Comparison of radiopacity of current restorative CAD/CAM blocks with digital radiography.
While the radiopacity of restorative material affects the radiographic diagnosis of the teeth, there is no data about the radiopacity of current restorative computer-aided design (CAD)/computer-aided manufacturing (CAM) materials. Therefore, the present study compared the radiopacity values of current restorative CAD/CAM blocks to facilitate the material choice within such a wide variety of materials. Specimens were prepared from 13 different restorative CAD/CAM blocks to compare with enamel and dentin. The specimens placed on the occlusal phosphor plate were imaged with aluminum step wedge and tooth section. The radiopacity values were calculated using the Image J program. The radiopacity values of the specimens were converted to mmAl values with the Curve Expert 1.4 program. The difference between the radiopacity values of dentin and e.max CAD was not significant, however, they exhibited a significant difference from the other 14 groups (P < .05). Enamel and Obsidien, Suprinity, and Celtra Duo had greater radiopacity values with significant differences from the other 12 materials whereas the difference within these groups was not significant (P > .05). The evaluated restorative CAD/CAM materials have significantly different radiopacity values. Among these permanent restoration blocks, the highest radiopacity value was observed in Celtra Duo, the lowest in Block HC. Cerasmart, Lava Ultimate, Obsidian, Vita Suprinity, Celtra Duo Blocks have adequate radiopacity for inlay, onlay and crown restorations, however, the use of Vita Enamic, Vita Mark II, GC LRF blocks and the others which have lower radiopacity value than dentin for the same kind of restorations depend on the radiopacity of the luting cement for the purpose of recurrent caries detection.


Experimental and Numerical Investigation of a Photoacoustic Resonator for Solid Samples: Towards a Non-Invasive Glucose Sensor.
T-cell resonators have been used lately for non-invasive blood glucose measurements for photoacoustic spectroscopy on skin samples. A resonator has a significant role in determining the strength of the measured signal and the overall sensitivity of the sensor. Here we present results of the measurement of the photoacoustic signal of such a T-cell resonator. The signal is also modelled using the amplitude mode expansion method, which is based on eigenmode expansion and the introduction of losses in the form of loss factors. The measurement reproduced almost all the calculated resonances from the numerical models with fairly good agreement. The cause of the differences between the measured and the simulated resonances are explained. In addition, the amplitude mode expansion simulation model is established as a faster and computationally less demanding photoacoustic simulation alternative to the viscothermal model. The resonance frequencies from the two models differ by less than 1.8%. It is noted that the relative height of the amplitudes from the two models depends on the location of the antinodes within the different parts of the resonator. The amplitude mode expansion model provides a quick simulation tool for the optimization and design of macro resonators.


Engineering atomic-scale magnetic fields by dysprosium single atom magnets.
Atomic scale engineering of magnetic fields is a key ingredient for miniaturizing quantum devices and precision control of quantum systems. This requires a unique combination of magnetic stability and spin-manipulation capabilities. Surface-supported single atom magnets offer such possibilities, where long temporal and thermal stability of the magnetic states can be achieved by maximizing the magnet/ic anisotropy energy (MAE) and by minimizing quantum tunnelling of the magnetization. Here, we show that dysprosium (Dy) atoms on magnesium oxide (MgO) have a giant MAE of 250 meV, currently the highest among all surface spins. Using a variety of scanning tunnelling microscopy (STM) techniques including single atom electron spin resonance (ESR), we confirm no spontaneous spin-switching in Dy over days at ≈ 1 K under low and even vanishing magnetic field. We utilize these robust Dy single atom magnets to engineer magnetic nanostructures, demonstrating unique control of magnetic fields with atomic scale tunability.


Secreted Glycoside Hydrolase Proteins as Effectors and Invasion Patterns of Plant-Associated Fungi and Oomycetes.
During host colonization, plant-associated microbes, including fungi and oomycetes, deliver a collection of glycoside hydrolases (GHs) to their cell surfaces and surrounding extracellular environments. The number and type of GHs secreted by each organism is typically associated with their lifestyle or mode of nutrient acquisition. Secreted GHs of plant-associated fungi and oomycetes serve a number of different functions, with many of them acting as virulence factors (effectors) to promote microbial host colonization. Specific functions involve, for example, nutrient acquisition, the detoxification of antimicrobial compounds, the manipulation of plant microbiota, and the suppression or prevention of plant immune responses. In contrast, secreted GHs of plant-associated fungi and oomycetes can also activate the plant immune system, either by acting as microbe-associated molecular patterns (MAMPs), or through the release of damage-associated molecular patterns (DAMPs) as a consequence of their enzymatic activity. In this review, we highlight the critical roles that secreted GHs from plant-associated fungi and oomycetes play in plant-microbe interactions, provide an overview of existing knowledge gaps and summarize future directions.


Feeding Deterrence to Reticulitermes speratus (Kolbe) by Fibroporia radiculosa (Peck) Parmasto 1968.
Brown rot fungus Fibroporia radiculosa (Peck) Parmasto grown in decayed wood and non-wood material, potato dextrose agar (PDA) media, deterred Reticulitermes speratus (Kolbe) feeding. Decayed wood and PDA media were extracted and tests were performed to assess termite feeding behavior towards the extracts. We found that the extract from PDA media also suppressed termite feeding, although it did not induce mortality. Using gas chromatography and mass spectrometry analysis, two bioactive compounds were detected from the decayed wood extract, and one was detected from the PDA extract. Based on National Institute of Science and Technology (USA) Mass Spectral library match and compound fragmentation, both of the compounds belong to the sesquiterpenes family.


How a low tissue O2 strategy could be conserved in early crustaceans: the example of the podocopid ostracods.
An adaptation strategy whereby O(2) partial pressure, P(O(2)), in the tissues is maintained within a low, narrow range of 1-3 kPa, largely independent of the inspired P(O(2)), has been reported in water- and air-breathing poikilotherms and in homeotherms. Based on the postulate that this basic cellular mechanism has been established since the early stages of evolution, it has been hypothesized that it could be the consequence of an early adaptation strategy to maintain cellular oxygenation within the same low and primitive range. To test this hypothesis we studied the basic mechanisms of oxygen regulation in podocopid ostracods, minute crustaceans that have existed on earth for at least 500 million years. Podocopids lack any regulatory mechanism for adapting their ventilation to cope with changes in water oxygenation, and instead adjust their tissue oxygenation status by migrating through the O(2) gradient to sediment layers where the P(O(2)) of the water is 3-5 kPa. Experimental manipulation of the O(2) profile induced their vertical migration to follow this precise water P(O(2)) and demonstrates the existence of a regulation strategy. This strategy must be associated with the lower P(O(2)) values within the animal's carapace valves, showing that podocopids can actively regulate their tissue P(O(2)) at constant but even lower values than the water. In conclusion, the low tissue P(O(2)) strategy could have existed in early crustaceans and, by extension, in early animals.


Empathy in Dentistry: How Attitudes and Interaction With Older Adults Make a Difference.
The development of empathy and positive attitudes are essential elements of professional education. This study explored the nature of empathy and its association with attitudes about, and exposure to older patients in a sample of dental students. Students completed an adapted version of the Jefferson Scale of Physician Empathy (JSPE), the Aging Semantic Differential (ASD) and answered questions about their exposure to older people. Factor analysis was used to identify four factors: (1) Empathy is Valuable, (2) Empathy is Demonstrated, (3) Empathy is not Influential, and (4) Empathy is Difficult to Accomplish. Higher empathy scores were related to the ASD subscale attitude of acceptability of aging and to greater exposure to older adults outside of clinical practice. There were no demographic predictors of higher empathy scores.


Chemokine CCL2 and chemokine receptor CCR2 in early active multiple sclerosis.
The chemokine monocyte chemoattractant protein (MCP)-1/CCL2 and its receptor CCR2 have been strongly implicated in disease pathogenesis in experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis (MS), whereas data on the CCL2-CCR2 axis are scarce in MS. We studied the expression of CCR2 on leukocytes in blood and cerebrospinal fluid (CSF) from patients with monosymptomatic optic neuritis and MS, and the concentration of CCL2 in the CSF from these patients. Results were compared with the results in non-inflammatory neurological controls and were correlated with other parameters (magnetic resonance imaging and CSF data). Our findings suggest a limited role for CCL2/CCR2 in early active MS.


Cysteine biosynthesis in Saccharomyces cerevisiae: mutation that confers cystathionine beta-synthase deficiency.
The cys2-1 mutation of Saccharomyces cerevisiae was originally thought to confer cysteine dependence through a serine O-acetyltransferase deficiency. In this study, we show that cys2-1 strains lack not only serine O-acetyltransferase but also cystathionine beta-synthase. However, a prototrophic strain was found to be serine O-acetyltransferase deficient because of a mutation allelic to cys2-1. Moreover, revertants obtained from cys2-1 strains had serine O-acetyltransferase but not cystathionine beta-synthase, whereas transformants obtained by treating a cys2-1 strain with an S. cerevisiae genomic library had cystathionine beta-synthase but not serine O-acetyltransferase. From these observations, we conclude that cys2-1 (serine O-acetyltransferase deficiency) accompanies a very closely linked mutation that causes cystathionine beta-synthase deficiency and that these mutations together confer cysteine dependence. This newly identified mutation is named cys4-1. These results not only support our previous hypothesis that S. cerevisiae has two functional cysteine biosynthetic pathways but also reveal an interesting gene arrangement of the cysteine biosynthetic system.


Streamlined procedure for gene knockouts using all-in-one adenoviral CRISPR-Cas9.
CRISPR-Cas9 is a powerful gene editing technique that can induce mutations in a target gene of interest in almost any mammalian cell line. However, its practicality can be limited if target cell lines are difficult to transfect and do not proliferate. In the current study, we have developed a streamlined approach for CRISPR-based gene knockouts with three key advantages, which allows phenotypic assay of gene knockouts without clonal selection and expansion. First, it integrates into a single, all-in-one vector transgenes for Cas9, sgRNA, and a fluorescence marker. Second, we used the Gateway system to rapidly clone specific sgRNAs into the all-in-one vector through PCR and in vitro recombination, without conventional enzyme digestion and ligation. Third, it uses adenovirus for the capacity to package the all-in-one vector, and for its high efficiency of transduction. We tested the all-in-one adenoviral CRISPR-Cas9 in a circadian clock model cell line U2OS, and demonstrated that essential clock genes such as Bmal1 and Per1 were knocked out so efficiently that functional assays could be performed from the heterogenic population without any clonal selection and expansion. This streamlined approach may prove invaluable for rapid functional assays of candidate genes in diverse biological pathways, including the circadian clock.


