Let's See a Boy and a Balloon: Argument Labels and Syntactic Frame in Verb Learning.
It is by now well established that toddlers use the linguistic context in which a new word-and particularly a new verb-appears to discover aspects of its meaning. But what aspects of the linguistic context are most useful? To begin to investigate this, we ask how 2-year-olds use two sources of linguistic information that are known to be useful to older children and adults in verb guessing tasks: syntactic frame, and the semantic content available in the noun phrases labeling the verb's arguments. We manipulate the linguistic contexts in which we present novel verbs to see how they use these two sources of information, both separately and in combination, to acquire the verb's meaning. Our results reveal that like older children and adults, toddlers make use of both syntactic frame and semantically contentful argument labels to acquire verb meaning. But toddlers also require these two sources of information to be packaged in a particular way, into a single sentence that identifies 'who did what to whom.'


Normalization in mouse primary visual cortex.
When multiple stimuli appear together in the receptive field of a visual cortical neuron, the response is typically close to the average of that neuron's response to each individual stimulus. The departure from a linear sum of each individual response is referred to as normalization. In mammals, normalization has been best characterized in the visual cortex of macaques and cats. Here we study visually evoked normalization in the visual cortex of awake mice using optical imaging of calcium indicators in large populations of layer 2/3 (L2/3) V1 excitatory neurons and electrophysiological recordings across layers in V1. Regardless of recording method, mouse visual cortical neurons exhibit normalization to varying degrees. The distributions of normalization strength are similar to those described in cats and macaques, albeit slightly weaker on average.


Validation of the Chinese version of the gambling motivation scale (C-GMS).
The French items of the Gambling Motivation Scale (GMS) were first developed and validated by Chantal and colleagues in 1994. The scale then became one of the most widely used motivational scales in the gambling literature of the West. The present study recruited 932 Chinese university students in order to validate the Chinese version of the Gambling Motivation Scale (C-GMS). The results of a confirmatory factor analysis of the Chinese data supported the 7-factor model as proposed by Chantal et al. (Soc Leis 17:189-212, 1994). This study also found a second-order model with three major factors, which corresponded to three types of gambling motivation including self-determined motivation (for knowledge, for accomplishment, for stimulation, and due to identified regulation), non self-determined motivation (due to introjected regulation and external regulation), and amotivation. All subscales demonstrated satisfactory internal consistency, and showed significant correlations with gambling correlates such as problem gambling symptoms and gambling intention. In sum, the C-GMS showed adequate psychometric properties and can be extended for use with Chinese populations.


Electrochemical Behavior of the Fe(III) Complexes of the Cyclic Hydroxamate Siderophores Alcaligin and Desferrioxamine E.
The redox behavior of Fe(III) complexes of the cyclic hydroxamate siderophores alcaligin and desferrioxamine E was investigated by cyclic voltammetry. The limiting, pH independent redox potential (E(1/2) vs NHE) is -446 mV for alcaligin above pH 9 and -477 mV for ferrioxamine E above pH 7.5. At lower pH values, the redox potential for both complexes shifts positive, with a loss of voltammetric reversibility which is interpreted to be the consequence of a secondary dissociation of Fe(II) from the reduced form of the complexes. These observations are of biological importance, since they suggest the possibility of a reductive mechanism in microbial cells which utilize these siderophores to acquire Fe. For comparison purposes, cyclic voltammograms were obtained for Fe(III) complexes with trihydroxamic acids of cyclic (ferrioxamine E) and linear (ferrioxamine B) structures, with dihydroxamic acids of cyclic (alcaligin) and linear (rhodotorulic and sebacic acids) structures, and with monohydroxamic acids (acetohydroxamic and N-methylacetohydroxamic acids) at identical conditions. The observed redox potentials allow us to estimate the overall stability constants for fully coordinated Fe(II) complexes as log beta(II)(Fe(2)alcaligin(3)) = 24.6 and log beta(II)(ferrioxamine E) = 12.1. A linear correlation between E(1/2) and pM was found, and the basis for this relationship is discussed in terms of structural (denticity and cyclic/acyclic) and electronic differences among the {alkyl-NOH-CO-alkyl} type of hydroxamic acid ligands studied.


