Spiralian gastrulation: germ layer formation, morphogenesis, and fate of the blastopore in the slipper snail Crepidula fornicata.
Gastrulation is a critical step in bilaterian development, directly linked to the segregation of germ layers, establishment of axes, and emergence of the through-gut. Theories about the evolution of gastrulation often concern the fate of the blastopore (site of endomesoderm internalization), which varies widely in a major branch of bilaterians, the Spiralia. In this group, the blastopore has been said to become the mouth, the anus, both, or neither. Different developmental explanations for this variation exist, yet no modern lineage tracing study has ever correlated the position of cells surrounding the blastopore with their contribution to tissues of the mouth, foregut, and anus in a spiralian. This is the first study to do so, using the gastropod Crepidula fornicata. Crepidula gastrulation occurs by epiboly: the first through third quartet micromeres form an epithelial animal cap that expands to cover vegetal endomesodermal precursors. Initially, descendants of the second and third quartet micromeres (2a-2d, 3a-3d) occupy a portion of the blastopore lip. As the blastopore narrows, the micromeres' progeny exhibit lineage-specific behaviors that result in certain sublineages leaving the lip's edge. Anteriorly, cells derived from 3a(2) and 3b(2) undergo a unique epithelial-to-mesenchymal transition involving proliferation and a collective movement of cells into the archenteron. These cells make a novel spiralian germ layer, the ectomesoderm. Posteriorly, cells derived from 3c(2) and 3d(2) undergo a form of convergence and extension that involves zippering of cells and their intercalation across the ventral midline. During this process, several of these cells, as well as the 2d clone, become displaced posteriorly, away from the blastopore. Progeny of 2a-2c and 3a-3d make the mouth and foregut, and the blastopore becomes the opening to the mouth. The anus forms days later, as a secondary opening within the 2d(2) clone, and not from the classically described "anal cells", which we identify as the 3c(221) and 3d(221) cells. Our analysis of Crepidula gastrulation constitutes the first description of blastopore lip morphogenesis and fates using lineage tracing and live imaging. These data have profound implications for hypotheses about the evolution of the bilaterian gut and help explain observed variation in blastopore morphogenesis among spiralians.


Improving the Assessment of Mild Cognitive Impairment in Advanced Age With a Novel Multi-Feature Automated Speech and Language Analysis of Verbal Fluency.
Introduction: Clinically relevant information can go uncaptured in the conventional scoring of a verbal fluency test. We hypothesize that characterizing the temporal aspects of the response through a set of time related measures will be useful in distinguishing those with MCI from cognitively intact controls. Methods: Audio recordings of an animal fluency test administered to 70 demographically matched older adults (mean age 90.4 years), 28 with mild cognitive impairment (MCI) and 42 cognitively intact (CI) were professionally transcribed and fed into an automatic speech recognition (ASR) system to estimate the start time of each recalled word in the response. Next, we semantically cluster participant generated animal names and through a novel set of time-based measures, we characterize the semantic search strategy of subjects in retrieving words from animal name clusters. This set of time-based features along with standard count-based features (e.g., number of correctly retrieved animal names) were then used in a machine learning algorithm trained for distinguishing those with MCI from CI controls. Results: The combination of both count-based and time-based features, automatically derived from the test response, achieved 77% on AUC-ROC of the support vector machine (SVM) classifier, outperforming the model trained only on the raw test score (AUC, 65%), and well above the chance model (AUC, 50%). Conclusion: This approach supports the value of introducing time-based measures to the assessment of verbal fluency in the context of this generative task differentiating subjects with MCI from those with intact cognition.


Placebo response and antidepressant response.
Much of the response to an antidepressant is the result of placebo response. The placebo response embedded in drug response confounds studies seeking to identify brain mechanisms essential for pharmacologic response. Exclusion of patients who fail to meet entry criteria at the end of a placebo lead-in phase has been inadequate to reduce the effect of placebo during pharmacologic trials. This study focused on the placebo lead-in phase and examines whether change in severity of depression during placebo lead-in predicts change in depressive symptoms during antidepressant treatment. The subjects were patients aged 60-85 years with nonpsychotic unipolar major depression not attributed to dementing disorders, medical illnesses, or drugs causing depression and had a 24-item Hamilton Depression Rating Scale score of 18 or greater. After a two-week placebo lead-in, subjects with Hamilton Depression Rating Scale score of 18 or greater received 10 mg escitalopram for 12 weeks. Worsening or limited change in depression during placebo treatment predicted improvement in depressive symptoms during escitalopram treatment. Limited change in anxiety, melancholia, helplessness, and paranoia during placebo treatment were the strongest predictors of improvement in depression while on escitalopram. These findings, if replicated, may be used to characterize depressed older patients likely to respond to the pharmacologic action of antidepressants rather than the placebo response embedded in drug trials and thus improve the methodology of biomarker studies of antidepressant response. On a clinical level, depressed older patients who improve during a prolonged evaluation may be candidates for nonpharmacologic treatments because their drug response may be limited.


Predictors of outcome after endovascular treatment of cerebral vasospasm.
Angioplasty and intra-arterial papaverine are promising treatments for severe symptomatic vasospasm after subarachnoid hemorrhage (SAH), but there is little information on the clinical factors that predict treatment outcome. We sought to determine variables for predicting functional outcome in this setting. We reviewed 81 consecutive patients with symptomatic cerebral vasospasm from aneurysmal SAH treated with percutaneous balloon angioplasty or selective intra-arterial papaverine infusion between 1990 and 2000 (105 procedures). Logistic regression analysis was used to assess the effect of various clinical and angiographic factors on outcome. Mean patient age was 54 years (range, 29-88 years). Twenty-nine patients (36%) presented with poor-grade (World Federation of Neurologic Surgeons [WFNS] grade IV or V) SAH. Clinical deficits were global in 55 patients (70%), and angiographic vasospasm was diffuse in 53 (65%). Endovascular treatment consisted of transluminal angioplasty alone (18 procedures, 17%), intra-arterial papaverine infusion (65 procedures, 62%), or both (22 procedures, 21%). Unequivocal arterial dilatation was achieved in all but two patients, and major complications occurred in 2% of the procedures. Ten patients (12%) died in the hospital, and 36 (44%) recovered poorly. Permanent deficits attributable to cerebral vasospasm were present in 37 patients (52% of survivors). On multivariate logistic regression analysis, advanced age and poor WFNS grade at presentation were predictive of poor clinical outcome. Advanced age and poor clinical status at the time of SAH onset are predictive of poor clinical outcome despite endovascular treatment with angioplasty or intra-arterial papaverine in patients with symptomatic vasospasm.


Spectrum and hematological profile of hereditary anemia in North Indians: SGPGI experience.
Thalassemia and other hemoglobinopathies together with red cell enzymopathies are a common cause of anemia, which can be prevented by population screening and genetic counseling. This study was designed to screen the anemic patients for thalassemia, structural hemoglobin variants and red cell enzymopathies. A total of 17047 cases were evaluated from 2009 to 2018 for thalassemia, hemoglobin variants, glucose 6 phosphate dehydrogenase deficiency, pyruvate kinase deficiency and hereditary spherocytosis. Patients' records were entered in a Microsoft excel sheet and a spectrum of disorders was evaluated. Year wise spectrum was also analyzed to see the difference in incidence at different time periods. Incidence of beta thalassemia trait and thalassemia major was found in 11.0% and 3.4% respectively, whereas other hemoglobinopathies were observed in 3.2% of the cases. G6PD deficient cases were 0.2 % and 0.4% had hereditary spherocytosis. No significant difference was observed in incidence of thalassemia and other hemoglobinopathies at different time points. This study provided a health burden and detailed spectrum and prevalence of hemoglobinopathies in North Indians high risk population which contribute toward the development of prevention strategies for better management of hemoglobinopathies. In view of high incidence of thalassemia a routine hematological screening at a primary health center may be introduced as a prospective premarital screening under a thalassemia control program. Moreover rapid and easy quantification of hemoglobin variants (Hb variants) make Cation exchange - High Performance Liquid Chromatography (CE-HPLC) a suitable diagnostic test for the routine investigation of genetic causes of anemia.


Ultrafast Synthetic Transmit Aperture Imaging Using Hadamard-Encoded Virtual Sources With Overlapping Sub-Apertures.
The development of ultrafast ultrasound imaging offers great opportunities to improve imaging technologies, such as shear wave elastography and ultrafast Doppler imaging. In ultrafast imaging, there are tradeoffs among image signal-to-noise ratio (SNR), resolution, and post-compounded frame rate. Various approaches have been proposed to solve this tradeoff, such as multiplane wave imaging or the attempts of implementing synthetic transmit aperture imaging. In this paper, we propose an ultrafast synthetic transmit aperture (USTA) imaging technique using Hadamard-encoded virtual sources with overlapping sub-apertures to enhance both image SNR and resolution without sacrificing frame rate. This method includes three steps: 1) create virtual sources using sub-apertures; 2) encode virtual sources using Hadamard matrix; and 3) add short time intervals (a few microseconds) between transmissions of different virtual sources to allow overlapping sub-apertures. The USTA was tested experimentally with a point target, a B-mode phantom, and in vivo human kidney micro-vessel imaging. Compared with standard coherent diverging wave compounding with the same frame rate, improvements on image SNR, lateral resolution (+33%, with B-mode phantom imaging), and contrast ratio (+3.8 dB, with in vivo human kidney micro-vessel imaging) have been achieved. The f-number of virtual sources, the number of virtual sources used, and the number of elements used in each sub-aperture can be flexibly adjusted to enhance resolution and SNR. This allows very flexible optimization of USTA for different applications.


Live Leishmania promastigotes can directly activate primary human natural killer cells to produce interferon-gamma.
Natural killer (NK) cells have been implicated in the natural protection and healing of leishmaniasis by their ability to secrete the macrophage activating cytokine interferon (IFN)gamma. Previous studies have demonstrated that early production of interleukin (IL)-12 triggers IFN gamma secretion by NK cells. Here we report that live Leishmania promastigotes (the form that is injected by the vector) can directly induce human peripheral blood NK cells from healthy donors to IFN gamma secretion in the absence of IL-12 and professional antigen presenting cells. Killing of promastigotes abolishes this property. This novel mechanism of activation of the innate immune response may be relevant for establishment of infection and thus also the design of vaccines against leishmaniasis.