Comparison of neem or oregano with thiram on organic matter decomposition of a sand loam soil amended with compost, and on soil biological activity.
Oregano essential oil and neem have been reported to be effective against soil-borne pathogens and nematodes. The possibility of having an effect on soil properties was the aim of this investigation. Moreover, thiram, a common widely used chemical pesticide, was used for comparison. The effects of all three above-mentioned substances were investigated during an incubation experiment, for 15 weeks. Crushed neem pellets were added at 1.0, 2.0 or 3.0 g per 50 g of soil. Oregano dry matter was applied at 0.2, 0.4 or 0.6 g per 50 g of soil. Finally, thiram was applied at 0.1, 0.2 and 0.3 g per 50 g of soil. The addition of neem resulted in a decrease in organic carbon mineralisation, higher than that of oregano, but not as prominently as thiram. The addition of neem resulted in an increase in the content of nitrate, organic P, bioavailable P, bioavailable K and bioavailable Mn. Oregano had a slight negative effect on organic matter biodegradation, but caused an increase of nutritional mineral elements. Microbe colonies in soil were increased by the addition of neem cake granules, whereas oregano and thiram treatments caused a decrease. The addition of thiram reduced nitrate and available Mn contents, but at the lowest dose increased organic P and available K. Thiram reduced prominently available Cu at the upper rates, but it increased these forms at the lowest rate, while available Zn content was increased in two lowest rates. The results of this study indicated that neem or oregano could be applied to the soil without any extremely negative effect on the available forms of nutritional mineral elements, as thiram does.


Blessed by the Holy People.
Men-women and women-men have a long tradition in Diné (Navajo) culture where they were, and sometimes still are, held in high esteem. Their supernatural prototypes figure prominently in parts of the Diné Origin Story. It is in this cosmological worldview and tradition of acceptance that Carrie, a multi-dualistic spirit, grew up to be a female-bodied man supported and respected by his/her family and community. He/she has worked in various professions defined as "masculine" in Diné and Western contexts, such as fire fighter, heavy equipment operator, truck driver, and building contractor. In addition, Carrie is a keeper of Diné traditions, and has also been active educating Native and Non-Native audiences on 2SpiritLesbianGayBiTransIntersexQueer issues.


Panfacial Fat Injection Approach in Young Asian Patients for Facial Contouring: A Goal-Oriented Classification Method Based on the Facial Fat Compartments Theory.
In Asia, young patients tend to undergo panfacial fat injection for facial contouring rather than for rejuvenation. Although the procedure is widely performed, there is no single approach that has been shown to have consistent results. One personalized approach based on the facial fat compartment theory is presented here. Under the theory of facial fat compartments, the injection area is classified into functional zones and transition zones. A retrospective review of 105 cases over 6 years was performed to evaluate the effectiveness of this approach. Follow-up ranged from 6 to 47 months. Satisfactory results were achieved in 96 percent of cases. Typical cases were also reviewed. In the authors' view, this approach is safe and easy to learn, and produces aesthetically satisfactory results. The approach is stable, and satisfactory clinical results were achieved under the guidance of the facial fat compartment theory, which in turn verified the accuracy of this theory. Therapeutic, V.


Mechanical and Structural Remodeling of Cardiac Muscle after Aerobic and Resistance Exercise Training in Rats.
Aerobic and resistance exercise training results in distinct structural changes of the heart. The mechanics of how cardiac cells adapt to resistance training and the benefits to cells when combining aerobic and resistance exercise remains largely unknown. The purpose of this study was to compare mechanical adaptations of skinned cardiac fiber bundles after chronic resistance, aerobic and combined exercise training in rats. We hypothesized that differences in mechanical function on the fiber bundle level coincide with differences previously reported in the structure of the heart. Twelve-week-old rats were assigned to (i) an aerobic running group (n = 6), (ii) a ladder climbing resistance group (n = 6), (iii) a combination group subjected to aerobic and resistance training (n = 6), or (iv) a sedentary (control) group (n = 5). Echocardiography was used to measure cardiac structural remodeling. Skinned cardiac fiber bundles were used to determine active and passive force properties, maximal shortening velocity, and calcium sensitivity. Aerobically trained animals had 43%-49% greater ventricular volume and myocardial thickness, and a 4%-17% greater shortening velocity and calcium sensitivity compared with control group rats. Resistance-trained rats had 37%-71% thicker ventricular walls, a 56% greater isometric force production, a 9% greater shortening velocity, and a 4% greater calcium sensitivity compared with control group rats. The combination exercise-trained rats had 25%-43% greater ventricular volume and myocardial wall thickness, a 55% greater active force production, a 7% greater shortening velocity, and a 60% greater cross-bridge cooperativity compared with control group rats. The heart adapts differently to each exercise modality, and a combination of aerobic and resistance training may have the greatest benefit for cardiac health and performance.


Analytical Quality Requirements in Human Biomonitoring Programs: Trace Elements in Human Blood.
Human biomonitoring (HBM) programs consist of several interrelated and equally important steps. Of these steps, the study design must answer a specific question: How many individuals must be recruited in order to define the spatial or temporal trends of exposure to environmental pollutants in a given HBM study? Two components must be considered at this stage: the population variability of the expected exposure and the performance characteristics of the analytical methods used. The objective of the present study was to quantify the contribution to the required sample size arising from (i) measurement uncertainty and (ii) inter-laboratory measurement variability. For this purpose, the sample size was calculated using the measurement uncertainty of one laboratory, inter-laboratory comparison exercise data, and population variability for commonly studied metals (mercury, cadmium, and lead) in blood. Measurement uncertainty within one laboratory proved to have little influence on the sample size requirements, while the inter-laboratory variability of the three metals increased the requirements considerably, particularly in cases of low population variability. The multiple laboratories approach requires that laboratory variability be considered as early as the planning stage; a single-laboratory approach is thus a cost-effective compromise in HBM to reduce variability due to the participation of different laboratories.


Quantification of Lipid Metabolism in Living Cells through the Dynamics of Lipid Droplets Measured by Stimulated Raman Scattering Imaging.
Dysregulation of lipid metabolism is associated with many diseases including cancer. Lipid droplet (LD), a ubiquitous organelle in mammalian cells, serves as a hub for lipid metabolism. Conventional assays on the measurement of lipid metabolism rely on the quantification of the lipid composition or amount. Such methods cannot distinguish LDs having different biofunctionalities in living cells, and thus could be inaccurate in measuring the instantaneous lipogenesis of the living cells. We applied label-free stimulated Raman scattering microscopy to quantify the LDs' spatial-temporal dynamics, which showed direct links to cellular lipid metabolisms and can separate LDs involved in different metabolic events. In human cancer cells, we found that changes in the maximum displacement of LDs reflected variations in cellular lipogenic activity, and changes in the average speed of LDs revealed alterations in LD size. The LD dynamics analysis allowed for more accurate measurement in the lipogenesis and LD dimensions, and can break the optical diffraction limit to detect small variation in lipid metabolism that was conventionally undetectable. By this method, we revealed changes in the lipogenic activity and LD sizes during glucose starvation of HeLa cells and transforming growth factor beta-induced epithelial-to-mesenchymal transition of SKOV-3 cells. This method opens a way to quantify lipid metabolism in living cells during cellular development and transition.


Review on Bioorganometallic Chemistry and New Outcomes in the Synthesis and Substitution of Tetracarbonyl(pyrrolylimine) Complexes of Rhenium with Organophosphorus Ligands.
: After a short review dealing with bioorganometallic chemistry, the synthesis of tetracarbonyl (pyrrolylimine) complexes of rhenium bearing chirality on the pyrrolyl ligands was reported.The reactivity of these compounds towards the substitution of one carbonyl ligand with triphenyl phosphine, tricyclohexyl phosphine and trimethyl phosphite was studied. The rhenium becoming a stereogenic center in that transformation, the resulting tricarbonyl species were obtained as mixtures of diastereomers, with diastereomeric excesses varying from 8 to 84%, according to the reaction conditions and the relative steric hindrances of the pyrrolylimine and the organophosphorus ligands. These bioorganometallics are potential CO releasing molecules that could be used in the field of medicinal chemistry.


[Effects on development of the testicle in diet-induced obesity rats].
We established diet-induced obesity animal model and observed the influence of obesity on testicle of the male rats. Rats in model group were fed with fat-enriched diet for 6 weeks. We measured the body weight and the testis weight and observed the development of testicle by microscope, and compared with those of control group. Meanwhile, we measured the serum levels of testosterone (T) and estradiol (E2). The average body weight in model group rats was higher 29% than that in control group at 6 weeks old, 30% at 9 weeks old. We observed under microscope that most of the convoluted seminiferous tubule developed no good in model group, the 4 layer cells was sparse and the arrangement of them was not in order. The testosterone to estradiol ratios(T/E2) in model group was significant lower then that of control group (P < 0.05). The fat-enriched fat diet can induce obesity. It influenced the development of the testicle. This probably related to decrease of the testosterone to estradiol ratios.


Characterization of the gene encoding glyoxalase II from Leishmania donovani: a potential target for anti-parasite drugs.
The glyoxalase system is a ubiquitous detoxification pathway that protects against cellular damage caused by highly reactive oxoaldehydes such as methylglyoxal which is mainly formed as a by-product of glycolysis. The gene encoding GLOII (glyoxalase II) has been cloned from Leishmania donovani, a protozoan parasite that causes visceral leishmaniasis. DNA sequence analysis revealed an ORF (open reading frame) of approximately 888 bp that encodes a putative 295-amino-acid protein with a calculated molecular mass of 32.5 kDa and a predicted pI of 6.0. The sequence identity between human GLOII and LdGLOII (L. donovani GLOII) is only 35%. The ORF is a single-copy gene on a 0.6-Mb chromosome. A approximately 38 kDa protein was obtained by heterologous expression of LdGLOII in Escherichia coli, and homogeneous enzyme was obtained after affinity purification. Recombinant L. donovani GLOII showed a marked substrate specificity for trypanothione hemithioacetal over glutathione hemithioacetal. Antiserum against recombinant LdGLOII protein could detect a band of anticipated size approximately 32 kDa in promastigote extracts. By overexpressing the GLOII gene in Leishmania donovani using Leishmania expression vector pspalphahygroalpha, we detected elevated expression of GLOII RNA and protein. Overexpression of the GLOII gene will facilitate studies of gene function and its relevance as a chemotherapeutic target. This is the first report on the molecular characterization of glyoxalase II from Leishmania spp. The difference in the substrate specificity of the human and Leishmania donovani glyoxalase II enzyme could be exploited for structure-based drug design of selective inhibitors against the parasite.


