Effect of multiple prolyl isomerization reactions on the stability and folding kinetics of the notch ankyrin domain: experiment and theory.
Studies on the folding kinetics of the Notch ankyrin domain have demonstrated that the major refolding phase is slow, the minor refolding phase is limited by the isomerization of prolyl peptide bonds, and that unfolding is multiexponential. Here, we explore the relationship between prolyl isomerization and folding heterogeneity using a combination of experiment and simulation. Proline residues were replaced with alanine, both singly and in various combinations. These destabilizing substitutions combine to eliminate the minor refolding phase, although unfolding heterogeneity persists even when all seven proline residues are replaced. To test whether prolyl isomerization influences the major refolding phase, we modeled folding and prolyl isomerization as a system of sequential reactions. Simulations that use rate constants of the major folding phase of the Notch ankyrin domain to represent intrinsic folding indicate that even with seven prolyl isomerization reactions, only two significant phases should be observed, and that the fast observed phase provides a good approximation of the intrinsic folding in the absence of prolyl isomerization. These results indicate that the major refolding phase of the Notch ankyrin domain reflects an intrinsically slow folding transition, rather than coupling of fast folding events with slow prolyl isomerization steps. This is consistent with the observation that the single observed refolding phase of a construct in which all proline residues are replaced remains slow. Finally, the simulation fails to produce a second unfolding phase at high urea concentrations, indicating that prolyl isomerization does not play a role in the three-state mechanism that leads to this heterogeneity.


Constraints on the Hubble constant from supernova Refsdal's reappearance.
The gravitationally lensed supernova Refsdal appeared in multiple images produced through gravitational lensing by a massive foreground galaxy cluster. After the supernova appeared in 2014, lens models of the galaxy cluster predicted that an additional image of the supernova would appear in 2015, which was subsequently observed. We use the time delays between the images to perform a blinded measurement of the expansion rate of the Universe, quantified by the Hubble constant (H0). Using eight cluster lens models, we infer [Formula: see text]. Using the two models most consistent with the observations, we find [Formula: see text]. The observations are best reproduced by models that assign dark-matter halos to individual galaxies and the overall cluster.


Selectively triggering photoelectromacro for CO2to CH4reduction over SrTiO3{110} facet with dual-metal sites.
In this article, the roles of surface-active sites in dominating photoelectron selectivity for CO2reduction products are well demonstrated over photocatalyst models of SrTiO3{100} and {110} facets. On the easily exposed {100} facets terminated with Sr-O atoms, photoelectrons are of 8 mol % for CH4and 92 mol % for CO generation. The Sr-O-Ti configuration in the {110} facets could enrich the surface charge density due to the lower interface resistance for higher photocatalytic efficiency (1.6 fold). The dual sites of Ti and adjacent Sr atoms are active for strong adsorption and activation of the generated CO* species from primary CO2reduction on the surface, thus kinetically favoring the activity of photoelectrons (73 mol %) in hydrogenation for CH2* species and hence CH4product. Inversely, the poor CH4selectivity is due to difficulty in the subsequent photoelectron reduction reaction by the weak adsorption of CO* at the single-Sr site on the {100} facets, independent of the electron and proton concentration. Our results may offer some illuminating insights into the design of a highly efficient photocatalyst for selective CO2reduction.


Kinetics of human chromosome loss from 3T3-human hybrid cells.
Human chromosomes were lost from populations of 3T3-HeLa and 3T3-diploid human fibroblast (HF) hybrids with similar first-order kinetics. Whereas loss began immediately in 3T3-HF hybrids, there was a lag of 5-10 cell divisions before chromosome loss began in 3T3-HeLa hybrids. Human chromosome loss was not affected by aminopterin selection, the use of polyethylene glycol rather than Sendai virus as fusagen, or by the presence of one or two 3T3 genomes. However, when cell division was retarded by growing 3T3-HF hybrids in low serum or at low temperatures, fewer human chromosomes were lost. This suggests that cell cycle traverse is important in chromosome loss. The distribution of human chromosomes among hybrid metaphases indicated that gradual chromosome loss occurred in all hybrids rather than extensive loss from a portion of the hybrids. During the period of chromosome loss, increased numbers of individual asynchronously condensed human chromosomes were randomly distributed among hybrid metaphases.


Platelets and vascular smooth muscle: abnormalities of phosphodiesterase, aggregation, and cell growth in experimental and human diabetes.
Platelets appear to be involved in both the maintenance of homeostasis and the regulation of proliferation of vascular smooth muscle cells. Anomalies of platelet function may be responsible in part for the pathogenesis of vascular disease in experimental and human diabetes. In a search for an appropriate animal model, we have studied platelet function and the properties of platelet cyclic NCL-PDE in rats with streptozocin-induced diabetes, spontaneous diabetes, and human insulin-dependent diabetes mellitus (IDDM). It appears that opposite abnormalities in both aggregation and phosphodiesterase activity exist in the two animal models. In human IDDM, similar abnormalities to those seen in the BB model were observed. Vascular smooth muscle cells in culture can be used as a model for studies of the effect of circulating growth factors in animals and humans. A growth inhibitory factor was found in plasma and serum from STZ and human IDDM but not BB. In humans we observed increased growth-promoting activity of diabetic platelets, but this phenomenon was absent in both animal models. It remains to be evaluated whether these differences may account for the fact that diabetic rats appear resistant to development of vascular complications. It also remains to be established which animal model is the best choice for studying growth abnormalities in diabetes.


A novel approach to determining physicochemical and absorption properties of 6-fluoroquinolone derivatives: experimental assessment.
The ToSS MoDe approach is used to estimate the n-octanol/buffer partition coefficient, the apparent intestinal absorption rate constant and intestinal permeability from a 6-fluoroquinolone data set. Improved in silico methods for predicting a drug's ability to be transported across biological membranes and other biopharmaceutical properties is highly desirable to optimize new drug development. The physicochemical property (Log P) of 26 6-fluoroquinolone derivatives and the absorption properties (Log K(a) and Log P(eff)) of 21 derivatives were well described by the present approach. The models obtained confirm the important role of lipophilicity in the absorption process and its relation with the piperazinyl ring spectral moment and general local spectral moment. The normalized group contributions to each property, at the R4 and R5 positions of a 6-fluoroquinolone framework, were calculated. Principal factor analysis between these contributions and the Hammett and Hansch constant, molar refractivity and sterimol parameters was also carried out. Three principal factors explained 78% of the total variance and the correlation coefficients were higher than 0.98. The isocontribution zone analysis for the Log P and Log K(a) of Sarafloxacin and Sparfloxacin, used as external corroboration compounds, was carried out. The absorption rate constants (in situ rat gut technique) for these drugs were also evaluated, and the results were compared with the values predicted by theoretical models for evaluating predictive performance. The present approach proved to be a good method for studying the oral absorption of drug candidates in drug development studies.


Breast cancer and religion in greater Bombay women: an epidemiological study of 2130 women over a 9-year period.
The resident female population of Greater Bombay consists of women professing different religious faiths, between which the frequency of breast cancer varies to a great extent. During the 9-year period 1964 to 1972 inclusive, a total of 2130 women with breast cancer were seen, with break-down by religion as follows: Hindus (1259), Muslims (306), Christians (264), Parsi (Zoroastrians) (226), Jains (25), Buddhists (26) and others (24). The average annual age-adjusted (world population) incidence rates, however, were found to be 48.5 and 18.2 per 100,000 in the Parsis and non-Parsis respectively, with an average of 19.9 per 100,000 for the total population. For reasons not yet clear, in every age group the incidence rate in Parsis was 2 to 3 times higher than in the non-Parsis. Time-trend analyses of our data do not reveal any statistically significant increase or decrease in the incidence of breast cancer in any particular age group. Data from death certificates for the same 9-year period show that the age-adjusted mortality rate (world population) is 9.2 per 100,000/year.


Discovery of Unknown Posttranslational Modifications by Top-Down Mass Spectrometry.
Protein encoding genes can undergo modifications posttranscriptionally and posttranslationally, yielding many different "proteoforms." The chemical diversity of such modifications is known to be important biomarkers of function within biological systems but is not completely understood. Top-down mass spectrometry is a valuable tool for the characterization of proteoforms, especially for histones that have complex combinations of posttranslational modifications (PTMs). In this chapter, we present a top-down liquid chromatography-mass spectrometry experimental and data analysis workflow for the identification of novel, unexpected modifications on histones. Proteoforms of interest are first discovered using the "open" modification search in TopPIC. Then target proteoforms are manually confirmed using the data visualization tool-LcMsSpectator, part of the Informed-Proteomics package. The workflow can be very helpful in targeted PTM analysis and can be expanded to other types of proteins for discovery of unknown PTMs.


Effect of static magnetic field treatment on γ-aminobutyric acid content and sensory characteristics of germinated brown rice cake.
This study aimed to improve γ-aminobutyric acid (GABA) content and sensory characteristics of brown rice (BR) cake by static magnetic field (SMF)-assisted germination. BR was pre-treated by SMF (10 mT, 60 min, 25 °C), germinated for 36 h, and then germinated BR (GBR) was used to prepare rice cake. The optimal formula was: 60 % GBR, 40 % white rice, 1 % yeast, 20 % sugar, and 55 % water. SMF significantly increased the GABA content by stimulating glutamate decarboxylase, with the values increasing from 28.17 to 32.43 mg/100 g and from 2.50 to 6.27 mg/100 g in GBR (36 h) and GBR cake, respectively. SMF also altered the swelling power and water solubility of GBR flour by promoting the hydrolysis of starch, protein, and fiber, thus improving the texture, flavor, and storage stability of GBR cake. Overall, SMF could be a prospective technique for improving the nutritional and sensory qualities of whole-grain food.


Microplastics and linear alkylbenzene levels in oysters Crassostrea gigas driven by sewage contamination at an important aquaculture area of Brazil.
The levels of linear alkylbenzenes (LABs) and the occurrence of microplastics (MPs) in the oysters Crassostrea gigas were evaluated in six farming areas in southern Brazil. The results revealed higher concentrations of LABs in oyster tissue from the Serraria (1977 ± 497.7 ng g-1) and Imaruim (1038 ± 409.9 ng g-1) sites. Plastic microfibers were found in oysters from all locations with values from 0.33 to 0.75 MPs per oyster (0.27-0.64 MPs per gram) showing the ubiquitous presence of this contaminant in the marine environment, which could be considered a threat to farming organisms. In addition, elements such as Ti, Al, Ba, V, Rb, Cr, and Cu were found in the chemical composition of the microfibers, suggesting MPs as vectors of inorganic compounds. A positive correlation between LABs and thermotolerant coliforms suggests that sewage discharges are the main source of contamination in these oysters cultured for human consumption. The present study highlights the need for efficient wastewater treatment plants and the implementation of depuration techniques in oysters from farming areas.


