Evolutionary innovation in behavior and speciation: opportunities for behavioral neuroethology.
Populations of animals differentiate by speciation. How speciation takes place, for long a puzzle for evolutionary biologists, should be regarded as a research opportunity for neuroethologists and comparative behaviorists. It is now clear that behavior may play an important part in the process of speciation. The existence of sexually dimorphic anatomy and behavior in many animals has provided a rich subject for investigation by comparative neurobiologists and behaviorists, who emphasize analysis of proximate mechanisms that generate dimorphism in phenotype. However, sexual dimorphisms also figure prominently in the theory of sexual selection. Sexual selection is viewed as a primary behavioral mechanism in the process of speciation. Some examples of 'explosive' speciation are presented, and the putative role that sexual selection plays in these cases is reviewed. A consideration of any evolutionary selective process must include genetics, and I will briefly summarize studies that indicate that even complex behavioral systems such as sexual selection, which sociobiologists refer to as a 'strategy', can have a relatively simple genetic basis.


[Initial research of customized aberration correction with adaptive optics technique].
To analyze the effects of different wavefront-guided aberration ablation patterns on visual function using adaptive optics technique and explore the effective and feasible pattern of customized refractive surgery. Control study. Under dark condition, six volunteers'right eyes were fully dilated with 5 g/L phenyl ephedrine. The myopia and astigmatism were first fully corrected with optical lens, and then 100% contrast visual acuity of different wavefront correction patterns was detected using adaptive optics system in through a 6.0 mm pupil optical path. All of the eight different aberration correction strategies obtained excellent correction effect. On the basis of this correction, visual acuity test was performed. The results showed that all aberration correction strategies improved the visual acuity compared with static correction. Correction of high order aberrations and full correction of aberrations could obtain better visual acuity than that of only correction of low order aberrations. Correction of aberrations from second to third order and fourth order spherical aberration second only to full correction of aberration. As for low myopia patients, eliminating ocular high order aberrations could improve visual acuity. Correction of aberration from second to third order and fourth spherical aberration might be an effective and feasible pattern of customized refractive surgery.


Preparation of Catechin Nanoemulsion from Oolong Tea Leaf Waste and Its Inhibition of Prostate Cancer Cells DU-145 and Tumors in Mice.
Anti-cancer activity of catechin nanoemulsions prepared from Oolong tea leaf waste was studied on prostate cancer cells DU-145 and DU-145-induced tumors in mice. Catechin nanoemulsions composed of lecithin, Tween-80 and water in an appropriate proportion was prepared with high stability, particle size of 11.3 nm, zeta potential of -67.2 mV and encapsulation efficiency of 83.4%. Catechin nanoemulsions were more effective than extracts in inhibiting DU-145 cell growth, with the IC50 being 13.52 and 214.6 μg/mL, respectively, after 48 h incubation. Furthermore, both catechin nanoemulsions and extracts could raise caspase-8, caspase-9 and caspase-3 activities for DU-145 cell apoptosis, arresting the cell cycle at S and G2/M phases. Compared to control, catechin nanoemulsion at 20 μg/mL and paclitaxel at 10 μg/mL were the most effective in reducing tumor volume by 41.3% and 52.5% and tumor weight by 77.5% and 90.6% in mice, respectively, through a decrease in EGF and VEGF levels in serum.


Knowledge of first-aid measures of avulsion and replantation of teeth: an interview of 221 Kuwaiti schoolchildren.
The prognosis of replantation of an avulsed tooth is determined by which first-aid measures are taken during the first 15 min after avulsion. Knowledge of the correct first-aid measures is therefore crucial to successful replantation. The aims of this study were (i) to assess the present knowledge level of emergency measures for tooth avulsion in Kuwaiti schoolchildren, and (ii) to design and test an interview form with structured standardized questions. A total of 221 Kuwaiti schoolchildren (aged 7-15 years old) were interviewed by professionals using a standardized method to score several areas of knowledge about tooth avulsion and replantation. Earlier experience of first-aid information and subjection to dental trauma was registered. The following fields of knowledge were assessed: general body injury treatment principles, tooth avulsion and replantation principles, avulsed permanent/primary teeth, cleaning of avulsed tooth before replantation, extra alveolar time and storage media. The form for interviewing children proved to be sufficiently structured in performing the interviews and data management. The results of the interviews showed that 30.3% of the children had been exposed to dental trauma in the past. Among children 7-9 years of age, 25% had received information on general first aid as compared with 75% in children 10 years and older. Children 10 years and older, in general, had a high knowledge level of general principles of how to manage injuries to the body. Regardless of age group, there were generally a low knowledge level regarding tooth avulsion, replantation, extra-alveolar time and storage media. We conclude that first-aid knowledge in Kuwaiti schoolchildren is low on avulsion and replantation of teeth despite a high knowledge level of body injuries. The knowledge level of first-aid measures on avulsion and replantation of teeth could be increased through intervention programs.


Effects of finerenone in people with chronic kidney disease and type 2 diabetes are independent of HbA1c at baseline, HbA1c variability, diabetes duration and insulin use at baseline.
To evaluate the effect of finerenone by baseline HbA1c, HbA1c variability, diabetes duration and baseline insulin use on cardiorenal outcomes and diabetes progression. Composite efficacy outcomes included cardiovascular (cardiovascular death, non-fatal myocardial infarction, non-fatal stroke or hospitalization for heart failure), kidney (kidney failure, sustained ≥ 57% estimated glomerular filtration rate decline or renal death) and diabetes progression (new insulin initiation, increase in antidiabetic medication, 1.0% increase in HbA1c from baseline, new diabetic ketoacidosis diagnosis or uncontrolled diabetes). In 13 026 participants, risk reductions in the cardiovascular and kidney composite outcomes with finerenone versus placebo were consistent across HbA1c quartiles (P interaction .52 and .09, respectively), HbA1c variability (P interaction .48 and .10), diabetes duration (P interaction .12 and .75) and insulin use (P interaction .16 and .52). HbA1c variability in the first year of treatment was associated with a higher risk of cardiovascular and kidney events (hazard ratio [HR] 1.20; 95% confidence interval [CI] 1.07-1.35; P = .0016 and HR 1.36; 95% CI 1.21-1.52; P < .0001, respectively). There was no effect on diabetes progression with finerenone or placebo (HR 1.00; 95% CI 0.95-1.04). Finerenone was well-tolerated across subgroups; discontinuation and hospitalization because of hyperkalaemia were low. Finerenone efficacy was not modified by baseline HbA1c, HbA1c variability, diabetes duration or baseline insulin use. Greater HbA1c variability appeared to be associated with an increased risk of cardiorenal outcomes.


Mortality predictors in hospitalized elderly patients.
To find out which of the two predictors, Charlson co-morbidity index or vitamin B12, better estimates the risk of in-hospital mortality in seriously ill patients. Electronic hospital records of 1509 elderly patients aged 65 and older were retrospectively surveyed. Albumin, age and elevated vitamin B12 levels were significantly associated with increased in-hospital mortality. Charlson co-morbidity index was not significantly associated with death. The highest mortality (24.3%) was found in the group of patients who were concomitantly in the lowest albumin quartile and the highest vitamin B12 levels quartile. In this group, mortality increased significantly with age. By elasticity calculation, vitamin B12 capability to predict mortality was higher by ≈ 3 times than that of Charlson co-morbidity index. In view of the fact that vitamin B12 levels have been found to predict mortality, they should be measured in geriatric practice, in addition to albumin levels, as a practical and reliable tool for identifying high risk elderly hospitalized patients. Probably, a combination of two or more available and inexpensive routinely taken tests can give a better estimation of mortality than some complicated tools, like Charlson co-morbidity index.


Transport of the neonate.
A regionalization program, initiated in 1973 at Vanderbilt University Medical Center, has included community hospital site visits, formal nursing courses, and neonatal transport. The neonatal transport program was designed not only to give initial medical care to the sick newborn, but also to develop the educational aspects of the regionalization program. Analysis of data from the first two years (614 transports) revealed significant educational interactions occurring during transport. Improved perinatal care the second year was reflected in decreased regional neonatal mortality and improved neonatal management.


Spatial expression of DNA topoisomerase I genes during cell proliferation in Daucus carota.
The spatial expression of carrot (Daucus carota L.) top1 genes encoding the two isoforms of the enzyme DNA topoisomerase I (EC 188.8.131.52) was investigated. In situ hybridization analysis performed with a probe recognizing both top1 transcripts provided evidence that in explanted hypocotyls induced to proliferate in vitro by the addition of the growth regulator 2,4-dichlorophenoxyacetic acid (2,4-D), the mRNA accumulation parallels the proliferation of provascular cells of the stelar cylinder. During somatic embryogenesis, the histological distribution of top1 transcripts was strongly evident at the stage of torpedo-shaped embryos, but gene expression was not only restricted to meristematic regions. When the spatial localization was extended to carrot vegetative apices and the investigation was carried out with specific probes for top1alpha and top1beta, both transcripts preferentially accumulated in tissues having mitotic activity.


Thrombectomy with or without Bridging Thrombolysis for Anterior Circulation Stroke.
Currently, there is still no clear consensus on bridging thrombolysis (BT) before mechanical thrombectomy (MT). In this study, we aimed to compare clinical and procedural outcomes and complication rates of BT versus direct mechanical thrombectomy (d-MT) in anterior circulation stroke. A total of 359 consecutive anterior circulation stroke patients who received d-MT or BT in our tertiary stroke center between January 2018 and December 2020 were retrospectively analyzed. The patients were divided into two groups as Group d-MT (n = 210) and Group BT (n = 149). The primary outcome was the impact of BT on clinical and procedural outcomes, whereas the secondary outcome was the safety of BT. The incidence of atrial fibrillation was higher in the d-MT group (p = 0.010). The median duration of the procedure was significantly higher in Group d-MT than in Group BT (35 vs 27 min, respectively; P = 0.044). The number of patients achieving good and excellent outcomes was significantly higher in Group BT (p = 0.006 and P = 0.03). The edema/malign infarction rate was higher in the d-MT group (p = 0.003). Successful reperfusion, first-pass effects, symptomatic intra-cranial hemorrhage, and mortality rates were similar between the groups (p > 0.05). In this study, BT seems to yield better clinical and procedural outcomes with lower complication rates than d-MT. These findings may support the additional value of intravenous alteplase in anterior system strokes. Further large-scale, prospective, randomized-controlled studies will clarify the gray lines in this consensus, but this paper is important for reflecting the real-world data in developing countries.