Activation of latent collagenase purified from human leukocytes.
Latent human leukocyte collagenase was isolated to apparent homogeneity by a simple and rapid method. Isolation was accomplished by gel filtration on Sepharose 6B and ion exchange chromatography on QAE Sephadex A-50 followed by affinity chromatography on Cibacron Blue Sepharose. The purified latent enzyme exhibits an apparent molecular weight of 70 kD as estimated by SDS-polyacrylamide gel electrophoresis. Reduction with dithiothreitol does not change the mobility of the latent human leukocyte collagenase on SDS-polyacrylamide gel electrophoresis, indicating that the enzyme consists of a single polypeptide chain. The enzyme could be activated by trypsin and thiol reagents such as phenylmercuric chloride and N-ethylmaleimide. Upon activation by trypsin a 54 kD polypeptide was formed from the 70 kD latent enzyme. Concomitant with the activation by thiol reagents, no loss of molecular weight was detected. Inactivated trypsin, i.e. phenylmethyl sulfonyl-trypsin or soybean trypsin inhibitor treated trypsin, was not able to activate latent human leukocyte collagenase. The results support the concept that latent human leukocyte collagenase exists as a proenzyme and thiol-dependent activation occurs through conformational perturbation in the proenzyme molecule.


Loop-Mediated Isothermal Amplification for Detection of Plant Pathogens in Wheat (Triticum aestivum).
Wheat plants can be infected by a variety of pathogen species, with some of them causing similar symptoms. For example, Zymoseptoria tritici and Parastagonospora nodorum often occur together and form the Septoria leaf blotch complex. Accurate detection of wheat pathogens is essential in applying the most appropriate disease management strategy. Loop-mediated isothermal amplification (LAMP) is a recent molecular technique that was rapidly adopted for detection of plant pathogens and can be implemented easily for detection in field conditions. The specificity, sensitivity, and facility to conduct the reaction at a constant temperature are the main advantages of LAMP over immunological and alternative nucleic acid-based methods. In plant pathogen detection studies, LAMP was able to differentiate related fungal species and non-target strains of virulent species with lower detection limits than those obtained with PCR. In this review, we explain the amplification process and elements of the LAMP reaction, and the variety of techniques for visualization of the amplified products, along with their advantages and disadvantages compared with alternative isothermal approaches. Then, a compilation of analyses that show the application of LAMP for detection of fungal pathogens and viruses in wheat is presented. We also describe the modifications included in real-time and multiplex LAMP that reduce common errors from post-amplification detection in traditional LAMP assays and allow discrimination of targets in multi-sample analyses. Finally, we discuss the utility of LAMP for detection of pathogens in wheat, its limitations, and current challenges of this technique. We provide prospects for application of real-time LAMP and multiplex LAMP in the field, using portable devices that measure fluorescence and turbidity, or facilitate colorimetric detection. New technologies for detection of plant pathogen are discussed that can be integrated with LAMP to obtain elevated analytical sensitivity of detection.


Measuring Excess Heat Capacities of Deoxyribonucleic Acid (DNA) Folding at the Single-Molecule Level.
Measurements of the thermodynamic properties of biomolecular folding (ΔG°, ΔH°, ΔS°, etc.) provide a wealth of information on the folding process and have long played a central role in biophysical investigation. In particular, the excess heat capacity of folding (ΔCP) is crucial, as typically measured in bulk ensemble studies by differential scanning calorimetry (DSC) and isothermal titration calorimetry (ITC). Here, we report the first measurements of ΔCP at the single-molecule level using the single-molecule fluorescence resonance energy transfer (smFRET) as well as the very first measurements of the heat capacity change associated with achieving the transition state (ΔC‡P) for nucleic acid folding. The deoxyribonucleic acid (DNA) hairpin used in these studies exhibits an excess heat capacity for hybridization (ΔCP = -340 ± 60 J/mol/K per base pair) consistent with the range of literature expectations (ΔCP = -100 to -420 J/mol/K per base pair). Furthermore, the measured activation heat capacities (ΔC‡P) for such hairpin unfolding are consistent with a folding transition state containing few fully formed base pairs, in agreement with prevailing models of DNA hybridization.


Isolation and characterization of a new member of the scavenger receptor superfamily, glycoprotein-340 (gp-340), as a lung surfactant protein-D binding molecule.
We have purified a previously unknown glycoprotein (designated gp-340) from human bronchioalveolar lung washings from a patient with alveolar proteinosis. gp-340 was identified by its calcium-dependent binding to lung surfactant protein D (SP-D) and by its molecular mass of 340 kDa in the reduced state on SDS-polyacrylamide gel electrophoresis (PAGE). gp-340 was purified from the 10,000 x g pellet of the lavage fluid by ion-exchange and gel permeation chromatography. On SDS-PAGE, gp-340 showed an apparent molecular mass of 290 kDa in the unreduced state. On gel chromatography under non-dissociating conditions, the apparent molecular mass of gp-340 was >1000 kDa. The presence of N-linked glycosylation was shown by digestion with N-glycosidase F, which reduced the apparent molecular mass of gp-340 under reducing condition to about 300 kDa. Partial amino acid sequence data showed the presence of scavenger-receptor type domains. Monoclonal and polyclonal antibodies were raised against gp-340, and their specificities were confirmed by Western blotting. The antibodies were used for immunohistochemical localization of gp-340 in the lung, where it was found on the surface of and within alveolar macrophages. Direct binding between gp-340 and SP-D took place at physiological ionic strength, required the presence of calcium, and was not inhibited by maltose. The binding between SP-D and mannan also required the presence of calcium, but this interaction was completely inhibited by maltose. The same binding pattern was seen between gp-340 and recombinant human SP-D composed of the trimeric neck region and three carbohydrate recognition domains. These findings indicate that the binding between gp-340 and SP-D is a protein-protein interaction rather than a lectin-carbohydrate interaction and that the binding to gp-340 takes place via the carbohydrate recognition domain of SP-D. We conclude that gp-340 is a new member of the scavenger-receptor superfamily and likely to be a truncated form of a receptor for SP-D.


Diagnosis of atypical hemolytic uremic syndrome and response to eculizumab therapy.
Atypical hemolytic uremic syndrome (aHUS) has a high mortality rate if not detected and treated early. While in the past, it was associated with renal failure in children, today, it has become increasingly identified among adults. Due to recent advances in the pathogenesis of aHUS and other major thrombotic microangiopathies (TMA), diagnosing it has become a lot easier. We present a case of a 62-year-old man who was initially thought to have thrombotic thrombocytopenic purpura (TTP), but after further evaluation was diagnosed with aHUS. We will discuss how to distinguish aHUS from other major TMA and the role of eculizumab in the management of aHUS.


Mechanistic studies on N-benzyladriamycin-14-valerate (AD 198), a highly lipophilic alkyl adriamycin analog.
AD 198, a novel lipophilic N-alkyl derivative of adriamycin (ADR) and a potential anticancer agent for preclinical development, was studied for its effects on the activities of DNA and RNA polymerases in vitro and its ability to bind DNA. AD 198, which contains a benzyl substituent on the glycosidic amine, was found to interact with DNA through drug-DNA binding to an extent less than its parent compound ADR as shown by fluorescent emission spectra studies. It had a preferential inhibitory effect against RNA vs. DNA synthesis in vitro by RNA or DNA polymerases from both E. coli and chicken leukemia cells. Preincubation studies indicated that AD 198 may inhibit the activity of E. coli RNA polymerase through drug-template interaction and that of leukemic RNA polymerase, which uses single stranded DNA as template, through enzyme inactivation.


Effects of gamma irradiation on the measurement of hepatitis B virus DNA in dentin harvested from chronically infected patients.
The manufacturing of the demineralized dentin matrix (DDM) has been proven to extensively reduce the presence of human hepatitis B viral DNA (HBV DNA). This study measured and compared HBV DNA in fresh dentin to that in gamma radiation (GR)-sterilized dentin extracted from HBV-infected patients. The application of GR as a means of terminal sterilization is hypothesized to inactivate or eliminate HBV within the dentin matrix. Dentin from 18 HBV-infected patients was collected and divided into three fragments. The first fragment was unaltered and used as the control group; the remaining two fragments were sterilized with gamma radiation doses of 15 or 25 kGy. DNA was extracted and purified from each fresh (control), and the GR-sterilized (experimental) dentin specimen and HBV DNA copy numbers were evaluated on the basis of the real-time polymerase chain reaction. The copy numbers were used to assess GR efficacy as a means of terminal sterilization for HBV inactivation or elimination. HBV DNA was detected in 66.67% of the fresh dentin specimens. The differences in HBV DNA levels between the fresh dentin and the GR-sterilized dentin were confirmed by the Wilcoxon signed-rank test for the doses of 15 and 25 kGy with P value of 0.012 and 0.010, respectively. Among the twelve HBV-DNA-positive fresh dentin samples, HBV DNA persisted in eleven after GR sterilization, yet the copy number was reduced to <10 (except for a single sample within each experimental group). The results suggest that 15 and 25 kGy of GR significantly reduced the HBV DNA levels in the fresh dentin matrix. Expansion of the possible clinical applications of allogenic grafts with the irradiated DDM will require additional studies, including validation of viral load inactivation to prevent infectious transmission and examination of GR exposure effects on the osteoinductivity of the matrix.


The importance of pulmonary complications as a cause of death in surgical patients.
The authors have retrospectively reviewed 390 autopsy reports from their surgical clinic, to establish the frequency of chronic and acute lung complications in surgical patients. The retrospective analysis revealed that pneumonia was the most frequent acute pulmonary complication among patients who died in the surgical department. In postoperative cases, pneumonia contributed to or was responsible for 65% of all deaths.


Calcium micro-depositions in jugular truncular venous malformations revealed by Synchrotron-based XRF imaging.
It has been recently demonstrated that the internal jugular vein may exhibit abnormalities classified as truncular venous malformations (TVMs). The investigation of possible morphological and biochemical anomalies at jugular tissue level could help to better understand the link between brain venous drainage and neurodegenerative disorders, recently found associated with jugular TVMs. To this end we performed sequential X-ray Fluorescence (XRF) analyses on jugular tissue samples from two TVM patients and two control subjects, using complementary energies at three different synchrotrons. This investigation, coupled with conventional histological analyses, revealed anomalous micro-formations in the pathological tissues and allowed the determination of their elemental composition. Rapid XRF analyses on large tissue areas at 12.74 keV showed an increased Ca presence in the pathological samples, mainly localized in tunica adventitia microvessels. Investigations at lower energy demonstrated that the high Ca level corresponded to micro-calcifications, also containing P and Mg. We suggest that advanced synchrotron XRF micro-spectroscopy is an important analytical tool in revealing biochemical changes, which cannot be accessed by conventional investigations. Further research on a larger number of samples is needed to understand the pathogenic significance of Ca micro-depositions detected on the intramural vessels of vein walls affected by TVMs.