Parietal-eye phototransduction components and their potential evolutionary implications.
The parietal-eye photoreceptor is unique because it has two antagonistic light signaling pathways in the same cell-a hyperpolarizing pathway maximally sensitive to blue light and a depolarizing pathway maximally sensitive to green light. Here, we report the molecular components of these two pathways. We found two opsins in the same cell: the blue-sensitive pinopsin and a previously unidentified green-sensitive opsin, which we name parietopsin. Signaling components included gustducin-alpha and Galphao, but not rod or cone transducin-alpha. Single-cell recordings demonstrated that Go mediates the depolarizing response. Gustducin-alpha resembles transducin-alpha functionally and likely mediates the hyperpolarizing response. The parietopsin-Go signaling pair provides clues about how rod and cone phototransduction might have evolved.


TUMOR INOCULATION INTO ORGANS AND THE ANALOGY BETWEEN HUMAN CANCER AND THE TUMORS OF WHITE MICE AND WHITE RATS.
The analysis of the experiments described above indicates that tumors of the white rat or white mouse inoculated into parenchymatous organs acquire a different biological character from those inoculated subcutaneously. The latter are a great deal more benign in their behavior than human cancer or spontaneous tumors in the same species of animals. Tumors inoculated into organs, on the other hand, are quite identical in their biological behavior with the malignant tumors of animal and man. A conclusion must then be drawn, even a priori, that the method of inoculation into organs is a very important aid in the experimental investigation of cancer. It is true that the method is a great deal more complicated and time-consuming than the ordinary subcutaneous inoculation. The subcutaneous method is satisfactory for a number of cancer problems. One of these is the study of general susceptibility and resistance of the organism of the host to the inoculation of the tumors, and this is a subject of paramount importance in cancer research. On the other hand, the investigations of the writer (10) have shown that an animal may be susceptible to a subcutaneous inoculation of a certain tumor and resist the inoculation of the same tumor into the testicle. Undoubtedly this method of inoculation will reveal the existence of a number of other phenomena. The discovery of specific therapeutic measures is certainly the greatest problem in cancer research. A great deal of work has been done already on the subject, and the latest investigations of Wassermann on the chemotherapy of experimental tumors seem to be of great promise. But here also the therapeutic methods must be tried on animals in which the inoculations of tumor cells have been made into parenchymatous organs before the growths thus treated will have any analogy to human cancer. In this connection one must bear in mind the fact that all the empirical so-called specific cancer remedies, which are continually being devised, are usually successful in treating localized skin cancers and fail utterly in the malignant growths of the internal organs. It is comparatively easy to produce a localized necrosis and softening in a circumscribed growth of the skin and subcutaneous tissue, but whether the same result will be produced on a diffuse and better nourished tumor growing inside of a parenchymatous organ cannot be decided a priori. To determine this it is necessary to have experimental proof on animals in which the tumor was inoculated into organs.


Impact of poor-quality medicines in the 'developing' world.
Since our ancestors began trading several millennia ago, counterfeit and substandard medicines have been a recurring problem, with history punctuated by crises in the supply of anti-microbials, such as fake cinchona bark in the 1600s and fake quinine in the 1800s. Unfortunately this problem persists, in particular afflicting unsuspecting patients in 'developing' countries. Poor-quality drugs are a vital (but neglected) public health problem. They contribute to a 'crevasse' between the enormous effort in therapeutic research and policy decisions and implementation of good-quality medicines.


Patient preferences for the delivery of disease management in chronic heart failure: a qualitative study.
Chronic heart failure (CHF) management programs (CHF-MPs) are applied in different ways including via face-to-face settings. However, we know little about consumer preferences when applying CHF-MPs via a patient's home or specialist hospital clinic. The aim of this pilot study was to explore CHF-MP characteristics that are considered desirable by patients with CHF. Semistructured interviews with a purposive sample of 12 CHF patients. Participants had a mean age of 61 (SD, 17) years, 3 were female, and the majority was of white background. Most were assessed as either functional New York Heart Association class III (n = 3) or IV (n = 6). Home- and clinic-based CHF-MPs were preferred by 5 and 7 participants, respectively. Key themes around patient preferences related to practical aspects of program delivery and social and peer support, as well as health-related benefits that translate to traditional outcomes in program evaluations. Participants identified transport, cost, and ill health as barriers to attending a clinic-based program. However, they also highlighted benefits (eg, the ability to share experiences with other patients) that may be difficult to provide with a home-based service unless specifically organized. These preliminary data suggest that patients value aspects of a program beyond those directly related to health outcomes. They also recognize a need for flexibility in program delivery, with potential preferences for home- or clinic-based programs depending largely on individual patient circumstances. More definitive studies are required to explore how best to cater for individual preferences while optimizing health outcomes.


Characterization of a decrease in muscarinic m2 mRNA in cerebellar granule cells by carbachol.
Studies involving carbachol (100 microM) treatment of cerebellar granule cells for 1, 3, 6, 9, 12 and 24 hr show a decrease in the mRNA encoding for the muscarinic m2 receptor. The response was transient, decreasing m2 mRNA by 25 to 50% in 6 and 9 hr, respectively. The data presented in this work were quantified by ribonuclease protection assay, using a [32P]-cRNA probe corresponding to nucleotide +1138 to 1650 of the rat m2 muscarinic receptor. Because cerebellar granule cells express muscarinic m2 and m3 receptors, we tested whether the carbachol-mediated decrease in m2 mRNA resulted from a homologous or heterologous activation of muscarinic receptors. At a 1 microM concentration, methoctramine specifically blocked the muscarinic m2 receptor and reversed carbachol's action. These data suggested that carbachol acts via a possible homologous activation of muscarinic m2 receptors. The half-life of the receptor mRNA measured in the presence of actinomycin D with and without carbachol were similar. Because carbachol treatments decrease the steady-state levels of m2 mRNA without changing the half-life of the message, we suggest that a carbachol treatment induces a decrease in the transcription of the gene for the muscarinic m2 receptor.


Interface modalities of technical aids used by people with disability.
The objective of this paper was to present some initial thoughts and concepts on the subject of the interface "mechanisms" used between individuals with disability and their technical aids. These interface modalities are considered to be of primary importance in matching adaptive aids to the needs of these individuals. With this in mind, a number of basic premises concerning technical aids, user needs, and user environments are discussed. In addition, an initial framework is presented that describes a series of different levels of interface types. It is hoped that this framework will promote a greater insight into the inherent complexity of interface modalities and thereby stimulate further study.


[In vitro antimicrobial activities of quinolones, rifamycins and macrolides against Mycobacterium tuberculosis and M.avium complex: attempt to establish new assay methods which accurately reflect therapeutic effects of test agents in vivo].
Profiles of expression of the antimicrobial activities of LVFX, KRM-1648 (KRM), and CAM against M. tuberculosis (MTB) and M. avium complex (MAC) residing in MONO-MAC-6 human macrophage like cells (MM6-M phi s) and A-549 human type II alveolar pneumocyte cells (A-549 cells) were determined. First, the antimicrobial activities of LVFX, KRM, and CAM against intracellular organisms of MTB Kurono and MAC N-444 strains were examined under conditions in which infected MM6-M phi s and A-549 cells were cultured for up to 7 days or longer in medium containing the antimicrobials at their Cmaxs in the blood, achievable after oral administration of clinical dosages of these drugs. The antimicrobial effects of LVFX and KRM against respectively MTB and MAC within A-549 cells were significantly less than the activities they displayed against the same organisms residing in MM6-M phi s. Notably, it was also found that KRM had a markedly larger MIC (0.25 microgram/ml) for MAC N-444 within A-549 cells than its MIC (0.008 microgram/ml) for the same strain residing in MM6-M phi s. Thus, the profiles of LVFX- and KRM-mediated killing or inhibition of intracellular MTB or MAC organisms in A-549 cells were markedly different from those observed for the organisms residing in MM6-M phi s. Second, invasive and multiplicative phenotypes of MTB and MAC organisms, which had been adapted to either an extracellular or intracellular environment (designated as E- and I-type organisms, respectively), were studied. In the case of MTB, I-type organisms (retrieved from infected MM6-M phi s after bacterial growth within the M phi s during 5-day cultivation) were less efficient than E-type organisms (prepared by cultivating the organisms in 7H9 medium) in entering MM6-M phi s, whereas I-type organisms were more efficient than E-type organisms in invading A-549 cells. On the other hand, in the case of MAC, infectivity of I-type organisms not only in MM6-M phi s but also in A-549 cells was larger than that of E-type organisms. Next, while I-type organisms of MTB and MAC displayed more vigorous replication within MM6-M phi s than E-type organisms, the growth rate of E-type organisms within A-549 cells was more rapid than that of I-type organisms residing in A-549 cells. These findings indicate that there are significant differences between E- and I-type organisms of MTB or MAC in ability to invade and multiply within M phi s (professional phagocytes) and alveolar epithelial cells (nonprofessional phagocytic cells).


The origin and evolution of Homo sapiens.
If we restrict the use of Homo sapiens in the fossil record to specimens which share a significant number of derived features in the skeleton with extant H. sapiens, the origin of our species would be placed in the African late middle Pleistocene, based on fossils such as Omo Kibish 1, Herto 1 and 2, and the Levantine material from Skhul and Qafzeh. However, genetic data suggest that we and our sister species Homo neanderthalensis shared a last common ancestor in the middle Pleistocene approximately 400-700 ka, which is at least 200 000 years earlier than the species origin indicated from the fossils already mentioned. Thus, it is likely that the African fossil record will document early members of the sapiens lineage showing only some of the derived features of late members of the lineage. On that basis, I argue that human fossils such as those from Jebel Irhoud, Florisbad, Eliye Springs and Omo Kibish 2 do represent early members of the species, but variation across the African later middle Pleistocene/early Middle Stone Age fossils shows that there was not a simple linear progression towards later sapiens morphology, and there was chronological overlap between different 'archaic' and 'modern' morphs. Even in the late Pleistocene within and outside Africa, we find H. sapiens specimens which are clearly outside the range of Holocene members of the species, showing the complexity of recent human evolution. The impact on species recognition of late Pleistocene gene flow between the lineages of modern humans, Neanderthals and Denisovans is also discussed, and finally, I reconsider the nature of the middle Pleistocene ancestor of these lineages, based on recent morphological and genetic data.This article is part of the themed issue 'Major transitions in human evolution'.