Improved recovery of fatty acid through direct transesterification without prior extraction or purification.
Methods currently in use for the quantitative measurement of fatty acids by gas-liquid chromatography after transesterification are usually lengthy and cumbersome. The technique described is a one-step reaction that is carried out in the same tube and bypasses all the extraction and purification steps. Recoveries of fatty acid and triglyceride standards (C6:0 to C24:1) were better than 96%. When the direct transesterification method was compared to the Folch extraction procedure, increases of fatty acid concentration of 11.4% and 15.8% were observed in human milk and adipose tissue, respectively. The method appears to be particularly advantageous for the recovery of the highly volatile medium chain triglycerides and there is no need to add an antioxidant to protect unsaturated lipids.


Tuberculosis 2004: challenges and opportunities.
Tuberculosis (TB) continues as a major public health challenge worldwide. HIV-TB coinfection is especially concerning as it accelerates progression of infection to active disease and amplifies spread of TB including drug resistant disease. Application of molecular biology and insights from classic microbiology to TB control have resulted in important innovations in diagnosis and treatment. Radiometric assay and, particularly, PCR, with nucleic acid probing, have reduced the time to diagnosis. Moreover, the sensitivity of these techniques is potentially log orders of magnitude more sensitive. Molecular techniques can be adapted to drug susceptibility testing. The differential activity and post-antibiotic effect of various drugs against TB have led to highly effective briefer regimens and to directly observed therapy. Insights into basic host defense against TB and description of the M. tuberculosis genome have created optimism for developing new treatments and effective vaccines in the years to come.


[A case of cryptogenic organizing pneumonia after postoperative irradiation for breast cancer].
The patient was a 69-year-old woman who had undergone breast-conserving surgery of the right breast, followed by tangential irradiation of 50 Gy to the remaining breast. Her subsequent progress was monitored by endocrine therapy with toremifene. Symptoms consisting of cough and slight fever developed 5 months after completion of radiation therapy. A chest X-ray image revealed an infiltrating shadow in the right middle and lower pulmonary fields, and the patient visited our hospital. BOOP was diagnosed by thoracoscopic biopsy. Spontaneous remission was observed during monitoring of the patient's progress. A shadow subsequently reappeared, for which a steroid was administered. Improvement was rapid. This case suggests that when tangential irradiation with or without endocrine therapy is provided, subsequent to breast-conserving treatment, patients should be monitored for the occurrence of BOOP.


[Patient with West Nile fever in the Netherlands].
A 45-year-old patient was admitted with high fever and inadequate behaviour. She had recently stayed in Israel. Extensive investigation suggested a viral infection, but the cause was not established. Her partner suggested the possibility of West Nile virus infection, based upon information he had found on the Internet. Serology was performed and specific IgM and IgG antibodies were demonstrated, but a paired serum sample was not available. The patient recovered spontaneously. The West Nile virus is endemic in Israel, Africa, Asia and the Balkans. It is transmitted by mosquitoes. The incubation time is 1-3 weeks. The disease is characterised by fever, malaise, maculopapular exanthema and lymphadenopathy. Complications are encephalitis, pancreatitis, hepatitis and myocarditis. However, these symptoms are present in less than 5% of all infections. A full-blown encephalitis has a mortality rate of 50%. There is no specific therapy, but prevention by means of controlling the mosquito population is feasible. The diagnosis is made by the detection of West Nile virus-specific IgM antibodies in serum or cerebrospinal fluid. However, in the acute stage viral RNA detection by a polymerase chain reaction in the serum is also possible.


Development of in vitro correlate assays of immunity to infection with Yersinia pestis.
Pneumonic plague is a severe, rapidly progressing disease for which there is no effective vaccine. Since the efficacy of new vaccines cannot be tested in humans, it is essential to develop in vitro surrogate assays that are valid predictors of immunity. The F1 capsule antigen stimulates a protective immune response to most strains of Yersinia pestis. However, strains of Y. pestis that are F1- but still virulent have been isolated, and an in vitro assay, the results which can predict protection against both F1+ and F1- strains, is needed. The virulence antigen (V) is an essential virulence factor of Y. pestis and stimulates protective antibodies. We investigated potential correlates of plague immunity that are based on anti-V antibody-mediated neutralization of Yersinia-induced macrophage cytotoxicity. The neutralizing activity of sera from mice vaccinated with an F1-V fusion candidate vaccine was determined. The decrease in the level of the apoptosis-specific enzyme caspase-3 significantly predicted survival in one- and two-dose vaccination experiments. Sera from F1-V-vaccinated nonhuman primates were evaluated with macrophage assays based on caspase-3 and on other markers manifested at the different stages in cell death. Using murine- and human-derived macrophages in microscopic and fluorescence-activated-cell-sorting-based live/dead staining assays of terminal necrosis, we demonstrated a strong association between in vitro neutralization of macrophage cytotoxicity induced by serum-treated Yersinia and in vivo protection against lethal infection. These results provide a strong base for the development of reliable in vitro correlate bioassays that are predictive of protective immunity to plague.


Single mage gene in the chicken genome encodes CMage, a protein with functional similarities to mammalian type II Mage proteins.
In mammals, the type II melanoma antigen (Mage) protein family is constituted by at least 10 closely related members that are expressed in different tissues, including the nervous system. These proteins are believed to regulate cell cycle withdrawal, neuronal differentiation, and apoptosis. However, the analysis of their specific function has been complicated by functional redundancy. In accordance with previous studies in teleosts and Drosophila, we present evidence that only one mage gene exists in genomes from protists, fungi, plants, nematodes, insects, and nonmammalian vertebrates. We have identified the chicken mage gene and cloned the cDNA encoding the chick Mage protein (CMage). CMage shares close homology with the type II Mage protein family, and, as previously shown for the type II Mage proteins Necdin and Mage-G1, it can interact with the transcription factor E2F-1. CMage is expressed in specific regions of the developing nervous system including the retinal ganglion cell layer, the ventral horn of the spinal cord, and the dorsal root ganglia, coinciding with the expression of the neurotrophin receptor p75 (p75(NTR)) in these regions. We show that the intracellular domain of p75(NTR) can interact with both CMage and Necdin, thus preventing the binding of the latter proteins to the transcription factor E2F-1, and facilitating the proapoptotic activity of E2F-1 in N1E-115 differentiating neurons. The presence of a single mage gene in the chicken genome, together with the close functional resemblance between CMage and Necdin, makes this species ideal to further analyze signal transduction through type II Mage proteins.


Cyclin D3 deficiency inhibits skin tumor development, but does not affect normal keratinocyte proliferation.
Rearrangement and amplification of the D-type cyclin genes have been reported in human cancer. Previous studies have demonstrated that Ras-mediated skin tumorigenesis depends on pathways that act through cyclin D1 and D2; however, the role of cyclin D3 remains unknown. The present study demonstrates that cyclin D3 ablation does not affect keratinocyte proliferation, but instead increases apoptosis levels in the bulge region of the hair follicle. Consequently, cyclin D3 ablation reduces skin papilloma development in a Ras-dependent carcinogenesis model. Previous results revealed that cyclin D3 preferentially binds to cyclin-dependent kinase 6 (CDK6) in mouse keratinocytes and transgenic expression of CDK6 (K5CDK6 mice) inhibits skin tumor development. Thus, we hypothesized that the inhibitory effect of CDK6 is dependent on cyclin D3 expression. To test this hypothesis, a mouse model that overexpresses CDK6 and does not express cyclin D3 (K5CDK6/cyclin D3-/- compound mouse) was developed. Biochemical analysis of the epidermis of K5CDK6/cyclin D3-/- mice revealed that cyclin D3 ablation resulted in increased expression of cyclin D1 protein, with a consequent elevation in the level of CDK6/cyclin D1 and CDK4/cyclin D1 complexes. These findings were concurrent with the increase skin papilloma malignant progression observed in K5CDK6/cyclin D3-/- mice. In summary the absence of cyclin D3 led to fewer number of papillomas in cyclin D3-ablated mice than in the wild-type owing to increased apoptosis, suggesting that alterations in cell survival are a crucial mechanism for crippling cellular defense against neoplasia. The results of the current study also suggest that although cyclin D3 expression does not alter the tumor suppressive role of CDK6 in skin carcinogenesis, the compensatory increase in cyclin D1 can shift the balance towards malignant progression.


A live auxotrophic vaccine confers mucosal immunity and protection against lethal pneumonia caused by Pseudomonas aeruginosa.
Pseudomonas aeruginosa is one of the leading causes of nosocomial pneumonia and its associated mortality. Moreover, extensively drug-resistant high-risk clones are globally widespread, presenting a major challenge to the healthcare systems. Despite this, no vaccine is available against this high-concerning pathogen. Here we tested immunogenicity and protective efficacy of an experimental live vaccine against P. aeruginosa pneumonia, consisting of an auxotrophic strain which lacks the key enzyme involved in D-glutamate biosynthesis, a structural component of the bacterial cell wall. As the amounts of free D-glutamate in vivo are trace substances in most cases, blockage of the cell wall synthesis occurs, compromising the growth of this strain, but not its immunogenic properties. Indeed, when delivered intranasally, this vaccine stimulated production of systemic and mucosal antibodies, induced effector memory, central memory and IL-17A-producing CD4+ T cells, and recruited neutrophils and mononuclear phagocytes into the airway mucosa. A significant improvement in mice survival after lung infection caused by ExoU-producing PAO1 and PA14 strains was observed. Nearly one third of the mice infected with the XDR high-risk clone ST235 were also protected. These findings highlight the potential of this vaccine for the control of acute pneumonia caused by this bacterial pathogen.


Cochlear helix and duct length identification - Evaluation of different curve fitting techniques.
Within the field of cochlear implantation (CIs), the role of utilizing patient-specific cochlear anatomy for choosing the optimal implant electrode is becoming increasingly important. Unfortunately, performing detailed anatomical measurements of a cochlea using clinical imaging data is rather time consuming and hence difficult to implement into the clinical routine. In order to accelerate clinical cochlear anatomy evaluations, previously developed mathematical models can be adjusted to the patient-specific anatomy by measuring just a few overall cochlear dimensions. However, the accuracy of model-based cochlear anatomy estimations is unclear, and incorrect evaluations may lead to false conclusions regarding the suitability of specific implant electrodes. Based on 10 cochleae, an error evaluation of various commonly used curve fitting approaches for cochlear shape and duct length approximation was conducted. Spline tracings of the cochlear contours were used as reference values for the various approximations. Parameterized average cochlear helix models and two of five analytical approaches were found to be suitable for reconstructing the cochlear helical shape and estimating its length. Spline curve reconstructions are the most accurate and reliable method for assessing patient-specific cochlear geometry, especially in the case of anatomical irregularities. The most accurate results within the group of model-based evaluations still resulted in mean overall cochlear length deviations of approximately 5%. Spline curve reconstructions appear to be the best option for anatomical diagnostics in clinical practice. Retrospective studies can be performed to further evaluate model-based evaluations.