Subspecialized radiology review at multidisciplinary pancreas conference: impact on patient management.
At our tertiary medical center, multidisciplinary subspecialists meet twice a week during a CME-accredited conference to discuss oncologic and non-oncologic patients with pancreatic diseases at which time a subspecialized abdominal staff radiologist reinterprets the patient's relevant imaging studies. This study assesses the changes in patient management due to imaging reinterpretation during multidisciplinary pancreas conference (MPC). In this retrospective, IRB-approved, HIPAA-compliant study, imaging studies of all patients discussed at MPC between July 1 and December 31, 2015 were assessed for technical adequacy, and original reports analyzed for congruency with reinterpretation. Management measures included change in diagnosis, clinical stage, treatment, or workup. Additional data were obtained on referring services affected and their resultant change in practice. Changes in surgical resectability, surgical approach, or delayed operative dates were noted for surgeons. Changes in chemotherapeutic or radiation oncology regimens as well as decisions for additional imaging, laboratory workup, or histologic evaluation were also recorded. A total of 252 patients were included (54.4% males, 45.6% females, mean age 63.71 years). Relevant imaging consisted of 142 abdominal CT scans, 112 abdominal MRI scans, 1 abdominal ultrasound, and 1 nuclear medicine octreotide study of which 69.4% were performed in-house. Image quality was deemed appropriate in 95.2%. Cases presented included solid pancreatic malignancies (n = 140; 55.6%), cystic pancreatic lesions (n = 41; 16.3%), acute and chronic inflammatory conditions (n = 52; 20.6%), and miscellaneous entities (n = 10; 4.0%); 9 (3.6%) cases were normal. Image reinterpretation was congruent with original reports in 56.7%, with minor, moderate, and major discrepancies occurring in 9.5, 26.2%, and 7.5% of cases, respectively. Incongruent reinterpretation was predominantly due to perceptional errors (false-negative reports due to missed findings) and interpretative errors (false-positive results due to over-reporting or misclassification of diagnoses). Services most commonly affected included surgical oncology, radiology, and gastroenterology at 16.7%, 13.1%, and 12.7% of cases, respectively. Management changes included a change in diagnosis in 8.7%, change in clinical stage in 8.7%, change in treatment in 17.9%, and further workup needed in 19.0% of patients, respectively. No change in management occurred in the remaining 61.5% of cases. Subspecialized abdominal radiologist reinterpretation in the context of more comprehensive patient information heavily impacts the multidisciplinary management of patients with pancreatic disorders. Further efforts are needed to solidify the abdominal radiologist's role in the multidisciplinary clinical setting.


Feeling older and wanting to be younger.
Two samples of middle-aged women were compared using a questionnaire adapted from Rossi (1980). One sample (N = 20) consisted of women who felt older than and desired to be younger than their own age. The second sample (N = 19) was comprised of women who felt and desired to be their own age. Statistically significant differences between the groups were that women who felt older reported more physical changes of aging, less satisfaction with their life context, and more worries than the women who felt their chronological age. Results of this study suggest that current life stress contributes to middle-aged women's perceptions of feeling older and desiring to be younger than their own age.


Transcriptional and metabolic data integration and modeling for identification of active pathways.
With the growing availability of omics data generated to describe different cells and tissues, the modeling and interpretation of such data has become increasingly important. Pathways are sets of reactions involving genes, metabolites, and proteins highlighting functional modules in the cell. Therefore, to discover activated or perturbed pathways when comparing two conditions, for example two different tissues, it is beneficial to use several types of omics data. We present a model that integrates transcriptomic and metabolomic data in order to make an informed pathway-level decision. Since metabolites can be seen as end-points of perturbations happening at the gene level, the gene expression data constitute the explanatory variables in a sparse regression model for the metabolite data. Sophisticated model selection procedures are developed to determine an appropriate model. We demonstrate that the transcript profiles can be used to informatively explain the metabolite data from cancer cell lines. Simulation studies further show that the proposed model offers a better performance in identifying active pathways than, for example, enrichment methods performed separately on the transcript and metabolite data.


Interaction between the physical form of the starter feed and straw provision on growth performance of Holstein calves.
Two experiments were conducted to assess the effect of physical form of a starter feed with or without straw supplementation on growth performance of Holstein calves. In experiment 1, a total of 32 calves were randomly assigned at 7 d of age to texturized starter feed (containing rolled barley, corn, and oats) without straw, texturized starter feed with chopped straw, and pelleted starter feed with chopped straw. All calves were offered 4 L of pasteurized whole milk twice daily from 7 to 35 d of age, 2 L of milk twice daily from 36 to 42 d of age, and 2 L of milk from 43 to 49 d of age. Animals were weaned at 50 d of age, and the study finished when calves were 63 d old. In experiment 2, a total of 60 calves (8 d of age) were randomly assigned to texturized starter feed (containing whole corn) without straw, pelleted starter feed without straw, and pelleted starter feed with chopped straw. All calves were offered the same milk replacer (MR; 23% crude protein and 19.5 fat) at 11% dry matter concentration, 4 L/d of MR until 14 d of age, 6 L/d of MR from 14 to 37 d, 3 L/d of MR from 38 to 44 d, and 1.5 L/d of MR from 45 to 52 d of age. The experiment finished when calves were 58 d old (1 wk after weaning). Rumen liquid pH was measured after weaning. In both studies, calves were individually housed in pens on sawdust bedding and starter feed and chopped straw were offered free choice in separate buckets. In experiment 1, starter feed and straw intake and growth did not differ among treatments. However, calves receiving straw showed a greater rumen pH compared with those not receiving straw. In experiment 2, pelleted started feed supplemented with straw fostered an increase in solid feed intake (as percentage of body weight) compared with a pelleted or texturized starter feed without straw supplementation. However, calves that received the texturized starter feed containing whole corn had rumen pH similar to those fed a pelleted starter feed with straw. Feeding a texturized starter feed containing rolled barley, corn, and oats (with or without straw provision) was not able to maintain rumen pH or promote growth and intake compared with offering a pelleted starter feed with chopped straw. However, when whole corn was used in the texturized starter feed, rumen pH was equivalent to that obtained with a pelleted starter feed and straw supplementation.


Ectopic expression of a genomic fragment containing a homeobox causes neural defects in the axolotl.
An axolotl (Ambystoma mexicanum) genomic fragment containing the Ahox1 homeobox was placed under the control of the mouse hsp68 promoter, which seems to function constitutively in the axolotl. The resulting construct was injected into fertilized axolotl eggs to see if it would perturb development. Of the injected embryos, 20% showed severe reduction of the anterior neural plate. Later in development, these embryos had small heads, no eyes, and appeared to lack the normal regionalization of the brain. An additional 35% of the embryos were less severely affected, but had reduced or missing eyes. Control embryos, including ones injected with a construct missing the DNA recognition helix of the homeobox, developed normally.


Potential Candidates for Emergency Department Initiated Extracorporeal Cardiopulmonary Resuscitation (ECPR) in a Canadian Institution.
Introduction Out-of-hospital cardiac arrest (OHCA) patients experience poor survival. The use of extracorporeal membrane oxygenation (ECMO), a form of heart-lung bypass, in the setting of cardiac arrest, termed extracorporeal cardiopulmonary resuscitation (ECPR), has promise in improving survival with good neurologic outcomes. The study objective was to determine the number of potential annual ECPR candidates among the OHCA population in a health region within the Atlantic Canadian province of Nova Scotia. Methods A retrospective chart review was conducted over a five-year period: January 1st, 2012 to December 31st, 2016. Consecutive non-traumatic OHCA and emergency department (ED) cardiac arrests occurring in a pre-determined catchment area (20-minute transport to ECMO center) defined by a geographic bounding box were identified. Criteria for ECPR were developed to identify candidates for activation of a "Code ECPR": (1) age 16-70, (2) witnessed arrest, (3) no flow duration (time to CPR, including bystander) <10 minutes, (4) resuscitation >10 minutes without return of spontaneous circulation (ROSC), (5) emergency medical service (EMS) transport to hospital <20 minutes, (6) no patient factors precluding ongoing resuscitation (do not resuscitate status (DNR), palliative care involvement, or metastatic cancer), and (7) initial rhythm not asystole. Candidates were stratified by initial rhythm. Candidates were considered ultimately ED ECPR eligible if they failed conventional treatment, defined by death or resuscitation >30 minutes. Clinical data related to candidacy was extracted by an electronic query from prehospital and ED electronic records and manual chart review by three researchers. Results Our search yielded 561 cases of EMS-treated OHCA or in-ED arrests. Of those 204/561 (36%; 95% CI 33-40%) met the criteria for activation of a "Code ECPR". Ultimately 79/204 (34%; 95% CI 28-41%) of those who met activation criteria were considered ED ECPR eligible; which is 14% (95% CI 11-17%) of the total number of arrests-of the total number of arrests, the initial rhythms were pulseless electrical activity (PEA) 33/79 (42%; 95% CI 32-53%) and shockable 46/79 (58%; 95% CI 47-69%). Conclusion Of all cardiac arrests in the area surrounding our ECMO center, approximately 41 per year met the criteria for a Code ECPR activation, with 16 per year ultimately being eligible for ED ECPR. This annual estimate varies based on the inclusion of initial rhythm. This provides insight into both prehospital and hospital implications of an ED ECPR program and will help guide the establishment of a program within our Nova Scotian health region. This study also provides a framework for similar investigation at other institutions contemplating ED ECPR program implementation.


A comparative study of Ag(I) adsorption on raw and modified spent grain: kinetic and thermodynamic aspects.
The capability of modified spent grain (MSG) to adsorb silver (I) [Ag(I)] from aqueous solution was investigated and compared with raw spent grain (RSG) regarding their adsorption isotherms, kinetics, and thermodynamics. The monolayer adsorption capacity was 30.28 mg/g for RSG and 158.23 mg/g for MSG according to the Langmuir isotherm. The Ag(I) adsorption on MSG was approximately 4 times higher than that on RSG. The pseudo-second-order kinetic model provided the best description of Ag(I) adsorption on the two adsorbents. The calculated activation energy (Ea) implies that the adsorption of Ag(I) on RSG is a physical adsorption and on MSG is of a chemical nature. Thermodynamic results suggest that the adsorption of Ag(I) is an exothermic process for RSG and an endothermic process for MSG, whereas both adsorption processes are spontaneous in nature.


