Contribution of myoglobin-induced increases in vascular resistance to shock decompensation in experimental Crush-syndrome in anesthetized rats.
Myoglobin is known to become nephrotoxic when released in greater amounts from skeletal muscle into the general circulation during shock. The present study deals with the question as to whether a myoglobin-induced increase in vascular tone additionally contributes to the detrimental role of this protein in hypovolemic shock. Anesthetized rats were subjected to 250 mg kg x h(-1) myoglobin infused i.v. during hemorrhagic hypotension of 50 mmHg. Shock survival time was measured, as were blood flow and vascular resistance in kidney, intestine, brain, and heart, using the microsphere method. Rats subjected to only myoglobin or hemorrhage survived a period of >120 min; in contrast, rats, exposed to both myoglobin and hemorrhage died at 68 +/- 9 min. When the animals subjected to only hemorrhage and to myoglobin/ hemorrhage were compared, significantly lower values were found in the latter group with respect to blood flow in the kidney (1.7 +/- 0.1 vs. 0.2 +/- 0.05 ml x min(-1) x g(-1)), small intestine (1.0 +/- 0.1 vs. 0.5 +/- 0.1 ml x min(-1) x g(-1)), cardiac output (112 +/- 5 vs. 62 +/- 10 ml(-1) x min(-1) x kg(-1)), and significantly higher values of total peripheral vascular resistance (0.45 +/- 0.02 vs. 0.81 +/- 0.12 mmHg x min x ml(-1) x kg) at 50 min of hypotension. It is assumed that these effects of myoglobin are induced by its ability to scavenge endogenous nitric oxide, because a modified, non-nitrosylable myoglobin was unable to induce such effects. The results support the view that a pathological release of myoglobin into the general circulation causes increases in vascular resistance of vital organs that may contribute to decompensation of tissue supply when occurring in hypovolemic shock.


Hypopituitarism and growth hormone deficiency (GHD) after traumatic brain injury (TBI).
In adults, hypopituitarism and growth hormone deficiency (GHD) should be suspected and diagnosed within an appropriate clinical context. It has been demonstrated that all patients with primary hypothalamic-pituitary diseases before and after any medical intervention (defined as neurosurgery, radiotherapy and medical therapy) are at obvious risk - more than just at high risk - for hypopituitarism (greater than 80% had severe GHD). The same obvious risk applies to patients diagnosed as having congenital or acquired GHD in childhood (between 30% and 50% of patients with severe GHD after retesting). Taking into account the fragility of the infundibular-hypothalamic structure, patients with other common pathological conditions of the central nervous system (CNS), such as traumatic brain injury (TBI), subarachnoid haemorrhage (SAH) or primary brain tumours (BT) with related medical intervention could be at risk for developing hypopituitarism, including GHD. GHD is the first and most common sign of pituitary impairment. Despite the risk of pituitary dysfunction and the results of some studies that these risks are greater than previously believed, neuroendocrine evaluations are still not routinely included as part of the clinical management plan for patients with TBI and SAH. Preliminary results of a multicenter study performed under the auspices of the Italian Society of Endocrinology underscore the high risk of hypopituitarism and GHD under these clinical conditions. Thus, careful screening of pituitary function should always be performed in patients following TBI and SAH.


Strategies to increase adoption of animal vaccines by smallholder farmers with focus on neglected diseases and marginalized populations.
Most smallholder farmers (SHFs) and marginalized populations (MPs) in Africa, Asia, and Latin America depend on livestock for their livelihoods. However, significant numbers of these animals do not achieve their potential, die due to disease, or transmit zoonotic diseases. Existing vaccines could prevent and control some of these diseases, but frequently the vaccines do not reach SHFs, especially MPs, making it necessary for specific vaccine adoption strategies. Several strategies that have the potential to increase the adoption of animal vaccines by SHFs and MPs have been identified depending on the type of vaccines involved. The strategies differed depending on whether the vaccines were aimed at diseases that cause economic losses, government-controlled diseases, or neglected diseases. The adoption of vaccines for neglected diseases presents a major challenge, because they are mostly for zoonotic diseases that produce few or no clinical signs in the animals, making it more difficult for the farmers to appreciate the value of the vaccines. Strategies can be aimed at increasing the availability of quality vaccines, so that they are produced in sufficient quantity, or aimed at increasing access and demand by SHFs and/or MPs. Some of the strategies to increase vaccine adoption might not provide a definite solution but might facilitate vaccine uptake by decreasing barriers. These strategies are varied and include technical considerations, policy components, involvement by the private sector (local and international), and innovation. Several strategies with the potential to reduce livestock morbidity and mortality, or prevent zoonoses in SHFs communities and MPs through vaccination, require the involvement of donors and international organisations to stimulate and facilitate sustainable adoption. This is especially the case for neglected zoonotic diseases. Support for national and regional vaccine manufacturers is also required, especially for vaccines against diseases of interest only in the developing world and public goods.


Tinnitus in Normal-Hearing Participants after Exposure to Intense Low-Frequency Sound and in Ménière's Disease Patients.
Tinnitus is one of the three classical symptoms of Ménière's disease (MD), an inner ear disease that is often accompanied by endolymphatic hydrops. Previous studies indicate that tinnitus in MD patients is dominated by low frequencies, whereas tinnitus in non-hydropic pathologies is typically higher in frequency. Tinnitus of rather low-frequency (LF) quality was also reported to occur for about 90 s in normal-hearing participants after presentation of intense, LF sound (120 dB SPL, 30 Hz, 90 s). LF sound has been demonstrated to also cause temporary endolymphatic hydrops in animal models. Here, we quantify tinnitus in two study groups with chronic (MD patients) and presumably transient endolymphatic hydrops (normal-hearing participants after LF exposure) with a psychophysical procedure. Participants matched their tinnitus either with a pure tone of adjustable frequency and level or with a noise of adjustable spectral shape and level. Sensation levels of matching stimuli were lower for MD patients (mean: 8 dB SL) than for normal-hearing participants (mean: 15 dB SL). Transient tinnitus after LF-exposure occurred in all normal-hearing participants (N = 28). About half of the normal-hearing participants matched noise to their tinnitus, the other half chose a pure tone with frequencies below 2 kHz. MD patients matched their tinnitus with either high-frequency pure tones, mainly above 3 kHz, or with a noise. Despite a significant proportion of MD patients matching low-pass (roaring) noises to their tinnitus, the range of matched stimuli was more heterogeneous than previous data suggested. We propose that in those participants with noise-like tinnitus, the percept is probably generated by increased spontaneous activity of auditory nerve fibers with a broad range of characteristic frequencies, due to an impaired ion balance in the cochlea. For tonal tinnitus, additional mechanisms are conceivable: focal hair cell loss can result in decreased auditory nerve firing and a central auditory overcompensation. Also, normal-hearing participants after LF-exposure experience alterations in spontaneous otoacoustic emissions, which may contribute to a transient tonal tinnitus.


Tumour antigen-targeted immunotherapy for chronic myeloid leukaemia: is it still viable?
In haematological cancers, malignant cells circulate in the blood and lymphatic system. This may make leukaemic cells easier to target by immunotherapy than in other types of cancer. Various immunotherapy strategies have been trialled in several leukaemias including chronic myeloid leukaemia (CML) and in general, these have been aimed at targeting tumour-associated antigens (TAA). There are numerous TAA expressed by CML patients including WT1, proteinase 3, BCR-ABL and HAGE amongst others. The immunogenicity of the CML-specific tumour antigen, BCR-ABL, has been the subject of much debate and its role in the development of the disease and its unique sequence spanning the breakpoint region make it an ideal target for immunotherapy. However, there are a limited number of immunogenic epitopes across the junctional region, which are restricted to only a few HLA types, namely A2, A3 and B7 (Clark et al. in Blood 98:2887-2893, 2001). The second CML-associated antigen is the helicase antigen HAGE, a cancer-testis antigen found to be over-expressed in more than 50% of myeloid leukaemias (Adams et al. in Leukaemia 16:2238-2242, 2002). Very little is known about the function of this antigen and its significance to CML. However, its membership of the DEAD-box family of ATP-dependent RNA helicases and the involvement of other members of this family in tumour cell proliferation (Eberle et al. in Br J Cancer 86:1957-1962, 2002; Yang et al. in Cell Signal 17:1495-504, 2005) suggest a crucial role in the RNA metabolism of tumour cells. For these reasons, HAGE also seems to be a good target for immunotherapy as it would be applicable for the majority of patients with CML. This review aims to discuss the potential of immunotherapy for the treatment of leukaemia, in particular CML, and the prospect of targeting three CML associated antigens: BCR, ABL and HAGE. During his career, Prof. Tony Dodi made a significant contribution in this area of leukaemia research, confirming the identity of immunogenic HLA-A3 and B7-restricted peptides as targets for CTL. Published, as a highlighted paper in Clark et al. (Blood 98:2887-2893, 2001), this study demonstrated the expression of MHC-peptide complexes on the surface of CML cells and the presence of tetramer-positive CTL activity in CML patients positive for these two HLA alleles. His drive and dedication for research excellence will be remembered by all who knew and worked with him.


Comparison of Cryoballoon and Hybrid surgical Posterior Wall Isolation for Persistent Atrial Fibrillation to conventional ablation.
Optimalablation strategy for persistent atrial fibrillation (AF) is unclear;PWI of the left atrium may improve outcome.Our aim was to compare outcomes of posterior wall isolation (PWI) ablation for persistent AF achieved by cryoballoon ablation (CRYO) or hybrid surgical ablation (HABL) to matched patients undergoing conventional radiofrequency ablation (CRA). In our single center retrospective study,patients underwent HABL and CRYO withcircumferential pulmonary vein ablation (CPVA),roof and floor lines to complete PW box lesion. CRA consisted of CPVA, roof line and lateral mitral isthmus line (MVI). Of 61 patients (mean duration of AF 1.3 ± 0.4 yrs)who underwent ablation, after follow-up of 366 ± 62 days, AF recurrence was 10.5% and 48% (p=0.001) and the need for repeat ablation 5% vs 30% (p=0.007) in PWI and CRA groups respectively, without a significant difference in incidence of AT/FL 18 vs 26 % or cardioversion 5.2 vs 7.1 %. Total procedure time and fluoroscopic time were 242 ±70 min vs 279 ±53 min (p=0.08) and 20±9 min vs 12 ± 4 min (p=0.003) for PWI and CRA respectively. CRYO had less AF recurrence and complications than HABL. Mean length of stayfor CRYO patients was 41 hrs compared to 145 hrs in HABL group, who underwent two procedures. PWI in persistent AF patients decreases recurrence of AF and need for repeat procedure compared to CRA; PWI by CRYO is superior to HABL due to less LOS and complications.