Not an 'iron pipeline', but many capillaries: regulating passive transactions in Los Angeles' secondary, illegal gun market.
California has strict firearm-related laws and is exceptional in its regulation of firearms retailers. Though evidence suggests that these laws can reduce illegal access to guns, high levels of gun violence persist in Los Angeles (LA), California. This research seeks to describe the sources of guns accessed by active offenders in LA, California and reports offenders' motivations for obtaining guns. Los Angeles County Jail (LACJ) system (four facilities). Random sampling from a screened pool of eligible participants was used to conduct qualitative semistructured interviews with 140 incarcerated gun offenders in one of four (LACJ) facilities. Researchers collected data on firearm acquisition, experiences related to gun violence, and other topics, using a validated survey instrument. Grounded theory guided the collection and analysis of data. Respondents reported possession of 77 specific guns (79.2% handguns) collectively. Social networks facilitate access to illegal guns; the majority of interviewees acquired their illegal guns through a social connection (85.7%) versus an outside broker/unregulated retailer (8.5%). Most guns were obtained through illegal purchase (n=51) or gift (n=15). A quarter of gun purchasers report engaging in a passive transaction, or one initiated by another party. Passive gun buyers were motivated by concerns for personal safety and/or economic opportunity. In LA's illegal gun market, where existing social relationships facilitate access to guns across a diffuse network, individuals, influenced by both fear and economic opportunity, have frequent opportunities to illegally possess firearms through passive transactions. Gun policies should better target and minimise these transactions.


Polyfunctional cytomegalovirus-specific CD4+ and pp65 CD8+ T cells protect against high-level replication after liver transplantation.
To determine whether polyfunctional CD4+ T-cell responses coupled with CD8+ T-cell responses against human cytomegalovirus (HCMV) are key to the control of HCMV replication we prospectively analyzed 29 liver transplant recipients for CD4+ T-cell responses against soluble HCMV antigen, pp65 and IE1 proteins, CD8+ T-cell responses against pp65 and IE1 proteins and a range of T helper (Th) 1 and Th2 cytokines. Eleven patients (38%) developed HCMV DNAemia at a median of 21 days post-liver transplantation (range 17-31 days). There was a significantly lower frequency and absolute number of total HCMV CD4+ T cells producing IFNgamma, IFNgamma+IL2 and IL2 and pp65-CD8+ T cells producing IFNgamma in patients with DNAemia. The quantities of Th1 and Th2 cytokines present during the first 20 days posttransplant were not predictive of DNAemia. Cut-off levels during the first 20 days posttransplant of 0.1% of lysate stimulated CD4+ T cells producing IL2, and pp65-CD8+ T cells producing IFNgamma above 0.4% had positive and negative predictive values for DNAemia of 54% and 100% and 50% and 92%, respectively. Measuring polyfunctional CD4+ T cells against HCMV early posttransplant may allow targeted intervention to minimize the occurrence and acute and long-term consequences of HCMV replication.


Transfected rat high-molecular-weight neurofilament (NF-H) coassembles with vimentin in a predominantly nonphosphorylated form.
A fully encoding cDNA for the high-molecular-weight rat neurofilament protein (NF-H) has been isolated from a lambda gt11 library, sequenced and subcloned into eukaryotic expression vectors. Sequence analysis shows that rat NF-H has an overall homology of 72 and 88% with human and mouse NF-H, respectively. The head and rod domains are almost entirely identical, and the divergences are due to differences in the long C-terminal extensions of the molecule. The consensus phosphorylation sequence for neurofilaments Lys-Ser-Pro (KSP) is present 52 times. The predicted molecular mass of the protein is 115 kDa, 42% lower than that observed by SDS-PAGE. Upon transfection into vimentin-containing fibroblasts, such as L tk-, L929, and 3T6 cells, NF-H is seen distributed with vimentin by light and electron microscopic examinations indicating that copolymers of NF-H and vimentin are formed in these cells. Only a negligible proportion of the cells is positive when stained with a number of antibodies directed against phosphorylated NF-H epitopes. This is in contrast with the middle molecular weight NF protein (NF-M) transfected into L tk- and L929 cells, which can readily be detected by antibodies against phosphorylated neurofilament epitopes. The mobilities of the transfected protein on 1- and 2-dimensional gels confirm that NF-H is predominantly in a nonphosphorylated form. These results indicate that phosphorylation of NF-H, but not NF-M, on the KSP sequence is due to protein kinases, which are not present in fibroblasts and are presumably NF-H specific. The stable NF-H-expressing cell lines can therefore be used to study these putative neurofilament kinases in vitro and in vivo.