Anti-idiotypic antibodies to a polyomavirus monoclonal antibody recognize cell surface components of mouse kidney cells and prevent polyomavirus infection.
Anti-idiotypic antibodies have been successfully used to identify and isolate the receptor for several cell ligands. To prepare an immunologic probe for identification of the polyomavirus receptor on mouse kidney cells, polyclonal antisera against antipolyomavirus antibodies were prepared in rabbits. Fab fragments of the previously characterized monoclonal antibody E7, which neutralizes polyomavirus infection, were used for immunization (S. J. Marriott and R. A. Consigli, J. Virol. 56:365-372, 1985). Sera containing the greatest anti-idiotype activity were identified by enzyme-linked immunosorbent assay (ELISA) and purified by a series of affinity columns. The anti-idiotypic antibodies recognized the E7 idiotope in an ELISA, and anti-idiotype binding could be inhibited by preincubation of E7 monoclonal antibody with polyomavirus virions. When mixed with anti-idiotype immunoglobulin G (IgG), E7 was no longer capable of binding or immunoprecipitating polyomavirus virions or neutralizing polyomavirus infection. Direct immunofluorescence showed anti-idiotype IgG reactivity with a cell surface component of mouse kidney cells. Anti-idiotype F(ab')2 effectively competed with polyomavirus for binding to mouse kidney cells and displayed binding kinetics similar to those of polyomavirus. Virus infection of mouse kidney cells was blocked in a dose-dependent manner following treatment of the cells with anti-idiotype IgG. The anti-idiotype identified several proteins (95, 50, and 24 to 31 kilodaltons) in an immunoblot of mouse kidney cell membrane proteins but reacted predominantly with a single 50-kilodalton protein in a radioimmunoassay. The anti-idiotype failed to react with virus proteins in three assays, including ELISA, immunoprecipitation, and immunoblotting. The implications of this work for future identification and characterization of the polyomavirus receptor on mouse kidney cells are discussed.


Irradiation of peripheral blood mononuclear cells with 7.5 Gy X-rays prior to donor lymphocyte infusion inhibits proliferation while preserving cytotoxicity, and improves the effectiveness of HSCT in patients with hematological malignancies.
The aim of the present study was to explore the effect of different X-ray doses on the proliferation and cytotoxic activity of peripheral blood mononuclear cells (PBMCs), particularly lymphocytes, in order to assess whether this reduces the incidence of graft vs. host disease (GVHD) while preserving the graft vs. tumor (GVT) effect in patients with hematological malignancies following hematopoietic stem cell transplantation (HSCT). PBMCs from healthy donors were irradiated with X-rays at doses of 0, 2.5, 5, 7.5, 10, 15, 25 or 50 Gy, and their proliferative activity was determined using a WST-8 assay kit. The cytotoxic activity of non-irradiated PBMCs and PBMCs irradiated with 7.5 Gy X-rays was tested in the leukemic cell line K562 and its Adriamycin-resistant strain K562A using a lactate dehydrogenase assay. The clinical data of 7 patients who received 7.5 Gy X-ray-irradiated PBMC infusions following autologous HSCT were analyzed. PBMCs irradiated with ≥7.5 Gy X-rays exhibited a complete inhibition of proliferation. PBMCs irradiated with 7.5 Gy X-rays exhibited significantly increased cytotoxic activity towards K562 cells compared with K562A cells (P<0.05). There was no significant difference in cytotoxicity between irradiated and non-irradiated PBMCs, irrespective of the target cell, K562 or K562A (P>0.05). Based on the in vitro data, clinical data from patients who received 7.5 Gy X-ray-irradiated PBMC infusions following HSCT between January 2005 and January 2013 were assessed retrospectively. A total of 7 patients were included in the current study. The majority achieved various degrees of remission following donor lymphocyte infusion (DLI) and none suffered from GVHD. This indicates that 7.5 Gy-irradiated PMBCs have a potential application in DLI for the treatment of patients following HSCT. However, further studies on larger patient cohorts are required to assess the clinical potential of 7.5 Gy-irradiated PBMCs for preserving the GVT effect while avoiding GVHD following HSCT.


Demystifying patient price estimates. The advantages of transparency.
With the increase of high-deductible health plans, more consumers want to know the cost of their health care before they purchase services. A healthcare organization should formulate transparent price policies that: Fit with its intentions, processes, and goals. Ensure consumers are thoroughly educated about their financial responsibilities. Include the use of consumer pricing tools that help patients feel like empowered consumers. Reflect an enterprisewide culture of transparency.


New monocyte locomotion inhibitory factor analogs protect against cerebral ischemia-reperfusion injury in rats.
Monocyte locomotion inhibitory factor (MLIF) is an oligopeptide with anti-inflammatory properties. The carboxyl-terminal end group Cys-Asn-Ser serves as the pharmacophore of MLIF. The aim of this study was to investigate the neuroprotective effects of two new synthetic analogs, Arg-Cys-Asn-Ser and D-Cys-Asn-Ser, on focal cerebral ischemia, which were designed and synthesized to increase the penetrability and enzymatic stability of Cys-Asn-Ser. Ninety-one male Sprague-Dawley rats were randomly divided into six groups: I - Sham; II - Ischemia-reperfusion (I/R); III - Nimodipine; IV - Cys-Asn-Ser; V - D-Cys-Asn-Ser; and VI - Arg-Cys-Asn-Ser. The rats in groups II-VI were subjected to middle cerebral artery occlusion. After 24 hours of reperfusion, the neurological deficit, cerebral infarct volume, and levels of the pro-inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor-alpha in brain tissue homogenates were assessed. Compared with the sham group, the mean neurological deficit scores were significantly higher in groups II-VI (p ≤ 0.019 for all). The mean infarct volumes were significantly higher in I/R and Cys-Asn-Ser groups compared with the sham group (both p ≤ 0.046). The mean IL-1β level was significantly lower in D-Cys-Asn-Ser and Arg-Cys-Asn-Ser groups compared with I/R group (both p ≤ 0.046). In conclusion, the results showed that Arg-Cys-Asn-Ser and D-Cys-Asn-Ser have the potential for protective effects against focal cerebral ischemia injury.


Fever Through a Jaundiced Eye.
Pyogenic liver abscess (PLA) is an important clinical entity to consider in a patient with fever and abdominal pain. Previously, the condition was difficult to diagnose and treat, but with the introduction of widely available and reliable imaging techniques, its diagnosis has become more straightforward. Although uncommon, PLA should especially be considered in the differential diagnosis for patients with specific predisposing conditions such as underlying biliary tract disease, whether as a result of chronic inflammatory disease or malignancy. The introduction of percutaneous drainage has revolutionized the management of PLA, and thus, this disease has become largely correctable.


Screening Program of Abdominal Aortic Aneurysm.
In Europe, the prevalence of abdominal aortic aneurysms (AAAs) in the elderly population (≥65 year old) has declined in the past decades to <4%. Aneurysmal degeneration of the aorta is a serious and potentially life-threatening vascular disease. Abdominal aortic aneurysms typically develop subclinically and often only become symptomatic when complicated by impending rupture. Most AAAs are discovered incidentally while investigating for an unrelated pathology. Ruptured AAA is the tenth leading cause of death in Belgium (0.32% of all deaths in 2014). Health-care providers have emphasized the importance of early detection of AAA and elective repair when the rupture risk outweighs operative risk (usual diameter threshold of 55 mm). Routine AAA screening programs, consisting of a single abdominal ultrasonography at the age of 65 years, aim to reduce the number of AAA-related deaths. Does population-based ultrasound screening for AAA achieve its objective and is it cost-effective? This literature review tries to answer these challenging questions.


Freirean Conscientization With Critical Care Nurses to Reduce Moral Distress and Increase Perceived Empowerment: A Pilot Study.
Moral distress in nursing has been linked to a relative lack of power in the workplace. We designed and piloted a Freirean-based conscientization intervention to raise awareness and address disempowerment with 13 critical care nurses in the US Midwest who recently experienced moral distress. A transformative, mixed-methods, pre/post design was used with convergence of qualitative and quantitative data. Results showed improved moral distress and mixed results in perceived personal and group empowerment. Further study is warranted to test conscientization-based interventions that give nurses opportunity collectively to develop empowered responses to situations that lead to moral distress.


Concentration and purification of hepatitis B antigen with polyethylene glycol and polyelectrolyte 60, a cross-linked copolymer of isobutylene maleic anhydride.
A simple two-step procedure is described for the concentration and purification of hepatitis B antigen. The antigen is precipitated by polyethylene glycol and then adsorbed on and eluted from an insoluble polyelectrolyte, polyelectrolyte 60, which is a cross-linked copolymer of isobutylene maleic anhydride.


Criticism and interpretation: teaching the persuasive aspects of research articles.
Research articles are an excellent tool for promoting active learning about the scientific process. One difficulty in teaching research articles is that they address a professional audience and often seek to be persuasive as well as informative. This essay discusses pedagogical strategies that are intended to help students differentiate the purely informative aspects of research articles, such as descriptions of the methods and results, from the persuasive aspects, such as interpretation of results and critical evaluation of the work of other scientists.


Similar HCV viral load levels and genotype distribution among end-stage renal disease patients on hemodialysis and HCV-infected patients with normal renal function.
At present it is not clearly established if hepatitis C virus (HCV)-RNA levels and genotype distribution have any peculiar aspect in HCV-infected end-stage renal disease (ESRD) patients. The aim of this study was to evaluate in HCV-infected ESRD patients, the alanine aminotransferase (ALT) profile, HCV-RNA levels and genotype distribution, comparing them with HCV-infected patients with normal renal function. A cross-sectional study was performed in 66 hemodialysis patients (group 1) and 264 subjects with normal renal function (group 2). All participants in both groups had detectable HCV-RNA. Mean ALT levels were determined in all subjects as well the viral load and the genotype. Groups were similar according to gender and age. ALT was normal in 74% patients in group 1 and in 23% in group 2 (p<0.001). The median viral load was 5.3 x 10(5) IU/mL in group 1 and 6.6 x 10(5) IU/mL in group 2 (p=0.23). Genotype 1b was the most prevalent in both groups (56% vs. 57%; p=0.38). HCV-infected ESRD patients have lower ALT levels, but the viral load and the genotype distribution are similar to those observed in HCV-infected individuals with normal renal function.