Phylogenetic structure of European Salmonella Enteritidis outbreak correlates with national and international egg distribution network.
Outbreaks of Salmonella Enteritidis have long been associated with contaminated poultry and eggs. In the summer of 2014 a large multi-national outbreak of Salmonella Enteritidis phage type 14b occurred with over 350 cases reported in the United Kingdom, Germany, Austria, France and Luxembourg. Egg supply network investigation and microbiological sampling identified the source to be a Bavarian egg producer. As part of the international investigation into the outbreak, over 400 isolates were sequenced including isolates from cases, implicated UK premises and eggs from the suspected source producer. We were able to show a clear statistical correlation between the topology of the UK egg distribution network and the phylogenetic network of outbreak isolates. This correlation can most plausibly be explained by different parts of the egg distribution network being supplied by eggs solely from independent premises of the Bavarian egg producer (Company X). Microbiological sampling from the source premises, traceback information and information on the interventions carried out at the egg production premises all supported this conclusion. The level of insight into the outbreak epidemiology provided by whole-genome sequencing (WGS) would not have been possible using traditional microbial typing methods.


An ancestral bacterial division system is widespread in eukaryotic mitochondria.
Bacterial division initiates at the site of a contractile Z-ring composed of polymerized FtsZ. The location of the Z-ring in the cell is controlled by a system of three mutually antagonistic proteins, MinC, MinD, and MinE. Plastid division is also known to be dependent on homologs of these proteins, derived from the ancestral cyanobacterial endosymbiont that gave rise to plastids. In contrast, the mitochondria of model systems such as Saccharomyces cerevisiae, mammals, and Arabidopsis thaliana seem to have replaced the ancestral α-proteobacterial Min-based division machinery with host-derived dynamin-related proteins that form outer contractile rings. Here, we show that the mitochondrial division system of these model organisms is the exception, rather than the rule, for eukaryotes. We describe endosymbiont-derived, bacterial-like division systems comprising FtsZ and Min proteins in diverse less-studied eukaryote protistan lineages, including jakobid and heterolobosean excavates, a malawimonad, stramenopiles, amoebozoans, a breviate, and an apusomonad. For two of these taxa, the amoebozoan Dictyostelium purpureum and the jakobid Andalucia incarcerata, we confirm a mitochondrial localization of these proteins by their heterologous expression in Saccharomyces cerevisiae. The discovery of a proteobacterial-like division system in mitochondria of diverse eukaryotic lineages suggests that it was the ancestral feature of all eukaryotic mitochondria and has been supplanted by a host-derived system multiple times in distinct eukaryote lineages.


Inhalable antibiotic delivery using a dry powder co-delivering recombinant deoxyribonuclease and ciprofloxacin for treatment of cystic fibrosis.
To achieve efficient antibiotic delivery to the cystic fibrosis (CF) airway using a single inhalable powder co-encapsulating a mucolytic and an antibiotic. Inhalable dry powders containing deoxyribonuclease and/or ciprofloxacin (DNase, Cipro, and DNase/Cipro powders) were produced by spray-drying with dipalmitylphosphatidylcholine, albumin, and lactose as excipients, and their antibacterial effects were evaluated using the artificial sputum model. All powders showed mass median aerodynamic diameters below 5 microm. Both drugs were loaded in the dry powders without loss in quantity and activity. Dry powders containing DNase significantly decreased the storage modulus of the artificial sputum medium in less than 30 min. When applied to artificial sputum laden with Pseudomonas aeruginosa, Cipro/DNase powder showed better antibacterial activity than Cipro powder. The higher activity of the Cipro/DNase powder is attributable to the mucolytic activity of DNase, which promotes penetration of the dry powder into the artificial sputum and efficient dissolution and diffusion of ciprofloxacin. Inhalational delivery of antibiotics to the CF airway can be optimized when the sputum barrier is concomitantly addressed. Co-delivery of antibiotics and DNase using an inhalable particle system may be a promising strategy for local antipseudomonal therapy in the CF airway.


Etiology of troponin elevation in critically ill patients.
The aim of this study was to assess the etiology of cardiac troponin elevation among patients admitted to the intensive care unit (ICU) and to examine whether etiology affects mortality and length of stay. All patients admitted over 2 months underwent screening with troponin measurements and were included if 1 or more measurements were elevated. Two adjudicators retrospectively reviewed patient charts to determine the likely cause of troponin elevation. Of 103 patient admissions, 52 (50.5%) had 1 or more elevated troponin measurements, and 49 (94.2%) had medical charts available for review. Troponin elevation was adjudicated as myocardial infarction (MI) in 53.1% of patients, sepsis in 18.4%, renal failure in 12.2%, and other causes in 16.3%. Overall ICU mortality was 16.0%; 2.0% for patients with no troponin elevation, 23.1% in patients with MI, and 39.1% in patients with troponin elevation not due to MI. Having an elevated troponin level not due to MI was significantly associated with increased hospital mortality compared with having no troponin elevation. The most common cause of troponin elevation among critically ill patients was MI. Patients with elevated troponin had worse outcomes compared with patients without troponin elevation, and troponin elevation not due to MI was predictive of increased hospital mortality.


Molecular basis of altered red blood cell membrane properties in Southeast Asian ovalocytosis: role of the mutant band 3 protein in band 3 oligomerization and retention by the membrane skeleton.
Southeast Asian ovalocytosis (SAO) is an asymptomatic trait characterized by rigid, poorly deformable red cells that resist invasion by several strains of malaria parasites. The underlying molecular genetic defect involves simple heterozygous state for a mutant band 3 protein, which contains a deletion of amino acids 400 through 408, linked with a Lys 56-to-Glu substitution (band 3-Memphis polymorphism). To elucidate the contribution of the mutant SAO band 3 protein to increased SAO red blood cell (RBC) rigidity, we examined the participation of the mutant SAO band 3 protein in increased band 3 attachment to the skeleton and band 3 oligomerization. We found first that SAO RBC skeletons retained more band 3 than normal cells and that this increased retention preferentially involved the mutant SAO band 3 protein. Second, SAO RBCs contained a higher percentage of band 3 oligomer-ankyrin complexes than normal cells, and these oligomers were preferentially enriched by the mutant SAO protein. At the ultrastructural level, the increased oligomer formation of SAO RBCs was reflected by stacking of band 3-containing intramembrane particles (IMP) into longitudinal strands. The IMP stacking was not reversed by treating SAO RBCs in alkaline pH (pH 11), which is known to weaken ankyrin-band 3 interactions, or by removing the cytoplasmic domain of band 3 from SAO membranes with trypsin. Finally, we found that band 3 protein in intact SAO RBCs exhibited a markedly decreased rotational mobility, presumably reflecting the increased oligomerization and the membrane skeletal association of the SAO band 3 protein. We propose that the mutant SAO band 3 has an increased propensity to form oligomers, which appear as longitudinal strands of IMP and exhibit increased association with membrane skeleton. This band 3 oligomerization underlies the increase in membrane rigidity by precluding membrane skeletal extension, which is necessary for membrane deformation.


The interaction of beryllium with benzene and graphene: a comparative investigation based on DFT, MP2, CCSD(T), CAS-SCF and CAS-PT2.
The interaction of beryllium with benzene, graphene and graphitic compounds involves multi-reference electronic states, Jahn-Teller distortion, charge transfer and van der Waals interactions. This is investigated herein using periodic and molecular models at different levels of theory: (i) the second-order Møller-Plesset (MP2) perturbation theory, (ii) the coupled cluster method with inclusion of single double and perturbative triple excitations (CCSD(T)), (iii) the complete active space self-consistent field (CAS-SCF) and (iv) the complete active space with perturbation theory truncated at the 2nd order (CAS-PT2). Molecular and periodic Density Functional Theory (DFT) methods are also used. The two major failures of DFT are addressed with regard to the beryllium benzene and graphene interaction: the degeneracy problem is the source of no specific problem while the delocalization error causes DFT with the Perdew Burke, Ernzerhof functional plus the Grimme correction (DFT/PBE-D2) to be over-binding by about 0.4 eV at a short-range. The agreement between DFT/PBE-D2 and wave-function based methods is nevertheless good; DFT/PBE-D2 provides an accurate description of the electronic structure of the system. By the end of this work, we shall get a better insight into the mechanisms leading beryllium to physisorb on graphene and to chemisorb into the bilayer of graphite.


Grainyhead-like 3, a transcription factor identified in a microarray screen, promotes the specification of the superficial layer of the embryonic epidermis.
The Xenopus ectoderm consists of two populations of cells, superficial polarised epithelial cells and deep, non-epithelial cells. These two cell types differ in their developmental fate. In the neural ectoderm, primary neurons are derived only from the deep cells. In the epidermal ectoderm, superficial cells express high levels of differentiation markers, while most of the deep cells do not differentiate until later when they produce the stratified adult epidermis. However, few molecular differences are known between the deep and superficial cells. Here, we have undertaken a systematic approach to identify genes that show layer-restricted expression by microarray analysis of deep and superficial cells at the gastrula stage, followed by wholemount in situ hybridisation. We have identified 32 differentially expressed genes, of which 26 show higher expression in the superficial layer and 6 in the deep layer and describe their expression at the gastrula and neurula stage. One of the identified genes is the transcription factor Grhl3, which we found to be expressed in the superficial layer of the gastrula ectoderm and the neurula epidermis. By using markers identified in this work, we show that Grlh3 promotes superficial gene expression in the deep layer of the epidermis. Concomitantly, deep layer specific genes are switched off, showing that Grlh3 can promote deep cells to take on a superficial cell identity in the embryonic epidermis.