Effects of grazer presence on genetic structure of a phenotypically diverse diatom population.
Studies of predator-prey systems in both aquatic and terrestrial environments have shown that grazers structure the intraspecific diversity of prey species, given that the prey populations are phenotypically variable. Populations of phytoplankton have traditionally considered comprising only low intraspecific variation, hence selective grazing as a potentially structuring factor of both genetic and phenotypic diversity has not been comprehensively studied. In this study, we compared strain specific growth rates, production of polyunsaturated aldehydes, and chain length of the marine diatom Skeletonema marinoi in both grazer and non-grazer conditions by conducting monoclonal experiments. Additionally, a mesocosm experiment was performed with multiclonal experimental S. marinoi populations exposed to grazers at different levels of copepod concentration to test effects of grazer presence on diatom diversity in close to natural conditions. Our results show that distinct genotypes of a geographically restricted population exhibit variable phenotypic traits relevant to grazing interactions such as chain length and growth rates. Grazer presence affected clonal richness and evenness of multiclonal Skeletonema populations in the mesocosms, likely in conjunction with intrinsic interactions among the diatom strains. Only the production of polyunsaturated aldehydes was not affected by grazer presence. Our findings suggest that grazing can be an important factor structuring diatom population diversity in the sea and emphasize the importance of considering clonal differences when characterizing species and their role in nature.


Constraints on the off-shell Higgs boson signal strength in the high-mass ZZ and WW final states with the ATLAS detector.
Measurements of the ZZ and WW final states in the mass range above the [Formula: see text] and [Formula: see text] thresholds provide a unique opportunity to measure the off-shell coupling strength of the Higgs boson. This paper presents constraints on the off-shell Higgs boson event yields normalised to the Standard Model prediction (signal strength) in the [Formula: see text], [Formula: see text] and [Formula: see text] final states. The result is based on pp collision data collected by the ATLAS experiment at the LHC, corresponding to an integrated luminosity of 20.3 fb[Formula: see text] at a collision energy of [Formula: see text] TeV. Using the [Formula: see text] method, the observed 95 [Formula: see text] confidence level (CL) upper limit on the off-shell signal strength is in the range 5.1-8.6, with an expected range of 6.7-11.0. In each case the range is determined by varying the unknown [Formula: see text] and [Formula: see text] background K-factor from higher-order quantum chromodynamics corrections between half and twice the value of the known signal K-factor. Assuming the relevant Higgs boson couplings are independent of the energy scale of the Higgs boson production, a combination with the on-shell measurements yields an observed (expected) 95 [Formula: see text] CL upper limit on [Formula: see text] in the range 4.5-7.5 (6.5-11.2) using the same variations of the background K-factor. Assuming that the unknown [Formula: see text] background K-factor is equal to the signal K-factor, this translates into an observed (expected) 95 [Formula: see text] CL upper limit on the Higgs boson total width of 22.7 (33.0) MeV.


Mechanism generating endocochlear potential: role played by intermediate cells in stria vascularis.
The endocochlear DC potential (EP) is generated by the stria vascularis, and essential for the normal function of hair cells. Intermediate cells are melanocytes in the stria vascularis. To examine the contribution of the membrane potential of intermediate cells (E(m)) to the EP, a comparison was made between the effects of K(+) channel blockers on the E(m) and those on the EP. The E(m) of dissociated guinea pig intermediate cells was measured in the zero-current clamp mode of the whole-cell patch clamp configuration. The E(m) changed by 55.1 mV per 10-fold changes in extracellular K(+) concentration. Ba(2+), Cs(+), and quinine depressed the E(m) in a dose-dependent manner, whereas tetraethylammonium at 30 mM and 4-aminopyridine at 10 mM had no effect. The reduction of the E(m) by Ba(2+) and Cs(+) was enhanced by lowering the extracellular K(+) concentration from 3.6 mM to 1.2 mM. To examine the effect of the K(+) channel blockers on the EP, the EP of guinea pigs was maintained by vascular perfusion, and K(+) channel blockers were administered to the artificial blood. Ba(2+), Cs(+) and quinine depressed the EP in a dose-dependent manner, whereas tetraethylammonium at 30 mM and 4-aminopyridine at 10 mM did not change the EP. A 10-fold increase in the K(+) concentration in the artificial blood caused a minor decrease in the EP of only 10.6 mV. The changes in the EP were similar to those seen in the E(m) obtained at the lower extracellular K(+) concentration of 1.2 mM. On the basis of these results, we propose that the EP is critically dependent on the voltage jump across the plasma membrane of intermediate cells, and that K(+) concentration in the intercellular space in the stria vascularis may be actively controlled at a concentration lower than the plasma level.