Synthesis and biological properties of sequence-specific DNA-alkylating pyrrole-imidazole polyamides.
In recent years, many diseases including cancer and hereditary and viral diseases have been understood at the DNA sequence level. Direct control of the expression level of a specific gene would provide a promising approach for knowledge-based therapy. N-Methylpyrrole (Py) and N-methylimidazole (Im) polyamides are a new type of small compound that precisely bind to the minor groove of the DNA duplex in a sequence-specific fashion and recruit alkylating agents to the target sequence. We designed and synthesized a series of sequence-specific alkylating Py-Im polyamide conjugates that selectively alkylate predetermined DNA sequences. We have demonstrated that sequence-specific alkylating agents possess gene-silencing activities and a promising potency against human cancer cell lines as well as against xenografts of human cancer cell lines. In this Account, we focus on recent progress in alkylating Py-Im polyamides with regard to sequence specificity and biological activities and the future direction of rational molecular design of genetic switches in the postgenome era is described.


Funisitis as a Risk Factor for Adverse Neonatal Outcomes in Twin Neonates with Spontaneous Preterm Birth: A Retrospective Cohort Study.
Funisitis, inflammation of the umbilical cord, is considered a strong risk factor for adverse neonatal outcomes; however, a clinical definition of funisitis has not been established. In this study, we aimed to determine the clinical significance of funisitis in twin neonates with spontaneous preterm birth. The study included preterm twin neonates (<35 weeks) delivered after spontaneous preterm labor and/or preterm premature rupture of amniotic membranes. The presence of funisitis was examined in the umbilical cord of each twin. We analyzed the risk of adverse neonatal outcomes according to the presence and absence of funisitis. Adverse neonatal outcomes were defined as the occurrence of neonatal mortality, significant morbidity, or both. Among 474 preterm neonates (237 twin pairs) included in this study, the frequency of funisitis was 6.5% (31 cases). Funisitis was significantly associated with neonatal mortality and adverse neonatal outcomes after adjustment for confounding variables [neonatal mortality, odds ratio (OR) 9.043, 95% confidence interval (CI) 2.620-31.204; adverse neonatal outcome, OR 2.445, 95% CI 1.017-5.875]. The concordance rate of funisitis between the twins was 10.7%, and in the absence of funisitis in one twin, the risk of neonatal mortality or adverse neonatal outcome was not influenced by the presence of funisitis in the other twin. The presence of funisitis appears to be associated with an increased risk for adverse neonatal outcomes in twin neonates with spontaneous preterm birth.


Structural basis for the unusual carbohydrate-binding specificity of jacalin towards galactose and mannose.
Evidence is presented that the specificity of jacalin, the seed lectin from jack fruit (Artocarpus integrifolia), is not directed exclusively against the T-antigen disaccharide Galbeta1,3GalNAc, lactose and galactose, but also against mannose and oligomannosides. Biochemical analyses based on surface-plasmon-resonance measurements, combined with the X-ray-crystallographic determination of the structure of a jacalin-alpha-methyl-mannose complex at 2 A resolution, demonstrated clearly that jacalin is fully capable of binding mannose. Besides mannose, jacalin also interacts readily with glucose, N-acetylneuraminic acid and N-acetylmuramic acid. Structural analyses demonstrated that the relatively large size of the carbohydrate-binding site enables jacalin to accommodate monosaccharides with different hydroxyl conformations and provided unambiguous evidence that the beta-prism structure of jacalin is a sufficiently flexible structural scaffold to confer different carbohydrate-binding specificities to a single lectin.


[Antioxidants of Contracaecum rudolphii (Nematoda)].
Contracaecum rudolphii is the parasitic nematode of fish-eating birds. In the extracts from female, male and larvae L3 and L4 isolated from the alimentary tracts of black cormorants the activity of five antioxidant enzymes: superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione transferase (GST), glutathione reductase (GR), catalase (CAT) and the content of ascorbate and total antioxidative status (TAS) were determined. They can be put in order according to the activity growth: GPX, SOD, GST, CAT and GR. The activity of GPX were very low in the nematodes' extracts (1.23-7.67 microU/mg). CAT had higher activity (0.47-0.72 U/mg). The activity of GR was the highest (50.51-69.88 U/mg). SOD activity in the female was higher by ca. 50% than in the male while GST activity was at similar levels. GR and CAT activities were higher by ca. 30% in the male than in the female nematodes. GST and GPX activity and TAS in larvae L3 were significantly lower than in the adult nematodes or in L4 larvae. The activity of GPX, GR and CAT was lower in L4 larvae than in the adult male (p<0.05). The content of ascorbate was almost the same in all stages of parasite development (0.21-0.38 mg/g). The above results indicate differences in antioxidant systems related to both the sex and the developmental stage of C. rudolphii.


Involvement of cellular protrusions in gamete interactions.
Gamete fusion is of considerable importance in reproductive events, as it determines the gamete pairs or chromosomes that the next generation will inherit. To preserve species specificity with an appropriate karyotype, the fusion between gametes requires regulatory mechanisms to ensure limited fusion competency. In many organisms, gamete surfaces are not smooth, but present constitutive or transient cellular protrusions suggested to be involved in gamete fusion. However, the molecular mechanisms and the factors essential for the membrane-membrane fusion process and cellular protrusion involvement have remained unclear. Recent advances in the identification and functional analysis of the essential factors for gamete interaction have revealed the molecular mechanisms underlying their activity regulation and dynamics. In homogametic fertilization, dynamic regulation of the fusion core machinery on cellular protrusions was precisely uncovered. In heterogametic fertilization, oocyte fusion competency was suggested to correlate with the compartmentalization of the fusion essential factor and protrusion formation. These findings shed light on the significance of cellular protrusions in gamete fusion as a physically and functionally specialized site for cellular fusion. In this review, we consider the developments in gamete interaction research in various species with different fertilization modes, highlighting the commonalities in the relationship between gamete fusion and cellular protrusions.


THE ROLE OF EMPLOYERS IN FACILITATING SMOKING CESSATION AMONG STAFF: PERSPECTIVE OF DRIVERS IN A NIGERIAN UNIVERSITY.
Several interventions have been instituted to encourage smoking cessation among smokers. Many adults are currently working and spend several hours a day at work. Employers thus have a role to play in encouraging staff who smoke to quit. This study investigated the perception of drivers employed in the University of Ibadan, Nigeria on the role of their employers in facilitating smoking cessation among staff who smoke. All 176 drivers employed by the University and who were available on the days of the interviews were interviewed. A semi-structured questionnaire was used to obtain information on drivers' opinion on the role of employers in facilitating smoking cessation among staff. All the drivers were male and had a mean age of 51.2 ± 5.8 years. Five (2.8%) drivers were current smokers. One hundred and thirty-six (77.3%) drivers were of the opinion that their employers had a role to play in encouraging smoking cessation among staff who smoke. They felt this could be done by organizing workshops to sensitize staff on the dangers of smoking 73 (53.7%), educating staff about the health effects of smoking 26 (19.1%) and arresting staff caught smoking during official hours 20 (14.7%). Many of the drivers felt that their employers had a role to play in encouraging smoking cessation among staff. The University authorities should build on this and take specific steps to institute a comprehensive workplace antitobacco policy which includes smoking cessation interventions to assist staff who smoke to quit.


[Contents of copper, zinc, iron and manganese in selected cheeses].
Contents of trace elements (copper, zinc, iron, and manganese) were determined in melted cheeses, mold-ripening semi-hard and soft types purchased in 2005 at local shops. The AAS technique was used. Material for study was produced by different Dairy Centers all over Poland in Lublin, Mazowsze, Opole, Podlasie, Silesia, and Wielkopolska regions. Zinc and iron levels in melted cheeses were significantly higher (p < or = 0.05) as compared to mold-ripening soft and semi-hard cheeses. The highest zinc and iron concentrations were found in the samples of Sertop and Jal cheese. The lowest contents were found in Mabelle cheese. Rycki Edam cheese was characterized by highest level of manganese, and Mabelle--by the lowest. However, copper, iron, and manganese contents in all cheese samples could be considered as low and not hazardous for consumer's.


Adrenal hypofunction and sudden death.
Adrenal insufficiency is an infrequent and easily overlooked cause of sudden death, often occurring in individuals being treated for other serious disease processes. Three cases are presented to demonstrate some of the clinical presentations encountered, and the symptoms and signs are discussed.


The big and intricate dreams of little organelles: Embracing complexity in the study of membrane traffic.
Compartmentalization of eukaryotic cells into dynamic organelles that exchange material through regulated membrane traffic governs virtually every aspect of cellular physiology including signal transduction, metabolism and transcription. Much has been revealed about the molecular mechanisms that control organelle dynamics and membrane traffic and how these processes are regulated by metabolic, physical and chemical cues. From this emerges the understanding of the integration of specific organellar phenomena within complex, multiscale and nonlinear regulatory networks. In this review, we discuss systematic approaches that revealed remarkable insight into the complexity of these phenomena, including the use of proximity-based proteomics, high-throughput imaging, transcriptomics and computational modeling. We discuss how these methods offer insights to further understand molecular versatility and organelle heterogeneity, phenomena that allow a single organelle population to serve a range of physiological functions. We also detail on how transcriptional circuits drive organelle adaptation, such that organelles may shift their function to better serve distinct differentiation and stress conditions. Thus, organelle dynamics and membrane traffic are functionally heterogeneous and adaptable processes that coordinate with higher-order system behavior to optimize cell function under a range of contexts. Obtaining a comprehensive understanding of organellar phenomena will increasingly require combined use of reductionist and system-based approaches.


Estimation of direct and maternal breed effects for prediction of expected progeny differences for birth and weaning weights in three multibreed populations.
Direct and maternal breed effects on birth and 200-d weights were estimated for nine parental breeds (Hereford [H], Angus [A], Braunvieh [B], Limousin [L], Charolais [C], Simmental [S], Gelbvieh [G], Red Poll [R], and Pinzgauer [P]) that contributed to three composite populations (MARC I = 1/4B, 1/4C, 1/4L, 1/8H, 1/8A; MARC II = 1/4G, 1/4S, 1/4H, 1/4A; and MARC III = 1/4R, 1/4P, 1/4H, 1/4A). Records from each population, the composite plus pure breeds and crosses used to create each composite, were analyzed separately. The animal model included fixed effects of contemporary group (birth year-sex-dam age), proportions of individual and maternal heterosis and breed inheritance as covariates, and random effects of additive direct genetic (a) and additive maternal genetic (m) with covariance (a,m), permanent environment, and residual. Sampling correlations among estimates of genetic fixed effects were large, especially between direct and maternal heterosis and between direct and maternal breed genetic effects for the same breed, which were close to -1. This resulted in some large estimates with opposite sign and large standard errors for direct and maternal breed genetic effects. Data from a diallel experiment with H, A, B, and R breeds, from grading up and from a top cross experiment were required to separate breed effects satisfactorily into direct and maternal genetic effects. Results indicate that estimation of direct and maternal breed effects needed to predict hybrid EPD for multibreed populations from field data may not be possible. Information from designed crossbreeding experiments will need to be incorporated in some way.