Reaction of monoclonal H-Y antibody in the ELISA.
Monoclonal H-Y antibody of demonstrated specificity was reacted with soluble H-Y antigen in a newly-developed enzyme-linked immunosorbent assay (ELISA). Typing of cell lines was accomplished by qualitative and quantitative absorption. In one case, the antibody was subdivided into equal portions, each of which was absorbed with 1, 3, 6 or 12 X 10(6) cells from male or female; after absorption, the cells were discarded and the antibody was titrated for residual activity in the ELISA. With increasing cell numbers, optical density scores for male-absorbed antibody approached baseline levels. Although male-female differences were pronounced and highly significant, there was also a fall in reactivity after absorption with female cells.


Comparison of Novel Immunoassay With Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS) for Therapeutic Drug Monitoring of Clozapine.
Clozapine is the most effective antipsychotic for treatment-resistant schizophrenia. Although serum clozapine levels can help guide treatment, they are underutilized owing to requirements for frequent venous blood draws and lack of immediate results. Clozapine levels measured with a novel immunoassay technology (which enables point-of-care development) were compared with those measured by standard liquid chromatography/tandem mass spectrometry (LC-MS/MS). Frozen serum aliquots of 117 samples (N = 48 patients with schizophrenia on clozapine; N = 24 patients with schizophrenia not on clozapine; N = 45 healthy controls) were sent to a national reference laboratory (NRL) for clozapine level determination by LC-MS/MS, and matching samples were subjected to novel immunoassay (3 runs). At a later date, another frozen aliquot from the same date was sent to the NRL for repeat testing. The NRL obtained 18 false-positive clozapine results (mean 42.39 ± 32.06, range 21-159 ng/mL) in participants not on clozapine (N = 3) and healthy controls (N = 15). The immunoassay showed no false-positive clozapine results. The clozapine levels were correlated between both assays (r = 0.84, P < 0.0001), despite 16% higher clozapine levels with immunoassay (482.08 ± 270.88 ng/mL immunoassay, 414.98 ± 186.29 ng/mL LC-MS/MS [P = 0.03]). Agreement analysis using concordance correlation coefficient (CCC) for LC-MS/MS of the 2 aliquots yielded CCC = 0.869; 95% confidence interval = 0.690-0.970, whereas higher agreement results were observed for the 3 runs of immunoassay (CCC = 0.99; 95% confidence interval = 0.979-0.997). The lack of false positives observed with immunoassay, higher repeat performance agreement, and good correlation with LC-MS/MS may indicate the more robust performance of immunoassay than that of LC-MS/MS clozapine-level determination.


Estimates of global and regional potential health gains from reducing multiple major risk factors.
Estimates of the disease burden due to multiple risk factors can show the potential gain from combined preventive measures. But few such investigations have been attempted, and none on a global scale. Our aim was to estimate the potential health benefits from removal of multiple major risk factors. We assessed the burden of disease and injury attributable to the joint effects of 20 selected leading risk factors in 14 epidemiological subregions of the world. We estimated population attributable fractions, defined as the proportional reduction in disease or mortality that would occur if exposure to a risk factor were reduced to an alternative level, from data for risk factor prevalence and hazard size. For every disease, we estimated joint population attributable fractions, for multiple risk factors, by age and sex, from the direct contributions of individual risk factors. To obtain the direct hazards, we reviewed publications and re-analysed cohort data to account for that part of hazard that is mediated through other risks. Globally, an estimated 47% of premature deaths and 39% of total disease burden in 2000 resulted from the joint effects of the risk factors considered. These risks caused a substantial proportion of important diseases, including diarrhoea (92%-94%), lower respiratory infections (55-62%), lung cancer (72%), chronic obstructive pulmonary disease (60%), ischaemic heart disease (83-89%), and stroke (70-76%). Removal of these risks would have increased global healthy life expectancy by 9.3 years (17%) ranging from 4.4 years (6%) in the developed countries of the western Pacific to 16.1 years (43%) in parts of sub-Saharan Africa. Removal of major risk factors would not only increase healthy life expectancy in every region, but also reduce some of the differences between regions. The potential for disease prevention and health gain from tackling major known risks simultaneously would be substantial.


[Unusual angiographic technic in a ruptured aortic aneurysm (author's transl)].
In a 65-year-old patient with amputation of both legs, a life-threatening acute abdominal hemorrhage necessitates aortography. Following surgical dissection of the right femoral artery a catheter could be inserted through the thrombosed vessel into the aortic lumen. This approach made the diagnosis of a perforated aortic aneurysm by angiography possible.


Angular oversampling with temporally offset layers on multilayer detectors in computed tomography.
Today's computed tomography (CT) scanners operate at an increasingly high rotation speed in order to reduce motion artifacts and to fulfill the requirements of dynamic acquisition, e.g., perfusion and cardiac imaging, with lower angular sampling rate as a consequence. In this paper, a simple method for obtaining angular oversampling when using multilayer detectors in continuous rotation CT is presented. By introducing temporal offsets between the measurement periods of the different layers on a multilayer detector, the angular sampling rate can be increased by a factor equal to the number of layers on the detector. The increased angular sampling rate reduces the risk of producing aliasing artifacts in the image. A simulation of a detector with two layers is performed to prove the concept. The simulation study shows that aliasing artifacts from insufficient angular sampling are reduced by the proposed method. Specifically, when imaging a single point blurred by a 2D Gaussian kernel, the method is shown to reduce the strength of the aliasing artifacts by approximately an order of magnitude. The presented oversampling method is easy to implement in today's multilayer detectors and has the potential to reduce aliasing artifacts in the reconstructed images.


Patients With Diabetes Mellitus Have a Higher Risk of Tendon Retear After Arthroscopic Rotator Cuff Repair: A Meta-analysis.
Retear of a repaired rotator cuff tendon is a major issue for shoulder surgeons. It is possible that diabetes mellitus (DM) is associated with a greater risk of tendon retear after arthroscopic rotator cuff repair. To determine whether patients with DM have a higher tendon retear risk after arthroscopic rotator cuff repair. Systematic review; Level of evidence, 4. A systematic review was conducted following the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines using the Web of Science, PubMed, and Embase databases. Inclusion criteria were articles written in the English language that included patients undergoing arthroscopic rotator cuff repair surgeries, reported the numbers of patients with and those without DM, and reported the number of rotator cuff retears. Data relevant to this study were extracted and statistically analyzed. Random-effects models were used to generate pooled odds ratio estimates and CIs. A total of 160 studies were identified from the initial search, and 5 of them met the inclusion criteria. A total of 1065 patients (207 patients with DM and 858 patients without DM) were included. The pooled results showed that the patients in the DM group had a significantly higher tendon retear risk than did those in the non-DM group (relative risk, 2.25; 95% CI, 1.14-4.45; P = .02). Patients with DM have a 2.25 times higher risk of tendon retear after arthroscopic rotator cuff repair compared with patients without DM.


Relationship between the resistance of crossbred cattle to ticks, Boophilus microplus (Canestrini, 1887) and Hyalomma anatolicum anatolicum (Koch, 1844).
Studies were undertaken to determine the effect of repeated pure infestations with Boophilus microplus on susceptibility to subsequent pure infestations with Hyalomma anatolicum anatolicum, and the effects of pure infestations with both species of tick on susceptibility to a series of mixed infestations. Crossbred (Bos indicus X Bos taurus) calves were infested with Boophilus microplus (17 times), H. a. anatolicum (four times), followed by five mixed infestations of B. microplus and H. a. anatolicum. The decline in B. microplus engorgement from a mean yield of 274.4 +/- 60.3 ticks per host after the first exposure, to a mean yield of 9 +/- 4.6 per animal after the seventeenth exposure, was observed in animals exposed to only B. microplus. This might be due to acquired resistance. However, these animals were found to be as susceptible to H. a. anatolicum as animals which had never been exposed to ticks of either species. A decline in the yield of H. a. anatolicum from a mean yield of 92.1 +/- 10.7 after the first exposure to 54.7 +/- 11.3 after the fourth exposure, indicated that the cattle could also acquire resistance to repeated pure infestations with this species. After repeated pure infestations with both tick species, cattle reacted to five mixed infestations showing a high degree of resistance to B. microplus and low resistance to H. a. anatolicum (mean yield for B. microplus was only 10 +/- 8.1 ticks per host after the first mixed exposure and declined to 1.3 +/- 1.7 after the fifth, whereas the mean yield for H. a. anatolicum was 71.4 +/- 11.3 ticks per host following the first exposure and declined to 37.3 +/- 7.8 after the fifth). Host responses elicited to one species do not provide cross-resistance to the second species used in this study.


Conducting In-Home Functional Analyses of Aggression and Self-Injury Exhibited by Boys with Fragile X Syndrome.
Boys with fragile X syndrome (FXS) are at increased risk for exhibiting aggression and self-injury that can be extremely disruptive to families. Few studies, however, have examined the extent to which social-environmental factors can influence these behaviors, particularly in home settings. Caregivers of 32 boys with FXS, aged 3 to 10 years, were coached by a board-certified behavior analyst to implement a functional analysis (FA) with their child in their home over 2 days. The functions identified in the FAs were validated by behavioral interventions conducted through telehealth over 12 weeks. Social functions for aggression and self-injury were identified in 29 (90.6%) cases, with the most common functions identified being escape from demands/transitions and/or tangible reinforcement. Subsequent function-based behavioral interventions (i.e., functional communication training) implemented for 22 cases by telehealth resulted in decreased rates of aggression and self-injury by 74% to 100%. These data underscore the significant impact that social-environmental factors can have on the development and maintenance of aggression and self-injury in FXS. Conducting early home-based behavioral interventions designed to increase functional communication skills may therefore be an effective solution for the treatment of aggression and self-injury in FXS in the long term.


Intraspinal transplantation of neurogenin-expressing stem cells generates spinal cord neural progenitors.
Embryonic stem (ES) cells and their derivatives are an important resource for developing novel cellular therapies for disease. Controlling proliferation and lineage selection, however, are essential to circumvent the possibility of tumor formation and facilitate the safe translation of ES-based therapies to man. Expression of appropriate transcription factors is one approach to direct the differentiation of ES cells towards a specific lineage and stop proliferation. Neural differentiation can be initiated in ES cells by expression of Neurogenin1 (Ngn1). In this study we investigate the effects of controlled Ngn1 expression on mouse ES (mES) cell differentiation in vitro and following grafting into the rat spinal cord. In vitro, Ngn1 expression in mES cells leads to rapid and specific neural differentiation, and a concurrent decrease in proliferation. Similarly transplantation of Ngn1-expressing mES cells into the spinal cord lead to in situ differentiation and spinal precursor formation. These data demonstrate that Ngn1 expression in mES cells is sufficient to promote neural differentiation and inhibit proliferation, thus establishing an approach to safely graft ES cells into the spinal cord.