Difficulty of discrimination modulates attentional capture by regulating attentional focus.
Attentional capture for distractors is enhanced by increasing the difficulty of discrimination between the standard and the target in the three-stimulus oddball paradigm. In this study, we investigated the cognitive mechanism of this modulation of attentional capture. Event-related brain potentials were recorded from participants while they performed a visual three-stimulus oddball paradigm (frequent standard, rare target, and rare distractor). The discrimination difficulty between standard and target was manipulated in the central location. Distractor stimuli were presented in the central or surrounding locations. The P3a component was elicited by distractor stimuli and was used as a measure of attentional capture. The results revealed that discrimination difficulty had opposite effects on the P3a response between central and surrounding locations. With an increase in the difficulty of discrimination, the P3a response was enhanced when distractor stimuli were presented in the central location. In contrast, the P3a response was reduced when distractor stimuli were presented in a surrounding location. This finding suggests that spatial attention was focused by the difficulty of discrimination, and deviant processing was increased within its focus but decreased outside its focus. Therefore, attentional capture for deviant distractors is modulated by top-down controlled attentional focus.


Glucocorticoid excess during adolescence leads to a major persistent deficit in bone mass and an increase in central body fat.
Endogenous Cushing's syndrome (CS) in children causes growth retardation, decreased bone mass, and increased total body fat. No prospective controlled studies have been performed in children to determine the long-term sequelae of CS on peak bone mass and body composition. A 15-year-old girl with Cushing disease (CD), and her healthy identical co-twin, were followed for 6 years after the CD was cured. At the 6-year follow-up both twins had areal bone mineral density (BMD) and body composition determined by dual-energy X-ray absorptiometry (DXA) and three-dimensional quantitative computed tomography (3DQCT). Z scores for height, weight, and body mass index (BMI) were -2.3, -0.8 and 0.2, and 1.2, 0.2, and -0.6, in the twin with CD and her co-twin, respectively. In the twin with CD, areal BMD and bone mineral apparent density (BMAD) at different sites varied from 0.7 to 3 SD below her co-twin. Volumetric lumbar spine bone density Z score was -0.75 and 1.0, and total body, abdominal visceral, and subcutaneous fat (%) was 42, 10, and 41 versus 26, 4, and 17 in the twin with CD and her co-twin, respectively. The relationship between total body fat and L2-L4 BMAD was inverse in the twin with CD (p < 0.05), which by contrast in her co-twin was opposite and direct (p < 0.001). In the twin with CD, despite cure, there was a persistent deficit in bone mass and increase in total and visceral body fat. These observations suggest that hypercortisolism (exogenous or endogenous) during adolescence may have persistent adverse effects on bone and fat mass.


Granulocyte-macrophage colony-stimulating factor (GM-CSF) as an adjunct to autologous hemopoietic stem cell transplantation for lymphoma.
To determine the hemopoietic effects of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) in patients having autologous hemopoietic stem cell transplantation for Hodgkin or non-Hodgkin lymphoma. Placebo or GM-CSF was administered after bone marrow or peripheral blood stem cell transplantation or both in a randomized, double-blind phase III trial by daily intravenous infusion (10 micrograms/kg body weight) until absolute neutrophil counts reached greater than or equal to 1000/mm3 on 3 consecutive days. Bone marrow transplantation unit in a university hospital. Sixty-nine consecutive patients with Hodgkin or non-Hodgkin lymphoma received GM-CSF (36 patients) or placebo (33 patients). Patients who received GM-CSF achieved absolute neutrophil counts greater than or equal to 500/mm3 (median, 12 compared with 16 days, P = 0.02) and absolute neutrophil counts greater than or equal to 1000/mm3 (median, 15 compared with 24 days, P less than 0.001) more quickly than patients who received placebo. Multivariate analysis indicated that use of GM-CSF, peripheral blood stem cells, and unpurged bone marrow were the strongest predictors for early neutrophil recovery greater than 500/mm3. Bacterial infections were significantly reduced in the GM-CSF group (P = 0.04). Delayed engraftment (neutrophils less than 500/mm3 at day 30) occurred in 26% and 17% of the placebo and GM-CSF groups, respectively, and correlated with the absence of detectable myeloid progenitor cells (colony-forming units-granulocyte macrophage, CFU-GM) (P less than 0.001) in marrow aspirate specimens obtained on day 15. Time to platelet independence, duration of hospital stay, severe adverse reactions, relapse, and disease-free survival rates did not differ significantly between the two groups. Administration of GM-CSF after autologous hemopoietic stem cell transplantation in patients with lymphoma resulted in accelerated myeloid recovery, particularly in patients who received peripheral blood stem cells and nonpurged bone marrow, and was associated with a decreased incidence of bacterial infections.