Binding of fluorescent and spin-labeled C-terminal hirudin analogs to thrombin.
Synthetic peptides based on the sequence of the negatively charged carboxyl tail of hirudin exhibit anticoagulant activity. Several antithrombin agents are being developed by chemical and structural optimization of these "hirupeptides". The present work demonstrates the design and use of novel spin-labeled and fluorescent-labeled C-terminal hirudin analogs to study the interactions of these antithrombin agents with thrombin in solution. Three labeled hirulabels were synthesized based upon the amino acid sequence of the antithrombin agent MDL 28050, X-NH-(CH2)7-CO-Asp-Tyr-Glu-Pro-Ile-Pro-Glu-Glu-Ala-Cha-D-Glu-OH, where X = anthraniloyl, 1,5-dansyl, or 3-carbamoyl-2,2,5,5-tetramethyl-3-pyrrolin-1-oxyl. The modifications did not significantly alter the potency of these inhibitors which showed Ki values of 100 nM. Their interactions with human and bovine thrombin were studied by ESR and fluorescence techniques. The spin-labeled hirupeptide was able to discern subtle differences in binding to human versus bovine thrombin. The 8-aminooctanoic acid spacer arm placed the nitroxide moieties near the active site, near regions of the autolysis loops which differentiates between human alpha- and gamma-thrombin. It was also able to discern paramagnetic quenching and fluorescence energy transfer interactions, respectively, between covalently attached spin labels and fluorescent probes at the active site Ser 195 and the fluorophore on the hirupeptide.


Preparing for practice: Nursing intern and faculty perceptions on clinical experiences.
Clinical experience and exposure to real patients are required elements of nursing education. Trainees in nursing are expected to be prepared adequately for the hard-working environment, increasing patient complexity, and higher-level competencies. This study investigates differences between nursing interns and clinical faculty on actual and perceived importance of educational preparation and development of clinical competencies, focusing on the nursing curriculum and transition to practice. A convenient sampling technique with a mixed-methods design was used to collect quantitative and qualitative data, by surveying and interviewing nursing interns and faculty members from King Abdul-Aziz University in Saudi Arabia; data collection occurred in December 2015. The survey (23 items) and focused interviews measured perceptions of clinical instruction and experience. Descriptive statistics and t-tests were used to analyze differences in mean ratings between actual and perceived importance. Themes collected from narrative interview data were summarized. Significant differences were found between nursing interns (n = 46) and faculty (n = 29) perceptions of actual clinical teaching and experiences and its importance including the clinical teaching and the development of clinical competence, p < .01. Moreover, nursing interns rated actual experiences of knowledge base and skills significantly lower than faculty perceptions, p = .001. Narrative data provided in-depth information on factors contributing and hindering the learning and teaching environment. Findings from this study call for clinical instruction and experiences to take a step further to meet current practice standards and to improve patient safety in the health professions education of nurses.


Importance of local positive charges on cytochrome f for electron transfer to plastocyanin and potassium ferricyanide.
To study the relationship between the electron transfer rate and the net or local charge of protein, chemically modified cytochrome f, in which positively charged amino groups are replaced with negatively charged carboxyl groups, has been prepared by using an arylating reagent 4-chloro-3,5-dinitrobenzoic acid. Four distinct species of chemically modified cytochrome f, having 1 to 4 mol of modified amino residues per mol of cytochrome f, were separated by preparative polyacrylamide gel electrophoresis. The rate of electron transfer from the reduced singly substituted cytochrome f to the oxidized spinach plastocyanin was only about 50% of that of the native unmodified cytochrome f. The reaction rate further decreased about 50% upon the modification of each amino residue. The biphasic oxidation of cytochrome f by plastocyanin was observed when more than 2 mol of amino residues were modified. The rate of the second phase also decreased with an increasing number of modified amino residues. On the other hand, the oxidation of chemically modified cytochrome f by potassium ferricyanide was clearly monotonic. The rate decreased about 30% upon the modification of each amino residue. The midpoint potentials of chemically modified cytochrome f were almost the same as that of the native protein. These results clearly indicate the importance of local positive charges on cytochrome f, since the overall net charge of cytochrome f is negative at neutral pH. The theory of electrostatic corrected outer-sphere electron transfer of Marcus explained the effect of charge on cytochrome f for the reaction with the small molecule of ferricyanide well, but not the reaction with the protein of plastocyanin.


Parent-Infant Attachment Insecurity and Emotional Eating in Adolescence: Mediation through Emotion Suppression and Alexithymia.
Emotional eating (EE), the propensity to eat in response to emotions, is thought to have its origins in the early parent-infant relationship. This study tested the hypothesis that infant attachment insecurity results in EE in adolescence through the increased use of the emotion regulation strategy suppression of emotions and subsequent alexithymia. At the age of 15 months, parent-infant attachment security (n = 129) was observed with two abbreviated attachment measures: the shortened strange situation procedure (SSSP), and the shortened attachment Q-set (S-AQS). At the age of 12 years, children completed self-report questionnaires to assess the suppression of emotions, alexithymia, and EE. At the age of 16 years, EE was measured again. The mediation models indicated that lower parent-infant attachment security predicted increased use of suppression of emotions, which was related to increased alexithymia, and in turn more EE at the age of 12 years. These results were similar and significant for both attachment measures, and also (marginal) significant with EE at the age of 16 years as an outcome. Lastly, when parental caregiving quality was included, the models with the SSSP as predictor remained significant, but the models with the S-AQS became insignificant. These results indicated that to a certain extent, infant attachment security could predict adolescent EE above and beyond parental caregiving quality.


Changing blood transfusion practice in elective joint arthroplasty: a nursing initiative.
The use of re-infusion drains on 99 consecutive patients undergoing total knee arthroplasty surgery at a large hospital was analysed. As a control group the records of 99 patients treated without re-infusion were analysed retrospectively. The primary aim was to ascertain the cost effectiveness of the drains. Secondary aims were to assess safety of the drains, whether or not they reduced the need for allogeneic blood transfusion and whether they decreased the length of stay in hospital. The direct cost of consumables increased for the evaluation period. There was a smaller proportion of allogeneic blood transfusion (27% vs 38%) and a smaller mean number of units transfused (0.92 vs 0.54) in the re-infusion group compared to the control group. Patients benefited directly in that the mean length of stay was also significantly shorter in the re-infusion group. We anticipate more direct cost saving with experience and best practice and conclude that the use of re-infusion drains is a cost effective blood saving method in total knee joint arthroplasty.


Retroposed elements as archives for the evolutionary history of placental mammals.
Reconstruction of the placental mammalian (eutherian) evolutionary tree has undergone diverse revisions, and numerous aspects remain hotly debated. Initial hierarchical divisions based on morphology contained many misgroupings due to features that evolved independently by similar selection processes. Molecular analyses corrected many of these misgroupings and the superordinal hierarchy of placental mammals was recently assembled into four clades. However, long or rapid evolutionary periods, as well as directional mutation pressure, can produce molecular homoplasies, similar characteristics lacking common ancestors. Retroposed elements, by contrast, integrate randomly into genomes with negligible probabilities of the same element integrating independently into orthologous positions in different species. Thus, presence/absence analyses of these elements are a superior strategy for molecular systematics. By computationally scanning more than 160,000 chromosomal loci and judiciously selecting from only phylogenetically informative retroposons for experimental high-throughput PCR applications, we recovered 28 clear, independent monophyly markers that conclusively verify the earliest divergences in placental mammalian evolution. Using tests that take into account ancestral polymorphisms, multiple long interspersed elements and long terminal repeat element insertions provide highly significant evidence for the monophyletic clades Boreotheria (synonymous with Boreoeutheria), Supraprimates (synonymous with Euarchontoglires), and Laurasiatheria. More importantly, two retropositions provide new support for a prior scenario of early mammalian evolution that places the basal placental divergence between Xenarthra and Epitheria, the latter comprising all remaining placentals. Due to its virtually homoplasy-free nature, the analysis of retroposon presence/absence patterns avoids the pitfalls of other molecular methodologies and provides a rapid, unequivocal means for revealing the evolutionary history of organisms.


[Research advances in molecular genetics of epilepsies].
Last years brought a large number of papers shedding light on the molecular and genetic background of epilepsy. The aim of the present work was to review recent literature concerning this issue. In the first section, the authors presented biochemical and functional characteristics of receptors and ion channels. Mutations in their genes can lead to epileptic seizure development. Particularly, the role of GABA receptors as well as voltage-gated sodium, calcium and potassium channels in epilepsy pathogenesis was discussed. The second part of the work focuses on clinical implications of these receptors and channels disturbances. Epileptic syndromes with proved genetic defect are presented on the basis of recent literature and OMIM (Online Mendelian Inheritance in Man) database. The authors point out that reviewed results show vast heterogeneity of many known epileptic syndromes. On the other hand, defects in the same locus can result in heterogeneous phenotypes. This can indicate multifactorial pathogenesis of epilepsy and implicate further revision of epilepsies and epileptic seizures classification.


Macrophages and Stem Cells-Two to Tango for Tissue Repair?
Macrophages (MCs) are present in all tissues, not only supporting homeostasis, but also playing an important role in organogenesis, post-injury regeneration, and diseases. They are a heterogeneous cell population due to their origin, tissue specificity, and polarization in response to aggression factors, depending on environmental cues. Thus, as pro-inflammatory M1 phagocytic MCs, they contribute to tissue damage and even fibrosis, but the anti-inflammatory M2 phenotype participates in repairing processes and wound healing through a molecular interplay with most cells in adult stem cell niches. In this review, we emphasize MC phenotypic heterogeneity in health and disease, highlighting their systemic and systematic contribution to tissue homeostasis and repair. Unraveling the intervention of both resident and migrated MCs on the behavior of stem cells and the regulation of the stem cell niche is crucial for opening new perspectives for novel therapeutic strategies in different diseases.