Cyclin D1-specific cytotoxic T lymphocytes are present in the repertoire of cancer patients: implications for cancer immunotherapy.
Cyclin D1, a key cell cycle regulator, is overexpressed in multiple types of cancer. Such tumor-associated genes may be useful targets for cancer immunotherapy. Nevertheless, it had previously been suggested that efficient T cells recognizing cyclin D1-derived epitopes are absent from the repertoire because of thymic deletion. We attempted to induce autologous CTL from healthy donors and patients with cyclin D1-overexpressing tumors using a highly efficient T-cell expansion system based on CD40-activated B cells as antigen-presenting cells. Cyclin D1-derived, HLA-A*0201-restricted epitopes were predicted by multiple computer algorithms, screened in HLA-A2-binding assays, and used for T-cell stimulation. The generated CTL lines and clones were analyzed by IFN-gamma enzyme-linked immunosorbent spot assay or cytolysis assay. After screening, at least two naturally processed and presented HLA-A*0201-binding cyclin D1 epitopes were identified. CTL specific for these epitopes could be successfully generated from HLA-A2(+) donors. T cells efficiently recognized target cells pulsed with the cognate peptide and cyclin D1-expressing tumor cell lines in an HLA-A*0201-restricted manner. More importantly, HLA-A*0201-matched, primary cyclin D1(+) tumor cells were efficiently recognized by cyclin D1-specific CTL. These CTL could be generated from patients with mantle cell lymphoma and cyclin D1(+) colon cancer. These results underscore that cyclin D1 needs to be considered as a target for broad-based antitumor immunotherapy.


Megakaryocytes and platelet homeostasis in diffuse alveolar damage.
Platelet homeostasis reflects a balance between the production of platelets via cytoplasmic fragmentation of megakaryocytes in the pulmonary microvasculature and their catabolism. Increased numbers of megakaryocytes are entrapped in the injured lung, potentially affecting circulating platelet counts. We enumerated pulmonary megakaryocytes and blood platelets in patients with diffuse alveolar damage (DAD) in order to determine their association with clinical outcome. Lung biopsies were examined from 21 patients with histologically documented DAD in its proliferative phase and secondary to a variety of causes. Blood platelet counts were determined within 24 h prior to lung biopsy, and CD61+ pulmonary megakaryocytes were localized in in situ immunohistochemical stains. The overall mortality in this series was 67%. Patients with DAD attributable to drug toxicity (DAD-D) had higher mortality (80%) and greater number of intrapulmonary CD61+ megakaryocytes than those with DAD due to other causes (23+/-7, 10+/-2, p<0.05). Patients with blood platelet counts =350 th/cm(3) showed increased survival (p<0.05). The findings support the hypothesis that abnormal platelet homeostasis is associated with increased mortality in acute lung injury and indicate that thrombocytosis in ARDS is associated with improved survival. The mechanisms of altered platelet homeostasis in DAD merit further investigation.


Blood burden of di(2-ethylhexyl) phthalate and its primary metabolite mono(2-ethylhexyl) phthalate in pregnant and nonpregnant rats and marmosets.
A comparison of the dose-dependent blood burden of di(2-ethylhexyl) phthalate (DEHP) and mono(2-ethylhexyl) phthalate (MEHP) in pregnant and nonpregnant rats and marmosets is presented. Sprague-Dawley rats and marmosets were treated orally with 30 or 500 mg DEHP/kg per day, nonpregnant animals on 7 (rats) and 29 (marmosets) consecutive days, pregnant animals on gestation days 14-19 (rats) and 96-124 (marmosets). In addition, rats received a single dose of 1000 mg DEHP/kg. Blood was collected up to 48 h after dosing. Concentrations of DEHP and MEHP in blood were determined by GC/MS. In rats, normalized areas under the concentration-time curves (AUCs) of DEHP were two orders of magnitude smaller than the normalized AUCs of the first metabolite MEHP. Metabolism of MEHP was saturable. Repeated DEHP treatment and pregnancy had only little influence on the normalized AUC of MEHP. In marmosets, most of MEHP concentration-time courses oscillated. Normalized AUCs of DEHP were at least one order of magnitude smaller than those of MEHP. In pregnant marmosets, normalized AUCs of MEHP were similar to those in nonpregnant animals with the exception that at 500 mg DEHP/kg per day, the normalized AUCs determined on gestation days 103, 117, and 124 were distinctly smaller. The maximum concentrations of MEHP in blood of marmosets were up to 7.5 times and the normalized AUCs up to 16 times lower than in rats receiving the same daily oral DEHP dose per kilogram of body weight. From this toxicokinetic comparison, DEHP can be expected to be several times less effective in the offspring of marmosets than in that of rats if the blood burden by MEHP in dams can be regarded as a dose surrogate for the MEHP burden in their fetuses.


Deep insights into fungal diversity in traditional Chinese sour soup by Illumina MiSeq sequencing.
Sour soup is a traditional condiment in Guizhou Province, China. The purpose of this study was to investigate the differences in the fungi present in 5 types of sour soup (tomato sour soup, chili sour soup, cherry tomato sour soup, spoiled tomato sour soup, and red sour soup made from blended tomato and chili sour soup subjected to secondary fermentation) and to determine the reasons for the deterioration of tomato sour soup by comparing the fungal communities in normal and deterioratedtomato sour soup. A total of 5 phyla were detected in all 5 samples, including Ascomycota (69.38%), Basidiomycota (7.63%), Zygomycota (1.59%), Chytridiomycota (0.01%) and unclassified phyla (21.39%). Ascomycota was the main phylum in each sample except the red sour soup made from blended tomato and chili sour soup subjected to secondary fermentation. That sour soup contained many unrecognized phyla. At the genus level, there were major differences among the different samples. Dekkera spp. and Pichia spp. were the main dominant fungus in tomato sour soup, Saccharomyces spp. and Pichia spp. were the dominant fungus in chili sour soup, and Pichia spp. were the dominant fungus in cherry tomato sour soup. When sour soup went bad, the fungus of sour soup changed greatly, and the unknown fungal genera, Cladospora spp., Saccharomyces spp. and Emericella spp. became the dominant fungal genera. In addition, after the secondary fermentation of tomato and chili sour soup mixed with garlic and ginger, the fungal genera of the base fermentation were replaced by unknown fungal genera. Moreover, there were various spoilage fungi in sour soup, which indicated that there were safety risks in naturally fermented sour soup and should be further controlled. This study revealed the fungal flora in sour soup made from different vegetables and compared the fungal diversity of spoiled and normal tomato sour soup and thereby provided a basis for understanding the fungal diversity of sour soup in China and guiding the production of sour soup.


Incidence of plastic debris in Sooty Tern nests: A preliminary study on Trindade Island, a remote area of Brazil.
Plastic is abundant in the oceans, reaching pelagic zones away from continents. Here we present the first recordings of plastic used as nest material in Sooty Tern nests, on a remote oceanic island. We describe our findings in terms of quantity, size and color of plastic debris. A total of 78 plastics were noted in 54 nests. Four color categories were found: Blue, White, Green and Red. Blue fragments were the most frequent color, present three times as much as white debris. This pattern was present despite blue fragments being smaller and lighter. The plastic debris of lowest frequency were the larger and heavier pieces (red). To our knowledge this is the first record of plastic in Sooty Tern nests. Trindade Island is on an oceanic zone expected to accumulate garbage due to the dynamic ocean currents. Such findings call for a closer inspection of pollution in the Atlantic Ocean.


Self-inhibition of an AV sequential demand (DVI) pulse generator due to polyurethane lead insulation disruption.
Self-inhibition of bipolar DVI AV-sequential pacemakers has been attributed to electrode malposition, although the potential for self-inhibition due to current leakage has been recognized. Previous cases of current leakages due to insulation defects in bipolar VVI ventricular demand systems have usually presented as abnormal sensing or pacing function without a change to actual unipolar sensing and pacing. This article describes a case of disruption of the insulation of polyurethane bipolar atrial and ventricular leads by silk ligatures, resulting in unipolar atrial and ventricular pacing and self-inhibition of ventricular output of a DVI pulse generator. This case emphasizes the importance of avoiding any application of suture material directly to polyurethane leads. This case also emphasizes the importance of monitoring pacemaker artifact amplitude and axis in patient follow-up.


Fused high b-value diffusion weighted and T2-weighted MR images in staging of pediatric Hodgkin's lymphoma: A pilot study.
To evaluate the value of fused high b-value diffusion weighted and T2-weighted MRI compared to T1-weighted imaging, T2-weighted imaging and DWI for staging pediatric Hodgkin's lymphoma. 21 consecutive pediatric patients who underwent whole-body MRI at Hodgkin's lymphoma staging were included. Fused, colorized DWI/T2-weighted images were created. Image sets consisting of (a) T1-weighted, T2-weighted and DWI images and (b) T1-weighted, T2-weighted, DWI and DWI/T2-weighted fused images were reviewed by a radiologist using a cross-over design with blinding and randomization. Scoring was performed using a standardized form, based on detection, characterization and reading time, using a FDG-PET/CT based reference standard. Test characteristics, test agreement to a FDG-PET/CT based reference standard and reading times were calculated. Agreement for whole-body MRI without fused images and FDG-PET/CT was very good for nodal staging (κ = 0.86, 95% CI 0.78-0.93) and extra-nodal staging (κ = 0.90, 95% CI 0.71-1.09). Agreement improved with the addition of the fused DWI/T2-weighted images (κ = 0.92 95% CI 0.87-0.97 (nodal staging), κ = 0.92 95% CI 0.76-1.08 (extra-nodal staging). Sensitivity and specificity for staging nodal disease were 99 % and 95% respectively for whole-body MRI including DWI/T2-weighted fused images (versus 88 % and 97 % without fused images) and 100 % and 99 % for extra-nodal disease (83 % and 100 % without fused images). Disease stage for MRI including fused DWI/T2-weighted images agreed with the reference standard in 18 out of 21 patients. The addition of DWI/T2-weighted fusion images to T1-weighted, T2-weighted and DWI whole-body MRI might shorten reading times and might improve the diagnostic performance of whole-body MRI in staging pediatric Hodgkin's lymphoma.