Extending Miscanthus Cultivation with Novel Germplasm at Six Contrasting Sites.
Miscanthus is a genus of perennial rhizomatous grasses with C4 photosynthesis which is indigenous in a wide geographic range of Asian climates. The sterile clone, Miscanthus × giganteus (M. × giganteus), is a naturally occurring interspecific hybrid that has been used commercially in Europe for biomass production for over a decade. Although, M. × giganteus has many outstanding performance characteristics including high yields and low nutrient offtakes, commercial expansion is limited by cloning rates, slow establishment to a mature yield, frost, and drought resistance. In this paper, we evaluate the performance of 13 novel germplasm types alongside M. × giganteus and horticultural "Goliath" in trials in six sites (in Germany, Russia, The Netherlands, Turkey, UK, and Ukraine). Mean annual yields across all the sites and genotypes increased from 2.3 ± 0.2 t dry matter ha-1 following the first year of growth, to 7.3 ± 0.3, 9.5 ± 0.3, and 10.5 ± 0.2 t dry matter ha-1 following the second, third, and fourth years, respectively. The highest average annual yields across locations and four growth seasons were observed for M. × giganteus (9.9 ± 0.7 t dry matter ha-1) and interspecies hybrid OPM-6 (9.4 ± 0.6 t dry matter ha-1). The best of the new hybrid genotypes yielded similarly to M. × giganteus at most of the locations. Significant effects of the year of growth, location, species, genotype, and interplay between these factors have been observed demonstrating strong genotype × environment interactions. The highest yields were recorded in Ukraine. Time needed for the crop establishment varied depending on climate: in colder climates such as Russia the crop has not achieved its peak yield by the fourth year, whereas in the hot climate of Turkey and under irrigation the yields were already high in the first growing season. We have identified several alternatives to M. × giganteus which have provided stable yields across wide climatic ranges, mostly interspecies hybrids, and also Miscanthus genotypes providing high biomass yields at specific geographic locations. Seed-propagated interspecific and intraspecific hybrids, with high stable yields and cheaper reliable scalable establishment remain a key strategic objective for breeders.


Grasshopper community response to climatic change: variation along an elevational gradient.
The impacts of climate change on phenological responses of species and communities are well-documented; however, many such studies are correlational and so less effective at assessing the causal links between changes in climate and changes in phenology. Using grasshopper communities found along an elevational gradient, we present an ideal system along the Front Range of Colorado USA that provides a mechanistic link between climate and phenology. This study utilizes past (1959-1960) and present (2006-2008) surveys of grasshopper communities and daily temperature records to quantify the relationship between amount and timing of warming across years and elevations, and grasshopper timing to adulthood. Grasshopper communities were surveyed at four sites, Chautauqua Mesa (1752 m), A1 (2195 m), B1 (2591 m), and C1 (3048 m), located in prairie, lower montane, upper montane, and subalpine life zones, respectively. Changes to earlier first appearance of adults depended on the degree to which a site warmed. The lowest site showed little warming and little phenological advancement. The next highest site (A1) warmed a small, but significant, amount and grasshopper species there showed inconsistent phenological advancements. The two highest sites warmed the most, and at these sites grasshoppers showed significant phenological advancements. At these sites, late-developing species showed the greatest advancements, a pattern that correlated with an increase in rate of late-season warming. The number of growing degree days (GDDs) associated with the time to adulthood for a species was unchanged across the past and present surveys, suggesting that phenological advancement depended on when a set number of GDDs is reached during a season. Our analyses provide clear evidence that variation in amount and timing of warming over the growing season explains the vast majority of phenological variation in this system. Our results move past simple correlation and provide a stronger process-oriented and predictive framework for understanding community level phenological responses to climate change.


Dual-wavelength laser speckle imaging to simultaneously access blood flow, blood volume, and oxygenation using a color CCD camera.
We developed a dual-wavelength laser speckle imaging system using a single industrial-grade color CCD camera with Bayer filters to simultaneously image changes in blood flow, blood volume, and oxygenation. One frame of a color image recorded with dual-wavelength laser illumination provides not only the intensity fluctuation of the speckle pattern, but also the dual-wavelength optical reflectance signal. The method was validated using a tissue phantom and cuff ischemia experiments in the human arm. This system achieves complete time synchronization, unlike conventional time-sharing systems. Compared with a multicamera system, it also avoids the problem of image registration and can be less expensive.


Nurses' reflections on pain management in a nursing home setting.
Achieving optimal and safe pain-management practices in the nursing home setting continues to challenge administrators, nurses, physicians, and other health care providers. Several factors in nursing home settings complicate the conduct of clinical process improvement research. The purpose of this qualitative study was to explore the perceptions of a sample of Colorado nursing home staff who participated in a study to develop and evaluate a multifaceted pain-management intervention. Semistructured interviews were conducted with 103 staff from treatment and control nursing homes, audiotaped, and content analyzed. Staff identified changes in their knowledge and attitudes about pain and their pain-assessment and management practices. Progressive solutions and suggestions for changing practice include establishing an internal pain team and incorporating nursing assistants into the care planning process. Quality improvement strategies can accommodate the special circumstances of nursing home care and build the capacity of the nursing homes to initiate and monitor their own process-improvement programs using a participatory research approach.


Identifying context-specific transcription factor targets from prior knowledge and gene expression data.
Numerous methodologies, assays, and databases presently provide candidate targets of transcription factors (TFs). However, TFs rarely regulate their targets universally. The context of activation of a TF can change the transcriptional response of targets. Direct multiple regulation typical to mammalian genes complicates direct inference of TF targets from gene expression data. We present a novel statistic that infers context-specific TF regulation based upon the CoGAPS algorithm, which infers overlapping gene expression patterns resulting from coregulation. Numerical experiments with simulated data showed that this statistic correctly inferred targets that are common to multiple TFs, except in cases where the signal from a TF is negligible relative to noise level and signal from other TFs. The statistic is robust to moderate levels of error in the simulated gene sets, identifying fewer false positives than false negatives. Significantly, the regulatory statistic refines the number of TF targets relevant to cell signaling in gastrointestinal stromal tumors (GIST) to genes consistent with the phosphorylation patterns of TFs identified in previous studies. As formulated, the proposed regulatory statistic has wide applicability to inferring set membership in integrated datasets. This statistic could be naturally extended to account for prior probabilities of set membership or to add candidate gene targets.


Seizures presenting as apnoea.
Between the ages of 3 and 6 months a baby boy presented with repeated, non-specific episodes of cyanosis, apnoea, bradycardia, and abnormal movements of the limbs. The episodes were severe and required resuscitation and several admissions to hospital. Initial investigations showed only signs of oesophagitis. Despite treatment of the oesophagitis the symptoms recurred, and electroencephalography and polygraphy eventually showed evidence of minor seizures. Severe epilepsy with tonic-clonic seizures developed when he was 6 months old.


Predicting efficiency of post-induction mask ventilation based on demographic and anatomical factors.
Failure to ventilate patients by mask leads to serious complications especially if associated with difficult intubation. Previous studies have used subjective and indirect measures to evaluate difficulty in mask ventilation, which are associated with high inter-observer discrepancies. In this study, we have defined and used efficiency of mask ventilation (EMV) as an objective and direct surrogate for ease of mask ventilation in patients undergoing GA and mask ventilation using neuromuscular relaxation. 1050 adult patients prospectively were evaluated with respect to different patients demographic and physical factors and EMV. EMV was defined as the ratio of minute ventilation via anesthesia mask to that via tracheal tube expressed as percentage. Edentolous patients were ventilated using lip-over-mask techniques. Separate analyses were done for edentolous and non-edentolous patients. EMV in edentolous patients (n=269) using the lip-over-mask method was relatively high (90.9 ± 14.3%, 60.14-128.57 range). The result of multiple regression analysis in patients with normal denture determined receded chin, presence of beard, male gender, high Mallampati classes, high neck circumference, low inter-incisors gap, and old age as independent factors for estimating EMV. A regression formula for predicting EMV was developed which had an acceptable R-square value with a good model fit. Using EMV is an easy and reliable tool for measuring efficiency of mask ventilation. Based on the result of this study, EMV can be estimated from patient's demographic and physical factors. In edentolous patients, using the lip-over-mask method results in adequate ventilation of lungs.


Identification of FBXL4 as a Metastasis Associated Gene in Prostate Cancer.
Prostate cancer is the most common cancer among western men, with a significant mortality and morbidity reported for advanced metastatic disease. Current understanding of metastatic disease is limited due to difficulty of sampling as prostate cancer mainly metastasizes to bone. By analysing prostate cancer bone metastases using high density microarrays, we found a common genomic copy number loss at 6q16.1-16.2, containing the FBXL4 gene, which was confirmed in larger series of bone metastases by fluorescence in situ hybridisation (FISH). Loss of FBXL4 was also detected in primary tumours and it was highly associated with prognostic factors including high Gleason score, clinical stage, prostate-specific antigen (PSA) and extent of disease, as well as poor patient survival, suggesting that FBXL4 loss contributes to prostate cancer progression. We also demonstrated that FBXL4 deletion is detectable in circulating tumour cells (CTCs), making it a potential prognostic biomarker by 'liquid biopsy'. In vitro analysis showed that FBXL4 plays a role in regulating the migration and invasion of prostate cancer cells. FBXL4 potentially controls cancer metastasis through regulation of ERLEC1 levels. Therefore, FBXL4 could be a potential novel prostate cancer suppressor gene, which may prevent cancer progression and metastasis through controlling cell invasion.