[Alcoholism in the elderly.].
The author discusses the problem of alcoholism in the elderly. In the first part he describes the general situation of old people and the problems they most frequently encounter. He puts forward the hypothesis that the specific problems related to old age are a determining factor in the on-set or aggravation of an alcool abuse problem. In the second part he presents a brief survey of the literature on the subject and mentions the difficulties relative to a request for help by an elderly person grappling with an alcool abuse problem. Finally, he examines the nature of the treatment to be provide to such a person.


Gene-specific random mutagenesis of Escherichia coli in vivo: isolation of temperature-sensitive mutations in the acyl carrier protein of fatty acid synthesis.
Acyl carrier proteins (ACPs) are very small acidic proteins that play a key role in fatty acid and complex lipid synthesis. Moreover, recent data indicate that the acyl carrier protein of Escherichia coli has a large protein interaction network that extends beyond lipid synthesis. Despite extensive efforts over many years, no temperature-sensitive mutants with mutations in the structural gene (acpP) that encodes ACP have been isolated. We report the isolation of three such mutants by a new approach that utilizes error-prone PCR mutagenesis, overlap extension PCR, and phage lambda Red-mediated homologous recombination and that should be generally applicable. These mutants plus other experiments demonstrate that ACP function is essential for the growth of E. coli. Each of the mutants was efficiently modified with the phosphopantetheinyl moiety essential for the function of ACP in lipid synthesis, and thus lack of function at the nonpermissive temperature cannot be attributed to a lack of prosthetic group attachment. All of the mutant proteins were largely stable at the nonpermissive temperature except the A68T/N73D mutant protein. Fatty acid synthesis in strains that carried the D38V or A68T/N73D mutations was inhibited upon a shift to the nonpermissive temperature and in the latter case declined to a small percentage of the rate of the wild-type strain.


The Association of Loneliness and Non-prescribed Opioid Use in Patients With Opioid Use Disorder.
To investigate the relationship between loneliness and non-prescribed opioid use in patients diagnosed with opioid use disorder (OUD). This was a cohort study conducted at a federally qualified health center (FQHC) in New Haven, CT. Patients who were treated for OUD by health center providers and prescribed buprenorphine were eligible. Participants were asked to complete the UCLA-Loneliness Scale Version 3. From the electronic medical record, we collected patient demographic and clinical characteristics as well as the results of biological fluid testing obtained throughout their treatment course since entry into care. Multivariable logistic regression was performed to identify independent predictors of the detection of non-prescribed opioids within biological fluid. Of the 82 patients enrolled in the study, 33 (40.3%) of the participants had at least 1 biological fluid test positive for non-prescribed opioids after maintenance onto buprenorphine treatment. A higher loneliness score was associated with increased odds of non-prescribed opioids (adjusted odds ratio 1.16; 95% confidence interval 1.06-1.27). Patient age, the number of problems on the problems list, and cocaine use were also positively associated with the presence of non-prescribed opioids whereas having diabetes was negatively associated. Among the individuals being treated with buprenorphine for OUD, loneliness may be a risk factor for the use of non-prescribed opioids or treatment failure.


Risk factors of severe hand, foot and mouth disease complicated with cardiopulmonary collapse.
The aim of this study was to investigate the risk factors for severe hand, foot and mouth disease (HFMD) complicated with cardiopulmonary collapse in children. In total, 176 children aged 6-45 months with severe HFMD from March 2013 to May 2014 were enrolled in the study and divided into two groups, one with cardiopulmonary collapse and the other without. Univariate analysis and multiple logistic regression analysis were used to analyze the risk factors for severe HFMD complicated with cardiopulmonary collapse. The univariate analysis showed that there was no significant correlation between age, body temperature, consciousness disorders, blood glucose level and severe HFMD complicated with cardiopulmonary collapse. The multiple logistic regression analysis demonstrated that vomiting, circulatory disturbance, enterovirus 71 infection, dysfunction of respiratory rhythm and high level of brain natriuretic peptide were five independent risk factors for severe HFMD complicated with cardiopulmonary collapse. Children with HFMD and the above five risk factors may be at risk for cardiopulmonary collapse and poor prognosis.


Elucidation of causal direction between asthma and obesity: a bi-directional Mendelian randomization study.
Observational associations between asthma and obesity are well established, but inferring causality is challenging. We leveraged publicly available summary statistics to ascertain the causal direction between asthma and obesity via Mendelian randomization in European-ancestry adults. We performed two-sample bi-directional Mendelian randomization analysis using publicly available genome-wide association studies summary statistics. Single nucleotide polymorphisms associated with asthma and body mass index at genome-wide significance were combined using a fixed effect meta-analysis in each direction. An extensive sensitivity analysis was considered. There was evidence in support of increasing causal effect of body mass index on risk of asthma (odds ratio 1.18 per unit increase, 95% confidence interval (CI) (1.11, 1.25), P = 2 × 10-8. No significant causal effect of asthma on adult body mass index was observed [estimate -0.004, 95% CI (-0.018, 0.009), P = 0.553]. Our results confirmed that in European-ancestry populations, adult body mass index is likely to be causally linked to the risk of asthma; yet the effect of asthma on body mass index is small, if present at all.


First-passage and first-exit times of a Bessel-like stochastic process.
We study a stochastic process X(t) which is a particular case of the Rayleigh process and whose square is the Bessel process, with various applications in physics, chemistry, biology, economics, finance, and other fields. The stochastic differential equation is dX(t)=(nD/X(t))dt+√(2D)dW(t), where W(t) is the Wiener process. The drift term can arise from a logarithmic potential or from taking X(t) as the norm of a multidimensional random walk. Due to the singularity of the drift term for X(t)=0, different natures of boundary at the origin arise depending on the real parameter n: entrance, exit, and regular. For each of them we calculate analytically and numerically the probability density functions of first-passage times or first-exit times. Nontrivial behavior is observed in the case of a regular boundary.


Malignant Pleural Effusion As the Initial Presentation of Renal Cell Carcinoma: A Case Report and Literature Review.
Renal cell carcinoma is the most common renal neoplasm. Its presentation is often very occult, and it may be discovered incidentally. It may present with the classic symptoms of back pain, flank pain, hematuria, or hypertension. Renal cell carcinoma may also present with malignant pleural effusion at diagnosis; however, it is very rare. In this case report and literature review, we describe a 77-year-old male who was diagnosed with renal cell carcinoma after presenting with a malignant pleural effusion - an extremely rare phenomenon. An analysis of the literature yielded 13 case reports, including ours, where the diagnostic presentation of renal cell carcinoma was a malignant pleural effusion. Our patient presented with left-sided chest pain. Imaging was suggestive of pleural effusion. CT and MRI imaging demonstrated masses in the upper and lower poles of the right kidney suggestive of renal cell carcinoma. CT imaging also showed lung nodules that were suggestive of pulmonary metastases. Biopsy and immunostaining of pleural tissue were positive for clear cell renal cell carcinoma. Therapeutic thoracentesis was performed. Despite this, the patient developed recurrent large-volume pleural effusions requiring drainage and placement of a pleural catheter. Our patient's extremely rare presentation of malignant pleural effusion as the diagnostic presentation of renal cell carcinoma along with recurrent, large-volume effusions requiring drainage has only been reported in the form of case reports in the literature.


A New Frontier in Studying Dietary Phytochemicals in Cancer and in Health: Metabolic and Epigenetic Reprogramming.
Metabolic rewiring and epigenetic reprogramming are closely inter-related, and mutually regulate each other to control cell growth in cancer initiation, promotion, progression, and metastasis. Epigenetics plays a crucial role in regulating normal cellular functions as well as pathological conditions in many diseases, including cancer. Conversely, certain mitochondrial metabolites are considered as essential cofactors and regulators of epigenetic mechanisms. Furthermore, dysregulation of metabolism promotes tumor cell growth and reprograms the cells to produce metabolites and bioenergy needed to support cancer cell proliferation. Hence, metabolic reprogramming which alters the metabolites/epigenetic cofactors, would drive the epigenetic landscape, including DNA methylation and histone modification, that could lead to cancer initiation, promotion, and progression. Recognizing the diverse array of benefits of phytochemicals, they are gaining increasing interest in cancer interception and treatment. One of the significant mechanisms of cancer interception and treatment by phytochemicals is reprogramming of the key metabolic pathways and remodeling of cancer epigenetics. This review focuses on the metabolic remodeling and epigenetics reprogramming in cancer and investigates the potential mechanisms by which phytochemicals can mitigate cancer.


Steady-state expression of self-regulated genes.
Regulatory gene networks contain generic modules such as feedback loops that are essential for the regulation of many biological functions. The study of the stochastic mechanisms of gene regulation is instrumental for the understanding of how cells maintain their expression at levels commensurate with their biological role, as well as to engineer gene expression switches of appropriate behavior. The lack of precise knowledge on the steady-state distribution of gene expression requires the use of Gillespie algorithms and Monte-Carlo approximations. In this study, we provide new exact formulas and efficient numerical algorithms for computing/modeling the steady-state of a class of self-regulated genes, and we use it to model/compute the stochastic expression of a gene of interest in an engineered network introduced in mammalian cells. The behavior of the genetic network is then analyzed experimentally in living cells. Stochastic models often reveal counter-intuitive experimental behaviors, and we find that this genetic architecture displays a unimodal behavior in mammalian cells, which was unexpected given its known bimodal response in unicellular organisms. We provide a molecular rationale for this behavior, and we implement it in the mathematical picture to explain the experimental results obtained from this network.


Effect of a clinical evidence technology on patient skin disease outcomes in primary care: a cluster-randomized controlled trial.
Providers' use of clinical evidence technologies (CETs) improves their diagnosis and treatment decisions. Despite these benefits, few studies have evaluated the impact of CETs on patient outcomes. The investigators evaluated the effect of one CET, VisualDx, on skin problem outcomes in primary care. A cluster-randomized controlled pragmatic trial was conducted in outpatient clinics at an academic medical center in the northeastern United States. Participants were primary care providers (PCPs) and their adult patients seen for skin problems. The intervention was VisualDx, as used by PCPs. Outcomes were patient-reported time from index clinic visit to problem resolution, and the number of follow-up visits to any provider for the same problem. PCPs who were randomly assigned to the intervention agreed to use VisualDx as their primary evidence source for skin problems. Control group PCPs agreed not to use VisualDx. Investigators collected outcome data from patients by phone at thirty-day intervals. Cox proportional hazards models assessed time to resolution. Wilcoxon-rank sum tests and logistic regression compared the need for return appointments. Thirty-two PCPs and 433 patients participated. In proportional hazards modelling adjusted for provider clusters, the time from index visit to skin problem resolution was similar in both groups (hazard ratio=0.92; 95% confidence interval [CI]=0.70, 1.21; p=0.54). Patient follow-up appointments did not differ significantly between groups (odds ratio=1.26; CI=0.94, 1.70; p=0.29). This pragmatic trial tested the effectiveness of VisualDx on patient-reported skin disease outcomes in a generalizable clinical setting. There was no difference in skin problem resolution or number of follow-up visits when PCPs used VisualDx.