Increased Dickkopf-1 in Recent-onset Rheumatoid Arthritis is a New Biomarker of Structural Severity. Data from the ESPOIR Cohort.
Rheumatoid arthritis (RA) is the most common chronic inflammatory rheumatic condition over the world. RA is potentially disabling because chronic inflammation of the joints leads to joint destruction. To date, the best predictor of radiographic progression for patients with early RA is the presence of radiographic erosions at baseline, but a limited number of predictive biomarkers of structural progression are currently used in daily practice. Here, we investigated Dickkopf-1 (DKK-1) and sclerostin (SOST) serum levels in patients with recent inflammatory arthritis from the ESPOIR cohort. This cohort is a prospective, multicenter French cohort of 813 patients with early arthritis. We observed that mean baseline DKK-1 level was higher among RA patients with than without radiological progression within the first 2 years of evolution. DKK-1 level was still associated with radiographic progression in a model including other main predictors of severity (erosions at baseline, and anti-CCP antibody positivity). This study demonstrates that increased DKK-1 level at baseline predicted structural progression after 2-year follow-up and suggests that DKK-1 might be a new structural biomarker for early RA.


Molecular interaction study of novel indoline derivatives with EGFR-kinase domain using multiple computational analysis.
Epidermal growth factor receptors are constitutively overexpressed in breast cancer cells, which in turn stimulate many downstream signaling pathways that are involved in many carcinogenic processes. This makes EGFR a striking target for cancer therapy. This study focuses on the EGFR kinase domain inactivation by novel synthesized indoline derivatives. The compounds used are N-(2-hydroxy-5-nitrophenyl (4'-methyl phenyl) methyl) indoline (HNPMI), alkylaminophenols - 2-((3,4-Dihydroquinolin-1(2H)-yl) (p-tolyl) methyl) phenol (THTMP) and 2-((1, 2, 3, 4-Tetrahydroquinolin-1-yl) (4 methoxyphenyl) methyl) phenol (THMPP). To get a clear insight into the molecular interaction of EGFR and the three compounds, we have used ADME/Tox prediction, Flexible docking analysis followed by MM/GB-SA, QM/MM analysis, E-pharmacophore mapping of the ligands and Molecular dynamic simulation of protein-ligand complexes. All three compounds showed good ADME/Tox properties obeying the rules of drug-likeliness and showed high human oral absorption. Molecular docking was performed with the compounds and EGFR using Glide Flexible docking mode. This showed that the HNPMI was best among the three compounds and had interactions with key residue Lys 721. The protein-ligand complexes were stable when simulated for 100 ns using Desmond software. The interactions were further substantiated using QM/MM analysis and MM-GB/SA analysis in which HNPMI was scored as the best molecule. All the analyses were carried out with a reference molecule-Gefitinib which is a known standard inhibitor of EGFR. Thus, the study elucidates the potential role of the indoline derivatives as an anti-cancer agent against breast cancer by effectively inhibiting EGFR.Communicated by Ramaswamy H. Sarma.


Nucleoporin 75 is involved in the ethylene-mediated production of phytoalexin for the resistance of Nicotiana benthamiana to Phytophthora infestans.
Mature Nicotiana benthamiana shows stable resistance to the oomycete pathogen Phytophthora infestans. Induction of phytoalexin (capsidiol) production is essential for the resistance, which is upregulated via a mitogen-activated protein kinase (MAPK) cascade (NbMEK2-WIPK/SIPK) followed by ethylene signaling. In this study, NbNup75 (encodes a nuclear pore protein Nucleoporin75) was identified as an essential gene for resistance of N. benthamiana to P. infestans. In NbNup75-silenced plants, initial events of elicitor-induced responses such as phosphorylation of MAPK and expression of defense-related genes were not affected, whereas induction of later defense responses such as capsidiol production and cell death induction was suppressed or delayed. Ethylene production induced by either INF1 or NbMEK2 was reduced in NbNup75-silenced plants, whereas the expression of NbEAS (a gene for capsidiol biosynthesis) induced by ethylene was not affected, indicating that Nup75 is required for the induction of ethylene production but not for ethylene signaling. Given that nuclear accumulation of polyA RNA was increased in NbNup75-silenced plants, efficient export of mRNA from nuclei via nuclear pores would be important for the timely upregulation of defense responses. Collectively, Nup75 is involved in the induction of a later stage of defense responses, including the ethylene-mediated production of phytoalexin for the resistance of N. benthamiana to P. infestans.