Glasdegib in combination with cytarabine and daunorubicin in patients with AML or high-risk MDS: Phase 2 study results.
Glasdegib is a Hedgehog pathway inhibitor. This ongoing, open-label, phase 2 study (NCT01546038) evaluated glasdegib plus cytarabine/daunorubicin in patients with untreated acute myeloid leukemia (AML) or high-risk myelodysplastic syndromes (MDS). Patients received glasdegib 100 mg orally, once daily in continuous 28-day cycles from day -3, with intravenous cytarabine 100 mg/m2 on days 1-7 and daunorubicin 60 mg/m2 on days 1-3. Patients in remission then received consolidation therapy (2-4 cycles of cytarabine 1 g/m2 twice daily on days 1, 3, 5 of each cycle), followed by maintenance glasdegib (maximum 6 cycles). Primary endpoint was complete remission (CR) in patients aged ≥55 years. Secondary endpoints included overall survival (OS), safety and outcome by mutational status. Patients had a median (range) age of 64.0 (27-75) years, 60.0% were male, and 84.5% were white. In 69 evaluable patients, 46.4% (80% confidence interval [CI]: 38.7-54.1) achieved investigator-reported CR. Among patients ≥55 years old (n = 60), 40.0% (80% CI 31.9-48.1) achieved CR. Among all 69 patients, median OS was 14.9 (80% CI 13.4-19.3) months, with 12-month survival probability 66.6% (80% CI 58.5-73.4). The most common treatment-related adverse events (≥50% patients) were diarrhea and nausea. There were no significant associations between mutational status (12 genes) and clinical response, suggesting potential benefit across diverse molecular profiles. Glasdegib plus cytarabine/daunorubicin was well tolerated and associated with clinical activity in patients with untreated AML or high-risk MDS. A randomized phase 3 trial of glasdegib in combination with chemotherapy (7 + 3 schedule) is ongoing.


Making the middle classes on shifting ground? Residential status, performativity and middle-class subjectivities in contemporary London.
This paper argues that shifts in access to housing - both in relation to rental and ownership - disrupt middle-class reproduction in ways that fundamentally influence class formation. While property ownership has had a long association with middle-class identities, status and distinction, an increasingly competitive rental market alongside inflated property prices has impacted on expectations and anxieties over housing futures. In this paper, we consider two key questions: (1) What happens to middle-class identities under the conditions of this wider structural change? (2) How do the middle classes variously manoeuvre within this? Drawing on empirical research conducted in London, we demonstrate that becoming an owner-occupier may be fractured along lines of class but also along the axes of age, wealth and timing, particularly as this relates to the housing market. It builds on understandings of residential status and place as central to the formation of class, orienting this around the recognition of both people and place as mutable, emphasizing that changing economic and social processes generate new class positionalities and strategies for class reproduction. We argue that these processes are writ large in practices of belonging and claims to place, with wider repercussions within the urban landscape.


Targeted delivery of polyoxometalate nanocomposites.
Polyoxometalate/carboxymethyl chitosan nanocomposites with an average diameter of 130 nm are synthesized and labeled with fluorescein isothiocyanate (FITC) for a combined drug-carrier and cellular-monitoring approach. [Eu(β(2) -SiW(11) O(39) )(2) ](13-) /CMC nanospheres as a representative example do not display cytotoxicity for POM concentrations up to 2 mg mL(-1) . Cellular uptake of fluoresecently labelled {EuSiW(11) O(39) }/FITC-CMC nanoparticles is monitored with confocal laser scanning microscopy. Nanoparticle uptake occurs after incubation times of around 1 h and no cyctotoxic effects are observed upon prolonged treatment. The preferential location of the POM/CMC nanocomposites in the perinuclear region is furthermore verified with transmission electron microscopy investigations on unlabeled nanoparticles. Therefore, this approach is a promising dual strategy for the safe cellular transfer and monitoring of bioactive POMs.


Effect of lysine acetylation on the regulation of Trypanosoma brucei glycosomal aldolase activity.
Post-translational modifications provide suitable mechanisms for cellular adaptation to environmental changes. Lysine acetylation is one of these modifications and occurs with the addition of an acetyl group to Nε-amino chain of this residue, eliminating its positive charge. Recently, we found distinct acetylation profiles of procyclic and bloodstream forms of Trypanosoma brucei, the agent of African Trypanosomiasis. Interestingly, glycolytic enzymes were more acetylated in the procyclic, which develops in insects and uses oxidative phosphorylation to obtain energy, compared with the bloodstream form, whose main source of energy is glycolysis. Here, we investigated whether acetylation regulates the T. brucei fructose 1,6-bisphosphate aldolase. We found that aldolase activity was reduced in procyclic parasites cultivated in the absence of glucose and partial recovered by in vitro deacetylation. Similarly, acetylation of protein extracts from procyclics cultivated in glucose-rich medium, caused a reduction in the aldolase activity. In addition, aldolase acetylation levels were higher in procyclics cultivated in the absence of glucose compared with those cultivated in the presence of glucose. To further confirm the role of acetylation, lysine residues near the catalytic site were substituted by glutamine in recombinant T. brucei aldolase. These replacements, especially K157, inhibited enzymatic activity, changed the electrostatic surface potential, decrease substrate binding and modify the catalytic pocket structure of the enzyme, as predicted by in silico analysis. Taken together, these data confirm the role of acetylation in regulating the activity of an enzyme from the glycolytic pathway of T. brucei, expanding the factors responsible for regulating important pathways in this parasite.


The failed invasion of Harmonia axyridis in the Azores, Portugal: Climatic restriction or wrong population origin?
We tested two questions: (i) whether the climatic conditions of the Azorean Islands in Portugal may have restricted the invasion of Harmonia axyridis across this archipelago and (ii) determine what population of this species could have a higher probability of invading the islands. We used MaxEnt to project the climate requirements of different H. axyridis populations from three regions of the world, and the potential global niche of the species in the Azorean islands. Then we assessed the suitability of the islands for each of the three H. axyridis populations and global potential niche through histograms analysis, Principal Component Analysis (PCA) of climate variables, and a variable-by-variable assessment of the suitability response curves compared with the climatic conditions of the Azores. Climatic conditions of the Azores are less suitable for the U.S. and native Asian populations of H. axyridis, and more suitable for European populations and the global potential niche. The PCA showed that the climatic conditions of the islands differed from the climatic requirements of H. axyridis. This difference is mainly explained by precipitation of the wettest month, isothermality, and the minimum temperature of the coldest month. We concluded that the climatic conditions of the Azores could have influenced the establishment and spread of H. axyridis on these islands from Europe. Our results showed that abiotic resistance represented by the climate of the potentially colonizable zones could hinder the establishment of invasive insects, but it could vary depending of the origin of the colonizing population.


Cryptorchidism.
Cryptorchidism remains a major cause of male infertility and can be associated with germ cell tumors. Recent reports regarding cause, diagnosis, treatment and outcome of this disorder continue to inform our understanding of this common and important problem. The frequency of the problem makes cryptorchidism an area where diagnostic knowledge is particularly important for healthcare professionals. The literature reviewed in this article approach cryptorchidism from multiple aspects. Reports regarding cause include studies of molecular genetics, endocrine chemical disruptors, the association with galactosemia, the association with low birth weight, and for acquired cryptorchidism, the relationship to infant feeding. In regard to treatment, the benefit of surgical repair at 9 months of age and compliance with recommendations is demonstrated. Further reports continue to document the cryptorchidism's negative impact on fertility, the higher risk of future gonadal malignancy and the lack of function of the unrepaired unilateral cryptorchid testis in adulthood. Management considerations such as the benefit of testicular biopsy at orchiopexy are also reviewed. It was concluded from an analysis of data from the Danish national registry that this can be a valid research tool for future evaluation of the outcome after cryptorchidism. Early recognition and surgery, before 1 year of age, remain the most important interventions to reduce the negative impact of both unilateral and bilateral cryptorchidism. Further research is needed to better understand causes of cryptorchidism and the mechanisms by which it exerts its negative effects and to clarify outcome factors to direct the best clinical management of cryptorchidism.


Mutational analysis of the charge selectivity filter of the alpha7 nicotinic acetylcholine receptor.
In the alpha7 nicotinic acetylcholine receptors, we analyze the contribution of mutations E237A and V251T, together with the proline insertion P236', in the conversion of the charge selectivity from cationic to anionic. We show that the triple mutant exhibits spontaneous openings displaying anionic selectivity. Furthermore, at position 251, hydrophilic or even negatively charged residues are compatible with an anionic channel. In contrast, the additional proline yields an anionic channel only when inserted between positions 234 and 237; insertion before 234 yields a cationic channel and after 238 alters the receptor surface expression. The coiled 234-238 loop thus directly contributes to the charge selectivity filter of the alpha7 channel.


Bandpass sampling of high-frequency tissue motion.
The characterization of tissue viscoelastic properties requires the measurement of tissue motion over a region of interest at frequencies that significantly exceed the frame rates of conventional ultrasound systems. In this paper, we propose that the bandpass sampling technique be applied to tissue motion sampling. With this approach, high-frequency signals limited to a frequency band can be sampled and reconstructed without aliasing at a sampling frequency that is lower than the Nyquist rate. We first review this approach and discuss the selection of the tissue excitation frequency band and of the feasible sampling frequencies that allow signal reconstruction without aliasing. We then demonstrate the approach using simulations based on the finite element method and ultrasound simulations. Finally, we perform experiments on tissue-mimicking materials and demonstrate accurate motion estimation using a lower sampling rate than that required by the conventional sampling theorem. The estimated displacements were used to measure the elasticity and viscosity in a phantom in which an inclusion has been correctly delineated. Thus, with bandpass sampling, it is feasible to use conventional beamforming on diagnostic ultrasound systems to perform high-frequency dynamic elastography. The method is simple to implement because it does not require beam interleaving, additional hardware, or synchronization.


A comparison between human and machine labelling of image regions.
In previous work (Campbell et al, 1997 Pattern Recognition 30 555-563) a vision system was developed which is capable of classifying objects in outdoor scenes. The approach involves segmenting the image into regions, obtaining a feature-based description of each region, and then passing this description on to an artificial neural network (ANN) which has been trained to label the region with one of eleven possible object types. The question addressed here is: how important is each of these features to overall performance, both in human and machine vision? A set of experiments was conducted in which human subjects were trained in the same labelling task as the ANN. The stimuli, each depicting a single image region, were generated from a large database of urban and rural images. The subjects were then tested on both intact and degraded stimuli. The results suggest that certain features are particularly influential in mediating overall labelling performance. An equivalent experiment was carried out with the ANN. A method is presented which allows individual features to be corrupted in such a way as to simulate the loss of certain forms of visual information. The results, which are broadly similar to those found in the previous experiment, imply that the ANN can provide a useful model of human image region labelling. It is anticipated that the methodology, which draws on both computational and psychophysical techniques, will be of use to other areas of investigation.