Effect of Laryngeal Mask Air Way Insertion versus Endotracheal Intubation over Hemodynamic Responses in Pediatrics Patient Who Underwent Ophthalmic Surgery at Menelik II Hospital, Addis Ababa: A Prospective Observational Study Design.
The airway of an anesthetized child is usually maintained with an endotracheal tube or laryngeal mask airway. However, both are related with some level of pressor response which may be risky in several groups of patient. An institutional-based prospective observational study design was employed. A systematic random sampling technique was used to select study participants. Data were entered into Epi info version 7 and transported to SPSS version 20 for analysis. Normality of the data was checked using Shapiro-Wilk tests. An independent t test was used to determine the mean differences between the two groups while the paired sample t test was used to determine the mean differences within the groups. A p value of less than 0.05 was used as a cut-off point for the presence of association. The changes in systolic and diastolic blood pressure were returned to baseline values at five and three minutes in both groups, respectively. However, the changes in the heart rate and mean arterial pressure returned to baseline values in five minutes in the ETT group and three minutes in the LMA group. At baseline, the difference in systolic blood pressure between the two groups was not statistically significant (p=0.328). A significant hemodynamic pressor response was observed after the insertion of both LMA and ETT groups. However, the LMA group has less hemodynamic change as compared to the ETT group. Therefore, the practice of LMA insertion was strongly recommended.


Regression of aggressive corneal vascularization after photodynamic therapy, subconjunctival Avastin injections and topical cyclosporin-A 1% drops: A case report.
We describe a case of significant regression of aggressive, deep and superficial corneal vascularization using photodynamic therapy (PDT) with verteporfin, repeated subconjunctival injections of Avastin®, plus topical cyclosporin-A 1% drops. The area of the corneal vascularization was treated with PDT with verteporfin. The subconjunctival space - next to the area of vascularization - was injected with Avastin® (1.25 mg/0.05 ml), immediately after PDT, and then for two more times during the follow-up period. The patient was kept on topical cyclosporin and prednisolone acetate. The outcome was evaluated clinically and photographically for 13 months. Corneal vascularization regressed significantly. No adverse effects were observed, neither locally or systemically. PDT with verteporfin, repeated subconjunctival injections of Avastin®, and topical cyclosporin-A drops appear to be safe and effective in treating aggressive corneal vascularization.


Comparison of two methods of measuring tracheal mucous velocity in anesthetized beagle dogs.
Tracheal mucous velocity measurements were made in 6 beagle dogs using both Sackner's method of radiographing teflon discs and Chopra's method using a gamma camera to detect the movement of instilled labeled material. Dogs were anesthetized by intravenous injection of 30 mg of sodium pentobarbital/kg of body weight in case 1 after pretreatment with atropine and Innovar-Vet; in case 2, without the pretreatment. The mean velocities measured using the 2 techniques were significantly different in both cases: case 1, 3.7 +/- 3.3 mm/min for Sackner's methods and 7.5 +/- 3.7 mm/min for Chopra's method (p less than 0.001); case 2. 3.7 +/- 3.1 mm/min for Sackner's method, and 5.5 +/- 2.1 mm/min for Chopra's method (p less than 0.05). Systematic differences in velocities should be considered when comparing data from investigators using different techniques.


Synthesis and proteinase inhibitory properties of diphenyl phosphonate analogues of aspartic and glutamic acids.
The synthesis of diphenyl phosphonate analogues of aspartic and glutamic acid, and their inhibitory activity against S. aureus V8 protease and granzyme B, is described. The study has revealed difficulties with protecting group compatibility in the synthesis of these analogues. Two analogues, Acetyl. AspP (OPh)2 and Acetyl.GluP (OPh)2 were found to function as irreversible inactivators of V8 proteinase, yet exhibit no activity against granzyme B.


Home-based treatment of cellulitis with twice-daily cefazolin.
To assess the clinical outcome and pharmacokinetics of therapy with cefazolin for patients with cellulitis in a hospital-in-the-home (HIH) program. Observational study with outcome data compared with previously published reports of therapy for cellulitis. A university teaching hospital and HIH unit, July 1996-December 1997. Patients with cellulitis were eligible for inclusion provided their medical condition was stable, they did not require surgical intervention, and their social circumstances allowed home-based therapy. Cefazolin 2 g intravenously twice daily, with regular nursing and medical assessment. Clinical efficacy; peak and trough serum concentrations of cefazolin. Fifty-seven patients (37 were men) with a mean age of 48 years (range, 18-90 years) had 61 episodes of moderate to severe cellulitis (41, lower limb; 17, upper limb; and three, face). They received a median of 11 doses of cefazolin (range, 3-27 doses). Clinical outcomes were: cure in 54, improvement in one, treatment failure in three, and in the remaining three episodes the outcome was indeterminate. Cefazolin concentrations were measured in 27 patients. All peak concentrations were more than 40 micrograms/mL, while trough concentrations were all above the MIC90 of the expected pathogens: median, 3.2 micrograms/mL (range: 0.4-18.5 micrograms/mL). Cefazolin was well tolerated. Twice-daily cefazolin 2 g intravenously is a convenient and effective option for home-based treatment of patients with cellulitis. Its clinical efficacy is comparable with other treatment regimens.


Leukocyte migration inhibition activity of nonimmune acute inflammatory pleural exudate.
Material with leuckocyte migration inhibition (LMI) activity has been demonstrated in various types of nonimmunologically induced acute pleural inflammatory exudates. This activity is present in inflammatory cell-free exudate and appears to involved the deposition of fibrin around the migrating leukocyte, resulting in "cell-trapping." This is supported by the fact that removal or inhibition of fibrin formation leads to loss of exudate LMI activity. Both fibrinogen and complement as well as vitamin K-dependent clotting factors appear to be required for LMI activity. The mechanism involved in the LMI reaction and its significance in nonimmune and cell-mediated immune inflammation are discussed.


Digoxin inhibits blood vessel density and HIF-1a expression in castration-resistant C4-2 xenograft prostate tumors.
Recent studies suggest a potential application for digoxin in the prevention and/or treatment of prostate cancer. However, the effect of digoxin on androgen receptor (AR)-positive prostate tumor in vivo is not clear. This study is designed to determine if digoxin can inhibit AR-positive xenograft prostate tumors. Athymic male nude mice were utilized to establish subcutaneous C4-2 castration-resistant prostate tumors. The animals were castrated and then treated with daily intraperitoneal (i.p.) injection of digoxin at 2 mg/kg along with vehicle controls for 7 consecutive days. Tumor growth was determined by measuring tumor volume changes, blood vessel density by immunostaining of CD31, and cell proliferation by BrdU labeling. The expression of HIF-1α in C4-2 tumors was measured by Western blot and real-time RT-PCR. Digoxin inhibited blood vessel density about fourfold and down-regulated HIF-1α expression at both mRNA and protein levels. However, digoxin did not inhibit C4-2 tumor growth. Digoxin is a potent inhibitor of HIF-1α signaling pathway and blood vessel formation in C4-2 castration-resistant prostate tumors.


Extremely sparse models of linkage disequilibrium in ancestrally diverse association studies.
Linkage disequilibrium (LD) is the correlation among nearby genetic variants. In genetic association studies, LD is often modeled using large correlation matrices, but this approach is inefficient, especially in ancestrally diverse studies. In the present study, we introduce LD graphical models (LDGMs), which are an extremely sparse and efficient representation of LD. LDGMs are derived from genome-wide genealogies; statistical relationships among alleles in the LDGM correspond to genealogical relationships among haplotypes. We published LDGMs and ancestry-specific LDGM precision matrices for 18 million common variants (minor allele frequency >1%) in five ancestry groups, validated their accuracy and demonstrated order-of-magnitude improvements in runtime for commonly used LD matrix computations. We implemented an extremely fast multiancestry polygenic prediction method, BLUPx-ldgm, which performs better than a similar method based on the reference LD correlation matrix. LDGMs will enable sophisticated methods that scale to ancestrally diverse genetic association data across millions of variants and individuals.


Loss of angiotensin-converting enzyme 2 leads to impaired glucose homeostasis in mice.
This study aimed to investigate the role of angiotensin-converting enzyme 2 (ACE2) in regulating glucose homeostasis. Immunohistochemistry was used to investigate ACE2 expression in the pancreas. Glucose tolerance test, insulin secretion test, and insulin tolerance test were performed in age-matched male ACE2 knockout (KO) and wild-type (WT) mice. We found that ACE2 was positively expressed in the pancreas. Male ACE2 KO mice displayed a selective decrease in first-phase insulin secretion in response to glucose and a progressive impairment of glucose tolerance compared with age- and sex-matched WT mice. On the other hand, insulin sensitivity of the peripheral tissue in age-matched ACE2 KO and WT mice showed no difference. These findings suggest that ACE2 might play an important role in glucose homeostasis as well as type 2 diabetes.


Assessing the biological significance of primary structure consensus patterns using sequence databanks. I. Heat-shock and glucocorticoid control elements in eukaryotic promoters.
We describe FORTRAN 77 software allowing for convenient searching of any segmented and ambiguous pattern in the currently available protein or nucleotide sequence databanks. For proteins, this software can be instrumental in defining conserved functional domains among non-homologous overall primary structures. For nucleic acids, it is used in detecting complex and/or low consensus structural or regulatory patterns. As first applications we have studied the distribution of short consensus sequences believed to characterize heat-shock and glucocorticoid regulated promoters. This analysis allowed an evaluation of the specificity, probable role and thus biological significance of various regions of these consensus. In addition, the expression of several known genes are predicted to be heat-shock or glucocorticoid sensitive.


Evaluation of radioactivity levels and radiological hazards of some endemic plants used as medicine in Ankara, Turkey.
In this study, natural radioactivity levels (226Ra, 232Th, and 4 K) of some medicinal plant samples with known anti-oxidative properties, which are frequently consumed by animals and humans, were obtained from Ankara province and its surroundings (Mamak, Kızılcahamam, Beypazarı, Kahramankazan, and Polatlı districts) were determined using a thallium-doped sodium iodide NaI(Tl) gamma spectrometry. By using the determined natural radioactivity concentrations in the collected plant samples, the number of radiological doses that people could be exposed by consuming these plants was calculated. As a result of the study, 226Ra, 232Th, and 4 K radioactivity concentration ranges of the plant samples were found be 14.69 ± 1.27-59.08 ± 3.12 Bq kg-1, 1.78 ± 0.04-50.05 ± 2.76 Bq kg-1 and 207.24 ± 34.09-826.13 ± 25.40 Bq kg-1, respectively. The highest 226Ra, 232Th, and 4 K activity concentrations were measured in Astragalus densifolius subsp. ayashensis (Kahramankazan), Astragalus kochakii (Kahramankazan) and Rumex patientia (Patience Dock) (Kahramankazan) plants, respectively. The lowest 226Ra, 232Th and,4 K activity concentration plants were determined respectively as Rumex patientia (Mamak), Lavandula angustifolia (Kızılcahamam), and Astragalus acikirensis (Polatlı). The establishment and routine repetition of environmental radioactivity monitoring programs in each region are important for human and animal health, and the results of this study gain importance for Ankara and its surroundings in terms of environmental health.