Virtual sensors for designing irrigation controllers in greenhouses.
Monitoring the greenhouse transpiration for control purposes is currently a difficult task. The absence of affordable sensors that provide continuous transpiration measurements motivates the use of estimators. In the case of tomato crops, the availability of estimators allows the design of automatic fertirrigation (irrigation + fertilization) schemes in greenhouses, minimizing the dispensed water while fulfilling crop needs. This paper shows how system identification techniques can be applied to obtain nonlinear virtual sensors for estimating transpiration. The greenhouse used for this study is equipped with a microlysimeter, which allows one to continuously sample the transpiration values. While the microlysimeter is an advantageous piece of equipment for research, it is also expensive and requires maintenance. This paper presents the design and development of a virtual sensor to model the crop transpiration, hence avoiding the use of this kind of expensive sensor. The resulting virtual sensor is obtained by dynamical system identification techniques based on regressors taken from variables typically found in a greenhouse, such as global radiation and vapor pressure deficit. The virtual sensor is thus based on empirical data. In this paper, some effort has been made to eliminate some problems associated with grey-box models: advance phenomenon and overestimation. The results are tested with real data and compared with other approaches. Better results are obtained with the use of nonlinear Black-box virtual sensors. This sensor is based on global radiation and vapor pressure deficit (VPD) measurements. Predictive results for the three models are developed for comparative purposes.


First Report of Rice yellow mottle virus on Rice in Burundi.
Since the mid-1980s, rice cultivation has expanded rapidly in Burundi to reach approximately 50,000 ha in 2011. In 2007, leaf mottling, reduced tillering, and stunting symptoms were observed on rice at Gatumba near Bujumbura, causing small patches in less than 10% of the fields. Rice yellow mottle virus (RYMV, genus Sobemovirus), which has seriously threatened rice cultivation in Africa (1) and was recently described in the neighboring Rwanda (3), was suspected to be involved because of similar symptoms. To identify the pathogen that caused the disease in Burundi, a survey was performed in the major rice-producing regions of Burundi and Rwanda. Six locations in Burundi and four in Rwanda were investigated in April and October 2011. Disease incidence in the fields was estimated to be 15 ± 5%. Symptomatic leaves of 24 cultivated rice plants were collected and tested by double antibody sandwich-ELISA with polyclonal antibodies raised against the RYMV isolate Mg1 (2). All tested samples reacted positively. Four isolates were inoculated on susceptible Oryza sativa cultivar IR64 (2). The typical symptoms of RYMV were reproduced 7 days after inoculation, whereas the noninoculated controls remained healthy. Total RNA was extracted by the RNeasy Plant Mini kit (QIAGEN, Hilden, Germany) from 12 samples. The RYMV coat protein gene was amplified by RT-PCR with primers 5'CGCTCAACATCCTTTTCAGGGTAG3' and 5'CAAAGATGGCCAGGAA3' (3). The sequences were deposited in GenBank (Accession Nos. HE654712 to HE654723). To characterize the isolates, the sequences of the tested samples were compared in a phylogenic tree including a set of 45 sequences of isolates from Rwanda, Uganda, western Kenya, and northern Tanzania (2,3). Six isolates from western Burundi, namely Bu1, Bu2, Bu4, Bu7, Bu10, and Bu13 (Accession Nos. HE654712 to HE654716 and HE654718), and the isolate Rw208 (HE654720) from southwestern Rwanda, belonged to strain S4-lm previously reported near Lakes Malawi and Tanganyika. They fell within the group gathering isolates from the western Bugarama plain of Rwanda (3). The isolates Bu16 (HE654719) and Bu17 (HE654717) from Mishiha in eastern Burundi belonged to strain S4-lv previously reported around Lake Victoria. However, they did not cluster with isolates from the eastern and southern provinces of Rwanda. They were genetically more closely related to isolates of strain S4-lv from northern Tanzania. Overall, the phylogeography of RYMV in Burundi and Rwanda region was similar. In the western plain of the two countries, the isolates belonged to the S4-lm lineage, whereas at the east of the two countries at midland altitude, they belonged to the S4-lv lineage. The presence of RYMV in Burundi should be considered in the future integrative pest management strategies for rice cultivation in the country. References: (1) D. Fargette et al. Annu. Rev. Phytopathol. 44:235, 2006. (2) Z. L. Kanyeka et al. Afr. Crop Sci. J. 15:201, 2007. (3) I. Ndikumana et al. New Dis. Rep. 23:18, 2011.


Molecular mechanisms of p21 and p27 induction by 3-methylcholanthrene, an aryl-hydrocarbon receptor agonist, involved in antiproliferation of human umbilical vascular endothelial cells.
We previously reported that 3-methylcholanthrene (3MC), an aryl-hydrocarbon receptor (AhR) agonist, inhibits the proliferation of human umbilical vascular endothelial cells (HUVECs; Juan et al., 2006, Eur J Pharmacol 530: 1-8). Herein, pretreatment of HUVECs with p21 or p27 small interfering (si)RNA reduced 3MC-induced elimination of [(3)H]thymidine incorporation, demonstrating their essential roles in the antiproliferation of HUVECs. The molecular mechanisms of p21 and p27 involved in the antiproliferative effects of 3MC were elucidated in this study. 3MC time- and concentration-dependently increased p21 and p27 levels, and decreased the protein level of CDK2 with no apparent alteration of p53. Interestingly, 3MC-mediated p21 and p27 inductions were eliminated by resveratrol, an AhR antagonist, suggesting their AhR dependency, further confirmed by AhR siRNA. Among the relevant pathways, p38MAPK activation sustained the levels of p21 and p27 induced by 3MC, which was eliminated by AhR antagonists and N-acetylcysteine (NAC), an antioxidant. 3MC concentration-dependently enhanced not only the consensus dioxin-responsive element (DRE)-driven luciferase activity, but also the binding activity of the AhR to the putative DRE derived from the p21 and p27 promoters. A deletion of the DRE (-285/-270) in p21 (-2,300/+8) only partially alleviated the 3MC-induced luciferase activity unless NAC was added, suggesting that there may be a DRE-independent mechanism associated with oxidative stress. However, a deletion of the DRE (-660/-645) in p27 (-1,358/-100) almost completely abrogated the activation. Our study demonstrated that both the functional DRE and the phosphorylation of p38MAPK are essential for the induction of p21 and p27, resulting in the antiproliferative action of 3MC in HUVECs.


Assessing coral reef health across onshore to offshore stress gradients in the US Virgin Islands.
Managing the effects of anthropogenic disturbance on coral reefs is highly dependant on effective strategies to assess degradation and recovery. We used five years of field data in the US Virgin Islands to investigate coral reef response to a potential gradient of stress. We found that the prevalence of old partial mortality, bleaching, and all forms of coral health impairment (a novel category) increased with nearshore anthropogenic processes, such as a five-fold higher rate of clay and silt sedimentation. Other patterns of coral health, such as recent partial mortality, other diseases, and benthic cover, did not respond to this potential gradient of stress or their response could not be resolved at the frequency or scale of monitoring. We suggest that persistent signs of disturbance are more useful to short-term, non-intensive (annual) coral reef assessments, but more intensive (semi-annual) assessments are necessary to resolve patterns of transient signs of coral health impairment.


Flexible alteration of optical nonlinearities of iodine charge-transfer complexes in solutions.
The type of donor used is found to influence greatly the optical nonlinearities of iodine charge-transfer (CT) complexes in solutions for several solvents. The magnitude and sign changes of nonlinear absorption of iodine CT complexes for different solvents as donors are observed, which implies the transition from saturable absorption to reverse saturable absorption. Compared with C60 in toluene, iodine CT complexes with toluene and benzene demonstrate larger reverse saturable absorption. Meanwhile, flexible alteration of optical nonlinearities is easily obtained by adjusting the mixture ratio of different solvents, yielding a prospective means of constructing a nonlinear medium with a large nonlinear coefficient.


Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors.
Differentiated cells can be reprogrammed to an embryonic-like state by transfer of nuclear contents into oocytes or by fusion with embryonic stem (ES) cells. Little is known about factors that induce this reprogramming. Here, we demonstrate induction of pluripotent stem cells from mouse embryonic or adult fibroblasts by introducing four factors, Oct3/4, Sox2, c-Myc, and Klf4, under ES cell culture conditions. Unexpectedly, Nanog was dispensable. These cells, which we designated iPS (induced pluripotent stem) cells, exhibit the morphology and growth properties of ES cells and express ES cell marker genes. Subcutaneous transplantation of iPS cells into nude mice resulted in tumors containing a variety of tissues from all three germ layers. Following injection into blastocysts, iPS cells contributed to mouse embryonic development. These data demonstrate that pluripotent stem cells can be directly generated from fibroblast cultures by the addition of only a few defined factors.


Effect of relative humidity on the migration of benzophenone from paperboard into the food simulant Tenax® and modelling hereof.
The aim of this study was to investigate the effect of relative humidity on the migration of benzophenone from paperboard into the food simulant Tenax®. Kinetic migration investigations were carried out with three relative humidities in the interval between 39% and >73%. All investigations were carried out at a constant temperature of 34°C. It was found that the migration of benzophenone after more than 30 days was 4.8 times higher at a relative humidity of 64%-71%, and 7.3 times higher at a relative humidity of >73%, compared with that at a relative humidity of 39%-49%. Diffusion and partition coefficients were derived from the results by using a software for modelling migration in multilayer materials. Both the diffusion coefficient and the partition coefficient, between paperboard and Tenax®, decrease with increasing relative humidity. The experimental results were correctly modelled only when the paperboard was regarded as a one-layer system as compared with a two-layer system: where the main part of the paperboard (B1) has a high diffusion rate and a thin part of the paperboard (B2) in contact with the foodstuff has a lower diffusion rate.


Sequence specificity of pausing by DNA polymerases.
We have constructed recombinant M13 DNA templates containing stretches of oligo (purines) and oligo (pyrimidines). Each of these inserts hinders the advancement of the large fragment of E. coli Pol I during DNA synthesis. The pattern of blockage is independent of changes in KCl or Mg2+ concentrations and pausing is moderately alleviated at lower pH. Blockage is not affected by either the concentration of template or by the position of the DNA primer. The pattern of pause sites is similar for calf thymus DNA polymerase-alpha, implying that replicative barriers are determined by the structure of the DNA at its growing point. There is a lack of correlation between the position of pause sites with different inserts and known alternate DNA structures. Thus, the homo-oligomeric inserts may possess a different structure when complexed with DNA polymerase. This concept accounts for the appearance of unique new upstream and downstream pause sites that result from the insertion of each oligonucleotide.