Modeling the effects of vasculature evolution on early brain tumor growth.
Mathematical modeling of both tumor growth and angiogenesis have been active areas of research for the past several decades. Such models can be classified into one of two categories: those that analyze the remodeling of the vasculature while ignoring changes in the tumor mass, and those that predict tumor expansion in the presence of a non-evolving vasculature. However, it is well accepted that vasculature remodeling and tumor growth strongly depend on one another. For this reason, we have developed a two-dimensional hybrid cellular automaton model of early brain tumor growth that couples the remodeling of the microvasculature with the evolution of the tumor mass. A system of reaction-diffusion equations has been developed to track the concentration of vascular endothelial growth factor (VEGF), Ang-1, Ang-2, their receptors and their complexes in space and time. The properties of the vasculature and hence of each cell are determined by the relative concentrations of these key angiogenic factors. The model exhibits an angiogenic switch consistent with experimental observations on the upregulation of angiogenesis. Particularly, we show that if the pathways that produce and respond to VEGF and the angiopoietins are properly functioning, angiogenesis is initiated and a tumor can grow to a macroscopic size. However, if the VEGF pathway is inhibited, angiogenesis does not occur and tumor growth is thwarted beyond 1-2mm in size. Furthermore, we show that tumor expansion can occur in well-vascularized environments even when angiogenesis is inhibited, suggesting that anti-angiogenic therapies may not be sufficient to eliminate a population of actively dividing malignant cells.


Lectin-like glycoprotein PsNLEC-1 is not correctly glycosylated and targeted in boron-deficient pea nodules.
Symbiosome development was studied in pea root nodules from plants growing in the absence of boron (B). Rhizobia released into the host cells of nodules from B-deficient plants developed to abnormal endophytic forms with an altered electrophoretic lipopolysaccharide pattern. Immunostaining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotting of nodule homogenates with antibodies that recognize glycoprotein components showed that two previously described lectin-like glycoproteins (PsNLEC-1A and PsNLEC-1B) did not harbor the carbohydrate epitope normally recognized by specific monoclonal antibodies. Material derived from B-deficient nodules, however, still contained three antigenic isoforms with similar electrophoretic mobilities to PsNLEC-1 isoforms A, B, and C. These could be detected following immunoblotting and immunostaining with a specific antiserum originating from the purified PsNLEC protein that had been heterologously expressed in Escherichia coli. Immunogold localization of PsNLEC-1 sugar epitopes in B-deficient nodules showed that they were associated mostly with cytoplasmic vesicles rather than normal localization in the symbiosome compartment of mature infected cells. These results suggest that a modification of the glycosyl-moieties of PsNLEC-1 and an alteration of vesicle targeting occur during the development of pea nodules in the absence of B, and that these changes are associated with the development of aberrant nonfunctional symbiosomes.


Multimodal wireless sensor network-based ambient assisted living in real homes with multiple residents.
Human activity recognition and behavior monitoring in a home setting using wireless sensor networks (WSNs) provide a great potential for ambient assisted living (AAL) applications, ranging from health and wellbeing monitoring to resource consumption monitoring. However, due to the limitations of the sensor devices, challenges in wireless communication and the challenges in processing large amounts of sensor data in order to recognize complex human activities, WSN-based AAL systems are not effectively integrated in the home environment. Additionally, given the variety of sensor types and activities, selecting the most suitable set of sensors in the deployment is an important task. In order to investigate and propose solutions to such challenges, we introduce a WSN-based multimodal AAL system compatible for homes with multiple residents. Particularly, we focus on the details of the system architecture, including the challenges of sensor selection, deployment, networking and data collection and provide guidelines for the design and deployment of an effective AAL system. We also present the details of the field study we conducted, using the systems deployed in two different real home environments with multiple residents. With these systems, we are able to collect ambient sensor data from multiple homes. This data can be used to assess the wellbeing of the residents and identify deviations from everyday routines, which may be indicators of health problems. Finally, in order to elaborate on the possible applications of the proposed AAL system and to exemplify directions for processing the collected data, we provide the results of several human activity inference experiments, along with examples on how such results could be interpreted. We believe that the experiences shared in this work will contribute towards accelerating the acceptance of WSN-based AAL systems in the home setting.


A systematic review of evidence-based treatments for prurigo nodularis.
Prurigo nodularis is a chronic dermatologic condition involving the development of multiple cutaneous nodules in the setting of intractable pruritus. Given emerging treatment options for this difficult-to-treat condition, a current review of therapeutics is needed. A systematic review was performed for clinical studies investigating prurigo nodularis treatment published from 1990 to present including ≥5 subjects. A total of 35 articles were assigned a level of evidence according to the Oxford Center for Evidence-based Medicine. All 5 studies investigating topical agents, including corticosteroids, calcineurin inhibitors, calcipotriol, and capsaicin, conveyed some beneficial effect with level of evidence 2b or higher. Six of 8 reports investigating photo- and photochemotherapy achieved levels of evidence 2b or greater and showed good partial response rates. Thalidomide was studied by 6 reports providing evidence of good symptom response, only 2 of which were rated level 2b or greater. Cyclosporine and methotrexate have demonstrated benefit in 4 combined studies, albeit with level 4 evidence. Pregabalin, amitriptyline, paroxetine, fluvoxamine, and neurokinin-1 receptor antagonists have demonstrated promising evidence in 5 level 2b studies. Higher-powered studies and additional randomized controlled trials are needed for the evaluation of safe and efficacious systemic treatment options for prurigo nodularis.


Decoding information about dynamically occluded objects in visual cortex.
During dynamic occlusion, an object passes behind an occluding surface and then later reappears. Even when completely occluded from view, such objects are experienced as continuing to exist or persist behind the occluder even though they are no longer visible. The contents and neural basis of this persistent representation remain poorly understood. Questions remain as to whether there is information maintained about the object itself (i.e. its shape or identity) or non-object-specific information such as its position or velocity as it is tracked behind an occluder, as well as which areas of visual cortex represent such information. Recent studies have found that early visual cortex is activated by "invisible" objects during visual imagery and by unstimulated regions along the path of apparent motion, suggesting that some properties of dynamically occluded objects may also be neurally represented in early visual cortex. We applied functional magnetic resonance imaging in human subjects to examine representations within visual cortex during dynamic occlusion. For gradually occluded, but not for instantly disappearing objects, there was an increase in activity in early visual cortex (V1, V2, and V3). This activity was spatially-specific, corresponding to the occluded location in the visual field. However, the activity did not encode enough information about object identity to discriminate between different kinds of occluded objects (circles vs. stars) using MVPA. In contrast, object identity could be decoded in spatially-specific subregions of higher-order, topographically organized areas such as ventral, lateral, and temporal occipital areas (VO, LO, and TO) as well as the functionally defined LOC and hMT+. These results suggest that early visual cortex may only represent the dynamically occluded object's position or motion path, while later visual areas represent object-specific information.


Genomic comparisons reveal biogeographic and anthropogenic impacts in the koala (Phascolarctos cinereus): a dietary-specialist species distributed across heterogeneous environments.
The Australian koala is an iconic marsupial with highly specific dietary requirements distributed across heterogeneous environments, over a large geographic range. The distribution and genetic structure of koala populations has been heavily influenced by human actions, specifically habitat modification, hunting and translocation of koalas. There is currently limited information on population diversity and gene flow at a species-wide scale, or with consideration to the potential impacts of local adaptation. Using species-wide sampling across heterogeneous environments, and high-density genome-wide markers (SNPs and PAVs), we show that most koala populations display levels of diversity comparable to other outbred species, except for those populations impacted by population reductions. Genetic clustering analysis and phylogenetic reconstruction reveals a lack of support for current taxonomic classification of three koala subspecies, with only a single evolutionary significant unit supported. Furthermore, ~70% of genetic variance is accounted for at the individual level. The Sydney Basin region is highlighted as a unique reservoir of genetic diversity, having higher diversity levels (i.e., Blue Mountains region; AvHecorr=0.20, PL% = 68.6). Broad-scale population differentiation is primarily driven by an isolation by distance genetic structure model (49% of genetic variance), with clinal local adaptation corresponding to habitat bioregions. Signatures of selection were detected between bioregions, with no single region returning evidence of strong selection. The results of this study show that although the koala is widely considered to be a dietary-specialist species, this apparent specialisation has not limited the koala's ability to maintain gene flow and adapt across divergent environments as long as the required food source is available.


Revising the Cannabis Use Disorders Identification Test (CUDIT) by means of Item Response Theory.
Cannabis use among adolescents and young adults has become a major public health challenge. Several European countries are currently developing short screening instruments to identify 'problematic' forms of cannabis use in general population surveys. One such instrument is the Cannabis Use Disorders Identification Test (CUDIT), a 10-item questionnaire based on the Alcohol Use Disorders Identification Test. Previous research found that some CUDIT items did not perform well psychometrically. In the interests of improving the psychometric properties of the CUDIT, this study replaces the poorly performing items with new items that specifically address cannabis use. Analyses are based on a sub-sample of 558 recent cannabis users from a representative population sample of 5722 individuals (aged 13-32) who were surveyed in the 2007 Swiss Cannabis Monitoring Study. Four new items were added to the original CUDIT. Psychometric properties of all 14 items, as well as the dimensionality of the supplemented CUDIT were then examined using Item Response Theory. Results indicate the unidimensionality of CUDIT and an improvement in its psychometric performance when three original items (usual hours being stoned; injuries; guilt) are replaced by new ones (motives for using cannabis; missing out leisure time activities; difficulties at work/school). However, improvements were limited to cannabis users with a high problem score. For epidemiological purposes, any further revision of CUDIT should therefore include a greater number of 'easier' items.