Trifluoroethanol stabilizes a helix-turn-helix motif in equine infectious-anemia-virus trans-activator protein.
The solution structure of the 75-amino-acid trans-activator (Tat) protein of the equine infectious-anemia virus in trifluoroethanol-containing solution was determined by two-dimensional and three-dimensional nuclear magnetic resonance spectroscopy, resulting in a total of 838 nuclear-Over-hauser-enhancement distance restraints, and restrained molecular-dynamics simulations. In contrast to the recently determined structure of this protein in trifluoroethanol-free pH 6.3 solution, the hydrophobic core and the adjacent basic RNA-binding region of the protein showed well-defined alpha-helical secondary structure in trifluoroethanol-containing solution. The helical regions comprise those parts of the molecule whose helix-forming tendencies were noted earlier in trifluoroethanol-free solution. Two helices (Gln38-Arg43 and Asp48-Ala64) are connected by a tight type-II turn centered at the strictly conserved Gly46 leading to a helix-turn-helix motif in the core and basic region of the protein. A third helix (Thr9-Asn13) is located in the less well defined N-terminal part of the protein. These observations may support the notion that the protein adopts a helical structure in the RNA-binding region on complex formation. Although the secondary-structure elements become better defined in trifluoroethanol-containing solution, the opposite is true for the hydrophobically stabilized tertiary structure. This adds a caveat to studies of protein structures in trifluoroethanol-containing solution in general.


Human cryptochrome exhibits light-dependent magnetosensitivity.
Humans are not believed to have a magnetic sense, even though many animals use the Earth's magnetic field for orientation and navigation. One model of magnetosensing in animals proposes that geomagnetic fields are perceived by light-sensitive chemical reactions involving the flavoprotein cryptochrome (CRY). Here we show using a transgenic approach that human CRY2, which is heavily expressed in the retina, can function as a magnetosensor in the magnetoreception system of Drosophila and that it does so in a light-dependent manner. The results show that human CRY2 has the molecular capability to function as a light-sensitive magnetosensor and reopen an area of sensory biology that is ready for further exploration in humans.


Effect of conceptus removal and intrauterine administration of conceptus tissue on luteal function in the cow.
Two experiments were conducted to determine the day on which the bovine embryo first affects luteal function. Experiment I was to determine the sole effect of flushing the uterine lumen and involved 12 non-inseminated heifers flushed 9, 14 and 16 days after estrus (4 animals per group). The same procedure was used to remove conceptuses from 15 pregnant heifers on days 9 (n = 4), 14 (n = 5) and 16 (n = 7) after the onset of estrus. Progesterone concentrations were measured daily throughout the cycle corresponding to conceptus removal (or flushing alone) and throughout the preceding cycle. In non-inseminated animals and when the conceptuses were removed on days 9 or 14, neither the progesterone pattern nor the inter-estrous interval was altered. By contrast, when the embryo was removed on day 16, the time of luteolysis was delayed by 4 to 7 days and the heifers were observed to be in estrus 26 to 29 days after AI. In experiment 2, 21 virgin heifers were distributed over four treatment groups. Animals in groups 2 to 4 received the following treatments twice a day between day 15 and day 19 of the estrous cycle in the horn ipsilateral to the corpus luteum: group 2 (n = 3), 0,25 ml saline; group 3 (n = 3), two 12-day-old conceptuses; group 4 (n = 5), one 16-day-old conceptus. progesterone concentrations were determined from day 13 of the cycle to 4 days after the following estrus. No antiluteolytic effect was found with intrauterine administration of either saline alone or day-12 embryos. In contrast, administration of day-16 conceptuses through the cervix (group 4) lengthened the estrous cycle by 6 to 7 days. The results of these experiments suggest that the antiluteolytic and/or luteotropic factor(s) originating from the conceptus is fully potent by day 16. Embryonic mortality occurring on day 16 or later may induce an extension of corpus luteum lifespan.


PepT1-mediated fMLP transport induces intestinal inflammation in vivo.
In the present study, the effect of H(+)/peptide transporter (PepT1)-mediated N-formylmethionyl-leucyl-phenylalanine (fMLP) transport on inflammation in vivo in the rat small intestine, which expresses high PepT1 levels, and in the rat colon, which does not express PepT1, were investigated using myeloperoxidase (MPO) activity and histological analysis. We found that 10 microM fMLP perfusion in the jejunum for 4 h significantly increased MPO activity and altered the architecture of jejunal villi. In contrast, 10 microM fMLP perfusion in the colon for 4 h did not induce any inflammation. In addition, we have shown that 50 mM Gly-Gly alone did not affect basal MPO activity but completely inhibited the MPO activity induced by 10 microM fMLP in the jejunum. Together, these experiments demonstrate that 1) the differential expression of PepT1 between the small intestine and the colon plays an important role in epithelial-neutrophil interactions and 2) the inhibition of fMLP uptake by jejunal epithelial cells (expressing PepT1) reduces the neutrophil ability to move across the epithelium, in agreement with our previously published in vitro study. This report constitutes the first in vivo study showing the implication of a membrane transporter (PepT1) in intestinal inflammation.


Optimal Paths for Variants of the 2D and 3D Reeds-Shepp Car with Applications in Image Analysis.
We present a PDE-based approach for finding optimal paths for the Reeds-Shepp car. In our model we minimize a (data-driven) functional involving both curvature and length penalization, with several generalizations. Our approach encompasses the two- and three-dimensional variants of this model, state-dependent costs, and moreover, the possibility of removing the reverse gear of the vehicle. We prove both global and local controllability results of the models. Via eikonal equations on the manifold R d × S d - 1 we compute distance maps w.r.t. highly anisotropic Finsler metrics, which approximate the singular (quasi)-distances underlying the model. This is achieved using a fast-marching (FM) method, building on Mirebeau (Numer Math 126(3):515-557, 2013; SIAM J Numer Anal 52(4):1573-1599, 2014). The FM method is based on specific discretization stencils which are adapted to the preferred directions of the Finsler metric and obey a generalized acuteness property. The shortest paths can be found with a gradient descent method on the distance map, which we formalize in a theorem. We justify the use of our approximating metrics by proving convergence results. Our curve optimization model in R d × S d - 1 with data-driven cost allows to extract complex tubular structures from medical images, e.g., crossings, and incomplete data due to occlusions or low contrast. Our work extends the results of Sanguinetti et al. (Progress in Pattern Recognition, Image Analysis, Computer Vision, and Applications LNCS 9423, 2015) on numerical sub-Riemannian eikonal equations and the Reeds-Shepp car to 3D, with comparisons to exact solutions by Duits et al. (J Dyn Control Syst 22(4):771-805, 2016). Numerical experiments show the high potential of our method in two applications: vessel tracking in retinal images for the case d = 2 and brain connectivity measures from diffusion-weighted MRI data for the case d = 3 , extending the work of Bekkers et al. (SIAM J Imaging Sci 8(4):2740-2770, 2015). We demonstrate how the new model without reverse gear better handles bifurcations.


Low Expression of BRCA1 as a Potential Relapse Predictor in B-Cell Acute Lymphoblastic Leukemia.
B-cell acute lymphoblastic leukemia (B-ALL) is the most common childhood hematological malignancy worldwide. Treatment outcomes have improved dramatically in recent years; despite this, relapse is still a problem, and the potential molecular explanation for this remains an important field of study. We performed microarray and single-cell RNA-Seq data mining, and we selected significant data with a P -value<0.05. We validated BRCA1 gene expression by means of quantitative (reverse transcription-polymerase chain reaction.) We performed statistical analysis and considered a P -value<0.05 significant. We identified the overexpression of breast cancer 1, early onset (BRCA1; P -value=2.52 -134 ), by means of microarray analysis. Moreover, the normal distribution of BRCA1 expression in healthy bone marrow. In addition, we confirmed the increases in BRCA1 expression using real-time (reverse transcription-polymerase chain reaction and determined that it was significantly reduced in patients with relapse ( P -values=0.026). Finally, we identified that the expression of the BRCA1 gene could predict early relapse ( P -values=0.01). We determined that low expression of BRCA1 was associated with B-cell acute lymphoblastic leukemia relapse and could be a potential molecular prognostic marker.


Anion-π interaction inside the polyanionic Mo132O372 cage with hydrophobic inner space.
In this paper, we provide experimental evidence to indicate that the polyanionic Mo132O372 cage with a hydrophobic inner nanospace has a unique capability to participate in anion-π interactions by showing a preference for electron-deficient mono-substituted benzenes over non-electron-deficient guests in inclusion.


High concordance of karyotype analysis and RT-PCR for CBF beta/MYH11 in unselected patients with acute myeloid leukemia. A single center study.
Identification of the inversion 16 in patients with acute myeloid leukemia (AML) is of great practical value since these patients have a relatively favorable prognosis, especially when treated with high-dose cytarabine. We compared the results of cytogenetic analysis and reverse transcriptase-polymerase chain reaction (RT-PCR) for core binding factor (CBF) beta/myosin heavy chain (MYH11) in 241 unselected cases of AML. In contrast with other studies, we found a high concordance between these 2 methods. Eighteen of 241 patients showed a cytogenetic anomaly of the chromosome 16. We detected the fusion transcript by RT-PCR in all 18 cases and in 2 additional patients with AML without any cytogenetic anomaly of chromosome 16. One patient had a normal diploid karyotype, and the second patient showed a trisomy 22 in karyotype analysis, which often is associated with inv(16). Only 8 of 20 CBF beta/MYH11-positive patients had M4Eo morphologic features. The much higher discrepancy between cytogenetic analysis and RT-PCR in other studies, especially in AMLs other than M4Eo, possibly indicates the necessity for PCR screening regardless of the French-American-British classification.