Children with high and intermediate imperforate anus: their experiences of hospital care.
The purpose of this study was to examine the experiences of children with high and intermediate imperforate anus (IA), and specifically their experiences of hospital care. Twenty-five children born with high and intermediate IA participated; 9 boys and 16 girls. The mean age was 10.5 years (range 8.0-13.6). Two control groups were involved in the study. A self-report questionnaire was used to gather the data concerning children's experiences of hospital care. Items were scored on a 5-point Likert scale. The children's responses on hospital care items scored high. The children with IA reported being less satisfied with the information given, and understood less why they needed to visit the hospital than did the children in the two control groups. The children's experiences of care seemed to be positive even though the children born with IA are subjected to invasive treatment. More research is called for in the unexplored area of information to the children, and particularly to the children born with a defect. Children's views are important and should always be considered, as their answers most certainly reflect a genuine experience, contributing to the further development of their specific care.


Examining trends in family planning among harder-to-reach women in Senegal 1992-2014.
Recent increases in family planning (FP) use have been reported among women of reproductive age in union (WRAU) in Senegal. However, trends have not been monitored among harder-to-reach groups (including adolescents, unmarried and rural poor women), key to understanding whether FP progress is equitable. We combined data from six Demographic and Health Surveys conducted in Senegal between 1992/93 and 2014. We examined FP trends over time among WRAU and subgroups, and trends in knowledge of FP and intention to use among women with unmet need for FP. Our results show that percent demand satisfied is lower among rural poor women and adolescents than WRAU, although higher among unmarried women. Marked recent increases have been observed in all subgroups, however fewer than 50% of women in need of FP use modern contraception in Senegal. Knowledge of FP has risen steadily among women with unmet need; however, intention to use FP has remained stable at around 40% since 2005 for all groups except unmarried women (75% of whom intend to use). Significant progress in meeting the need for FP has been achieved in Senegal, but more needs to be done particularly to improve acceptability of FP, and to strategically target interventions toward adolescents and rural poor women.


Maf1 ameliorates cardiac hypertrophy by inhibiting RNA polymerase III through ERK1/2.
Rationale: An imbalance between protein synthesis and degradation is one of the mechanisms of cardiac hypertrophy. Increased transcription in cardiomyocytes can lead to excessive protein synthesis and cardiac hypertrophy. Maf1 is an RNA polymerase III (RNA pol III) inhibitor that plays a pivotal role in regulating transcription. However, whether Maf1 regulates of cardiac hypertrophy remains unclear. Methods: Cardiac hypertrophy was induced in vivo by thoracic aortic banding (AB) surgery. Both the in vivo and in vitro gain- and loss-of-function experiments by Maf1 knockout (KO) mice and adenoviral transfection were used to verify the role of Maf1 in cardiac hypertrophy. RNA pol III and ERK1/2 inhibitor were utilized to identify the effects of RNA pol III and ERK1/2. The possible interaction between Maf1 and ERK1/2 was clarified by immunoprecipitation (IP) analysis. Results: Four weeks after surgery, Maf1 KO mice exhibited significantly exacerbated AB-induced cardiac hypertrophy characterized by increased heart size, cardiomyocyte surface area, and atrial natriuretic peptide (ANP) expression and by exacerbated pulmonary edema. Also, the deficiency of Maf1 causes more severe cardiac dilation and dysfunction than wild type (WT) mice after pressure overload. In contrast, compared with adenoviral-GFP injected mice, mice injected with adenoviral-Maf1 showed significantly ameliorated AB-induced cardiac hypertrophy. In vitro study has demonstrated that Maf1 could significantly block phenylephrine (PE)-induced cardiomyocyte hypertrophy by inhibiting RNA pol III transcription. However, application of an RNA pol III inhibitor markedly improved Maf1 knockdown-promoted cardiac hypertrophy. Moreover, ERK1/2 was identified as a regulator of RNA pol III, and ERK1/2 inhibition by U0126 significantly repressed Maf1 knockdown-promoted cardiac hypertrophy accompanied by suppressed RNA pol III transcription. Additionally, IP analysis demonstrated that Maf1 could directly bind ERK1/2, suggesting Maf1 could interact with ERK1/2 and then inhibit RNA pol III transcription so as to attenuate the development of cardiac hypertrophy. Conclusions: Maf1 ameliorates PE- and AB-induced cardiac hypertrophy by inhibiting RNA pol III transcription via ERK1/2 signaling suppression.


What is the heartland of psychiatry?
Psychiatry has long identified schizophrenia as its defining disorder, its heartland as it has been called. In the past 20 years, this has had a number of negative consequences for psychiatry as a medical specialty, which result from the uncertainty of diagnosis and an increasing emphasis on demedicalising services in an attempt to provide social care outside hospital. These changes have probably increased the stigma attached to psychiatric practice and threaten to deskill doctors. They have also meant that services for other disorders do not meet the needs of patients. To continue to allow schizophrenia to be the paradigm condition is against the interests of psychiatrists and their patients.


Primary vertebral tumors: a review of epidemiologic, histological, and imaging findings, Part I: benign tumors.
Primary vertebral tumors, although less common than metastases to the spine, make up a heterogeneous group of neoplasms that can pose diagnostic and treatment challenges. They affect both the adult and the pediatric population and may be benign, locally aggressive, or malignant. An understanding of typical imaging findings will aid in accurate diagnosis and help neurosurgeons appreciate anatomic subtleties that may increase their effective resection. An understanding of the histological similarities and differences between these tumors is imperative for all members of the clinical team caring for these patients. In this first review of 2 parts, we discuss the epidemiological, histological, and imaging features of the most common benign primary vertebral tumors-aneurysmal bone cyst, chondroma and enchondroma, hemangioma, osteoid osteoma, and osteoblastoma-and lesions related to eosinophilic granuloma and fibrous dysplasia. In addition, we discuss the basic management paradigms for each of these diagnoses. In combination with part II of the review, which focuses on locally aggressive and malignant tumors, this article provides a comprehensive review of primary vertebral tumors.


Region and Insurance Plan Type Influence Discharge Disposition After Hip and Knee Arthroplasty: Evidence From the Privately Insured US Population.
Little is known about regional variation in the use of postacute care services after elective procedures, such as total hip or knee arthroplasty (THA/TKA), and how insurance type may influence it. The goal of this study is to assess the influence of region and insurance arrangements on discharge disposition. A representative sample of the privately insured US population with THA or TKA in 2009 or 2010 was obtained from the MarketScan database applying individual-level weights from the Medical Expenditure Panel Survey. Multivariate logistic regression was used to predict the odds of being discharged to an extended care facility (ECF) compared with being discharged home. The model adjusted for region, insurance plan type, sociodemographic characteristics, comorbidities, and length of stay. Large variability was observed in ECF use across the US. Patients in the Northeast were 2.5 times more likely to receive care at an ECF compared with patients in the South (odds ratio [OR] = 2.51, 95% confidence interval [CI]: 1.97-3.19). Enrollees in noncapitated plans such as fee-for-service plans or exclusive provider organizations were less likely to be discharged to an ECF compared with health maintenance organizations/preferred provider organizations with capitation enrollees (OR = 0.74, 95% CI: 0.57-0.94; OR = 0.49, 95% CI: 0.34-0.74, respectively). Region and private insurance plan arrangements are related to extended care use among THA and TKA patients. Understanding regional variation in discharge disposition provides policy makers with important information as to where to focus new tests of hip and knee procedures such as same day arthroplasty.


Neutralisation of the anti-coagulant effects of heparin by histones in blood plasma and purified systems.
Neutrophil extracellular traps (NETs) composed primarily of DNA and histones are a link between infection, inflammation and coagulation. NETs promote coagulation and approaches to destabilise NETs have been explored to reduce thrombosis and treat sepsis. Heparinoids bind histones and we report quantitative studies in plasma and purified systems to better understand physiological consequences. Unfractionated heparin (UFH) was investigated by activated partial thromboplastin time (APTT) and alongside low-molecular-weight heparins (LMWH) in purified systems with thrombin or factor Xa (FXa) and antithrombin (AT) to measure the sensitivity of UFH or LMWH to histones. A method was developed to assess the effectiveness of DNA and non-anticoagulant heparinoids as anti-histones. Histones effectively neutralised UFH, the IC50 value for neutralisation of 0.2 IU/ml UFH was 1.8 µg/ml histones in APTT and 4.6 µg/ml against 0.6 IU/ml UFH in a purified system. Histones also inhibited the activities of LMWHs with thrombin (IC50 6.1 and 11.0 µg/ml histones, for different LMWHs) or FXa (IC50 7.8 and 7.0 µg/ml histones). Direct interactions of UFH and LMWH with DNA and histones were explored by surface plasmon resonance, while rheology studies showed complex effects of histones, UFH and LMWH on clot resilience. A conclusion from these studies is that anticoagulation by UFH and LMWH will be compromised by high affinity binding to circulating histones even in the presence of DNA. A complete understanding of the effects of histones, DNA and heparins on the haemostatic system must include an appreciation of direct effects on fibrin and clot structure.


[The neurobiological dimension of meditation--results from neuroimaging studies].
Meditation in general can be understood as a state of complete and unintentional silent and motionless concentration on an activity, an item or an idea. Subjectively, meditative experience is said to be fundamentally different from "normal" mental states and is characterized by terms like timelessness, boundlessness and lack of self-experience. In recent years, several fMRI- and PET-studies about meditation which are presented in this paper have been published. Due to different methods, especially different meditation types, the results are hardly comparable. Nevertheless, the data suggest the hypothesis of a "special" neural activity during meditative states being different from that during calm alertness. Main findings were increased activation in frontal, prefrontal and cingulate areas which may represent the mental state of altered self-experience. In the present studies, a considerable lack of scientific standards has to be stated making it of just casuistic value. Today's improved neurobiological examination methods - especially neuroimaging techniques - may contribute to enlighten the phenomenon of qualitatively different states of consciousness.