Characterization of the recD gene of Neisseria gonorrhoeae MS11 and the effect of recD inactivation on pilin variation and DNA transformation.
Pilin antigenic variation in Neisseria gonorrhoeae may result following intrachromosomal recombination between homologous pil genes. Despite extensive study, recA is the only previously characterized gene known to be involved in this process. In this study, the gonococcal recD gene, encoding one subunit of the putative RecBCD holoenzyme, was characterized and its role in pilin variation assessed. The complete recD gene of N. gonorrhoeae MS11 was cloned and its nucleotide sequence determined. The gonococcal recD gene complemented a defined Escherichia coli recD mutant, based on plaque formation of bacteriophage lambda and the restoration of ATP-dependent nuclease activity. Inactivation of the gonococcal recD gene had no measurable effect on cell viability or survival following UV exposure, but did decrease the frequency of DNA transformation approximately threefold. The frequency at which non-parental pilin phenotypes were spawned was 12-fold greater in MS11 recD mutants compared with the parental MS11 rec+ strain. Similar results were obtained using recD mutants that were not competent for DNA transformation. Complementation of the MS11 recD mutant with a wild-type recD gene copy restored the frequency of pilin phenotypic variation to approximately wild-type levels. The nucleotide changes at pilE in the recD mutants were confined to the variable regions of the gene and were similar to changes previously attributed to gene conversion.


Effects of chemotherapy on osseointegration of implants: a case report.
A patient underwent mandibular resection for high-grade osteosarcoma with immediate reconstruction with a microvascular fibula free bone graft and simultaneous placement of osseointegrated implants. Following initial healing, she underwent six cycles of chemotherapy and had further revision surgery prior to implant exposure and construction of a prosthesis. The chemotherapy appears to have had no deleterious effects on implant osseointegration or survival.


Sleep, daily activity rhythms and postpartum mood: A longitudinal study across the perinatal period.
Women with a diagnosis of bipolar and major depressive disorders are at higher risk to develop postpartum depression. The primary objective of this longitudinal study was to determine whether daily activity rhythms and sleep parameters differ between women with and without a history of a mood disorder across the perinatal period. A secondary objective was to determine whether changes in these parameters were associated with postpartum mood. In total, 33 women were included in this study, 15 of which had a history of a mood disorder (high-risk group) and 18 who did not (low-risk group). Sleep and daily rhythms were assessed subjectively and objectively during the third trimester (≥26 weeks gestation) and again at 6-12 weeks postpartum. Mood was also assessed at both time points. Women in the high-risk group showed greater subjective daily rhythms and sleep disturbances across the perinatal period. Objective sleep efficiency was worse in the high-risk group in the postpartum period. Changes in both subjective daily rhythms and objective sleep efficiency were predictive of changes in depressive symptoms across the perinatal period. These findings encourage the development of preventative therapeutics to ensure circadian rhythm and sleep stability throughout the perinatal period.


Comparable rest-related promotion of spatial memory consolidation in younger and older adults.
Flexible spatial navigation depends on cognitive mapping, a function that declines with increasing age. In young adults, a brief period of postnavigation rest promotes the consolidation and integration of spatial memories into accurate cognitive maps. We examined (1) whether rest promotes spatial memory consolidation and integration in older adults; and (2) whether the magnitude of the rest benefit changes with increasing age. Young and older adults learned a route through a virtual environment, followed by a 10-minute delay comprising either wakeful rest or a perceptual task, and a subsequent cognitive mapping task, requiring the pointing to landmarks from different locations. Pointing accuracy was lower in the older than younger adults. However, there was a comparable rest-related enhancement in pointing accuracy in the 2 age groups. Together our findings suggest that (1) the age-related decline in cognitive mapping cannot be explained by increased consolidation interference in older adults; and (2) as we grow older, rest continues to support the consolidation and integration of spatial memories.


A temporal dependency account of attentional inhibition in oculomotor control.
We used concurrent electroencephalogram (EEG) and eye tracking to investigate the role of covert attentional mechanisms in the control of oculomotor behavior. Human participants made speeded saccades to targets that were presented alongside salient distractors. By subsequently sorting trials based on whether the distractor was strongly represented or suppressed by the visual system - as evident in the accuracy (Exp. 1) or quality of the saccade (Exp. 2) - we could characterize and contrast pre-saccadic neural activity as a function of whether oculomotor control was established. Results show that saccadic behavior is strongly linked to the operation of attentional mechanisms in visual cortex. In Experiment 1, accurate saccades were preceded by attentional selection of the target - indexed by a target-elicited N2pc component - and by attentional suppression of the distractor - indexed by early and late distractor-elicited distractor positivity (Pd) components. In Experiment 2, the strength of distractor suppression predicted the degree to which the path of slower saccades would deviate away from the distractor en route to the target. However, results also demonstrated clear dissociations of covert and overt selective control, with saccadic latency in particular showing no relationship to the latency of covert selective mechanisms. Eye movements could thus be initiated prior to the onset of attentional ERP components, resulting in stimulus-driven behaviour. Taken together, the results indicate that attentional mechanisms play a role in determining saccadic behavior, but that saccade timing is not contingent on the deployment of attention. This creates a temporal dependency, whereby attention fosters oculomotor control only when attentional mechanisms are given sufficient opportunity to impact stimuli representations before an eye movement is executed.


Lipolytic Effects of 3-Iodothyronamine (T1AM) and a Novel Thyronamine-Like Analog SG-2 through the AMPK Pathway.
3-Iodothyronamine (T1AM) and its synthetic analog SG-2 are rapidly emerging as promising drivers of cellular metabolic reprogramming. Our recent research indicates that in obese mice a sub-chronic low dose T1AM treatment increased lipolysis, associated with significant weight loss independent of food consumption. The specific cellular mechanism of T1AM's lipolytic effect and its site of action remains unknown. First, to study the mechanism used by T1AM to gain entry into cells, we synthesized a fluoro-labeled version of T1AM (FL-T1AM) by conjugating it to rhodamine (TRITC) and analyzed its cellular uptake and localization in 3T3-L1 mouse adipocytes. Cell imaging using confocal microscopy revealed a rapid intercellular uptake of FL-T1AM into mitochondria without localization to the lipid droplet or nucleus of mature adipocytes. Treatment of 3T3-L1 adipocytes with T1AM and SG-2 resulted in decreased lipid accumulation, the latter showing a significantly higher potency than T1AM (10 µM vs. 20 µM, respectively). We further examined the effects of T1AM and SG-2 on liver HepG2 cells. A significant decrease in lipid accumulation was observed in HepG2 cells treated with T1AM or SG-2, due to increased lipolytic activity. This was confirmed by accumulation of glycerol in the culture media and through activation of the AMPK/ACC signaling pathways.


Ligand Depot: a data warehouse for ligands bound to macromolecules.
Ligand Depot is an integrated data resource for finding information about small molecules bound to proteins and nucleic acids. The initial release (version 1.0, November, 2003) focuses on providing chemical and structural information for small molecules found as part of the structures deposited in the Protein Data Bank. Ligand Depot accepts keyword-based queries and also provides a graphical interface for performing chemical substructure searches. A wide variety of web resources that contain information on small molecules may also be accessed through Ligand Depot. Ligand Depot is available at http://ligand-depot.rutgers.edu/. Version 1.0 supports multiple operating systems including Windows, Unix, Linux and the Macintosh operating system. The current drawing tool works in Internet Explorer, Netscape and Mozilla on Windows, Unix and Linux.


Accurate sampling of high-frequency motions in proteins by steady-state (15)N-{(1)H} nuclear Overhauser effect measurements in the presence of cross-correlated relaxation.
The steady-state {(1)H}-(15)N NOE experiment is used in most common NMR analyses of backbone dynamics to accurately ascertain the effects of the fast dynamic modes. We demonstrate here that, in its most common implementation, this experiment generates an incorrect steady state in the presence of CSA/dipole cross-correlated relaxation leading to large errors in the characterization of these high-frequency modes. This affects both the quantitative and qualitative interpretation of (15)N backbone relaxation in dynamic terms. We demonstrate further that minor changes in the experimental implementation effectively remove these errors and allow a more accurate interpretation of protein backbone dynamics.


Neural indicators of articulator-specific sensorimotor influences on infant speech perception.
While there is increasing acceptance that even young infants detect correspondences between heard and seen speech, the common view is that oral-motor movements related to speech production cannot influence speech perception until infants begin to babble or speak. We investigated the extent of multimodal speech influences on auditory speech perception in prebabbling infants who have limited speech-like oral-motor repertoires. We used event-related potentials (ERPs) to examine how sensorimotor influences to the infant's own articulatory movements impact auditory speech perception in 3-mo-old infants. In experiment 1, there were ERP discriminative responses to phonetic category changes across two phonetic contrasts (bilabial-dental /ba/-/ɗa/; dental-retroflex /ɗa/-/ɖa/) in a mismatch paradigm, indicating that infants auditorily discriminated both contrasts. In experiment 2, inhibiting infants' own tongue-tip movements had a disruptive influence on the early ERP discriminative response to the /ɗa/-/ɖa/ contrast only. The same articulatory inhibition had contrasting effects on the perception of the /ba/-/ɗa/ contrast, which requires different articulators (the lips vs. the tongue) during production, and the /ɗa/-/ɖa/ contrast, whereby both phones require tongue-tip movement as a place of articulation. This articulatory distinction between the two contrasts plausibly accounts for the distinct influence of tongue-tip suppression on the neural responses to phonetic category change perception in definitively prebabbling, 3-mo-old, infants. The results showing a specificity in the relation between oral-motor inhibition and phonetic speech discrimination suggest a surprisingly early mapping between auditory and motor speech representation already in prebabbling infants.


