% potassium hydroxide identified the presence of Demodex folliculorum. Further ocular examination also revealed blepharitis, dysfunction of Meibomian gland, cicatrix, and corneal neovascularization. The ANA titer was positive (1:320), while the anti-dsDNA was negative. The patient was treated according to standard treatment for rosacea. The patient showed a satisfactory response following 2 weeks of therapy. Signs of recurring red patches with papules, pustules, telangiectasia, and identification of D. folliculorum on skin scraping examination led to the diagnosis of papulopustular rosacea. A positive ANA test may also be present in other diseases, e.g. acute CLE. Therefore, the diagnosis of rosacea remains a challenge. Thorough observation and examination must be done in order to yield an accurate diagnosis of rosacea.


Combining 16S Sequencing and qPCR Quantification Reveals Staphylococcus aureus Driven Bacterial Overgrowth in the Skin of Severe Atopic Dermatitis Patients.
Atopic dermatitis (AD) is an inflammatory skin disease with a microbiome dysbiosis towards a high relative abundance of Staphylococcus aureus. However, information is missing on the actual bacterial load on AD skin, which may affect the cell number driven release of pathogenic factors. Here, we combined the relative abundance results obtained by next-generation sequencing (NGS, 16S V1-V3) with bacterial quantification by targeted qPCR (total bacterial load = 16S, S. aureus = nuc gene). Skin swabs were sampled cross-sectionally (n = 135 AD patients; n = 20 healthy) and longitudinally (n = 6 AD patients; n = 6 healthy). NGS and qPCR yielded highly inter-correlated S. aureus relative abundances and S. aureus cell numbers. Additionally, intra-individual differences between body sides, skin status, and consecutive timepoints were also observed. Interestingly, a significantly higher total bacterial load, in addition to higher S. aureus relative abundance and cell numbers, was observed in AD patients in both lesional and non-lesional skin, as compared to healthy controls. Moreover, in the lesional skin of AD patients, higher S. aureus cell numbers significantly correlated with the higher total bacterial load. Furthermore, significantly more severe AD patients presented with higher S. aureus cell number and total bacterial load compared to patients with mild or moderate AD. Our results indicate that severe AD patients exhibit S. aureus driven increased bacterial skin colonization. Overall, bacterial quantification gives important insights in addition to microbiome composition by sequencing.


Paraquat toxicity.
Paraquat (1,1'-dimethyl-4,4'-bipyridylium dichloride) is marketed as a contact herbicide. Although it has proved safe in use there have been a number of cases of poisoning after the intentional swallowing of the commercial product. The most characteristic feature of poisoning is lung damage, which causes severe anoxia and may lead to death. The specific toxicity to the lung can be explained in part by the accumulation of paraquat into the alveolar type I and type II epithelial cells by a process that has been shown to accumulate endogenous diamines and polyamines. When accumulated, paraquat undergoes an NADPH-dependent, one-electron reduction to form its free radical, which then reacts avidly with molecular oxygen to reform the cation and produce superoxide anion, which in turn will dismutate to form H2O2. This may lead to the formation of more reactive (and hence toxic) radicals which have the potential to cause lipid peroxidation and lead to cell death. Biochemical changes provoked by paraquat in the lung suggest that it causes a rapid, pronounced and prolonged oxidation of NADPH that initiates compensatory biochemical processes in the lung. NADPH may be further depleted as it is consumed in an attempt to detoxify H2O2 or lipid hydroperoxides. Thus it is possible that with toxic levels of paraquat in the cell, compensatory biochemical processes are insufficient to maintain levels of NADPH consistent either with cell survival or with the ability to detoxify H2O2 or prevent lipid peroxidation.