Description of seven candidate species affiliated with the phylum Actinobacteria, representing planktonic freshwater bacteria.
Actinobacteria comprise a substantial fraction of the bacterioplankton in freshwater lakes and streams. Numerous cultivation-independent investigations have retrieved actinobacterial 16S rRNA gene sequences from such habitats. The taxa detected in freshwater habitats are usually absent from terrestrial and marine systems. So far, none of the indigenous freshwater lineages is represented by a taxon with a validly published name. The seven organisms for which Candidatus status is described here were isolated from freshwater lakes and ponds located in tropical, subtropical and temperate climatic zones. Phylogenetic analyses demonstrated that they are affiliated with one of the actinobacterial lineages indigenous to freshwater bacterioplankton. The seven novel taxa could only be cultivated to date as mixed cultures that also contain non-actinobacterial strains. Due to the lack of pure cultures, I propose to establish the candidate species 'Candidatus Planktoluna difficilis', 'Candidatus Aquiluna rubra', 'Candidatus Flaviluna lacus', 'Candidatus Rhodoluna limnophila', 'Candidatus Rhodoluna planktonica', 'Candidatus Rhodoluna lacicola' and 'Candidatus Limnoluna rubra' for these taxa.


Superconductivity in diamond.
Diamond is an electrical insulator well known for its exceptional hardness. It also conducts heat even more effectively than copper, and can withstand very high electric fields. With these physical properties, diamond is attractive for electronic applications, particularly when charge carriers are introduced (by chemical doping) into the system. Boron has one less electron than carbon and, because of its small atomic radius, boron is relatively easily incorporated into diamond; as boron acts as a charge acceptor, the resulting diamond is effectively hole-doped. Here we report the discovery of superconductivity in boron-doped diamond synthesized at high pressure (nearly 100,000 atmospheres) and temperature (2,500-2,800 K). Electrical resistivity, magnetic susceptibility, specific heat and field-dependent resistance measurements show that boron-doped diamond is a bulk, type-II superconductor below the superconducting transition temperature T(c) approximately 4 K; superconductivity survives in a magnetic field up to Hc2(0) > or = 3.5 T. The discovery of superconductivity in diamond-structured carbon suggests that Si and Ge, which also form in the diamond structure, may similarly exhibit superconductivity under the appropriate conditions.


Depression in older people: visual impairment and subjective ratings of health.
The aim of this study was to establish the prevalence of depression in a sample of older adults with impaired vision and investigate associations between physical and visual disability and depression. We analyzed cross-sectional baseline data from 391 participants aged >or=75 years with visual acuity of 6/24 (20/80) or less, recruited for a randomized controlled trial of interventions to prevent falls (the VIP trial). Measures included the geriatric depression scale (GDS-15), the state-trait anxiety index, activities of daily living (Nottingham extended ADL scale), physical activity (human activity profile), an index of visual functioning (VF-14), health-related quality of life (SF-36), objective measures of physical ability, and a measure of visual acuity. Regression models were developed to investigate the association between depression scores and physical, psychological, and visual disability. About 29.4% (115 of 391) of participants were identified as potentially depressed (GDS-15 score >4). Physical function, physical activity, physical ability, visual function, anxiety, and self-reported physical and mental health were significantly worse for those with depressive symptomatology. Physical, visual, and psychological factors collectively explained 41% of the variance in the depression score in a linear regression model (R=0.421, adjusted R=0.410, F (7,382)=39.680, p<0.001). Depression was not related to age, gender, living situation, ethnicity, or number of prescription or antidepressant medications taken. Depression was common in this population of older adults with severe visual impairment. Impaired visual and physical functions were associated with symptoms of depression. The effect of visual disability was independent of the effect of physical disability. The strength of this relationship, and the results of the regression analyses, indicate that a person who is visually or physically disabled is more likely to suffer from depression.


The effect of a myocardial infarction on the normalized time-varying elastance curve.
It has been suggested that the shape of the normalized time-varying elastance curve [E(n)(t(n))] is conserved in different cardiac pathologies. We hypothesize, however, that the E(n)(t(n)) differs quantitatively after myocardial infarction (MI). Sprague-Dawley rats (n = 9) were anesthetized, and the left anterior descending coronary artery was ligated to provoke the MI. A sham-operated control group (CTRL) (n = 10) was treated without the MI. Two months later, a conductance catheter was inserted into the left ventricle (LV). The LV pressure and volume were measured and the E(n)(t(n)) derived. Slopes of E(n)(t(n)) during the preejection period (alpha(PEP)), ejection period (alpha(EP)), and their ratio (beta = alpha(EP)/alpha(PEP)) were calculated, together with the characteristic decay time during isovolumic relaxation (tau) and the normalized elastance at end diastole (E(min)(n)). MI provoked significant LV chamber dilatation, thus a loss in cardiac output (-33%), ejection fraction (-40%), and stroke volume (-30%) (P < 0.05). Also, it caused significant calcium increase (17-fold), fibrosis (2-fold), and LV hypertrophy. End-systolic elastance dropped from 0.66 +/- 0.31 mmHg/microl (CTRL) to 0.34 +/- 0.11 mmHg/microl (MI) (P < 0.05). Normalized elastance was significantly reduced in the MI group during the preejection, ejection, and diastolic periods (P < 0.05). The slope of E(n)(t(n)) during the alpha(PEP) and beta were significantly altered after MI (P < 0.05). Furthermore, tau and end-diastolic E(min)(n) were both significantly augmented in the MI group. We conclude that the E(n)(t(n)) differs quantitatively in all phases of the heart cycle, between normal and hearts post-MI. This should be considered when utilizing the single-beat concept.


Evaluation of antibiotic-loaded calcium phosphate bone cement in an cranium-infected experimental model.
Treatment of calvarial defects has remained a challenge in reconstruction surgery, especially because of infection at these sites. We produced a bactericidal biomaterial for treating infected bone defects by using calcium phosphate bone cement mixed with antibiotics. We evaluated the usefulness of this material mixed with the antibiotic vancomycin in a cranium-infected rat model. The concentration of vancomycin used was 5.0 wt%, as reported in our previous study. In order to establish the rat model, a cranium defect (diameter, 5 mm) was made that was infected with methicillin-resistant Staphylococcus aureus (MRSA). Thirty-six rats were divided into 6 groups depending on whether an autologous graft or bone cement with or without antibiotic was used for the defect. After 1 and 4 weeks, abscess formation was checked, tissue bacterial counts were determined, and pathological examination was performed. At both 1 and 4 weeks, no MRSA was detected on tissue bacterial culture or pathological examination in groups that received bone cement with antibiotics. In groups that received bone cement without antibiotic, MRSA was detected, and the bone cement had compromised and disintegrated into several slices. In conclusion, bone cement that contains antibiotics appears to be effective not only for reconstruction in cases of cranial defect, but also in terms of preventing infection.


Environmental contaminants in freshwater fish and their risk to piscivorous wildlife based on a national monitoring program.
Organochlorine chemical residues and elemental concentrations were measured in piscivorous and benthivorous fish at 111 sites from large U.S. river basins. Potential contaminant sources such as urban and agricultural runoff, industrial discharges, mine drainage, and irrigation varied among the sampling sites. Our objectives were to provide summary statistics for chemical contaminants and to determine if contaminant concentrations in the fish were a risk to wildlife that forage at these sites. Concentrations of dieldrin, total DDT, total PCBs, toxaphene, TCDD-EQ, cadmium, chromium, mercury, lead, selenium, and zinc exceeded toxicity thresholds to protect fish and piscivorous wildlife in samples from at least one site; most exceedences were for total PCBs, mercury, and zinc. Chemical concentrations in fish from the Mississippi River Basin exceeded the greatest number of toxicity thresholds. Screening level wildlife risk analysis models were developed for bald eagle and mink using no adverse effect levels (NOAELs), which were derived from adult dietary exposure or tissue concentration studies and based primarily on reproductive endpoints. No effect hazard concentrations (NEHC) were calculated by comparing the NOAEL to the food ingestion rate (dietary-based NOAEL) or biomagnification factor (tissue-based NOAEL) of each receptor. Piscivorous wildlife may be at risk from a contaminant if the measured concentration in fish exceeds the NEHC. Concentrations of most organochlorine residues and elemental contaminants represented no to low risk to bald eagle and mink at most sites. The risk associated with pentachloroanisole, aldrin, Dacthal, methoxychlor, mirex, and toxaphene was unknown because NOAELs for these contaminants were not available for bald eagle or mink. Risk differed among modeled species and sites. Our screening level analysis indicates that the greatest risk to piscivorous wildlife was from total DDT, total PCBs, TCDD-EQ, mercury, and selenium. Bald eagles were at greater risk to total DDT and total PCBs than mink, whereas risks of TCDD-EQ, mercury, and selenium were greater to mink than bald eagle.


Dietary n-3 polyunsaturated fatty acids, fish intake and healthy ageing.
Seafood consumption and dietary intake of n-3 polyunsaturated fatty acids (PUFA) protect from cardiovascular disease, muscle wasting and mortality; however, their role in delaying unhealthy ageing is uncertain. A prospective cohort study with 1592 community-dwelling individuals aged ≥60 years recruited in 2008-2010, and followed through 2015. Intake of seafood and n-3 PUFA [eicosapentanoic acid (EPA) and docosahexanoic acid (DHA)] was estimated using a validated diet history and food composition tables. Unhealthy ageing was assessed across the domains of physical and cognitive function, mental health, morbidity, self-rated health and vitality. Higher blue fish consumption was negatively associated with unhealthy ageing in 2015 [multivariate ß (95% confidence interval) per interquartile range (IQR) increase of fish: -0.49 (-0.90; -0.08)] and, specifically, was associated with lower accumulation of functional impairments [-1.00 (-1.71; -0.28)] and morbidities [-0.30 (-0.73; 0.13)]. Individuals with high intakes of EPA (≥0.47 g/day) and DHA (≥0.89 g/day) presented the highest reductions in age-related deficits accumulation: -1.61 (-3.01; -0.22) and -1.34 (-2.76; 0.08), respectively. Intake of EPA and DHA were negatively associated with the accumulation of functional deficits [results per IQR increase: -0.79 (-1.64; -0.06) and -0.84 (-1.67; -0.01), respectively] and morbidities [-0.60 (-1.10; -1.11) and -0.60 (-1.09; -0.11)]. No associations were observed between n-3 PUFA and self-rated or mental health, or between white fish and unhealthy ageing. In this cohort of Spanish individuals with elevated intake of fish and a high adherence to the Mediterranean diet, consumption of blue fish and n-3 PUFA had a prospective protective association with deficits accumulation.