A novel pyridinium modified tetraphenylethene: AIE-activity, mechanochromism, DNA detection and mitochondrial imaging.
We report the design and synthesis of a novel cationic AIE-gen, i.e., (E)-TPEDEPy-DBz, which can be derived from a facile synthetic route. (E)-TPEDEPy-DBz is a stereo-pure isomer and shows evident AIE properties with yellow-greenish emission in the solid state (ΦF = 43.7%). Distinct from most of the ionic AIE-gens reported in the literature, which are usually exemplified by their water solubility and the capacity for interaction with charged biological species, (E)-TPEDEPy-DBz demonstrated moderate solvatochromism and evident mechanoluminochromism behaviours. The emission of (E)-TPEDEPy-DBz can be switched between yellow-greenish in the crystalline state to red-orange in the amorphous state by grinding and fuming treatments. Due to the AIE-characteristics, cationic modification and the thus-acquired water solubility, (E)-TPEDEPy-DBz performed as a sensitive fluorescent probe to detect DNA in Tris-HCl buffer solution. More importantly, (E)-TPEDEPy-DBz showed good membrane permeability and cell viability, and the potentiality to be used as a mitochondria-specific probe working in a fluorescence turn-on mode.


Aging and Financial Decision Making.
This study examines how cognitive changes associated with aging impact the financial decision making capability of older Americans. We find that a decrease in cognition is associated with a decrease in financial literacy. Decreases in episodic memory and visuospatial ability are associated with a decrease in numeracy, and a decrease in semantic memory is associated with a decrease in financial knowledge. A decrease in cognition also predicts a drop in self-confidence in general, but importantly, it is not associated with a drop in confidence in managing one's own finances. Participants experiencing decreases in cognition do show an increased likelihood of getting help with financial decisions; however, many participants experiencing significant drops in cognition still do not get help.


Career choices for ophthalmology made by newly qualified doctors in the United Kingdom, 1974-2005.
The paper aims to report trends in career choices for ophthalmology among UK medical graduates. Postal questionnaire surveys were undertaken of qualifiers from all UK medical schools in nine qualification years since 1974. Data were analysed by univariate cross-tabulation. The significance of comparisons between groups of doctors were calculated by the use of chi-squared tests and adjusted residuals. Ophthalmology was the first choice of long term career for 2.3% of men and 1.5% of women one year after qualification; 2.0% of men and 1.4% of women three years after; and 1.8% of men and 1.2% of women at five years. Comparing early choices with eventual destinations, 64% who chose ophthalmology in year one, 84% in year three, and 92% in year five eventually practised in the specialty. The concordance between year one choice and eventual destination was higher for ophthalmology than for most other specialties. 'Enthusiasm for and commitment to the specialty' was the most important single factor in influencing career choice. The prospect of good working hours and conditions was also an important influence: it influenced career choice a great deal for a higher percentage of those who chose ophthalmology (66% in the third year) than those who made other surgical choices (23%). Those choosing ophthalmology show a high level of commitment to it. Their commitment is strengthened by the prospect of attractive hours and working conditions. Many doctors who become ophthalmologists have already made their choice by the end of their first post-qualification year.


Metabolic profiling of hearts exposed to sevoflurane and propofol reveals distinct regulation of fatty acid and glucose oxidation: CD36 and pyruvate dehydrogenase as key regulators in anesthetic-induced fuel shift.
Myocardial energy metabolism is a strong predictor of postoperative cardiac function. This study profiled the metabolites and metabolic changes in the myocardium exposed to sevoflurane, propofol, and Intralipid and investigated the underlying molecular mechanisms. Sevoflurane (2 vol%) and propofol (10 and 100 microM) in the formulation of 1% Diprivan (AstraZeneca Inc., Mississauga, ON, Canada) were compared for their effects on oxidative energy metabolism and contractility in the isolated working rat heart model. Intralipid served as a control. Substrate flux through the major pathways for adenosine triphosphate generation in the heart, that is, fatty acid and glucose oxidation, was measured using [H]palmitate and [C]glucose. Biochemical analyses of nucleotides, acyl-CoAs, ceramides, and 32 acylcarnitine species were used to profile individual metabolites. Lipid rafts were isolated and used for Western blotting of the plasma membrane transporters CD36 and glucose transporter 4. Metabolic profiling of the hearts exposed to sevoflurane and propofol revealed distinct regulation of fatty acid and glucose oxidation. Sevoflurane selectively decreased fatty acid oxidation, which was closely related to a marked reduction in left ventricular work. In contrast, propofol at 100 microM but not 10 microM increased glucose oxidation without affecting cardiac work. Sevoflurane decreased fatty acid transporter CD36 in lipid rafts/caveolae, whereas high propofol increased pyruvate dehydrogenase activity without affecting glucose transporter 4, providing mechanisms for the fuel shifts in energy metabolism. Propofol increased ceramide formation, and Intralipid increased hydroxy acylcarnitine species. Anesthetics and their solvents elicit distinct metabolic profiles in the myocardium, which may have clinical implications for the already jeopardized diseased heart.


Face adaptation effects show strong and long-lasting transfer from lab to more ecological contexts.
A review on recent experiments on figural face aftereffects reveals that adaptation effects in famous faces can last for hours up to days. Such adaptations seem to be highly reliable regarding test-retest designs as well as regarding the generalizability of adaptation across different adaptation routines and adaptations toward different kinds of facial properties. However, in the studies conducted so far, adaptation and the subsequent test phase were carried out in typical laboratory environments. Under these circumstances, it cannot be ruled out that the observed effects are, in fact, episodic learn-test compatibility effects. To test for ecological validity in adaptation effects we used an adaptation paradigm including environmental and social properties that differed between adaptation and test phase. With matched samples (n1 = n2 = 54) we found no main effects of experimental setting compatibility resulting from varying where the tests where conducted (environmental condition) nor any interaction with effects of stimulus compatibility resulting from varying stimulus similarity between adaptation and test phase using the same picture, different pictures of the same person, or different persons (transfer). This indicates that these adaptation effects are not artificial or merely lab-biased effects. Adaptation to face stimuli may document representational adaptations and tuning mechanisms that integrate new visual input in a very fast, reliable, and sustainable way.


Rrp17p is a eukaryotic exonuclease required for 5' end processing of Pre-60S ribosomal RNA.
Ribosomal processing requires a series of endo- and exonucleolytic steps for the production of mature ribosomes, of which most have been described. To ensure ribosome synthesis, 3' end formation of rRNA uses multiple nucleases acting in parallel; however, a similar parallel mechanism had not been described for 5' end maturation. Here, we identify Rrp17p as a previously unidentified 5'-3' exonuclease essential for ribosome biogenesis, functioning with Rat1p in a parallel processing pathway analogous to that of 3' end formation. Rrp17p is required for efficient exonuclease digestion of the mature 5' ends of 5.8S(S) and 25S rRNAs, contains a catalytic domain close to its N terminus, and is highly conserved among higher eukaryotes, being a member of a family of exonucleases. We show that Rrp17p binds late pre-60S ribosomes, accompanying them from the nucleolus to the nuclear periphery, and provide evidence for physical and functional links between late 60S subunit processing and export.


Potential role of intermittent functioning of baroreflexes in the etiology of hypertension in spontaneously hypertensive rats.
The spontaneously hypertensive rat (SHR) is a genetic model of primary hypertension with an etiology that includes sympathetic overdrive. To elucidate the neurogenic mechanisms underlying the pathophysiology of this model, we analyzed the dynamic baroreflex response to spontaneous fluctuations in arterial pressure in conscious SHRs, as well as in the Wistar-Kyoto (WKY), the Dahl salt-sensitive, the Dahl salt-resistant, and the Sprague-Dawley rat. Observations revealed the existence of long intermittent periods (lasting up to several minutes) of engagement and disengagement of baroreflex control of heart rate. Analysis of these intermittent periods revealed a predictive relationship between increased mean arterial pressure and progressive baroreflex disengagement that was present in the SHR and WKY strains but absent in others. This relationship yielded the hypothesis that a lower proportion of engagement versus disengagement of the baroreflex in SHR compared with WKY contributes to the hypertension (or increased blood pressure) in SHR compared with WKY. Results of experiments using sinoaortic baroreceptor denervation were consistent with the hypothesis that dysfunction of the baroreflex contributes to the etiology of hypertension in the SHR. Thus, this study provides experimental evidence for the roles of the baroreflex in long-term arterial pressure regulation and in the etiology of primary hypertension in this animal model.


Turning down the heat: An enhanced understanding of the relationship between urban vegetation and surface temperature at the city scale.
Guiding urban planners on the cooling returns of different configurations of urban vegetation is important to protect urban dwellers from adverse heat impacts. To this end, we estimated statistical models that fused multi-temporal very fine spatial (20 cm) and vertical (1 mm) resolution imagery, that captures the complexity of urban vegetation, with remotely sensed temperature data to assess how urban vegetation configuration influences urban temperatures. Perth, Western Australia, was used as a case-study for this analysis. Panel regression models showed that within a location an increase in tree and shrub cover has a larger cooling effect than grass coverage. On average, holding all else equal, an approximate 1 km2 increase in shrub (tree) cover within a location reduces surface temperatures by 12 °C (5 °C). We included a range of robustness checks for the observed relationships between urban vegetation type and temperature. Geographically weighted regression models showed spatial variation in the cooling effect of different vegetation types; this indicates that i) unobserved factors moderate temperature-vegetation relationships across urban landscapes, and ii) that urban vegetation type and temperature relationships are complex. Machine learning models (Random Forests) were used to further explore complex and non-linear relationships between different urban vegetation configurations and temperature. The Random Forests showed that vegetation type explained 31.84% of the out-of-bag variance in summer surface temperatures, that increased cover of large vegetation within a location increases cooling, and that different configurations of urban vegetation structure can lead to cooling gains. The models in this study were trained with vegetation data capturing local detail, multiple time-periods, and entire city coverage. Thus, these models illustrate the potential to develop locally-detailed and spatially explicit tools to guide planning of vegetation configuration to optimise cooling at local- and city-scales.