The Akt/FoxO1/p27 pathway mediates the proliferative action of liraglutide in β cells.
Numerous studies have shown that liraglutide, a modified form of human glucagon-like peptide-1 (GLP-1), increases β-cell mass. However, the underlying molecular mechanisms remain unclear. In the present study, we investigated the role of Akt/FoxO1/p27 signaling in liraglutide-induced β-cell proliferation. INS-1 rat insulinoma cells were exposed to two different concentrations of liraglutide. MTT assay was performed to evaluate β-cell proliferation. The expression of Akt/FoxO1/p27 was detected by quantitative real-time PCR and Western blotting. The results revealed that in comparison to the non-treatment group, stimulating INS-1 cells with 10 and 100 nM liraglutide caused β-cell proliferation to be significantly enhanced. The mRNA levels of p27 in INS-1 cells declined upon treatment with liraglutide compared to the non-treatment group. Western blot analysis revealed that the phosphorylation of Akt and FoxO1 was markedly elevated following exposure to liraglutide. Moreover, LY294002, a phosphatidylinositol 3-kinase (PI-3K) inhibitor, significantly abrogated liraglutide-induced effects. Therefore, we conclude that liraglutide increased the β-cell mass by upregulating β-cell proliferation and that the proliferative action of liraglutide in β cells was mediated by activation of PI-3K/Akt, which resulted in inactivation of FoxO1 and decreased p27.


Response Surface Optimisation for the Production of Antioxidant Hydrolysates from Stone Fish Protein Using Bromelain.
Protein hydrolysates produced from different food sources exhibit therapeutic potential and can be used in the management of chronic diseases. This study was targeted to optimise the conditions for the hydrolysis of stone fish protein to produce antioxidant hydrolysates using central composite design (CCD) by response surface methodology (RSM). The stone fish protein was hydrolysed under the optimum predicted conditions defined by pH (6.5), temperature (54°C), E/S ratio (1.5%), and hydrolysis time (360 min). The hydrolysates were then evaluated for 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) scavenging activity and ferrous ion- (Fe2+-) chelating activity. Results validation showed no significant difference between the experimental values of DPPH• scavenging activity (48.94%) and Fe2+-chelating activity (25.12%) obtained at 54.62% degree of hydrolysis (DH) compared to their corresponding predicted values of 49.79% and 24.08% at 53.08% DH, respectively. The hydrolysates demonstrated non-Newtonian behavior (n < 1) with stronger shear-thinning effect and higher viscosities at increasing concentration. Thus, RSM can be considered as a promising strategy to optimise the production of stone fish protein hydrolysates containing antioxidant peptides. It is hoped that this finding will enhance the potential of stone fish protein hydrolysates (SHs) as therapeutic bioactive ingredient in functional foods development.


Consumption of dental services by ethnic groups in Israel.
This study analyzes the consumption of dental services in Israel and is based on data collected on households and their expenditures on dental health. Medical and social research in Israel generally take into account the significance of the ethnic dimenison to explain findings. In accordance with this established research mode, interview subjects were divided into two groups: households with both parents born in Asian or African countries, and others. Analysis of the data established a difference between the two groups in their expenditures on dental services. While several studies focused primarily on the sociologic aspects of the problem, little attempt has been made to isolate the influence of economic factors on dental care. This study demonstrates that income and other socioeconomic variables characterizing the various ethnic groups are the underlying causes for the difference in dental care expenditure.


Ultrafast Symmetry-Breaking Charge Separation in a Perylene Bisimide Dimer Enabled by Vibronic Coupling and Breakdown of Adiabaticity.
Perylene bisimides (PBIs) have received great attention in their applicability to optoelectronics. Especially, symmetry-breaking charge separation (SB-CS) in PBIs has been investigated to mimic the efficient light capturing and charge generation in natural light-harvesting systems. However, unlike ultrafast CS dynamics in donor-acceptor heterojunction materials, ultrafast SB-CS in a stacked homodimer has still been challenging due to excimer formation in the absence of rigidifying surroundings such as a special pair in the natural systems. Herein, we present the detailed mechanism of ultrafast photoinduced SB-CS occurring in a 1,7-bis(N-pyrrolidinyl) PBI dimer within a cyclophane. Through narrow-band and broad-band transient absorption spectroscopy, we demonstrate that ultrafast SB-CS in the dimer is enabled by the combination of (1) vibrationally coherent charge-transfer resonance-enhanced excimer formation and (2) breakdown of adiabaticity (formation of SB-CS diabats) in the excimer state via structural and solvent fluctuation. Quantum chemical calculations also underpin that the participation of strong electron-donating substituents in overall vibrational modes plays a crucial role in triggering the ultrafast SB-CS. Therefore, our work provides an alternative route to facilitate ultrafast SB-CS in PBIs and thereby establishes a novel strategy for the design of optoelectronic materials.


A community development approach to deal with public drug use in Box Hill.
The use of alcohol and other drugs in public space is one that generates much heat in the public discourse and in the media. Too often the responses called for to reduce the problems of public amenity involve punitive policing and other responses that aim to engineer (mostly) young people out of these public spaces. Often local retailers are a key stakeholder group calling loudest for punitive action. In this Harm Reduction Digest Rogers and Anderson describe a community development approach taken to address these problems in Box Hill in the City of Whitehorse, near Melbourne. This approach which aimed to develop 'bridging social capital' between community retailers and other stakeholders in the area appears to have been effective in reducing harm associated with public drug use. Moreover these changes have become institutionalised and the approach has been expanded to address other public amenity problems in the area. It is a very nice example of how drug related harm can be reduced by grass roots networks of local councils, business people, law enforcement and health and welfare service providers to address these issues.


A systems view and lessons from the ongoing Ebola Virus Disease (EVD) outbreak in West Africa.
This article analyses the on-going (2014) Ebola Virus Disease (EVD) outbreak in West Africa from a systems perspective; and draws out lessons for West Africa in general and Ghana in particular.


Glutamate uptake in synaptic plasticity: from mollusc to mammal.
A great deal of research has been directed toward understanding the cellular mechanisms underlying synaptic plasticity and memory formation. To this point, most research has focused on the more "active" components of synaptic transmission: presynaptic transmitter release and postsynaptic transmitter receptors. Little work has been done characterizing the role neurotransmitter transporters might play during changes in synaptic efficacy. We review several new experiments that demonstrate glutamate transporters are regulated during changes in the efficacy of glutamatergic synapses. This regulation occurred during long-term facilitation of the sensorimotor synapse of Aplysia and long-term potentiation of the Schaffer-collateral synapse of the rat. We propose that glutamate transporters are "co-regulated" with other molecules/processes involved in synaptic plasticity, and that this process is phylogenetically conserved. These new findings indicate that glutamate transporters most likely play a more active role in neurotransmission than previously believed.


Molecular cloning of cDNA for double-stranded RNA adenosine deaminase, a candidate enzyme for nuclear RNA editing.
We have cloned human cDNA encoding double-stranded RNA adenosine deaminase (DRADA). DRADA is a ubiquitous nuclear enzyme that converts multiple adenosines to inosines in double-helical RNA substrates without apparent sequence specificity. The A --> I conversion activity of the protein encoded by the cloned cDNA was confirmed by recombinant expression in insect cells. Use of the cloned DNA as a molecular probe documented sequence conservation across mammals and detected a single transcript of 7 kb in RNA of all human tissues analyzed. The deduced primary structure of human DRADA revealed a bipartite nuclear localization signal, three repeats of a double-stranded RNA binding motif, and the presence of sequences conserved in the catalytic center of other deaminases, including a cytidine deaminase involved in the RNA editing of apolipoprotein B. These structural properties are consistent with the enzymatic signature of DRADA, and strengthen the hypothesis that DRADA carries out the RNA editing of transcripts encoding glutamate-gated ion channels in brain.


Two-dimensional electronic transport and surface electron accumulation in MoS2.
Because the surface-to-volume ratio of quasi-two-dimensional materials is extremely high, understanding their surface characteristics is crucial for practically controlling their intrinsic properties and fabricating p-type and n-type layered semiconductors. Van der Waals crystals are expected to have an inert surface because of the absence of dangling bonds. However, here we show that the surface of high-quality synthesized molybdenum disulfide (MoS2) is a major n-doping source. The surface electron concentration of MoS2 is nearly four orders of magnitude higher than that of its inner bulk. Substantial thickness-dependent conductivity in MoS2 nanoflakes was observed. The transfer length method suggested the current transport in MoS2 following a two-dimensional behavior rather than the conventional three-dimensional mode. Scanning tunneling microscopy and angle-resolved photoemission spectroscopy measurements confirmed the presence of surface electron accumulation in this layered material. Notably, the in situ-cleaved surface exhibited a nearly intrinsic state without electron accumulation.


A modified PAP (peroxidase-anti-peroxidase) staining technique using sera from patients with dengue hemorrhagic fever (DHF): 4 step PAP staining technique.
BHK-21 cells infected with dengue virus type 1 were stained by a newly developed 4 step PAP (peroxidase-anti-peroxidase) technique using sera from patients with dengue hemorrhagic fever as anti-virus antibody. The intensity of staining of the sera was proportional to the hemagglutination inhibition and neutralization titers. With this new technique using sera from patients it should be possible to use the PAP technique of virus infections.


Identification and characterization of extrachromosomal circular DNA in alcohol induced osteonecrosis of femoral head.
Alcohol-induced osteonecrosis of the femoral head (AIONFH) is a complicated refractory bone disease seen in the clinic. The pathogenesis of AIONFH is still controversial. Extrachromosomal circular DNA (eccDNA) elements have been indicated ubiquitously exist in eukaryotic genomes. However, the characteristics and biological functions of eccDNAs remain unclear in AIONFH. In this study, eccDNAs from AIONFH samples (n = 7) and fracture of femoral neck samples as a control (n = 7) were purified by removing linear DNA and rolling circle amplification. High-throughput sequencing and bioinformatics analysis were performed to study the characterization and biofunction of eccDNAs. We identified more than 600,000 unique eccDNAs. The number of detected eccDNAs in AIONFH was less than that in the control, and eccDNA formation may be related to transcription or other characteristics of coding genes. The eccDNA lengths are mainly distributed between 0.1 kb and 1 kb, with a major peak in 0.358 kb. The bioinformatic analysis showed that 25 significant genes were detected, including MAP3K1, ADCY1, CACNA1S, and MACF1, which contributed to regulating bone formation. GO and KEGG analyses suggested that the related genes derived from exons mainly affected metabolic processes and signal transduction, and bone metabolism-related pathways, such as the MAPK pathway and TGF-β pathway, were enriched. EccDNAs in AIONFH are common and may play an important role in pathogenesis by regulating bone metabolism.