HIV testing in national population-based surveys: experience from the Demographic and Health Surveys.
To describe the methods used in the Demographic and Health Surveys (DHS) to collect nationally representative data on the prevalence of human immunodeficiency virus (HIV) and assess the value of such data to country HIV surveillance systems. During 2001-04, national samples of adult women and men in Burkina Faso, Cameroon, Dominican Republic, Ghana, Mali, Kenya, United Republic of Tanzania and Zambia were tested for HIV. Dried blood spot samples were collected for HIV testing, following internationally accepted ethical standards. The results for each country are presented by age, sex, and urban versus rural residence. To estimate the effects of non-response, HIV prevalence among non-responding males and females was predicted using multivariate statistical models for those who were tested, with a common set of predictor variables. Rates of HIV testing varied from 70% among Kenyan men to 92% among women in Burkina Faso and Cameroon. Despite large differences in HIV prevalence between the surveys (1-16%), fairly consistent patterns of HIV infection were observed by age, sex and urban versus rural residence, with considerably higher rates in urban areas and in women, especially at younger ages. Analysis of non-response bias indicates that although predicted HIV prevalence tended to be higher in non-tested males and females than in those tested, the overall effects of non-response on the observed national estimates of HIV prevalence are insignificant. Population-based surveys can provide reliable, direct estimates of national and regional HIV seroprevalence among men and women irrespective of pregnancy status. Survey data greatly enhance surveillance systems and the accuracy of national estimates in generalized epidemics.


Comparison of Stent-Assisted Coiling vs Coiling Alone in 563 Intracranial Aneurysms: Safety and Efficacy at a High-Volume Center.
Stent-assisted coiling has been used in both unruptured and ruptured aneurysms, but the safety and efficacy still remain controversial. To compare the safety and efficacy of stent-assisted coiling with coiling alone for intracranial aneurysms. We retrospectively reviewed 512 patients treated by endovascular coiling or stent-assisted coiling over a 1-year period. The patients' clinical and imaging information was recorded. Procedure-related complication rates, recurrence rates, and clinical outcomes were analyzed in both the total aneurysms and the subgroups of ruptured and unruptured aneurysms. A total of 243 patients were treated by coiling alone and 269 patients were treated by stent-assisted coiling. Procedure-related complications occurred in 6.2% of patients in the coiling-alone group compared with 6.3% in the stent-assisted coiling group. The procedural permanent morbidity and mortality rates were 1.6% (4/243) and 1.2% (3/243), respectively, in the coiling-alone group and 1.1% (3/269) and 1.5% (4/269), respectively, in the stent-assisted coiling group. A significantly lower recurrence rate was found in the stent-assisted coiling group compared with the coiling-alone group (5.2% vs 16.5%, P = .002). In a comparison of subgroups of ruptured and unruptured aneurysms, the procedural complications rates were comparable in the 2 groups, with a lower recurrence rate in the stent-assisted coiling group. Multivariate analysis showed that larger aneurysm size and higher Hunt and Hess grade were predictors of procedural morbidity; larger aneurysm size, ruptured aneurysm, anterior circulation aneurysms, initial incomplete occlusion, and lack of stent assistance were predictors of recurrence. Compared with coiling alone, stent-assisted coiling may achieve lower recurrence rates, with comparable procedure-related complications and clinical outcomes in both ruptured and unruptured aneurysms.


Value of CT-guided biopsy in the diagnosis of septic discitis.
To determine the role of CT-guided biopsy in the management of cases of infective discitis. Data were examined from a retrospective case series of CT-guided biopsies for the 5-year period ending June 2006. 98 CT-guided biopsies were performed in the study period on 103 patients. Malignancy was diagnosed in 49 episodes. Discitis and paravertebral abscess accounted for 27 cases. Culture was positive in nine of 25 (36%) samples received by the microbiology laboratory. Staphylococcus aureus (four cases) and Mycobacterium tuberculosis (three cases) were the most frequent organisms isolated, followed by group G streptococci and coagulase-negative staphylococci (one case each). Blood cultures were diagnostic in a further nine patients. The main reason for a negative culture was prior antimicrobial therapy. The biopsy changed management in 9/25 (36%) of cases. There were no reported adverse events. Septic discitis is a serious condition with a wide variety of infective causes. CT-guided biopsy is a useful tool when the diagnosis of infectious spinal infection is considered in terms of commencing and targeting therapy, and it is a safe and well-tolerated procedure.