Ethnic differences for developing rheumatic fever in a low-income group living in Hawaii.
The risk factors responsible for acute rheumatic fever (ARF) are complex, in part, because group A streptococcus (GAS) infection is a prerequisite for this disease. We attempted to differentiate socioeconomic from genetic risk factors by studying subjects in a Hawaii pediatric cardiology clinic who qualified for Medicaid. This ethnically diverse group was unique because they maintained a low socioeconomic but generally healthy lifestyle with more limited risks than those living in extremely impoverished conditions. Questionnaires were administered to consenting subjects in the clinic, who were divided into those diagnosed with ARF (n = 26) and those with other (primarily congenital) heart diseases (n = 41). The socioeconomic status of the ARF and non-ARF groups was lower than that of the Hawaii population in general, and little differences were noted between the groups. The ARF group, however, had slightly larger household sizes and more children than the non-ARF group. The greatest difference was in ethnicity. By the Fisher exact test, the number of Polynesians belonging to the ARF group was significantly greater than all other ethnicities (P = .005). Polynesians had an odds ratio > 4.80 of developing ARF, which increased to 6.33 when number of children per household was considered. The potential contribution of genetic predisposing factors for developing ARF was analyzed in subjects living in a homogeneously low socioeconomic level relative to the general Hawaii population. Polynesians were at highest risk when compared to other ethnicities living in similar socioeconomic conditions.


Effect of endotoxin contamination on the antigenic skin test response.
Bacterial lipopolysaccharides (LPS), the major part of endotoxin, are proinflammatory substances present in the natural environment. Lipopolysaccharides enhance allergen-induced histamine release from human basophils of allergic subjects. The aim of this study was to assess the LPS activity on the allergy skin test response. Twenty patients allergic to house dust mite (HDM) underwent skin prick tests with LPS (E coli 026: B6 serotype) at concentrations of 1, 10, 100 micrograms/mL, with HDM extracts at different dilutions, and with solutions of LPS mixed with HDM. Endotoxin alone did not induce any immediate skin response; LPS had a synergistic effect on the HDM skin response both in terms of flare [27.4 +/- 2.8 mm, 26.6 +/- 3.3 mm, 28.1 +/- 2.5 mm versus 20.7 +/- 3.6 mm, respectively in presence of LPS 1 (P less than .02), 10 (P less than .05), 100 (P less than .01) micrograms/mL] and wheal response [6.4 +/- 1.0 mm, in presence of LPS 100 micrograms/mL versus 4.9 +/- 1.1 mm in absence of LPS (P less than .05)]. On the other hand, a possible LPS synergistic action on the histamine skin response was evaluated in ten normal subjects but the results were not significant. These findings show that LPS increases the allergy skin response probably due to a mechanism including a potentiation of the IgE receptor activation rather than an increase of the skin response to the release mediators.


Immunohistochemical characterization of rat central and peripheral nerve tumors induced by ethylnitrosourea.
Ethylnitrosourea-induced central and peripheral nerve tumors in Sprague-Dawley rats were tested for GFAP (Glial Fibrillary Acidic Protein), S-100 protein, NSE (Neuron Specific Enolase) and Anti-Leu 7 (HNK-1) immunoreactivity utilizing the ABC method (avidin-biotin-complex) for GFAP, S-100 protein and NSE, and the PAP method (peroxidase-antiperoxidase) for Anti-Leu 7. Peripheral nerve neurinomas were consistently positive for S-100 protein and consistently negative for GFAP and Anti-Leu 7. Neurinomas would occasionally exhibit positive staining for NSE (2 of 55 tumors). The staining intensity for S-100 protein varied from strongly positive in differentiated neurinomas to weakly positive in anaplastic tumors. Neoplastic and reactive astrocytes exhibited positive staining for both S-100 protein and GFAP. Variation in the GFAP staining intensity of glial tumors correlated with the degree of differentiation as anaplastic tumors did not stain with the same intensity as their more differentiated counterparts. Oligodendrogliomas exhibited occasional immunoreactivity to S-100 protein (3 of 36 tumors). NSE reactivity in oligodendrogliomas was rarely observed (1 tumor in 36) and immunoreactivity against GFAP or Anti-Leu 7 was consistently absent. Anti-Leu 7 and NSE proved to be of little value in the classification of ENU-induced neural tumors.


Biochemical and antioxidant changes in plasma and erythrocytes of pentathlon horses before and after exercise.
Physical exercise in the horse induces a series of normal physiological and biochemical adaptations. Increasing metabolism and oxygen uptake may induce oxidative stress in various organs. The aim of this study was to examine exercise-induced changes in some plasma and RBC biochemical and antioxidant variables in pentathlon horses. Blood samples were taken from 14 horses before, immediately after, and 24 hours after competing in two 1-minute runs of intense exercise over jumps. The peak intensity periods were preceded by a 20-minute warm-up and separated by a 20-minute break. The following plasma biochemical analytes were determined: total protein, uric acid, and lactate concentrations, and lactate dehydrogenase (LDH) and creatine kinase (CK) activities. Total antioxidant status (TAS) and the ferric reducing ability of plasma (FRAP) also were measured. Thiobarbituric acid-reactive substances (TBARS), reduced glutathione (GSH), and total protein concentrations, and glutathione peroxidase (GSHPx) and superoxide dismutase (SOD) activities were determined in RBC hemolysates. Significantly increased concentrations of total protein, lactate, and FRAP, and increased activities of CK and LDH were observed immediately postexercise compared with pre-exercise samples (P<0.05). All results returned to approximately initial values after 24 hours of rest. RBC GSH and TBARS concentrations did not change immediately after exercise, but decreased after 24 hours of rest (P<0.05). Plasma uric acid and FRAP values were positively correlated in a linear model (r = 0.78). In summary, the type of exercise applied in this study, which can be considered quite usual for pentathlon horses, caused detectable biochemical and lipid peroxidative changes in plasma and RBCs. FRAP and TAS values changed in opposite directions, indicating that when antioxidant capacity is assessed using different methods, highly different results may be obtained.


Citrate does not change uranium chemical speciation in cell culture medium but increases its toxicity and accumulation in NRK-52E cells.
Uranium (U), as a heavy metal, is a strong chemical toxicant, which induces the damage to proximal tubule kidney cells. In order to reproduce U toxicity in vitro and to avoid precipitation, it is necessary to complex it with a strong ligand such as bicarbonate before dilution with cell culture medium. It was recently shown, in vitro on the NRK-52E normal renal tubular epithelial cells, that citrate increased the toxicity of U(VI)-bicarbonate complexes. This property was attributed to a change in U speciation, characterized by the occurrence of U(VI)-citrate complexes, which were supposed to be more toxic than U(VI)-bicarbonate. Here, we present the results of extended X-ray absorption fine structure spectroscopy (EXAFS) analyses of the media that were used to expose cells in vitro. Resulting data show that even when citrate is added to the exposure medium, the predominant species is U(VI)-bicarbonate. Nonetheless, citrate increases U(VI) toxicity and accelerates its intracellular accumulation kinetics, without inducing precipitation. This study emphasizes another parameter that modulates U(VI) toxicity for renal tubule cells and further characterizes the mechanisms of U(VI) toxicity.


Reducing Central Line-Associated Bloodstream Infections in a Burn Intensive Care Unit: Using a Business Framework for Quality Improvement.
Central line-associated bloodstream infections (CLABSIs) pose a unique risk in burn patients, with rates of infection 2-3 times that of other Intensive Care Unit (ICU) populations. Here we present a detailed account of our experience in reducing CLABSI rates utilizing a business framework called the Four Disciplines of Execution (4DX). The Burn ICU CLABSI rate had risen to the 90th percentile nationally when compared to other burn units on the National Healthcare Safety Network. We applied the 4DX framework. This is a four-step method which includes creating a Wildly Important Goal, establishing measurable and accomplishable process measures, creating a scoreboard, and using a weekly meeting to provide accountability. Process changes included both physician and nursing practices. The physicians changed the criteria for when to order blood cultures, as well as requiring attending approval for cultures. The nurses engaged in a peer-observation practice improvement for "scrub the hub" and line dressing conditions and improved their own expertise for peripheral IV placement. The multidisciplinary team initiated a daily review of line indications to ensure removal as soon as possible. Overall, the CLABSI rate decreased from 7.39 infections per 1000 line days to 2.29 infections per 1000 line days over 1 year. We subsequently achieved over 635 days without a CLABSI. In conclusion, the 4DX was a successful quality improvement technique in our healthcare context. Because of the simplicity of implementation, we think it is broadly applicable in the healthcare setting.


Lipopolysaccharide is a frequent and significant contaminant in microglia-activating factors.
Lipopolysaccharide (LPS/endotoxin) is a potent immunologic stimulant. Many commercial-grade reagents used in research are not screened for LPS contamination. LPS induces a wide spectrum of proinflammatory responses in microglia, the immune cells of the brain. Recent studies have demonstrated that a broad range of endogenous factors including plasma-derived proteins and bioactive phospholipids can also activate microglia. However, few of these studies have reported either the LPS levels found in the preparations used or the effect of LPS inhibitors such as polymyxin B (PMX) on factor-induced responses. Here, we used the Limulus amoebocyte lysate assay to screen a broad range of commercial- and pharmaceutical-grade proteins, peptides, lipids, and inhibitors commonly used in microglia research for contamination with LPS. We then characterized the ability of PMX to alter a representative set of factor-induced microglial activation parameters including surface antigen expression, metabolic activity/proliferation, and NO/cytokine/chemokine release in both the N9 microglial cell line and primary microglia. Significant levels of LPS contamination were detected in a number of commercial-grade plasma/serum- and nonplasma/serum-derived proteins, phospholipids, and synthetic peptide preparations, but not in pharmaceutical-grade recombinant proteins or pharmacological inhibitors. PMX had a significant inhibitory effect on the microglia-activating potential of a number of commercial-, but not pharmaceutical-grade, protein preparations. Novel PMX-resistant responses to alpha(2)-macroglobulin and albumin were incidentally observed. Our results indicate that LPS is a frequent and significant contaminant in commercial-grade preparations of previously reported microglia-activating factors. Careful attention to LPS levels and appropriate controls are necessary for future studies in the neuroinflammation field.