Duloxetine does not relieve painful physical symptoms in depression: a meta-analysis.
Duloxetine inhibits both serotonin and norepinephrine reuptake and is marketed as a treatment for both the core emotional symptoms and painful physical complaints that often accompany depression. Some studies have found that duloxetine is efficacious in treating painful symptoms associated with depression but these findings have been inconsistent. Several narrative review articles have reached positive conclusions about the efficacy of duloxetine as an analgesic in depression but there has been no quantitative systematic review regarding the impact of duloxetine on pain among this population. A meta-analysis of data pertaining to duloxetine's purported analgesic effects on depressed patients was thus undertaken. Studies were selected through searching Medline and Cochrane Trial databases as well as examining Lilly's public clinical trial database. A random effects model was used. Across five trials, the results indicate a very small (d = 0.115) and statistically nonsignificant (p = 0.057) analgesic effect for duloxetine. Additionally, some of the relevant data on duloxetine's effects have not been reported fully, making it likely that the obtained results reflect an overestimate of its true impact on painful physical symptoms in depression. The current analysis is based on a small number of studies; further trials may yield significant results favoring duloxetine. Based upon the currently available evidence, the marketing of duloxetine as an antidepressant with analgesic properties for people with depression does not appear to be adequately supported.


Effects of intensive mariculture on sediment biochemistry.
The exponential growth of off-shore mariculture that has occurred worldwide over the last 10 years has raised concern about the impact of the waste produced by this industry on the ecological integrity of the sea bottom. Investigations into this potential source of impact on the biochemistry of the sea floor have provided contrasting results, and no compelling explanations for these discrepancies have been provided to date. To quantify the impact of fish-farm activities on the biochemistry of sediments, we have investigated the quantity and biochemical composition of sediment organic matter in four different regions in the temperate-warm Mediterranean Sea: Akrotiri Bay (Cyprus), Sounion Bay (Greece), Pachino Bay (Italy), and the Gulf of Alicante (Spain). In these four study regions, the concentrations of phytopigments, proteins, carbohydrates, and lipids in the sediments were measured, comparing locations receiving wastes from fish farms to control locations in two different habitats: seagrass beds and soft nonvegetated substrates. Downward fluxes were also measured in all of the regions, up to 200 m from the fish farms, to assess the potential spatial extent of the impact. In all four regions, with the exception of seagrass sediments in Spain, the biochemistry of the sediments showed significant differences between the control and fish-farm locations. However, the variables explaining the differences observed varied among the regions and between habitats, suggesting idiosyncratic effects of fish-farm waste on the biochemistry of sediments. These are possibly related to differences in the local physicochemical variables that could explain a significant proportion of the differences seen between the control and fish-farm locations. Biodeposition derived from the fish farms decreased with increasing distance from the fish-farm cages, but with different patterns in the four regions. Our results indicate that quantitative and qualitative changes in the organic loads of the sediments that arise from intensive aquaculture are dependent upon the ecological context and are not predictable only on the basis of fish-farm attributes and hydrodynamic regimes. Therefore, the siting of fish farms should only be allowed after a case-by-case assessment of the ecological context of the region, especially in terms of the organic matter load and its biochemical composition.


Reannotation of the CELO genome characterizes a set of previously unassigned open reading frames and points to novel modes of host interaction in avian adenoviruses.
The genome of the avian adenovirus Chicken Embryo Lethal Orphan (CELO) has two terminal regions without detectable homology in mammalian adenoviruses that are left without annotation in the initial analysis. Since adenoviruses have been a rich source of new insights into molecular cell biology and practical applications of CELO as gene a delivery vector are being considered, this genome appeared worth revisiting. We conducted a systematic reannotation and in-depth sequence analysis of the CELO genome. We describe a strongly diverged paralogous cluster including ORF-2, ORF-12, ORF-13, and ORF-14 with an ATPase/helicase domain most likely acquired from adeno-associated parvoviruses. None of these ORFs appear to have retained ATPase/helicase function and alternative functions (e.g. modulation of gene expression during the early life-cycle) must be considered in an adenoviral context. Further, we identified a cluster of three putative type-1-transmembrane glycoproteins with IG-like domains (ORF-9, ORF-10, ORF-11) which are good candidates to substitute for the missing immunomodulatory functions of mammalian adenoviruses. ORF-16 (located directly adjacent) displays distant homology to vertebrate mono-ADP-ribosyltransferases. Members of this family are known to be involved in immuno-regulation and similiar functions during CELO life cycle can be considered for this ORF. Finally, we describe a putative triglyceride lipase (merged ORF-18/19) with additional domains, which can be expected to have specific roles during the infection of birds, since they are unique to avian adenoviruses and Marek's disease-like viruses, a group of pathogenic avian herpesviruses. We could characterize most of the previously unassigned ORFs pointing to functions in host-virus interaction. The results provide new directives for rationally designed experiments.


Synthesis and biological evaluation of novel indoline-2,3-dione derivatives as antitumor agents.
A new series of 1,5-disubstituted indolin-2,3-diones was synthesized and their inhibition of the growth of a human acute promyelocytic leukemia (HL-60) cell line was evaluated. These compounds had promising inhibition of HL-60 cell growth in vitro. Results indicated that compounds with a benzyl substituent at the N-1 position on the indolin-2,3-dione ring had more potent antiproliferative activity than those with a (4-fluorobenzyl) amino-2-oxoethyl substituent at the N-1 position. Among the compounds synthesized, compound 8l inhibited half of cell growth at a concentration of 0.07 μM and compound 8p did so at a concentration of 0.14 μM. These compounds may serve as lead compounds for further optimization in order to develop novel anticancer agents.


The euryarchaeotes, a subdomain of Archaea, survive on a single DNA polymerase: fact or farce?
Archaea is now recognized as the third domain of life. Since their discovery, much effort has been directed towards understanding the molecular biology and biochemistry of Archaea. The objective is to comprehend the complete structure and the depth of the phylogenetic tree of life. DNA replication is one of the most important events in living organisms and DNA polymerase is the key enzyme in the molecular machinery which drives the process. All archaeal DNA polymerases were thought to belong to family B. This was because all of the products of pol genes that had been cloned showed amino acid sequence similarities to those of this family, which includes three eukaryal DNA replicases and Escherichia coli DNA polymerase II. Recently, we found a new heterodimeric DNA polymerase from the hyperthermophilic archaeon, Pyrococcus furiosus. The genes coding for the subunits of this DNA polymerase are conserved in the euryarchaeotes whose genomes have been completely sequenced. The biochemical characteristics of the novel DNA polymerase family suggest that its members play an important role in DNA replication within euryarchaeal cells. We review here our current knowledge on DNA polymerases in Archaea with emphasis on the novel DNA polymerase discovered in Euryarchaeota.


A comparison of the nursing practice environment in mental health and medical-surgical settings.
To examine the differences between characteristics of the work environment of nurses working in mental health and general acute inpatient nursing settings. Secondary analysis of data collected on 96 randomly selected medical and surgical (general) wards and six mental health wards in 24 public acute general hospitals across two Australian states between 2004 and 2006. All nurses on the participating wards were asked to complete a survey that included the Practice Environment Scale of the Nursing Work Index (NWI-PES). Responses were received from 2,556 nurses (76.3% response rate). Using the five-domain structure, comparisons were made between mental health and general nurses. Across the entire sample of nurses, those working in mental health settings scored more highly in regard to nurse-doctor relationships and staffing adequacy. Nurses in general wards reported more participation in hospital affairs, stronger leadership, and the presence of more of the foundations of nursing quality care such as access to continued education. Differences between the groups on each of the domains was statistically significant at p=.05 or greater, but not for the composite practice environment scale. A wide range of responses was seen when data were aggregated to the ward level. The work environment of mental health nurses is different from that of their colleagues working in general settings. Specific areas of the mental health environment, such as participation in the hospital, leadership, and the foundations of quality, may be enhanced to improve nurses' job satisfaction and, potentially, other nurse and patient outcomes. Factors in the medical and surgical nursing practice environment have been established as significant influences on nurse and patient outcomes. It is important to understand the existence and potential impact of these factors in mental health inpatient settings.


Guidance for Evaluating the Safety of Experimental Releases of Mosquitoes, Emphasizing Mark-Release-Recapture Techniques.
Experimental releases of mosquitoes are performed to understand characteristics of populations related to the biology, ability to transmit pathogens, and ultimately their control. In this article, we discuss considerations related to the safety of experimental releases of living mosquitoes, applying principles of good practice in vector biology that protect human health and comfort. We describe specific factors of experimental releases of mosquitoes that we believe are critical to inform institutional biosafety committees and similar review boards to which proposals to conduct mosquito release experiments have been submitted. In this study, "experimental releases" means those that do not significantly increase vector capacity or nuisance biting relative to the unperturbed natural baseline. This document specifically does not address releases of mosquitoes for ongoing control programs or trials of new control methods for which broader assessments of risk are required. It also does not address releases of transgenic or exotic (non-native) mosquito species, both of which require particular regulatory approval. Experimental releases may include females and males and evaluation must consider their effects based on the number released, their genotype and phenotype, the environment into which they are released, and postrelease collection activities. We consider whether increases of disease transmission and nuisance biting might result from proposed experimental releases against the backdrop of natural population size variation. We recommend that experimental releases be conducted in a manner that can be reasonably argued to have insignificant negative effects. Reviewers of proposals for experimental releases should expect applicants to provide such an argument based on evidence from similar studies and their planned activities. This document provides guidance for creating and evaluating such proposals.


2-{[(Dimethyl-amino)-methyl-idene]amino}-5-nitro-benzonitrile.
The title mol-ecule, C10H10N4O2, is almost planar and adopts an E configuration of the azomethine [C=N = 1.298 (2) Å] double bond. The benzene ring is attached to an essentially planar (r.m.s. deviation = 0.0226 Å) amidine moiety (N=CN/Me2), the dihedral angle between the two mean planes being 18.42 (11)°. The cyano group lies in the plane of the benzene ring [the C and N atoms deviating by 0.030 (3) and 0.040 (3) Å, respectively], while the nitro group makes a dihedral angle 5.8 (3)° with the benzene ring. There are two distinct inter-molecular hydrogen bonds, C-H⋯O and C-H⋯N, that stabilize the crystal structure; the former inter-actions result in centrosymmetric dimers about inversion centers resulting in ten-membered rings, while the later give rise to chains of mol-ecules running parallel to the b axis.