Substrate-tuning of correlated spin-orbit oxides revealed by optical conductivity calculations.
We have systematically investigated substrate-strain effects on the electronic structures of two representative Sr-iridates, a correlated-insulator Sr2IrO4 and a metal SrIrO3. Optical conductivities obtained by the ab initio electronic structure calculations reveal that the tensile strain shifts the optical peak positions to higher energy side with altered intensities, suggesting the enhancement of the electronic correlation and spin-orbit coupling (SOC) strength in Sr-iridates. The response of the electronic structure upon tensile strain is found to be highly correlated with the direction of magnetic moment, the octahedral connectivity, and the SOC strength, which cooperatively determine the robustness of Jeff = 1/2 ground states. Optical responses are analyzed also with microscopic model calculation and compared with corresponding experiments. In the case of SrIrO3, the evolution of the electronic structure near the Fermi level shows high tunability of hole bands, as suggested by previous experiments.


The frontotemporal dementias in a tertiary referral center: classification and demographic characteristics in a series of 232 cases.
Frontotemporal lobar degeneration (FTLD) comprises of behavioral variant frontotemporal dementia (bvFTD) and primary progressive aphasia (PPA) with its 3 main variants, namely nonfluent/agrammatic (naPPA), semantic (svPPA) and logopenic (lvPPA). Recently a clinical syndrome with predominant right temporal atrophy was recognized (rvFTD). FTLD often overlaps with parkinsonism plus syndromes such as corticobasal degeneration (CBD) and progressive supranuclear palsy (PSP), as well as with motor neuron disease (FTD-MND). While FTLD syndromes were thought to be rare and difficult to diagnose ante mortem, revised diagnostic criteria as well as recent studies highlighted the plausibility of accurate clinical diagnosis. 232 FTLD patients were assessed from January 1, 2003 to December 31, 2010 in the Neurology Department of a tertiary referral center. Patients were classified as bvFTD, naPPA, svPPA, lvPPA, CBD/PSP and rvFTD and their demographic characteristics were analyzed. From the 232 patients, 111 (47.8%) were diagnosed with bvFTD, 56 (24.1%) with naPPA, 21 (9.1%) with svPPA, 6 (2.6%) with lvPPA, 20 (8.6%) with CBD or PSP and 18 (7.8%) with rvFTD. 44% of the patients were under 65 years old at onset of symptoms, while only 4.3% reported family history of dementia. FTLD subgroups did not differ with respect to demographic characteristics, but early onset cases had higher educational level. FTLD represents a syndrome with different but clinically distinguishable phenotypes. Cultural, educational and socioeconomic status differences might regulate patients' access to medical care and therefore influence age of reported onset and prevalence of FTLD in clinical studies. High clinical alertness and sensitive neuropsychological tests could lead to timely clinical diagnosis in a common presenile type of dementia.


Ganglioneuromatosis of the Gallbladder in a Dog with Cholecystitis and Cholestasis.
A 2-year-old entire female mixed-breed dog was presented with signs of cholestasis. Abdominal ultrasonography revealed a gallbladder with a thickened and hyperechoic wall and luminal calculi. Exploratory laparotomy with cholecystectomy was performed and histopathological examination of the specimens from the cholecystectomy demonstrated extensive proliferation of large, prominent nerves containing ganglion cells with no atypia mainly located in the gallbladder mucosa. The neural nature of these components was confirmed by immunohistochemical labelling with antibodies specific for synaptophysin, glial fibrillary acidic protein, S100 protein and neurofilaments. Based on these findings, the lesion was diagnosed as ganglioneuromatosis. Reports of ganglioneuromatosis in animals have so far been restricted to the intestine. This is the first case of ganglioneuromatosis affecting the gallbladder in an animal in which no intestinal involvement was apparent.


[A method for the evaluation of nitroblue tetrazolium reduction in human platelets (author's transl)].
A long term incubation in NBT-solution leads not only to NBT-reduction by polymorphnuclear leukocytes and monocytes, but also in washed human platelets. The reaction is enhanced by ADP and to a smaller and very variable extent also by Thrombin and Ristocetin. It is inhibited by incubation in plasma environment as well by DBcAMP and PGE1. After a detailed demonstration of the applied method, some possible biochemical mechanisms of the NBT-reduction in human platelets are discussed.


Identification and Preclinical Evaluation of SC144, a Novel Pyrroloquinoxaline Derivative with Broad-Spectrum Anticancer Activity.
Design and discovery of new classes of anticancer agents with unique mechanisms of action is an urgent medical need. During the past several years, a series of salicylhydrazide class of compounds were reported to possess remarkable potency in a large panel of cancer cell lines from different tumor origins. In particular, the optimized lead compound, SC144, was further investigated and selected as a valuable drug candidate endowed with favorable pharmacokinetic and antiproliferative properties in various in vitro and in vivo xenograft models. This lead compound is active in cells resistant to conventional chemotherapies, synergistic with several standard-of-care drugs, and possesses an unique mechanism acting through the inhibition of the gp130-STAT3-survivin axis. Because of this novel mechanism, clinical development of SC144 will provide new therapeutic options for diverse cancers.


Circular RNA circCUL3 Accelerates the Warburg Effect Progression of Gastric Cancer through Regulating the STAT3/HK2 Axis.
The Warburg effect is a significant hallmark of gastric cancer (GC), and increasing evidence emphasizes the crucial role of circular RNAs (circRNAs) in GC tumorigenesis. However, the precise molecular mechanisms by which circRNAs drive the GC Warburg effect are still elusive. The present study was designed to unveil the roles of circRNAs and the corresponding potential mechanism. High-regulated expression of circCUL3 was observed in both GC tissues and cell lines. Clinically, the high expression of circCUL3 was closely correlated with advanced clinical stage and overall survival in GC patients. Functionally, cellular experimental investigations demonstrated that circCUL3 promoted the proliferation, glucose consumption, lactate production, ATP quantity, and extracellular acidification rate (ECAR) of GC cells. In vivo, circCUL3 knockdown repressed tumor growth. Mechanistic analysis demonstrated that circCUL3 promoted signal transducer and activator of transcription (STAT)3 expression through sponging miR-515-5p; moreover, transcription factor STAT3 accelerated the transcriptional level of hexokinase 2 (HK2). In summary, the present findings provide mechanistic insights into circCUL3/miR-515-5p/STAT3/HK2 axis regulation on the GC Warburg effect, providing a novel possibility for an understanding of GC pathogenesis.


Linking ecotoxicological effects on biodiversity and ecosystem functions to impairment of ecosystem services is a challenge: an illustration with the case of plant protection products.
There is growing interest in using the ecosystem services framework for environmental risk assessments of chemicals, including plant protection products (PPPs). Although this topic is increasingly discussed in the recent scientific literature, there is still a substantial gap between most ecotoxicological studies and a solid evaluation of potential ecotoxicological consequences on ecosystem services. This was recently highlighted by a collective scientific assessment (CSA) performed by 46 scientific experts who analyzed the international science on the impacts of PPPs on biodiversity, ecosystem functions, and ecosystem services. Here, we first point out the main obstacles to better linking knowledge on the ecotoxicological effects of PPPs on biodiversity and ecological processes with ecosystem functions and services. Then, we go on to propose and discuss possible pathways for related improvements. We describe the main processes governing the relationships between biodiversity, ecological processes, and ecosystem functions in response to effects of PPP, and we define categories of ecosystem functions that could be directly linked with the ecological processes used as functional endpoints in investigations on the ecotoxicology of PPPs. We then explore perceptions on the possible links between these categories of ecosystem functions and ecosystem services among a sub-panel of the scientific experts from various fields of environmental science. We find that these direct and indirect linkages still need clarification. This paper, which reflects the difficulties faced by the multidisciplinary group of researchers involved in the CSA, suggests that the current gap between most ecotoxicological studies and a solid potential evaluation of ecotoxicological consequences on ecosystem services could be partially addressed if concepts and definitions related to ecological processes, ecosystem functions, and ecosystem services were more widely accepted and shared within the ecotoxicology community. Narrowing this gap would help harmonize and extend the science that informs decision-making and policy-making, and ultimately help to better address the trade-off between social benefits and environmental losses caused by the use of PPPs.


Recent advances in salivary cancer diagnostics enabled by biosensors and bioelectronics.
There is a high demand for a non-invasive, rapid, and highly accurate tool for disease diagnostics. Recently, saliva based diagnostics for the detection of specific biomarkers has drawn significant attention since the sample extraction is simple, cost-effective, and precise. Compared to blood, saliva contains a similar variety of DNA, RNA, proteins, metabolites, and microbiota that can be compiled into a multiplex of cancer detection markers. The salivary diagnostic method holds great potential for early-stage cancer diagnostics without any complicated and expensive procedures. Here, we review various cancer biomarkers in saliva and compare the biomarkers efficacy with traditional diagnostics and state-of-the-art bioelectronics. We summarize biomarkers in four major groups: genomics, transcriptomics, proteomics, and metabolomics/microbiota. Representative bioelectronic systems for each group are summarized based on various stages of a cancer. Systematic study of oxidative stress establishes the relationship between macromolecules and cancer biomarkers in saliva. We also introduce the most recent examples of salivary diagnostic electronics based on nanotechnologies that can offer rapid, yet highly accurate detection of biomarkers. A concluding section highlights areas of opportunity in the further development and applications of these technologies.