Mitigation of Membrane Biofouling in MBR Using a Cellulolytic Bacterium, Undibacterium sp. DM-1, Isolated from Activated Sludge.
Biofilm formation on the membrane surface results in the loss of permeability in membrane bioreactors (MBRs) for wastewater treatment. Studies have revealed that cellulose is not only produced by a number of bacterial species but also plays a key role during formation of their biofilm. Hence, in this study, cellulase was introduced to a MBR as a cellulose-induced biofilm control strategy. For practical application of cellulase to MBR, a cellulolytic (i.e., cellulase-producing) bacterium, Undibacterium sp. DM-1, was isolated from a lab-scale MBR for wastewater treatment. Prior to its application to MBR, it was confirmed that the cell-free supernatant of DM-1 was capable of inhibiting biofilm formation and of detaching the mature biofilm of activated sludge and cellulose-producing bacteria. This suggested that cellulase could be an effective anti-biofouling agent for MBRs used in wastewater treatment. Undibacterium sp. DM-1-entrapping beads (i.e., cellulolytic-beads) were applied to a continuous MBR to mitigate membrane biofouling 2.2-fold, compared with an MBR with vacant-beads as a control. Subsequent analysis of the cellulose content in the biofilm formed on the membrane surface revealed that this mitigation was associated with an approximately 30% reduction in cellulose by cellulolytic-beads in MBR.


Fall prediction with wearable sensors--an empirical study on expert opinions.
Falls represent a considerable problem for many developed societies with a large growth in the elderly population, and they inflict a sizable personal and social cost. Assessment of an individual's risk of falling, facilitated by advanced movement sensing technologies, has been heralded as a potential means to target fall prevention interventions to those most at need. This paper presents the summary of a questionnaire-based survey of expert opinions collected from three major international conference meetings on the topic of fall risk assessment and fall prediction. Specifically, current problems are discussed, both in terms of technology as well as in study design, in order to harmonize and guide future research activities. The analysis of this survey confirms that sensor-based fall prediction is both relevant and promising, and that large-scale prospective studies are needed as well as further harmonization of research methods.


Influence of cranberry proanthocyanidins on formation of biofilms by Streptococcus mutans on saliva-coated apatitic surface and on dental caries development in vivo.
Cranberry crude extracts, in various vehicles, have shown inhibitory effects on the formation of oral biofilms in vitro. The presence of proanthocyanidins (PAC) in cranberry extracts has been linked to biological activities against specific virulence attributes of Streptococcus mutans, e.g. the inhibition of glucosyltransferase (Gtf) activity. The aim of the present study was to determine the influence of a highly purified and chemically defined cranberry PAC fraction on S. mutans biofilm formation on saliva-coated hydroxyapatite surface, and on dental caries development in Sprague-Dawley rats. In addition, we examined the ability of specific PAC (ranging from low-molecular-weight monomers and dimers to high-molecular-weight oligomers/polymers) to inhibit GtfB activity and glycolytic pH drop by S. mutans cells, in an attempt to identify specific bioactive compounds. Topical applications (60-second exposure, twice daily) with PAC (1.5 mg/ml) during biofilm formation resulted in less biomass and fewer insoluble polysaccharides than the biofilms treated with vehicle control had (10% ethanol, v/v; p < 0.05). The incidence of smooth-surface caries in rats was significantly reduced by PAC treatment (twice daily), and resulted in less severe carious lesions compared to the vehicle control group (p < 0.05); the animals treated with PAC also showed significantly less caries severity on sulcal surfaces (p < 0.05). Furthermore, specific A-type PAC oligomers (dimers to dodecamers; 0.1 mg/ml) effectively diminished the synthesis of insoluble glucans by GtfB adsorbed on a saliva-coated hydroxyapatite surface, and also affected bacterial glycolysis. Our data show that cranberry PAC reduced the formation of biofilms by S. mutans in vitro and dental caries development in vivo, which may be attributed to the presence of specific bioactive A-type dimers and oligomers.


Temporal representation of clinical algorithms using expert-system and database tools.
The HyperLipid Advisory System combines a rule-based implementation of a clinical algorithm (the NIH Cholesterol Education Program Expert Panel recommendations) with a temporal representation that facilitates reasoning over time while maintaining efficient storage in a standard database. The temporal representation consists of objects that model point events such as visits and interval events such as specific therapies. These objects are combined into abstractions called phases, which correspond to higher level clinical concepts such as a diet or drug treatment. The time-oriented data objects are referenced in the rules using a custom-tailored operator query language. Between user sessions relevant clinical data are stored in external files. When the advisory system is reconsulted, this information is retrieved and mapped back into an object-oriented format. Use of a commercially available expert-system shell for such tasks allows algorithm implementation in standard personal computing environments.