Assessment of the Quality of Life of Women after Osteoporotic Vertebral Fracture with Consideration of Socio-Demographic Characteristics and Selected Factors Concerning the State of Health.
Introduction: Fractures of the vertebral bodies are a frequent complication of osteoporosis, hospitalization, decline in physical fitness and, in consequence, deterioration in the quality of life. Objective: The aim of the study was assessment of the quality of life according to the QUALEFFO-41 questionnaire in patients who had undergone fractures of the vertebral bodies, and presentation of the relationships between the quality of life, socio-demographic characteristics, and selected factors concerning the state of health. Materials and Method: The study included 243 women with osteoporotic vertebral fractures, and was conducted in the Outpatient Departments for the Treatment of Osteoporosis in the city of Lublin (eastern Poland). For the purposes of the study, the Quality of Life Questionnaire (QUALEFFO-41) and the author's questionnaire were employed, and Spearman's rank correlation coefficient, t-Student test, and Tukey test were used, along with analysis of variance (ANOVA). The level of statistical significance was set at α = 0.05. Results: The quality of life of woman with vertebral compression fractures remains on a mediocre level. Significant relationships were observed between the respondents' quality of life and certain socio-demographic characteristics, duration of the disease, and complaints related with osteoporosis. Conclusions: It is important to implement appropriate therapy and provide comprehensive, holistic care to women after fractures.


Did adolescents in Norway respond to the elimination of copayments for general practitioner services?
Copayments for primary care services may lead to decreased access to and underconsumption of necessary health care for vulnerable patient groups, such as adolescents. In Norway, in 2010, adolescents aged 12 to 15 years were exempted from copayments for general practitioner (GP) services, and the aim of this study is to estimate whether being exempted from copayments led to increases in GP visits. We apply the synthetic control method using the elastic net regression as a data-driven approach to construct a relevant counterfactual from our pool of age groups not affected by the reform. Data on the number of GP consultations for males and females from 2006 to 2013 is obtained from the Norwegian Health Economics Administration. Our findings suggest that exempting adolescents from copayments increased the number of per person GP consultations by 22.1% among females and 13.8% among males. This indicates that adolescents between the ages of 12 and 15 were sensitive to copayments before the reform and females more so than males.


Cholangiocarcinoma in Wilson's Disease - a Case Report.
It has been suggested that hepatobiliary carcinomas are less frequent in Wilson's disease (WD) than in liver diseases of other etiology. However, the protective role of copper against malignancies is debated. Only a few cases of cholangiocarcinoma (CCC) in WD have been published. Here we report on a case of a 47-year-old male H1069Q homozygous, Kayser-Fleischer ring positive WD patient with a low ceruloplasmin level who was followed up and treated with chelating agents throughout nine years. The patient presented with neurological symptoms and liver cirrhosis at diagnosis. Clinical symptoms regressed after the treatment initiation. Rapidly developed tumour metastases were found in the bones, lung and liver (without jaundice). Autopsy revealed cholangiocarcinoma as the primary tumour confirmed by strong CK7 positivity and glypican-3 negativity. The curiosity of the presented case is the very rapid development of CCC despite continuous chelating agent therapy.


[Types of sensitivity (author's transl)].
Pathogen-host relationships are interactions which, on the part of the host, in contrast to in vitro infections not only depend on the available substrate but require a specific sensitivity to the individual pathogen. According to the species of the pathogen, the host responds with either a local or generalized sensitivity. Whereas local sensitivity persists throughout life, a generalized sensitivity is changed to a local sensitivity after the initial manifestation, by the development of a general immunity. Phylogenetic arguments support the assumption that sensitivity and its differences are genetically determined.


Effects of diterpene forskolin on the release reaction and protein phosphorylation of human platelets.
The effects of diterpene forskolin on the human platelet release reaction and on platelet protein phosphorylation were studied. This drug is shown to have the same effects as other agents which increase cAMP levels, namely, it inhibits the secretory response to diverse agonists and causes changes in the phosphorylation of several specific proteins. An increase of the 32P content is seen in the MW 47 000, 24 000 and 21 000 polypeptides while a decrease is observed in the MW 41 000 and 27 000 and 20 000 species. Forskolin also inhibits the changes in protein phosphorylation pattern induced by the powerful platelet secretagogue, thrombin. Our results relate the effects of antagonists of platelet secretion such as prostaglandins more closely to changes in cAMP levels and in protein phosphorylation than to other possible effects of the receptor-ligand interaction, which is by-passed by the use of forskolin. Our results also provide additional evidence involving these changes in the mechanisms which regulate the secretory process in platelets.


Influence of computer navigation on TKA revision rates.
We performed this study to determine whether the use of imageless navigation reduces revision rates after total knee arthroplasty (TKA). Data of 1,121 consecutive primary TKA with a follow-up of one to six years were retrospectively analysed. Following the conversion of the standard technique from conventional to navigated procedures, these data included the last 342 conventional and first 779 navigated procedures performed in our clinic. Demographic and perioperative covariates were recorded. All patients were asked by post to report instances of revisions. Data of 1,054 patients (94 %) were complete. Mean follow-up was 3.9 years for conventional and 2.4 years for navigated operations. Cumulative revision rate averaged 4.7 % for conventional and 2.3 % for navigated procedures. Cox's proportional hazard model was used to assess the effect of covariates on survival, resulting in significantly lower revision rates for older patients (p < 0.001) and for the navigated technique (p = 0.012). The reduced revision rate for navigated operations was mainly caused by a significantly reduced rate of aseptic implant loosening (1.9 % vs. 0.1 %, p = 0.024). Our study showed lower revision rates when computer navigation was used. However, due to the retrospective uncontrolled design, further prospective trials will be necessary to further evaluate this effect.


Surface-Tension-Driven Flows in a Thin Layer of a Water-n-Heptanol Solution.
A model for Marangoni flows that we observed on plane free surfaces of thin layers of aqueous fatty alcohol solutions is developed by considering evaporation-condensation of alcohol. The velocities of surface-tension-driven flows caused by gradients in temperature and in concentration are evaluated for water-n-heptanol solutions, which have a minimum of the equilibrium surface tension near 40 degrees C. Theoretical results are discussed on the basis of experimental observations. Copyright 2001 Academic Press.


Trends in major risk factors for cardiovascular disease among adults in the Mississippi Delta region, Mississippi Behavioral Risk Factor Surveillance System, 2001-2010.
The prevalences of major modifiable risk factors for cardiovascular disease (CVD) are disproportionately high in the 18-county Mississippi Delta region, and many of these risk factors disproportionately affect blacks. Temporal trends in the prevalence of CVD risk factors in the Mississippi Delta have not been determined. We examined trends in CVD risk factors from 2001 to 2010 in the region. Longitudinal trends in prevalence of high blood pressure, high cholesterol, diabetes, obesity, physical inactivity, and current smoking were investigated using self-reported data from the Behavioral Risk Factor Surveillance System. Joinpoint regression models were used to examine annual percentage change (APC) in the prevalence of these risk factors. Overall, from 2001 to 2010, we observed significant increases in the prevalence of high cholesterol (APC, 4.22%), obesity (APC, 3.65%), and diabetes (APC, 3.54%). Among blacks, we found significant increases in the prevalence of high cholesterol (APC, 3.41%), obesity (APC, 3.48%), and diabetes (APC, 4.96%). Among whites, we found significant increases in high blood pressure (APC, 2.18%), high cholesterol (APC, 4.78%), obesity (APC, 4.18%), and physical inactivity (APC, 3.06%). We also observed a significant decrease in smoking among whites (APC, -1.99%). From 2001 to 2010, we found a significant increase in the prevalence of high cholesterol, diabetes, and obesity in the Mississippi Delta. We also observed racial differences in those prevalences.


Sleep-Disordered Breathing in 2 Pediatric Patients on Peritoneal Dialysis.
Sleep-disordered breathing (SDB) is prevalent in children with chronic kidney disease (CKD), and has the potential to worsen vascular and neuro-cognitive health and quality of life. We present 2 children with CKD who experience central sleep apnea and nocturnal hypoventilation and discuss the possible underlying mechanisms in relation to CKD and dialysis.


Glucose tolerance and hyperinsulinaemia in obese women: role of adipose tissue distribution, muscle fibre characteristics and androgens.
The separate independent statistical contribution of abdominal distribution of fat, hyperandrogenicity and muscle morphology to glucose intolerance and hyperinsulinaemia was analysed in 88 obese women. In univariate analyses the waist/hip circumference ratio (WHR), body fat and lean body mass were all positively associated, and SHBG levels were negatively associated with insulin and glucose values. Muscle fibre areas were positively correlated with insulin but not with glucose concentrations. Adjustment for other variables did not remove the positive association between WHR and fasting insulin and glucose concentrations. SHBG, free testosterone and type IIb fibre areas were, however, significant confounding factors in the relationship between WHR and summed insulin and glucose concentrations. We conclude that fat distribution in obese women is associated with fasting hyperglycaemia and hyperinsulinaemia, independently of androgens and muscle fibre morphology, but that reduced SHBG concentrations and increased type IIb fibre areas may partly explain increased glucose and insulin responses to an oral glucose load in abdominally obese women.


Distribution of neutral lipids in the lipid droplet core.
Cholesteryl esters (CEs) are a form of cholesterol (CHOL) storage in the living cells, as opposed to free CHOL. CEs are major constituents of low density lipoprotein particles. Therefore, CEs are implicated in provoking atherosclerosis. Arranged into cytoplasmic lipid droplets (LDs), CEs are stored intracellularly. They can also be transported extracellularly by means of lipoproteins. In this work, large-scale molecular dynamics (MD) simulations are used to characterize the molecular structure of LDs containing various fractions (10-50 mol %) of cholesteryl oleate (CO) with respect to triolein (TO) fraction. The simulated LDs were covered by a phospholipid monolayer formed by a mixture of 1-palmitoyl-2-oleoylphosphatidylcholine, POPC (75 mol %), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, POPE (25 mol %), molecules. We report that most CO molecules are located within the hydrophobic core of LDs, whereas a small fraction (0.3-1.9 mol %) penetrates the monolayer. The solubility of CO in the phospholipid monolayer is relatively small. Due to a good miscibility with TO molecules, CO forms a liquid phase inside LD at 333 K. There is long-range order in the liquid TO-CO droplet core up to 8 nm from the phospholipid interface, resulting from the structuring of hydrophilic groups. This structuring slowly decays in the direction toward the LD center of mass. No sorting of TO and CO is detected, irrespective of the molar fractions simulated. The distribution of CO within the LDs is significant in determining the rate of their hydrolysis by surface-bound enzyme lipases, and thus has a subsequent impact on the levels of CO in plasma and LDLs.