Randomised phase 2 study of pembrolizumab plus CC-486 versus pembrolizumab plus placebo in patients with previously treated advanced non-small cell lung cancer.
Preclinical and early clinical studies suggest that combining epigenetic agents with checkpoint inhibitors can potentially improve outcomes in patients with previously treated advanced non-small cell lung cancer (NSCLC). This phase 2 trial examined second-line pembrolizumab + CC-486 (oral azacitidine) in patients with advanced NSCLC. Patients with one prior line of platinum-containing therapy were randomised in a ratio of 1:1 to CC-486 or placebo, on days 1-14, in combination with pembrolizumab on day 1 of a 21-day cycle. The primary end-point was progression-free survival (PFS). Key secondary end-points included overall survival (OS), overall response rate (ORR) and safety. Among 100 patients randomised (pembrolizumab + CC-486: 51; pembrolizumab + placebo: 49), most were male (57.0%), were white (87.0%) and had Eastern Cooperative Oncology Group performance status 1 (68.0%). No significant difference in PFS was observed between the pembrolizumab + CC-486 and pembrolizumab + placebo arms (median, 2.9 and 4.0 months, respectively; hazard ratio [HR], 1.374; 90% confidence interval [CI], 0.926-2.038; P = 0.1789). Median OS was 11.9 months versus not estimable (HR, 1.375; 90% CI, 0.830-2.276; P = 0.2968); ORR was 20% versus 14%. Median treatment duration was shorter (15.0 versus 24.1 weeks), and the number of cycles was lower (5.0 versus 7.0) with pembrolizumab + CC-486 versus pembrolizumab + placebo. No new safety signals for CC-486 or pembrolizumab were detected. Treatment-emergent adverse events were more common in the pembrolizumab + CC-486 arm, particularly gastrointestinal, potentially impacting treatment feasibility. No improvement in PFS was observed with pembrolizumab + CC-486 versus pembrolizumab + placebo. Decreased treatment exposure due to adverse events may have impacted efficacy with pembrolizumab + CC-486.


Efficacy of combined cataract extraction and endoscopic cyclophotocoagulation for the reduction of intraocular pressure and medication burden.
To report on the efficacy of combined endoscopic cyclophotocoagulation (ECP) and phacoemulsification cataract extraction (PCE) with intraocular lens placement for reduction of intraocular pressure (IOP) and medication burden in glaucoma. A retrospective case review of 91 eyes (73 patients) with glaucoma and cataract that underwent combined PCE/ECP surgery was performed. Baseline demographic and ocular characteristics were recorded, as well as intraocular pressure, number of glaucoma medications, and visual acuity postoperatively with 12-month follow-up. Treatment failure was defined as less than 20% reduction in IOP from baseline on two consecutive visits (at 1, 3, 6, or 12mo postoperatively), IOP ≥21 mm Hg or ≤5 mm Hg on two consecutive visits, or additional glaucoma surgery performed within 12mo after PCE/ECP. Overall, mean medicated IOP was reduced from 16.65 mm Hg at baseline to 13.38 mm Hg at 12mo (P<0.0001). Mean number of glaucoma medications was reduced from 1.88 medications at baseline to 1.48 medications at 12mo (P=0.0003). At 3mo postoperatively, the success rate was 73.6% (95%CI: 63.3, 81.5), 57.1% at 6mo (95% CI: 46.3, 66.6), and 49.7% at 12mo (95%CI: 38.9, 59.6). Patient demographic characteristics were not associated with treatment success. The only ocular characteristic associated with treatment success was a higher baseline IOP. Combined PCE/ECP surgery is an effective surgical option for the reduction of IOP and medication burden in glaucoma patients. Patients with higher baseline IOP levels are most likely to benefit from this procedure.


Unconventional T Cell Targets for Cancer Immunotherapy.
Most studies on the immunotherapeutic potential of T cells have focused on CD8 and CD4 T cells that recognize peptide antigens (Ag) presented by polymorphic major histocompatibility complex (MHC) class I and MHC class II molecules, respectively. However, unconventional T cells, which interact with MHC class Ib and MHC-I like molecules, are also implicated in tumor immunity, although their role therein is unclear. These include unconventional T cells targeting MHC class Ib molecules such as HLA-E and its murine ortholog Qa-1b, natural killer T (NKT) cells, mucosal associated invariant T (MAIT) cells, and γδ T cells. Here, we review the current understanding of the roles of these unconventional T cells in tumor immunity and discuss why further studies into the immunotherapeutic potential of these cells is warranted.


Refined Composite Multiscale Dispersion Entropy and its Application to Biomedical Signals.
We propose a novel complexity measure to overcome the deficiencies of the widespread and powerful multiscale entropy (MSE), including, MSE values may be undefined for short signals, and MSE is slow for real-time applications. We introduce multiscale dispersion entropy (DisEn-MDE) as a very fast and powerful method to quantify the complexity of signals. MDE is based on our recently developed DisEn, which has a computation cost of O(N), compared with O(N2) for sample entropy used in MSE. We also propose the refined composite MDE (RCMDE) to improve the stability of MDE. We evaluate MDE, RCMDE, and refined composite MSE (RCMSE) on synthetic signals and three biomedical datasets. The MDE, RCMDE, and RCMSE methods show similar results, although the MDE and RCMDE are faster, lead to more stable results, and discriminate different types of physiological signals better than MSE and RCMSE. For noisy short and long time series, MDE and RCMDE are noticeably more stable than MSE and RCMSE, respectively. For short signals, MDE and RCMDE, unlike MSE and RCMSE, do not lead to undefined values. The proposed MDE and RCMDE are significantly faster than MSE and RCMSE, especially for long signals, and lead to larger differences between physiological conditions known to alter the complexity of the physiological recordings. MDE and RCMDE are expected to be useful for the analysis of physiological signals thanks to their ability to distinguish different types of dynamics. The MATLAB codes used in this paper are freely available at http://dx.doi.org/10.7488/ds/1982.


[Evaluation of the efficacy of harmful agents].
A method is proposed to determine the efficacy of harmful agents, which is based on the assessment of a response of experimental animals by the mathematical-statistical methods formalizing the approaches accepted in hygienic toxicology. It makes it possible to introduce a more detailed classification of their efficacy in accordance with the respective rather strict criteria. This method unifies this procedure to a great extent, thus lowering the influence of subjective factors, which lets us correctly solve the task of establishing the hygienic regulations. The results of evaluation of the efficacy of two combinations of the concentrations of copper, iron, chromium, and zinc contained in the drinking water are given as an example.


Hepatitis E vaccine development: a 14 year odyssey.
The first prophylactic vaccine, Hecolin®, against hepatitis E virus (HEV) infection and the HEV associated disease was approved by China's State Food and Drug Administration (SFDA) in December 2011. Key milestones during the 14-year HEV vaccine development are summarized in this commentary. After years of innovative research the recombinant virus-like particle (VLP) based antigen with virion-like epitopes was successfully produced in E. coli production platform on a commercial scale. Safety and efficacy of this vaccine was demonstrated in a large scale phase III clinical trial.


Early-Life Nutritional Programming of Cognition-The Fundamental Role of Epigenetic Mechanisms in Mediating the Relation between Early-Life Environment and Learning and Memory Process.
The perinatal period is a window of heightened plasticity that lays the groundwork for future anatomic, physiologic, and behavioral outcomes. During this time, maternal diet plays a pivotal role in the maturation of vital organs and the establishment of neuronal connections. However, when perinatal nutrition is either lacking in specific micro- and macronutrients or overloaded with excess calories, the consequences can be devastating and long lasting. The brain is particularly sensitive to perinatal insults, with several neurologic and psychiatric disorders having been linked to a poor in utero environment. Diseases characterized by learning and memory impairments, such as autism, schizophrenia, and Alzheimer disease, are hypothesized to be attributed in part to environmental factors, and evidence suggests that the etiology of these conditions may date back to very early life. In this review, we discuss the role of the early-life diet in shaping cognitive outcomes in offspring. We explore the endocrine and immune mechanisms responsible for these phenotypes and discuss how these systemic factors converge to change the brain's epigenetic landscape and regulate learning and memory across the lifespan. Through understanding the maternal programming of cognition, critical steps may be taken toward preventing and treating diseases that compromise learning and memory.


HISTORICAL BIOGEOGRAPHY OF THE BANANAQUIT (COEREBA FLAVEOLA) IN THE CARIBBEAN REGION: A MITOCHONDRIAL DNA ASSESSMENT.
We analyzed mitochondrial DNA (mtDNA) restriction-site variation in bananaquit (Coereba flaveola; Aves, Coerebinae) populations sampled on 12 Caribbean islands and at 5 continental localities in Central America and northern South America. Multiple fixed restriction-site differences genetically defined several regional bananaquit populations. An mtDNA clade representing all Jamaican bananaquits was the most divergent; the estimated average sequence divergence (dxy ) between Jamaican and all other mtDNA haplotypes surveyed was 0.027. Three groups of populations, representing Central America, northern South America, and the eastern Antilles (Puerto Rico to Grenada) were nearly equally differentiated among themselves (average dxy = 0.014), and may represent a single, recent range expansion. Within the eastern Antilles, three geographically restricted haplotype groups were identified: Puerto Rico, north-central Lesser Antilles (U.S. Virgin Islands to St. Lucia), and Grenada-St. Vincent. The evolutionary relationships of these groups were not clear. Genetic homogeneity of the island populations from the U.S. Virgin Islands to St. Lucia suggested a recent spread of a specific north-central Lesser Antillean haplotype through most of those islands. Haplotype variation across this region indicated that this spread may have occurred in two waves, first through the southernmost islands of St. Lucia, Martinique, and Dominica, and more recently from Guadeloupe to the north. The geographic distribution of mtDNA haplotypes, and of bananaquit populations, suggests periods of invasiveness followed by relative geographic quiescence. Although most genetic studies of bird populations have revealed homogeneity over large geographic areas, our findings provide a remarkable counterexample of strong geographic structuring of mtDNA variation over relatively small distances. Furthermore, although the mtDNA data were consistent with several subspecific distinctions, it was clear that named subspecies do not define equally differentiated evolutionary entities.