Luteolin mediated targeting of protein network and microRNAs in different cancers: Focus on JAK-STAT, NOTCH, mTOR and TRAIL-mediated signaling pathways.
There has always been a keen interest of basic and clinical researchers to search for cancer therapeutics having minimum off-target effects and maximum anticancer activities. In accordance with this approach, there has been an explosion in the field of natural products research in the past few decades because of extra-ordinary list of natural extracts and their biologically and pharmacologically active constituents having significant medicinal properties. Apparently, luteolin-mediated anticancer effects have been investigated in different cancers but there is superfluousness of superficial data. Generalized scientific evidence encompassing apoptosis, DNA damage and anti-inflammatory effects has been reported extensively. However, how luteolin modulates deregulated oncogenic pathways in different cancers has not been comprehensively uncovered. In this review we have attempted to focus on cutting-edge research which has unveiled remarkable abilities of luteolin to modulate deregulated oncogenic pathways in different cancers. We have partitioned the review into various sections to separately discuss advancements in therapeutic targeting of oncogenic protein networks. We have provided detailed mechanistic insights related to JAK-STAT signaling and summarized how luteolin inhibited STAT proteins to inhibit STAT-driven gene network. We have also individually analyzed Wnt/β-catenin and NOTCH pathway and how luteolin effectively targeted these pathways. Mapping of the signaling landscape has revealed that NOTCH pathway can be targeted therapeutically. NOTCH pathway was noted to be targeted by luteolin. We have also conceptually analyzed how luteolin restored TRAIL-induced apoptosis in resistant cancers. Luteolin induced an increase in pro-apoptotic proteins and efficiently inhibited anti-apoptotic proteins to induce apoptosis. Luteolin mediated regulation of non-coding RNAs is an exciting and emerging facet. Excitingly, there is sequential and systematic accumulation of clues which have started to shed light on intricate regulation of microRNAs by luteolin in different cancers. Collectively, sophisticated information will enable us to develop a refined understanding of the multi-layered regulation of signaling pathways and non-coding RNAs by luteolin in different cancers.


Tracheobronchoplasty for resection of lung cancer arising from a tracheal bronchus.
Tracheal bronchus is a rare anomoly wherein a bronchus arises directly from the trachea and supplies the right upper lobe. We had a case of lung cancer arising from a tracheal bronchus with invasion into the trachea. Tracheobronchoplasty was needed for complete resection of the tumor. Because of the anatomical characteristics of the tracheal bronchus, special surgical techniques for resection of the neoplasm were needed.


Lithocarpus polystachyus (Sweet Tea) water extract promotes human hepatocytes HL7702 proliferation through activation of HGF/AKT/ERK signaling pathway.
Sweet Tea (ST), derived from the leaves of Lithocarpus polystachyus, is a Chinese folk medicine with wide pharmacological activities. However, the promotive effects of ST water extract on hepatocytes proliferation and its underlying mechanism remains still unknown. In the present study, the beneficial effects of ST water extract on human hepatocytes and its possible mechanism were investigated. MTT assay was used to detect the safety range of ST; HL7702 cells were divided into four groups: control group, ST low- (50 μg/mL), medium- (200 μg/mL) and high-concentration (800 μg/mL) groups; BrdU ELISA and EDU staining were used to observe DNA content and cell proliferation; Moreover, flow cytometry was applied to analyze the distribution of cell cycle. Furthermore, the expression of cyclin D1, CDK4, HGF/c-Met, Akt, Erk1/2 were detected by Western blot. It was found that ST water extract concentration-dependent promoted human hepatocytes HL7702 cell proliferation within 72 h through accumulating the cells in S phase and G2/M phase. Furthermore, ST water extract up-regulated expression of Cyclin D1 and CDK4 proteins. Moreover, ST water extract not only increased HGF expression and phosphorylation of c-Met level, but also activated the phosphorylation levels of AKT, ERK1/2. Interestingly, both of AKT inhibitor A6730 and ERK1/2 inhibitor U0126 reversed the promotive effects of ST water extract, which further confirmed that activation of AKT and ERK1/2 were involved. The findings reveal that ST water extract promoted HL7702 cells proliferation through the stimulation of cell cycle mediated by activating the AKT- and ERK1/2-related pathway.


Turkish Adaptation of the Moral Distress Scale-Revised for Pediatric Nurses: A Validity and Reliability Study.
Background and Purpose: Recent studies have demonstrated the adverse effects that moral distress experienced by nurses has on nurses, patients, and the healthcare system. This study aims to analyze the validity and reliability of the Turkish version of the Moral Distress Scale-Revised (MDS-R, pediatric). Methods: This study was conducted with 210 pediatric nurses. Results: Explanatory factor analysis was conducted, and a five-factor structure emerged. The Cronbach's α value of the scale was found to be 0.865, and the correlation-based item analysis showed that the values were within the acceptable range, and the discrimination of the items was adequate. Conclusions: Analyses conducted revealed that the Turkish version of the MDS-R (pediatric), consisting of 21 items and five subdimensions, is a valid and reliable measurement tool for the Turkish culture and language.


Simultaneous determination of harmine, harmaline and their metabolites harmol and harmalol in beagle dog plasma by UPLC-ESI-MS/MS and its application to a pharmacokinetic study.
Harmine (HAR) and harmaline (HAL) were metabolized by demethylation to form harmol (HOL) and harmalol (HAM) both in vivo and in vitro. It has been demonstrated tremendous value of HAR, HAL and their metabolites in the therapy of Alzheimer's disease. A rapid, selective and sensitive UPLC-ESI-MS/MS method was firstly developed and validated for the simultaneous determination of HAR, HAL, HOL, and HAM in beagle dog plasma with 9-aminoacridine as the internal standard (IS). After protein precipitation with acetonitrile, the analytes were separated within 4.5 min on an ACQUITY UPLC BEH C18 column with a gradient elution system composed of 0.1% formic acid and acetonitrile at a flow rate of 0.4 ml/min. Detection was performed using multiple reactions monitoring mode under a positive ionization condition. The calibration curves of four analytes showed good linearity (r(2)>0.9959) within the tested concentration ranges. The low limit of quantification for HAR, HAL, HOL, and HAM were all 1.00 ng/ml. The mean accuracy of the analytes was within the range of 94.56-112.23%, the R.S.D. values of intra-day and the inter-day precision were less than 6.26% and 7.51%, respectively. Matrix effects and extraction recoveries of the analytes from the beagle dog plasma were within the range of 94.48-105.77% and 89.07-101.44%, respectively. The validated method was successfully applied to a pharmacokinetic study of HAR, HAL, HOL, and HAM in beagle dogs after intravenous administration of HAR and HAL both of 1.0mg/kg. The main pharmacokinetic parameters of Cmax, Vd, CL, AUC and MRT, except Ke and t1/2 values, showed significant difference between the two parent drug HAR and HAL, respectively (p<0.05-0.001). Because of the different metabolic rate of HAR and HAL in vivo, the two metabolites, HOL and HAM, exhibited unique pharmacokinetic properties.


Characterization of Secondary Structure and Thermal Stability by Biophysical Methods of the D-alanyl,D-alanine Ligase B Protein from Escherichia coli.
Peptidoglycan (PG) is a key structural component of the bacterial cell wall and interruption of its biosynthesis is a validated target for antimicrobials. Of the enzymes involved in PG biosynthesis, D-alanyl,D-alanine ligase B (DdlB) is responsible for the condensation of two alanines, forming D-Ala-D-Ala, which is required for subsequent extracellular transpeptidase crosslinking of the mature peptidoglycan polymer. We aimed at the biophysical characterization of recombinant Escherichia coli DdlB (EcDdlB), considering parameters of melting temperature (Tm), calorimetry and Van't Hoff enthalpy changes of denaturation ( ΔHUcal and ΔHUvH ), as well as characterization of elements of secondary structure at three different pHs. DdlB was overexpressed in E. coli BL21 and purified by affinity chromatography. Thermal stability and structural characteristics of the purified enzyme were analyzed by circular dichroism (CD), differential scanning calorimetry and fluorescence spectroscopy. The stability of EcDdlB increased with proximity to its pI of 5.0, reaching the maximum at pH 5.4 with Tm and ΔHUvH U of 52.68 ºC and 484 kJ.mol-1, respectively. Deconvolutions of the CD spectra at 20 ºC showed a majority percentage of α-helix at pH 5.4 and 9.4, whereas for pH 7.4, an equal contribution of β-structures and α-helices was calculated. Thermal denaturation process of EcDdlB proved to be irreversible with an increase in β-structures that can contribute to the formation of protein aggregates. Such results will be useful for energy minimization of structural models aimed at virtual screening simulations, providing useful information in the search for drugs that inhibit peptidoglycan synthesis.


Immune reconstitution and survival of patients after allogeneic hematopoietic stem cell transplantation from older donors.
The impact of donor age on the immune reconstitution of patients with hematological malignancies who underwent hematopoietic cell transplantation (HCT) is unclear. We retrospectively compared the outcomes of 381 patients who underwent allogeneic peripheral blood stem cell transplantation (PBSCT) from 308 donors under 50 years of age and 73 donors over 50 years of age. IVIG was regularly supplemented for patients in the first 3 months post-HCT. The counts of CD8+CD45RA+ naïve T cells were significantly lower in patients of the older donor group than in the younger donor group in the first year after PBSCT (190.6 cells/μl vs. 239.6 cells/μl, p = .018). Patients in the older donor group had significantly fewer CD19+ B cells on day +270 (123.4 cells/μl vs. 183.5 cells/μl, p = .021) and day +365 (169 cells/μl vs. 271.1 cells/μl, p = .01) after PBSCT. Serum IgA (.76 g/L vs. .97 g/L, p < .001) and IgM levels (.75 g/L vs. 1.04 g/L, p < .001) were significantly lower in patients in the older donor group from day +60 to +365 after PBSCT. The EBV reactivation rate within the first 3 months after PBSCT was significantly higher in patients in the older donor group (48.6% vs. 38.3%, p = .034). However, the incidences of CMV reactivation, II-IV acute graft-versus-host disease (aGvHD), chronic GvHD (cGvHD), 3-year relapse rate, 3-year transplant-related mortality (TRM) and 3-year overall survival (OS) were not significantly different between the two groups. In conclusion, donors ≥50 years old were associated with inferior immune reconstitution and higher EBV reactivation in patients after PBSCT, but no change in OS.


Short- and long-term cardiovascular effects of mixed amphetamine salts extended release in children.
To assess the short- and long-term cardiovascular effects of once-daily treatment with a mixed amphetamine salts extended-release formulation (MAS XR; Adderall XR(R)) in children age 6 to 12 years with attention-deficit/hyperactivity disorder (ADHD). Short-term cardiovascular effects were assessed during a 4-week, double-blind, randomized, placebo-controlled, forced-dose-titration study of once-daily 10, 20, and 30 mg MAS XR (n = 580). Long-term cardiovascular effects were assessed in 568 subjects during a 2-year, open-label extension study of MAS XR (10 to 30 mg/day). Resting sitting blood pressure and pulse were measured at baseline and weekly during the short-term study, then monthly during long-term treatment. Changes in blood pressure, pulse, and QT interval corrected by Bazett's formula (QTcB) in children receiving MAS XR were not statistically significantly different than those changes seen in children receiving placebo during short-term treatment. Mean increases in blood pressure after 2 years of MAS XR treatment (systolic, 3.5 mm Hg; diastolic, 2.6 mm Hg) and pulse (3.4 bpm) were clinically insignificant, and there was no apparent dose-response relationship. Cardiovascular effects of short- and long-term MAS XR were minimal during short- and long-term MAS XR treatment at doses of </= 30 mg/day in otherwise healthy children.