Caries-preventive effect of glass ionomer and resin-based fissure sealants on permanent teeth: An update of systematic review evidence.
This article constitutes a partial update of the original systematic review evidence by Yengopal et al. from 15 January 2008 (published in the Journal of Oral Science in 2009) with primary focus on research quality in regard to bias risk in trials. Its aim is to update the existing systematic review evidence from the English literature as to whether caries occurrence on pits and fissures of teeth sealed with either GIC or resin is the same. In addition to the 12 trials included during the original systematic review, 5 new trials were identified during the database search (up to 26 August 2010) and 2 further trials were included from a hand search and reference check. Of these, 3 trials were excluded and 16 were accepted for data extraction and quality assessment. The quality of accepted trials was assessed, using updated quality criteria, and the risk of bias was investigated in more depth than previously reported. In addition, the focus of quantitative synthesis was shifted to single datasets that were extracted from the accepted trials. Twenty-six dichotomous and 4 continuous datasets were extracted. Meta-analysis and cumulative meta-analysis were used in combining clinically homogenous datasets. The overall outcome of the computed datasets suggest no difference between the caries-preventive effects of GIC- and resin-based fissure sealants. This overall outcome is in agreement with the conclusions of the original systematic review. Although the findings of the trials identified in this update may be considered to be less affected by attrition- and publication bias, their risk of selection- and detection-/performance bias is high. Thus, verification of the currently available results requires further high quality randomised control trials.


Extended V-Y latissimus dorsi musculocutaneous flap for anterior chest wall reconstruction.
Patients presenting advanced breast tumors are usually subject to major resections of the anterior chest wall tissue. Flaps taken from the abdominal wall, such as the TRAM, the external oblique flap, and the thoracoabdominal flap are frequently used for closure of this type of lesion. In this study, a different shape was planned for the skin island from the latissimus dorsi musculocutaneous flap with primary closure in V-Y for the correction of major lesions in the anterior chest wall after mastectomies occasioned by advanced breast cancer. The technique was used on eight female patients, between November of 1998 and July of 1999, victims of advanced breast cancer, who had been submitted to radical mastectomies with major resections of the cutaneous tegument. It was possible to make primary closure of lesions in the anterior chest wall, the preoperative areas of which varied between 15 x 15 and 29 x 14 cm (vertical x horizontal). This technical variant permitted use of the flap without the need to create tunnels for its advancement and rotation. It also proved to be easy to perform and presented a low morbidity rate, with three patients presenting minor complications that did not require correction through any further surgical intervention. Closure was obtained in the donor and recipient sites without the use of skin grafts or other more major procedures. According to the authors, this procedure is a viable alternative in repairing large defects in the anterior chest wall.


Ultrastructure of the Head of Okranema eileenae Greenslade and Nicholas, 1991 (Thoracostomopsidae: Nematoda).
Okranema eileenae is a marine nematode from Australian sandy beaches. The structure of its cephalic region is described by light, scanning, and transmission electron microscopy. Three large lips, separated by three deep clefts and surmounted by flexible liplets, surround the mouth. Transverse and longitudinal sections of the head have been used to investigate the ultrastructure of the lips, buccal cavity, mandibles, and cephalic sensilla. The pharyngeal muscles are attached to the external head cuticle over a broad band, forming the cephalic capsule, which terminates in the three connected cuticular cephalic arches-one in each lip. Three mandibles form the central core of each lip, cuticular structures from which two small teeth-denticles and one large central tooth, the onchium-project into the buccal cavity. The onchia are anterior extensions of the pharynx that contain the ducts of pharyngeal glands that discharge into the buccal cavity. Epidermal tissue extends anteriorly as far as the cephalic arch. Cephalic structures, apart from sensory setae, are formed from an expanded cephalic cuticle.The inner labial, outer labial, and cephalic setae each contain two dendritic processes. Aspects of amphidial structure are described for the first time in Thoracostomopsidae. The external apertures are illustrated by scanning electron microscopy and the internal structure in sections by transmission electron microscopy. Two bundles of about 100 dendritic processes are enclosed by the amphidial sheath cell, as well as a group of four other dendritic processes. Two amphidial duct cells are present on each side of the head, but without containing dendritic processes. However, the ultrastructural description of the amphids is incomplete.


Revisiting the link between breeding effort and oxidative balance through field evaluation of two sympatric sibling insect species.
The idea that oxidative stress could be a major force governing evolutionary trade-offs has recently been challenged by experimental approaches in laboratory conditions, triggering extensive debates centered on theoretical and methodological issues. Here, we revisited the link between oxidative stress and reproduction by measuring multiple antioxidant and oxidative damages in wild-caught females of two sibling weevil species (Curculio elephas, C. glandium). The strength of our study arised from (1) studied species that were sympatric and exploited similar resource, but displayed contrasting reproductive strategies and (2) individuals were sampled throughout adult life so as to relate oxidative status to breeding effort. We found that the short-lived C. elephas sacrifices red-ox homeostasis for immediate reproduction upon emergence as characterized by low antioxidant defenses and elevated oxidative damage. Comparatively, C. glandium massively invests in antioxidant and maintains low oxidative damage, which may contribute to their extended prereproductive period. Intriguingly, we also reveal, for the first time in a field study, an unexpected reactivation of antioxidant defenses with the onset of reproduction. Our results thus support the existence of a strong, but complex relationship between oxidative stress and life-history evolution and highlight the need for a finer-scale picture of antioxidant strategies.


Plant defence suppression is mediated by a fungal sirtuin during rice infection by Magnaporthe oryzae.
Crop destruction by the hemibiotrophic rice pathogen Magnaporthe oryzae requires plant defence suppression to facilitate extensive biotrophic growth in host cells before the onset of necrosis. How this is achieved at the genetic level is not well understood. Here, we report that a M. oryzae sirtuin, MoSir2, plays an essential role in rice defence suppression and colonization by controlling superoxide dismutase (SOD) gene expression. Loss of MoSir2 function in Δsir2 strains did not affect appressorial function, but biotrophic growth in rice cells was attenuated. Compared to wild type, Δsir2 strains failed to neutralize plant-derived reactive oxygen species (ROS) and elicited robust defence responses in rice epidermal cells that included elevated pathogenesis-related gene expression and granular depositions. Deletion of a SOD-encoding gene under MoSir2 control generated Δsod1 deletion strains that mimicked Δsir2 for impaired rice defence suppression, confirming SOD activity as a downstream output of MoSir2. In addition, comparative protein acetylation studies and forward genetic analyses identified a JmjC domain-containing protein as a likely target of MoSir2, and a Δsir2 Δjmjc double mutant was restored for MoSOD1 expression and defence suppression in rice epidermal cells. Together, this work reveals MoSir2 and MoJmjC as novel regulators of early rice cell infection.


Utilizing afterglow magnetization from cross-polarization magic-angle-spinning solid-state NMR spectroscopy to obtain simultaneous heteronuclear multidimensional spectra.
The time required for data acquisition and subsequent spectral assignment are limiting factors for determining biomolecular structure and dynamics using solid-state NMR spectroscopy. While strong magnetic dipolar couplings give rise to relatively broad spectra lines, the couplings also mediate the coherent magnetization transfer via the Hartmann-Hahn cross-polarization (HH-CP) experiment. This mechanism is used in nearly all backbone assignment experiments for carrying out polarization transfer between (1)H, (15)N, and (13)C. In this Article, we describe a general spectroscopic approach to use the residual or "afterglow" magnetization from the (15)N to (13)C selective HH-CP experiment to collect a second multidimensional heteronuclear data set. This approach allowed for the collection of two commonly used sequential assignment experiments (2D NCA and NCO or 3D NCACX and NCOCX) at the same time. Our "afterglow" technique was demonstrated with uniformly [(13)C,(15)N] and [1,3-(13)C] glycerol-labeled ubiquitin using instrumentation available on all standard solid-state NMR spectrometers configured for magic-angle-spinning. This method is compatible with several other sensitivity enhancement experiments and can be used as an isotopic filtering tool to reduce the spectral complexity and decrease the time needed for assignment.


Stroke risk in siblings with sickle cell anemia.
Cerebrovascular disease is a common cause of morbidity in sickle cell anemia (HbSS): approximately 10% of patients have a clinical stroke before 20 years of age, and another 22% have silent infarction on magnetic resonance imaging. The phenotypic variation among patients with HbSS suggests a role for modifier genes and/or environmental influences. To assess the familial component of clinical stroke in HbSS, we estimated the prevalence of clinical stroke among all patients and among HbSS sibling pairs at 9 pediatric centers. The sample included 3425 patients with sickle cell disease who were younger than 21 years, including 2353 patients with HbSS. The stroke prevalence was 4.9% for all genotypes; 7.1% for patients with HbSS; 1.1% for patients with HbSbeta(o) thalassemia; 0.6% for patients with Sbeta(+) thalassemia; and 0% for patients with HbSC. In 207 sibships, more than 1 child had HbSS. There were 42 sibships in which at least 1 sibling had a stroke, and in 10 of the 42, 2 siblings had a stroke. A permutation test indicated that the number of families in which 2 children had strokes was larger than the number expected if strokes were randomly distributed among children in sibships (P =.0012). There was no difference in stroke prevalence based on sex, nor was the mean age at stroke presentation significantly different between singletons and sibships with stroke. We conclude that there is a familial predisposition to stroke in HbSS. Attempts to identify genetic modifiers should be initiated with family-based studies.


Optimizing renal replacement therapy in older adults: a framework for making individualized decisions.
It is often difficult to synthesize information about the risks and benefits of recommended management strategies in older patients with end-stage renal disease since they may have more comorbidity and lower life expectancy than patients described in clinical trials or practice guidelines. In this review, we outline a framework for individualizing end-stage renal disease management decisions in older patients. The framework considers three factors: life expectancy, the risks and benefits of competing treatment strategies, and patient preferences. We illustrate the use of this framework by applying it to three key end-stage renal disease decisions in older patients with varying life expectancy: choice of dialysis modality, choice of vascular access for hemodialysis, and referral for kidney transplantation. In several instances, this approach might provide support for treatment decisions that directly contradict available practice guidelines, illustrating circumstances when strict application of guidelines may be inappropriate for certain patients. By combining quantitative estimates of benefits and harms with qualitative assessments of patient preferences, clinicians may be better able to tailor treatment recommendations to individual older patients, thereby improving the overall quality of end-stage renal disease care.