Activities of life: the COPD patient.
Chronic obstructive pulmonary disease (COPD) is a chronic, progressive and debilitating disease. With worsening disease, a COPD patient will progressively become less physically active, will have reduced social contacts, and this will create a downward vicious circle. This review discusses the significant sequelae of physical inactivity in the COPD patient, approaches to assessment and management, and the impact of COPD exacerbations on the activities of daily life. Patients with COPD are markedly inactive in daily life. Referral to pulmonary rehabilitation and self-management education to promote increase physical activities should be an important part of the management of COPD. Acute exacerbations in COPD patients are severely distressing events that impact greatly on health status, loss of symptom control and functional state with limitation on activities of daily living. If we are to progress in the treatment of acute exacerbation of COPD, it is evident that the effects of interventions on activities of daily living should be considered as an essential component of our goal of treatment.


Investigation of Impact Resistance of High-Strength Portland Cement Concrete Containing Steel Fibers.
Impact resistance of Portland cement concrete (PCC) is an essential property in various applications of PCC, such as industrial floors, hydraulic structures, and explosion-proof structures. Steel-fiber-fortified high-strength concrete testing was completed using a drop-weight impact assessment for impact strength. One mix was used to manufacture 320 concrete disc specimens cured in both humid and dry conditions. In addition, 30 cubic and 30 cylindrical specimens were used to evaluate the compressive and indirect tensile strengths. Steel fibers with hooked ends of lengths of 20, 30, and 50 mm were used in the concrete mixtures. Data on material strength were collected from impact testing, including the number of post-first-crack blows (INPBs), first-crack strength, and failure strength. Findings from the results concluded that all the steel fibers improved the mechanical properties of concrete. However, hooked steel fibers were more effective than crimped steel fibers in increasing impact strength, even with a smaller length-to-diameter ratio. Concrete samples containing hybrid fibers (hooked + crimped) also had lower compressive strength than the other fibers. Comparisons and analogies drawn between the test results and the static analyses (Kolmogorov-Smirnov and Kruskal-Wallis) show that the p-value of the analyses indicates a more normal distribution for curing in a humid environment. A significant difference was also observed between the energy absorptions of the reinforced mixtures into steel fibers.


Relative contributions of microbial and infrastructure heat at a crude oil-contaminated site.
Biodegradation of contaminants can increase the temperature in the subsurface due to heat generated from exothermic reactions, making temperature observations a potentially low-cost approach for determining microbial activity. For this technique to gain more widespread acceptance, it is necessary to better understand all the factors affecting the measured temperatures. Biodegradation has been occurring at a crude oil-contaminated site near Bemidji, Minnesota for 39 years, creating a quasi-steady-state plume of contaminants and degradation products. A model of subsurface heat generation and transport helps elucidate the contribution of microbial and infrastructure heating to observed temperature increases at this site. We created a steady-state, two-dimensional, heat transport model using previous-published parameter values for physical, chemical and biodegradation properties. Simulated temperature distributions closely match the observed average annual temperatures measured in the contaminated area at the site within less than 0.2 °C in the unsaturated zone and 0.4 °C in the saturated zone. The model results confirm that the observed subsurface heat from microbial activity is due primarily to methane oxidation in the unsaturated zone resulting in a 3.6 °C increase in average annual temperature. Another important source of subsurface heat is from the active, crude-oil pipelines crossing the site. The pipelines impact temperatures for a distance of 200 m and contribute half the heat. Model results show that not accounting for the heat from the pipelines leads to overestimating the degradation rates by a factor of 1.7, demonstrating the importance of identifying and quantifying all heat sources. The model results also highlighted a zone where previously unknown microbial activity is occurring at the site.


Sulfotransferase-mediated chlorination of 1-hydroxymethylpyrene to a mutagen capable of penetrating indicator cells.
Methylated polycyclic aromatic hydrocarbons are common in the human environment. Many of them are stronger carcinogens than their purely aromatic congeners. They may be metabolized to benzylic alcohols. We report here on biochemical and toxicological characteristics of 1-hydroxymethylpyrene (HMP), a typical representative of this class of compounds. Rat liver cytosol, fortified with 3'-phosphoadenosine-5'-phosphosulfate, converted HMP into its sulfate ester (HMPS), HMPS bound covalently to isolated DNA. In physiological buffer at 37 degrees C, HMPS had a half-life of 2 min, the major decomposition product being HMP. Thus, cyclic activation is possible. When Cl- anions were present at physiological concentrations, an additional reaction product of HMPS, 1-chloromethylpyrene (ClMP), could be identified on the basis of its chromatographic properties and its mass spectrum, using the authentic standard for comparison. ClMP was shorter-lived in buffer than HMPS. ClMP reacted with DNA, the adduct pattern in the 32P-postlabeling analysis being similar, or identical, to that of HMPS. ClMP proved to be a very potent mutagen in Salmonella typhimurium, whereas HMPS, and HMP in the presence of a sulfate-conjugating system, showed strong mutagenicity only when Cl- or Br- ions were present in the exposure buffer. It is concluded that HMPS is capable of reacting with DNA, but is hampered in its distribution by membrane barriers.(ABSTRACT TRUNCATED AT 250 WORDS)


NF546 [4,4'-(carbonylbis(imino-3,1-phenylene-carbonylimino-3,1-(4-methyl-phenylene)-carbonylimino))-bis(1,3-xylene-alpha,alpha'-diphosphonic acid) tetrasodium salt] is a non-nucleotide P2Y11 agonist and stimulates release of interleukin-8 from human monocyte-derived dendritic cells.
The G protein-coupled P2Y(11) receptor is involved in immune system modulation. In-depth physiological evaluation is hampered, however, by a lack of selective and potent ligands. By screening a library of sulfonic and phosphonic acid derivatives at P2Y(11) receptors recombinantly expressed in human 1321N1 astrocytoma cells (calcium and cAMP assays), the selective non-nucleotide P2Y(11) agonist NF546 [4,4'-(carbonylbis(imino-3,1-phenylene-carbonylimino-3,1-(4-methyl-phenylene)carbonylimino))-bis(1,3-xylene-alpha,alpha'-diphosphonic acid) tetrasodium salt] was identified. NF546 had a pEC(50) of 6.27 and is relatively selective for P2Y(11) over P2Y(1), P2Y(2), P2Y(4), P2Y(6), P2Y(12), P2X(1), P2X(2), and P2X(2)-X(3). Adenosine-5'-O-(3-thio)triphosphate (ATPgammaS), a nonhydrolyzable analog of the physiological P2Y(11) agonist ATP, and NF546 use a common binding site as suggested by molecular modeling studies and their competitive behavior toward the nanomolar potency antagonist NF340 [4,4'-(carbonylbis(imino-3,1-(4-methyl-phenylene)carbonylimino))bis(naphthalene-2,6-disulfonic acid) tetrasodium salt] in Schild analysis. The pA(2) of NF340 was 8.02 against ATPgammaS and 8.04 against NF546 (calcium assays). NF546 was further tested for P2Y(11)-mediated effects in monocyte-derived dendritic cells. Similarly to ATPgammaS, NF546 led to thrombospondin-1 secretion and inhibition of lipopolysaccharide-stimulated interleukin-12 release, whereas NF340 inhibited these effects. Further, for the first time, it was shown that ATPgammaS or NF546 stimulation promotes interleukin 8 (IL-8) release from dendritic cells, which could be inhibited by NF340. In conclusion, we have described the first selective, non-nucleotide agonist NF546 for P2Y(11) receptors in both recombinant and physiological expression systems and could show a P2Y(11)-stimulated IL-8 release, further supporting the immunomodulatory role of P2Y(11) receptors.


Subjective well-being and dental treatment in elderly African Americans.
This preliminary study examined the effect of dental treatment on psychological well-being in a sample of elderly African Americans (64 women, 43 men) seeking routine care at an urban medical/dental clinic. Scores on two scales, the Index of Self-esteem and the Generalized Contentment Scale, self-administered at initial and final dental appointments, yielded no significant differences between times of test. Dental treatment apparently was not associated in this sample with perceived psychological well-being.


Early Discontinuation of P2Y12 Antagonists and Adverse Clinical Events Post-Percutaneous Coronary Intervention: A Hospital and Primary Care Linked Cohort.
Background Early discontinuation of P2Y12 antagonists post-percutaneous coronary intervention may increase risk of stent thrombosis or nonstent recurrent myocardial infarction. Our aims were to (1) analyze the early discontinuation rate of P2Y12 antagonists post-percutaneous coronary intervention, (2) explore factors associated with early discontinuation, and (3) analyze the risk of major adverse cardiovascular events (death, acute coronary syndrome, revascularization, or stroke) associated with discontinuation from a prespecified prescribing instruction of 1 year. Method and Results We studied 2090 patients (2011-2015) who were recommended for clopidogrel for 12 months (+aspirin) post-percutaneous coronary intervention within a retrospective observational population cohort. Relationships between clopidogrel discontinuation and major adverse cardiac events were evaluated over 18-month follow-up. Discontinuation of clopidogrel in the first 4 quarters was low at 1.1%, 2.6%, 3.7%, and 6.1%, respectively. Previous revascularization, previous ischemic stroke, and age >80 years were independent predictors of early discontinuation. In a time-dependent multiple regression model, clopidogrel discontinuation and bleeding (hazard ratio=1.82 [1.01-3.30] and hazard ratio=5.30 [3.14-8.94], respectively) were independent predictors of major adverse cardiac events as were age <49 and ≥70 years (versus those aged 50-59 years), hypertension, chronic kidney disease stage 4+, previous revascularization, ischemic stroke, and thromboembolism. Furthermore, in those with both bleeding and clopidogrel discontinuation, hazard ratio for major adverse cardiac events was 9.34 (3.39-25.70). Conclusions Discontinuation of clopidogrel is low in the first year post-percutaneous coronary intervention, where a clear discharge instruction to treat for 1 year is provided. Whereas this is reassuring from the population level, at an individual level discontinuation earlier than the intended duration is associated with an increased rate of adverse events, most notably in those with both bleeding and discontinuation.