Peritoneal dialysis in the nursing home.
During the past few decades, the demographics of end stage renal disease have been changed significantly with the emerging predominance of elderly patients. Elderly dialysis patients are usually more dependent and may need long-term placement in a long-term care facility. Failure to meet the needs of these patients may have a significant impact on the peritoneal dialysis program. We report our experience of starting peritoneal dialysis program in a community-based Long Term Care Facility (LTCF). During the period of 2000-2001, after appropriate training of nursing home personnel, we admitted 8 peritoneal dialysis patients to one community-based nursing home. All information presented here has been collected through chart review. At the time of admission the average age of the 8 patients was 77.3 +/- 7.2 years (range 69 to 91 years). All patients had several comorbid diseases and six of the eight were bed-ridden. The patients stayed in the facility for a total of 29.57 patient months. One patient had three episodes of peritonitis within three months (all culture negative) and has been excluded from the analysis of the overall peritonitis rate. The peritonitis rate for the other seven patients was 1 per 7.54 patient month. Six patients were readmitted to hospital because of peritonitis, severe malnutrition and hip fracture. Four of them died in the hospital. One died in the nursing home. One patient remains in the nursing home at the present time. Our experience suggests that peritoneal dialysis can be achieved in a community-based nursing home. This requires a systematic training program for the LTCF personnel and the availability of a "dedicated" nephrology dialysis staff. This is crucial to the success of the program. It is important that patients, their families and ESRD care professionals are informed of the limited survival expectation particularly for very old and severely impaired patients.


Evolution in the clinic: Maladaptive units and "minor anomalies".
It is here argued that the application of the term "minor anomalies" is often imprecise and likely outdated. In the past, the designation was used indiscriminately to refer to a great variety of unrelated morphogenetic phenomena. Also, the term does not discriminate between mild qualitative defects of development (mild malformations) and quantitative variants of normal structure. The human face was formed by natural and sexual selection. Morphological and morphogenetic analyses have shown that the human face with its skin, muscles, nerves, arteries, veins, glands, and lymphatics is a complex structure made up of progeny of ectoderm and mesoderm. Holoprosencephaly demonstrates graphically how these embryonic derivatives fit together sequentially. These derivatives are the adaptive units of the human organism, the result of stringent evolutionary forces uniting essential function to a minimum of structure. Before an "unusual" facial appearance is diagnosed as "abnormal", phenotype analysis is required to determine if there is a family resemblance or if it is a pleiotropic structure. The facial structures of chimps and humans are homologous by virtue of descent from a common ancestor (Darwin, 1859). Differences in the appearance of these species reflect adaptive divergence over some 6-7 million years of evolution while retaining over 98-99% genetic identity. Both species may develop Down syndrome, evidence of similarly retained developmental plasticity. It has occurred to us that Dobzhansky's axiom ("Nothing in biology makes sense except in the light of evolution") applies not only to genetics, but to all of medicine.


Epithelial-myoepithelial carcinoma of floor of mouth: A case report with cytological, histological and immunohistochemical correlation.
A 61-year-old female presented with a 3-year-old swelling in the right floor of mouth. Clinical examination and fine needle aspiration cytology suggested a benign lesion. The mass was excised locally along with the involved sublingual and deep part of submandibular gland and duct. Post-operative histopathological examination revealed features of pleomorphic adenoma. However, on revision of histological sections, features were predominantly of a rare malignancy of the salivary glands, epithelial-myoepithelial carcinoma (EMC), along with focal areas of adenoid cystic carcinoma (Ad CC). The tumor was p-63, s-100 and smooth muscle actin positive but C-kit was negative, which ruled out Ad CC and the possibility of a hybrid carcinoma. The aim of this article is to describe a rare case of EMC in the floor of mouth and the confusing cytological picture that it created.


Oral morphine in chronic cancer pain.
Extensive clinical experience has been obtained in the use of opiates during the last decade in special units devoted to symptom control in advanced cancer. Important contradictions have emerged with the clinical pharmacological literature on opiates calling into question its relevance to the treatment of chronic pain. Specifically in the case of morphine it is clear that: it is a very effective analgesic given orally, dosage must be individualized, parenteral use or exotic analgesic 'cocktails' are usually unnecessary, and tolerance, dependence and respiratory depression are rarely common or serious problems which prevent effective pain control provided morphine is used appropriately in accordance with its pharmacological characteristics. Heroin is a suitable alternative to morphine (particularly for intramuscular administration) if differences in milligram potency are taken into account, but has no advantages in terms of either analgesic efficacy or side effects. This paper summarizes clinical experience in the use of oral morphine for cancer pain at St. Christopher's Hospice, any data from clinical investigations which support this approach, and comments on the areas of controversy which have emerged.


Fijac: fright and illness in highland Yemen.
Sudden fright, 'fijac', has played an important role in the traditional explanatory models of illness experiences in highland Yemen. Fijac is quite similar to other examples of the fright illness taxon in that it is a folk-illness category that is attributed to a wide variety of underlying conditions. It is argued that given the extremely labile symptomatology, fijac, like other examples of the fright illness taxon, does not constitute a culture-bound psychiatric syndrome. Based on analysis of case studies and preliminary survey data, fijac appears to be much more common among folk etiologies offered by Yemeni women than men. It is suggested that this social profile might be explained by changes in the distribution of medical knowledge in Yemen. With the rapid rate of social change and the increased exposure to cosmopolitan medicine resulting from internal development of cosmopolitan health care and international labor migration, men have supplemented their traditional explanatory models with alternatives drawn from cosmopolitan medicine. Succumbing to illness as a result of fright is contradictory to the male ideal of the courageous tribesman; alternative explanatory models that do not challenge this ideal self predominate. By contrast, the Yemeni value system defines women and children as vulnerable and weak; therefore, being subject to the impact of fright is consistent with youth and the cultural definition of the female self.


Quantitative and qualitative changes in AMPA receptor expression during spinal cord development.
Synaptic activity in early postnatal life is important for the acquisition of mature structural and functional properties of neurons. Previous studies indicate that the mature molecular features of spinal motor neurons emerge during a period of activity-dependent development in early postnatal life. Since glutamatergic synaptic transmission provides the major excitatory drive into motor neurons, glutamate receptors are likely to play a central role in motor neuron activity-dependent development. To gain insight into this process, we have used receptor autoradiography, immunoblotting and immunohistochemistry to determine the distribution, temporal expression and potential subunit composition of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid subtype glutamate receptors in the developing rat spinal cord. Using two different ligands, [3H]-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and [3H]-6-cyano-7-nitroquinoxaline-2,3-dione, we find that alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid binding sites in the adult are largely restricted to the substantia gelatinosa. In marked contrast, during early postnatal life, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid binding sites are transiently expressed at high levels in the ventral horn. This parallels previous findings on the developmental regulation of N-methyl-D-aspartate receptor expression. Using alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor subunit-specific antibodies we show by immunoblot analysis and immunohistology that, to varying degrees, the expression patterns of glutamate receptor subunit 1 and glutamate receptor subunits 2/3 are significantly developmentally regulated. The most conspicuous change is the downregulation of glutamate receptor 1 expression within motor neurons over the first three weeks of postnatal life. The qualitative and quantitative changes we observe in glutamate receptor expression in early postnatal life are likely to have a major impact on the electrophysiological properties of young motor neurons and thus may contribute to their activity-dependent development.


Novel intrinsic factor Yun maintains female germline stem cell fate through Thickveins.
Germline stem cells (GSCs) are critical for the reproduction of an organism. The self-renewal and differentiation of GSCs must be tightly controlled to avoid uncontrolled stem cell proliferation or premature stem cell differentiation. However, how the self-renewal and differentiation of GSCs are properly controlled is not fully understood. Here, we find that the novel intrinsic factor Yun is required for female GSC maintenance in Drosophila. GSCs undergo precocious differentiation due to de-repression of differentiation factor Bam by defective BMP/Dpp signaling in the absence of yun. Mechanistically, Yun associates with and stabilizes Thickveins (Tkv), the type I receptor of Dpp/BMP signaling. Finally, ectopic expression of a constitutively active Tkv (TkvQD) completely suppresses GSC loss caused by yun depletion. Collectively, these data demonstrate that Yun functions through Tkv to maintain GSC fate. Our results provide new insight into the regulatory mechanisms of how stem cell maintenance is properly controlled.


Staphylococcus aureus Nasopharyngeal Carriage and Antimicrobial Resistance among Adults with Sickle Cell Disease at the Korle Bu Teaching Hospital in Accra, Ghana.
Sickle cell disease (SCD) patients are an important risk group for Staphylococcus aureus (S. aureus) carriage and infections. Little is, however, known about the nasopharyngeal carriage epidemiology of the pathogen in this vulnerable population. The aim of this study was to evaluate S. aureus and methicillin-resistant S. aureus (MRSA) nasopharyngeal carriage prevalence, carriage determinants, and antimicrobial resistance among SCD adults in Ghana. Nasopharyngeal swabs, obtained from 200 SCD adults recruited at the Korle Bu Teaching Hospital, were cultured for S. aureus, and these isolates were subjected to antimicrobial susceptibility testing via the Kirby-Bauer method. The prevalence of S. aureus carriage was 41.5% (n = 83), and that of MRSA carriage was 1.0% (n = 2). Moreover, carriage of coagulase-negative Staphylococcus (CoNS) was the only determinant of S. aureus carriage identified (OR = 0.012, P < .0001). However, neither this variable nor the other features of the participants emerged as a determinant of MRSA carriage. The antimicrobial resistance rates decreased across penicillin (98.8%, n = 82), tetracycline (54.2%, n = 45), gentamicin (32.5%, n = 27), ciprofloxacin (21.7%, n = 18), erythromycin (18.1%, n = 15), clindamycin (10.8%, n = 9), amoxicillin-clavulanic acid (10.8%, n = 9), teicoplanin (1.2%, n = 1), and linezolid (0.0%, n = 0), and the multidrug resistance rate was 45.8% (n = 38). The nasopharyngeal carriage prevalence of S. aureus in the current study was high, while that of MRSA was low. The isolates were highly resistant to several of the antibiotics tested, but not teicoplanin and linezolid, making these antibiotics suitable for treatment of S. aureus infections among the SCD population.