MacroD1 Is a Promiscuous ADP-Ribosyl Hydrolase Localized to Mitochondria.
MacroD1 is a macrodomain containing protein that has mono-ADP-ribose hydrolase enzymatic activity toward several ADP-ribose adducts. Dysregulation of MacroD1 expression has been shown to be associated with the pathogenesis of several forms of cancer. To date, the physiological functions and sub-cellular localization of MacroD1 are unclear. Previous studies have described nuclear and cytosolic functions of MacroD1. However, in this study we show that endogenous MacroD1 protein is highly enriched within mitochondria. We also show that MacroD1 is highly expressed in human and mouse skeletal muscle. Furthermore, we show that MacroD1 can efficiently remove ADP-ribose from 5' and 3'-phosphorylated double stranded DNA adducts in vitro. Overall, we have shown that MacroD1 is a mitochondrial protein with promiscuous enzymatic activity that can target the ester bonds of ADP-ribosylated phosphorylated double-stranded DNA ends. These findings have exciting implications for MacroD1 and ADP-ribosylation within the regulation of mitochondrial function and DNA-damage in vivo.


Protonation-modulated localization of excess electrons in histidine aqueous solutions revealed by ab initio molecular dynamics simulations: anion-centered versus cation-centered localization.
In this work, we present an ab initio molecular dynamics simulation study on the interaction of an excess electron (EE) with histidine in its aqueous solution. Two different configurations of histidine (imidazole group protonated or not) are considered to reflect its different existing forms in neutral or slightly acidic surroundings. The simulation results indicate that localizations of EEs in different aqueous histidine solutions are quite different and are strongly affected by protonation of the side chain imidazole group and are thus pH-controlled. In neutral aqueous histidine solution, an EE localizes onto the carboxyl anionic group of the amino acid backbone after a relatively lengthy diffuse state, performing just like in an aliphatic amino acid solution. But in weakly acidic solution in which the side chain imidazole group is protonated, an EE undergoes a short lifetime diffuse state and finally localizes on the protonated imidazole group. We carefully examine these two different localization dynamics processes and analyze the competition between different dominating groups in their corresponding electron localization mechanisms. To explain the difference, we investigate the frontier molecular orbitals of these two systems and find that their energy levels and compositions are important to determine these differences. These findings can provide helpful information to understand the interaction mechanisms of low energy EEs with amino acids and even oligopeptides, especially with aromatic rings.


The Carotene and Retinol Efficacy Trial (CARET) to prevent lung cancer in high-risk populations: pilot study with asbestos-exposed workers.
Pilot studies are an essential component for major chemoprevention trials. Prior to initiating the multicenter Carotene and Retinol Efficacy Trial to assess the effectiveness of beta-carotene and retinol for preventing lung cancer, we conducted pilot studies in Seattle between 1985 and 1988 in two high risk populations: current and former heavy smokers and asbestos-exposed workers. The Asbestos Workers Pilot Study for the Carotene and Retinol Efficacy Trial demonstrated that recruitment of asbestos-exposed participants with relevant risk factors was feasible from identified sources. We documented negligible toxicity and high adherence with the protocol, schedule, and intervention. Results from the pilot led to extension of the placebo run-in period, changes in the eligibility criteria, improvements in recruitment strategies and scheduling, elimination of stratification by risk factors in randomization, modifications of study vitamin dosage and of side effects monitoring, and refinement of trial design parameters for Carotene and Retinol Efficacy Trial. The Smokers Pilot is reported in the accompanying article (G. E. Goodman et al., Cancer Epidemiol., Biomarkers & Prev., 2: 389-396, 1993).


SU-E-T-322: A Dosimetric Comparison of PBI Brachytherapy Techniques: SAVI, Contura, and Tube and Button Applicators.
To evaluate the dosimetry of partial breast irradiation brachytherapy techniques using the Strut Adjusted Volume Implant (SAVI), Contura, and Tube and Button (T&B) applicators. A total of 51 breast-cancer patients (23 SAVI, 6 Contura, and 22 T&B) were treated. The target was delineated following NSABP B-39 guidelines. 3D plans were optimized using the Inverse Planning Simulated Annealing algorithm to deliver 3.4 Gy per fraction to the target and minimize dose to organs at risk (OARs). Graphical optimization was then used to fine tune the final dose distribution. The minimum cavity-to-skin distance was measured. Target coverage (V90 and V95) and maximum dose (D0.1cc) to the OARs were evaluated. Dose homogeneity index (DHI = 1-V150/V100) was calculated. The average cavity-to-skin distances were 4.1 mm (0.5-9.6 mm, SAVI) and 11.7 mm (7.1-15.4 mm, Contura). The target-to-skin distance for the T&B cases was 8.7 mm (5.0-13.7 mm). The average V90 and V95 to the target were 96.8% and 94.5% (SAVI), 97.0% and 93.0% (Contura), 98.6% and 97.3% (T&B). The mean D0.1cc to the skin, ribs, and lung was 91.5%, 58.8%, 44.5% (SAVI), 93.1%, 51.3%, 40.5% (Contura), 69.1%, 41.5%, and 31.9% (T&B). The average V150 and V200 to the normal breast tissue were 30.4 cc and 14.9 cc (SAVI), 29.5 cc and 7.3 cc (Contura), 18.3 cc and 7.1 cc (T&B). The average DHI for the SAVI, Contura, and T&B cases was 0.55 (0.50-0.60), 0.70 (0.63-0.78), and 0.76 (0.74-0.79). All techniques provided clinically acceptable target coverage and dose to the OARs. The SAVI device provided a lower skin dose at close cavity-to-skin distances while providing excellent target coverage. However, the T&B and Contura applicators produced more homogeneous dose distribution (higher DHI) in the target than the SAVI. The correlations between dosimetric properties and follow-up mammogram results are under investigation.


An unusual natural history of a rare bladder tumor: Primitive neuroectodermal tumor.
A primary primitive neuroectodermal tumor (PNET) of the urinary bladder is a very rare bladder tumor. There are few described cases in the literature to date. We presented a 70-year-old man with primary nonmetastatic bladder PNET. The diagnosis was confirmed according to immunohistochemistry evaluation. The patient underwent bladder-sparing protocol using radiotherapy and standard systemic multidrug chemotherapy (vincristine + doxorubicin + cyclophosphamide). At the end of three years of follow-up, we performed radical cystoprostatectomy due to newly diagnosed high-grade transitional cell carcinoma of the bladder at the site far from the primary bladder PNET. In conclusion, multimodal bladder sparing protocol including radiotherapy and standard systemic chemotherapy can be used for the management of primary bladder PNET with acceptable oncological outcomes.


RNA polymerase mutations that facilitate replication progression in the rep uvrD recF mutant lacking two accessory replicative helicases.
We observed that cells lacking Rep and UvrD, two replication accessory helicases, and the recombination protein RecF are cryo-sensitive on rich medium. We isolated five mutations that suppress this Luria-Bertani (LB)-cryo-sensitivity and show that they map in the genes encoding the RNA polymerase subunits RpoB and RpoC. These rpoB (D444G, H447R and N518D) and rpoC mutants (H113R and P451L) were characterized. rpoB(H447R) and rpoB(D444G) prevent activation of the Prrn core promoter in rich medium, but only rpoB(H447R) also suppresses the auxotrophy of a relA spoT mutant (stringent-like phenotype). rpoC(H113R) suppresses the thermo-sensitivity of a greA greB mutant, suggesting that it destabilizes stalled elongation complexes. All mutations but rpoC(P451L) prevent R-loop formation. We propose that these rpo mutations allow replication in the absence of Rep and UvrD by destabilizing RNA Pol upon replication-transcription collisions. In a RecF(+) context, they improve growth of rep uvrD cells only if DinG is present, supporting the hypothesis that Rep, UvrD and DinG facilitate progression of the replication fork across transcribed sequences. They rescue rep uvrD dinG recF cells, indicating that in a recF mutant replication forks arrested by unstable transcription complexes can restart without any of the three known replication accessory helicases Rep, UvrD and DinG.


Polypoid esophageal adenocarcinoma--a clinicopathological variant of esophageal cancer?
Neoplastic polyps are common in the colon, but not elsewhere in the gastrointestinal tract. They are rare at the gastroesophageal junction, especially associated with Barrett's dysplasia or adenocarcinoma. We identify a subset of early esophageal adenocarcinomas that present as symptomatic polyps with gastrointestinal bleeding, reflux, and even dysphagia. Because little is known about polypoid esophageal adenocarcinomas, we examine the clinical presentation, pathological features, and postoperative outcomes of these lesions. We defined esophageal polyps as macroscopic, well-demarcated projections above the mucosa that project the greater percentage of their mass into the lumen without central ulceration. From a consecutive series of 400 patients who underwent esophagectomies for adenocarcinoma (1988 to 2000), 14 (4%) had polyps on presentation. Clinical records, pathology reports, photographs, and original slides of these resected specimens were reviewed. All patients with polyps had early staged esophageal adenocarcinomas. Lymph node involvement was found in only 4 patients, and no patients with distant metastases were found. All but 1 patient (13 of 14; 93%) were symptomatic at diagnosis. Most presented either with gastrointestinal bleeding or gastroesophageal reflux (11 of 14; 79%). Only 2 patients complained of mild dysphagia, and mean weight loss of the series was minimal at 1.3 +/- 4.0 kg (range, 0 to 15 kg). Barrett's was present in 10 patients (71%), and in 80% of these cases, high-grade dysplasia was found. The polyps were small 3.4 +/- 1.1 cm, and most contained poorly differentiated adenocarcinomas. No postoperative mortalities occurred, and follow-up was complete at 100%. Actuarial 2-year survival was 88%, with a mean survival of 61.3 months. Recurrent disease was identified in 4 patients. Polypoid adenocarcinomas of the distal esophagus are a subset of esophageal cancer that produce clear symptoms at an early tumor stage, which allows prompt detection and favorable prognosis. Their pathological features and demographics suggest an origin via the Barrett's esophagus-dysplasia-carcinoma sequence.