8-oxoguanine DNA glycosylase, but not Kin17 protein, is translocated and differentially regulated by estrogens in rat brain cells.
8-oxoguanine DNA glycosylase and Kin17 are proteins widely distributed and phylogenetically conserved in the CNS. 8-oxoguanine DNA glycosylase is a DNA repair enzyme that excises 7,8-dihydro-8-oxoguanine present in DNA damaged by oxidative stress. Kin17 protein is involved in DNA repair and illegitimate recombination in eukaryotic cells. The present study evaluates the effect of ovarian hormones on the expression of both proteins in the magnocellular paraventricular nucleus of the hypothalamus and the bed nucleus of the stria terminalis in female and male rat brains. In the paraventricular nucleus, ovariectomy induced a significant decrease in the number of 8-oxoguanine DNA glycosylase-positive nuclei as well as in their relative fluorescent intensity as compared with ovariectomized-estradiol treated and proestrous groups. Confocal microscopy observation demonstrated that oxoguanine DNA glycosylase protein is located in the Hoechst-dyed nuclei and cytoplasm in male and ovariectomized rats. Surprisingly, following estradiol administration to ovariectomized and proestrous rats, the 8-oxoguanine DNA glycosylase immunolabeling was observed in the nucleolus, the cytoplasm and the dendrites of cells, while Kin17 protein was always localized in the cell nuclei. In the bed nucleus of the stria terminalis, the number of 8-oxoguanine DNA glycosylase-positive nuclei during proestrous was significantly lower than the number obtained in males and ovariectomized rats and similar to the number of ovariectomized-estradiol-treated groups. In contrast to these observations, no significant differences were observed in the expression of kin17 protein. Our results suggest that estrogens differentially regulate the expression of 8-oxoguanine DNA glycosylase, but not that of Kin17 protein, in specific regions of the rat brain and that estradiol can translocate the 8-oxoguanine DNA glycosylase protein within nuclei and to other subcellular compartments.


Trial of early neonatal hydrocortisone: two-year follow-up.
Dexamethasone treatment is associated with an increased risk of cerebral palsy (CP). Early hydrocortisone (HC) treatment may decrease the incidence of bronchopulmonary dysplasia; however, the long-term effects are still under evaluation. Follow-up of randomized studies concerning early HC treatment is essential to confirm the long-term safety. We hypothesized that early HC treatment in very preterm infants does not impair the neurologic outcome. We report follow-up data from a randomized trial of early HC given for 10 days. Before the HC or placebo treatment, serum cortisol levels were measured. Receiver-operating characteristic was defined. Values below the median were classified as low endogenous cortisol and those above the median as high endogenous cortisol. A meta-analysis was performed. Altogether 98% of the 46 surviving infants participated in a follow-up study at a corrected age of 2 years. The growth characteristics were similar between the study groups. The developmental quotients (DQs) of the children with high endogenous cortisol and placebo treatment shortly after birth (100 +/- 13) and those with low endogenous cortisol and HC (97 +/- 7) were not lower than the DQs of the children with high endogenous cortisol and HC (92 +/- 3) or low cortisol and placebo (96 +/- 2). According to a meta-analysis of three available trials (411 children), the rate of CP and survival without neurosensory or cognitive impairment was not influenced by HC. Early low-dose HC administration had no adverse effects at 2 years of age. Further studies are required to define the target group for neonatal HC.


Cell surface receptors for lactate dehydrogenase-elevating virus on subpopulation of macrophages.
We examined the binding and internalization of unlabeled and 125I-labeled, purified lactate dehydrogenase-elevating virus (LDV) by peritoneal macrophages cultured in vitro. Upon incubation of the cells at 4 degrees C with greater than 100 ID50/cell, the virus was surface-bound on a small subpopulation of macrophages (about 5% of the total cells) as determined by electron microscopy, fluorescent antibody staining, and autoradiography of cells incubated with 125I-labeled LDV. At 37 degrees C, LDV particles were seen in intracellular endocytic vesicles also in about 5% of the cells, and the proportion of cells with virus-containing vesicles correlated with the proportion of cells which became productively infected with LDV as assessed by determining LDV RNA synthesis in individual cells and by fluorescent antibody staining. Pretreatment of the resident peritoneal macrophages with trypsin inhibited the binding of 125I-labeled LDV and the productive infection of the cells with the virus. After removal of the trypsin and incubation in complete medium, permissiveness for LDV reappeared after an 8-12 h lag, whereas Fc and C3 receptors reappeared more rapidly after trypsin treatment. Populations of resident peritoneal macrophages, starch-elicited peritoneal macrophages, splenic macrophages, and bone marrow macrophages contained a similar proportion of cells that could be productively infected with LDV. Little, if any, LDV replication was detected in cultures of lung, liver and peripheral blood macrophages as well as in thioglycollate-elicited and BCG-activated macrophages. We conclude that the permissiveness for LDV of resident peritoneal macrophages correlates with the presence of a trypsin-sensitive receptor present on a subpopulation of these cells. The identity of the receptor has not been definitively established. Treatment of macrophages with neuraminidase or various sugars had no significant effect on LDV replication. Lysis of I-A-positive macrophages with a monoclonal antibody and complement reduced the number of macrophages which could be productively infected by 50%, which suggests that macrophages lacking surface Ia can be productively infected with LDV in vitro.


Code teams and the review of cardiac arrests.
A new system for responding to Codes, or resuscitations of patients with cardiopulmonary arrest, was implemented at St Luke's Medical Center, Milwaukee. A teaching program was begun to establish the Advanced Cardiac Life Support (ACLS) guidelines as the standard of care. A task force defined the composition of the Code team and was then designated the Code Review Committee, meeting monthly to review all Codes. Indicators for review were established. Code review is followed by a letter to the physician Code team leader and/or registered nurse. Data for 18 months, still preliminary, show improved compliance with indicators for documentation and for adherence to ACLS guidelines.


Some statistical considerations for design of cancer prevention trials.
Carcinogenesis is believed to occur in at least two stages, initiation and promotion, followed by a preneoplastic lesion which develops into cancer. Cancer prevention trials can be classified as primary if the intervention precedes initiation, secondary if it occurs during promotion, and tertiary if it is applied to a preneoplastic lesion. Tertiary prevention trials resemble treatment trials, but primary and secondary prevention trials may be very different in size, duration, and cost. After reviewing some basic questions which must be addressed in designing any cancer prevention trial, some special design considerations appropriate for primary and secondary prevention trials are discussed. These include the use of factorial designs, group or cluster randomization, special sample size calculations needed for large-scale trials of long duration with cancer incidence as the endpoint, and the idea of the case-cohort approach for monitoring and for subsequent exploratory analysis of trial data.


An exploration of the social stigma of tuberculosis in five "municipios" of Nicaragua to reflect on local interventions.
The social stigma of tuberculosis is much less studied than those of other diseases such as AIDS or mental problems. However, it has important implications on the affected person's well being and on the epidemic's control. Our study aims at exploring this social stigma in five local health systems of Nicaragua, prior to implementing interventions to reduce it. Through in-depth interviews and focus groups involving stakeholders in the care of people affected by tuberculosis (PATBs), we analysed interactions between PATBs and family members, first line government health services' personnel, and community members. According to our results, the interaction between stakeholders and PATBs can be described as the intersection between two sets of contradictory feelings and attitudes: (a) feelings of affection and supportive attitudes toward PATBs opposed to the fear of being infected or that PATBs will infect others and, (b) confidence in PATBs considered to be unlucky opposed to mistrust of PATBs considered to be negligent. PATBs react against this mainly by hiding their condition which leads them to a, loss of confidence and depression. This intricate group of feelings and attitudes is influenced by two sets of determinants related to domination and power between stakeholders and issues of knowledge and information. Analysing tuberculosis-related social stigma as a social process enabled us to better understand some key social structural factors of health care system's organisation and identify locally acceptable interventions to reduce such stigma. The fact of analysing, in a more thorough study, some interventions in the currently changing social structural context of health care systems in Nicaragua will give us a better insight into the relevance of our analysis and the interventions' effectiveness in reducing the social stigma of tuberculosis.


Drug-eluting vs. bare metal stents for symptomatic vertebral artery stenosis.
To evaluate the immediate and long-term outcomes of drug-eluting stent (DES) vs. bare metal stent (BMS) for symptomatic vertebral artery stenosis (VAS). From 2003 to 2010, 206 consecutive patients (158 men; mean age 66.8 years) underwent DES (sirolimus-eluting or paclitaxel-eluting) or BMS placement for symptomatic extracranial and intracranial stenoses in 219 vertebral arteries. The technical success, clinical success, periprocedural complications, target vessel revascularization (TVR), and overall survival were compared between the DES and BMS groups. The technical success rate was 98.3% (119/121) for the DES group vs. 100% for the BMS group (p = 0.503). The clinical success rate was 95.5% (107/112) for the DES group vs. 97.9% (92/94) for the BMS group (p = 0.592). No periprocedural death or stroke occurred. The overall periprocedural complication rate was 2.7% (3/112) in the DES group vs. 4.3% (4/94) in BMS group (p = 0.813). The median follow-up was 43 months (range 3-95) for the DES group and 46 months (range 6-89) for BMS. At last follow-up, the TVR rate was 6.3% (7/112) for the DES group vs. 20.2% (19/94) for the BMS group (p = 0.003); 4 (3.6%) patients in the DES group and 8 (8.5%) patients in the BMS group experienced a VBS stroke (p = 0.132). By life-table analysis, the 5-year TVR rate was 4.5% (5/112) for the DES group vs. 19.1% (18/94) for the BMS group (p = 0.001). No difference was detected in the overall survival curves between the groups (p = 0.500). Both DES and BMS are feasible, safe, and effective for symptomatic VAS. However, DES can significantly decrease the TVR rate in the long term compared with BMS.


Using Genome Scale Mutant Libraries to Identify Essential Genes.
Identification of essential genes is key to understanding the required processes and gene products of organisms under one or more conditions. Transposon sequencing (Tn-seq) has been used to predict essential genes or ones that conditionally impact fitness in a wide variety of organisms. Here, we describe the generation of genome-scale mutant libraries and the analysis of Tn-seq data to identify essential genes from cultures grown in a single condition as well as those that are conditionally important by analyzing the behavior of these mutant libraries in different growth environments. While we illustrate the approach using data derived from Tn-seq analysis of the α-proteobacteria Rhodobacter sphaeroides and Zymomonas mobilis, the protocols and systems we describe should be generally applicable to a variety of organisms.