Projected spending for brand-name drugs in English primary care given US prices: a cross-sectional study.
To estimate additional spending if NHS England paid the same prices as US Medicare Part D for the 50 single-source brand-name drugs with the highest expenditure in English primary care in 2018. Retrospective analysis of 2018 drug prescribing and spending in the NHS England prescribing data and the Medicare Part D Drug Spending Dashboard and Data. We examined the 50 costliest drugs in English primary care available as brand-name-only in the US and England. We performed cost projections of NHS England spending with US Medicare Part D prices. We estimated average 2018 US rebates as 1 minus the quotient of net divided by gross Medicare Part D spending. England and US. NHS England and US Medicare systems. Total spending, prescriptions and claims in NHS England and Medicare Part D. All spending and cost measures were reported in 2018 British pounds. NHS England spent £1.39 billion on drugs in the cohort. All drugs were more expensive under US Medicare Part D than NHS England. The US-England price ratios ranged from 1.3 to 9.9 (mean ratio 4.8). Accounting for prescribing volume, if NHS England had paid US Medicare Part D prices after adjusting for estimated US rebates, it would have spent 4.6 times as much in 2018 on drugs in the cohort (£6.42 billion). Spending by NHS England would be substantially higher if it paid US Medicare Part D prices. This could result in decreased access to medicines and other health services.


Effect of Soil pH Increase by Biochar on NO, N2O and N2 Production during Denitrification in Acid Soils.
Biochar (BC) application to soil suppresses emission of nitrous- (N2O) and nitric oxide (NO), but the mechanisms are unclear. One of the most prominent features of BC is its alkalizing effect in soils, which may affect denitrification and its product stoichiometry directly or indirectly. We conducted laboratory experiments with anoxic slurries of acid Acrisols from Indonesia and Zambia and two contrasting BCs produced locally from rice husk and cacao shell. Dose-dependent responses of denitrification and gaseous products (NO, N2O and N2) were assessed by high-resolution gas kinetics and related to the alkalizing effect of the BCs. To delineate the pH effect from other BC effects, we removed part of the alkalinity by leaching the BCs with water and acid prior to incubation. Uncharred cacao shell and sodium hydroxide (NaOH) were also included in the study. The untreated BCs suppressed N2O and NO and increased N2 production during denitrification, irrespective of the effect on denitrification rate. The extent of N2O and NO suppression was dose-dependent and increased with the alkalizing effect of the two BC types, which was strongest for cacao shell BC. Acid leaching of BC, which decreased its alkalizing effect, reduced or eliminated the ability of BC to suppress N2O and NO net production. Just like untreated BCs, NaOH reduced net production of N2O and NO while increasing that of N2. This confirms the importance of altered soil pH for denitrification product stoichiometry. Addition of uncharred cacao shell stimulated denitrification strongly due to availability of labile carbon but only minor effects on the product stoichiometry of denitrification were found, in accordance with its modest effect on soil pH. Our study indicates that stimulation of denitrification was mainly due to increases in labile carbon whereas change in product stoichiometry was mainly due to a change in soil pH.


Food contaminant acrylamide increases expression of Cox-2 and nitric oxide synthase in breast epithelial cells.
Acrylamide has been discovered in foods cooked at high temperature. A potentially harmful effect of this dietary component has been suggested by data indicating its association with increased breast cancer. This study investigated the potential effects of acrylamide in nontumorigenic breast cells by assessing expression levels of inducible nitric oxide synthase (iNOS) and cycloogenase-2 (Cox-2) and NOS activity, which are known to be early molecular changes in disease formation. Treatment of cells with acrylamide increased levels of iNOS (both expression and activity) and Cox-2. Its potent metabolite, glycidamide, also induced both iNOS and Cox-2, with induction of iNOS occurring at a lower concentration. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), another food-borne carcinogen, was found to induce Cox-2 expression. Combining acrylamide with PhIP did not result in a further increase. These studies suggest that further research is needed to determine the role of carcinogens formed from cooking foods in inducing early molecular changes associated with breast cancer.


Spontaneous production of anti-IFN-alpha and anti-IL-12 autoantibodies by thymoma cells from myasthenia gravis patients suggests autoimmunization in the tumor.
Myasthenia gravis (MG) is mediated by autoantibodies to the acetylcholine receptor (AChR), expressed in muscle and rare thymic myoid cells. Most early-onset cases show thymic lymph node-type infiltrates, including pre-activated plasma cells spontaneously producing anti-AChR antibodies. Since these are not evident in the associated thymomas found in another 10% of MG patients, AChR-specific B cells must be autosensitized elsewhere. Unexpectedly, at diagnosis, >70% of MG/thymoma patients also have high-titer neutralizing autoantibodies to IFN-alpha, and >50% to IL-12; moreover, titers increase strikingly if the thymomas recur, indicating a closer tumor relationship than for anti-AChR. To investigate this, we have measured autoantibody production by cells cultured from thymomas, any available thymic remnants and blood, with or without the B cell stimulant pokeweed mitogen (PWM). To check autoantibody specificity and clonal origins, we isolated Fabs from two combinatorial libraries from producer thymus/thymoma cells. Surprisingly, thymoma cells spontaneously produced antibodies to IFN-alpha and/or IL-12 in >40% of seropositive cases, showing typical plasma cell behavior, whereas they produced anti-AChR only after PWM stimulation. We isolated 15 combinatorial Fabs to IFN-alpha (versus only one to AChR). Their strong binding in radio-immunoprecipitation and Western blots implies high affinities. The four Fabs tested neutralized anti-viral actions of IFN-alpha. The diverse V genes clearly showed ongoing antigen-driven selection. These results imply pre-activation in situ by native IFN-alpha/IL-12 expressed within a 'dangerous' tumor microenvironment. With these molecules, it should be easier to identify provoking cell type(s) that may give novel additional clues to autoimmunization against T-cell epitopes from the more complex AChR.


Stability analysis for postural control in a two-joint limb system.
The stability behavior of a multi-joint limb with electrically activated muscles provides important clues for postural control of motor tasks. The stability property of the musculoskeletal system can be characterized with its eigenvalues evaluated at operating postures in the workspace. A planar arm model with shoulder and elbow joints and three pairs of antagonistic muscles was constructed in ADAMS. Stability behavior of shoulder and elbow joints was analyzed using the loci of eigenvalues in the s-plane. In the analysis of open-loop cocontraction of antagonist muscles with increasing activation from 5% to 100%, the eigenvalues of the shoulder and elbow joints were confined within the left half of the s-plane in a stripe of +/- j 0.5, and moved toward left onto the real axis. The shoulder eigenvalues were generally nearer to the imaginary axis than the elbow ones, indicating a more oscillatory behavior at the shoulder joint than that at the elbow joint. The effects of joint configuration evaluated within the workspace from 40 degrees to 110 degrees for the elbow and from 40 degrees to 120 degrees for the shoulder showed that the elbow eigenvalues were more prone to configuration changes, particularly elbow angles. We also developed a simulation paradigm for sampled data FES control systems that contain a mixture of continuous time components and sampling and hold effects. This simulation paradigm is useful for realistic simulation of local feedback controller performance.


A laboratory-based Laue X-ray diffraction system for enhanced imaging range and surface grain mapping.
Although CCD X-ray detectors can be faster to use, their large-area versions can be much more expensive than similarly sized photographic plate detectors. When indexing X-ray diffraction patterns, large-area detectors can prove very advantageous as they provide more spots, which makes fitting an orientation easier. On the other hand, when looking for single crystals in a polycrystalline sample, the speed of CCD detectors is more useful. A new setup is described here which overcomes some of the limitations of limited-range CCD detectors to make them more useful for indexing, whilst at the same time making it much quicker to find single crystals within a larger polycrystalline structure. This was done by combining a CCD detector with a six-axis goniometer, allowing the compilation of images from different angles into a wide-angled image. Automated scans along the sample were coupled with image processing techniques to produce grain maps, which can then be used to provide a strategy to extract single crystals from a polycrystal.


Emerging severe and fatal infections due to Klebsiella pneumoniae in two university hospitals in France.
Severe infections caused by hypermucoviscous Klebsiella pneumoniae have been reported in Southeast Asian countries over the past several decades. This report shows their emergence in France, with 12 cases observed during a 2-year period in two university hospitals. Two clones (sequence type 86 [ST86] and ST380) of serotype K2 caused five rapidly fatal bacteremia cases, three of which were associated with pneumonia, whereas seven liver abscess cases were caused by K1 strains of ST23.


Membrane cholesterol modulates cochlear electromechanics.
Changing the concentration of cholesterol in the plasma membrane of isolated outer hair cells modulates electromotility and prestin-associated charge movement, suggesting that a similar manipulation would alter cochlear mechanics. We examined cochlear function before and after depletion of membrane cholesterol with methyl-β-cyclodextrin (MβCD) in an excised guinea pig temporal bone preparation. The mechanical response of the cochlear partition to acoustic and/or electrical stimulation was monitored using laser interferometry and time-resolved confocal microscopy. The electromechanical response in untreated preparations was asymmetric with greater displacements in response to positive currents. Exposure to MβCD increased the magnitude and asymmetry of the response, without changing the frequency tuning of sound-evoked mechanical responses or cochlear microphonic potentials. Sodium salicylate reversibly blocked the enhanced electromechanical response in cholesterol depleted preparations. The increase of sound-evoked vibrations during positive current injection was enhanced following MβCD in some preparations. Imaging was used to assess cellular integrity which remained unchanged after several hours of exposure to MβCD in several preparations. The enhanced electromechanical response reflects an increase in outer hair cell electromotility and may reveal features of cholesterol distribution and trafficking in outer hair cells.


HECTD3 mediates TRAF3 polyubiquitination and type I interferon induction during bacterial infection.
Lysine-63-linked (K63-linked) polyubiquitination of TRAF3 coordinates the engagement of pattern-recognition receptors with recruited adaptor proteins and downstream activator TBK1 in pathways that induce type I IFN. Whether autoubiquitination or other E3 ligases mediate K63-linked TRAF3 polyubiquitination remains unclear. We demonstrated that mice deficient in the E3 ligase gene Hectd3 remarkably increased host defense against infection by intracellular bacteria Francisella novicida, Mycobacterium, and Listeria by limiting bacterial dissemination. In the absence of HECTD3, type I IFN response was impaired during bacterial infection both in vivo and in vitro. HECTD3 regulated type I IFN production by mediating K63-linked polyubiquitination of TRAF3 at residue K138. The catalytic domain of HECTD3 regulated TRAF3 K63 polyubiquitination, which enabled TRAF3-TBK1 complex formation. Our study offers insights into mechanisms of TRAF3 modulation and provides potential therapeutic targets against infections by intracellular bacteria and inflammatory diseases.