Human and Murine Hematopoietic Stem Cell Aging Is Associated with Functional Impairments and Intrinsic Megakaryocytic/Erythroid Bias.
Aging within the human hematopoietic system associates with various deficiencies and disease states, including anemia, myeloid neoplasms and reduced adaptive immune responses. Similar phenotypes are observed in mice and have been linked to alterations arising at the hematopoietic stem cell (HSC) level. Such an association is, however, less established in human hematopoiesis and prompted us here to detail characteristics of the most primitive human hematopoietic compartments throughout ontogeny. In addition, we also attempted to interrogate similarities between aging human and murine hematopoiesis. Coupled to the transition from human cord blood (CB) to young and aged bone marrow (BM), we observed a gradual increase in frequency of candidate HSCs. This was accompanied by functional impairments, including decreased lymphoid output and reduced proliferative potential. Downstream of human HSCs, we observed decreasing levels of common lymphoid progenitors (CLPs), and increasing frequencies of megakaryocyte/erythrocyte progenitors (MEPs) with age, which could be linked to changes in lineage-affiliated gene expression patterns in aged human HSCs. These findings were paralleled in mice. Therefore, our data support the notion that age-related changes also in human hematopoiesis involve the HSC pool, with a prominent skewing towards the megakaryocytic/erythroid lineages, and suggests conserved mechanisms underlying aging of the blood cell system.


Associatively learned representations of taste outcomes activate taste-encoding neural ensembles in gustatory cortex.
Through learning processes, cues associated with emotionally salient reinforcing outcomes can come to act as substitutes for the reinforcer itself. According to one account of this phenomenon, the predictive cue associatively elicits a representation of the expected outcome by reactivating cells responsible for encoding features of the primary reinforcer. We tested this hypothesis by examining the role of neural ensembles in gustatory cortex (GC) during receipt of gustatory stimuli (sucrose and water) and cues associated with those stimuli using the immediate early genes (IEGs) Arc and Homer1a. Because these plasticity-related IEGs are expressed in the neuronal nucleus 5 and 30 min, respectively, after salient events, we examined how individual neurons encoded these stimuli in two separate behavioral epochs. In experiment 1, we showed that tasting identical sucrose solutions, but not tasteless water, in the two epochs increased both IEG activity and the degree of overlap between neural ensembles in GC. In experiment 2, odor cues associated with sucrose, but not water, evoked potentiation of IEG activity in GC similar to sucrose itself. Surprisingly, lesions of the basolateral amygdala had minimal effects on associative encoding in GC. Finally, these associatively driven representations of sucrose appeared to be outcome specific, as neural ensembles that were activated by the sucrose-associated cue were also activated by sucrose itself. This degree of overlap between associative and primary taste activity at the ensemble level suggests that GC neurons encode important information about anticipated outcomes. Such representations may provide outcome-specific information for guiding goal-directed behavior.


A transcriptomic and epigenomic cell atlas of the mouse primary motor cortex.
Single-cell transcriptomics can provide quantitative molecular signatures for large, unbiased samples of the diverse cell types in the brain1-3. With the proliferation of multi-omics datasets, a major challenge is to validate and integrate results into a biological understanding of cell-type organization. Here we generated transcriptomes and epigenomes from more than 500,000 individual cells in the mouse primary motor cortex, a structure that has an evolutionarily conserved role in locomotion. We developed computational and statistical methods to integrate multimodal data and quantitatively validate cell-type reproducibility. The resulting reference atlas-containing over 56 neuronal cell types that are highly replicable across analysis methods, sequencing technologies and modalities-is a comprehensive molecular and genomic account of the diverse neuronal and non-neuronal cell types in the mouse primary motor cortex. The atlas includes a population of excitatory neurons that resemble pyramidal cells in layer 4 in other cortical regions4. We further discovered thousands of concordant marker genes and gene regulatory elements for these cell types. Our results highlight the complex molecular regulation of cell types in the brain and will directly enable the design of reagents to target specific cell types in the mouse primary motor cortex for functional analysis.


Impact of simulated hyperopia on academic-related performance in children.
To investigate the impact of simulated hyperopia and sustained near work on children's ability to perform a range of academic-related tasks. Fifteen visually normal children (mean [±SD] age, 10.9 [±0.8] years; 10 male and 5 female) were recruited. Performance on a range of standardized academic-related outcome measures was assessed with and without 2.50 diopters of simulated bilateral hyperopia (administered in a randomized order), before and after 20 minutes of sustained near work, at two separate testing sessions. Academic-related measures included a standardized reading test (the Neale Analysis of Reading Ability), visual information processing tests (the Coding and Symbol Search subtests from the Wechsler Intelligence Scale for Children), and a reading-related eye movement test (the Developmental Eye Movement test). Simulated bilateral hyperopia and sustained near work each independently impaired reading, visual information processing, and reading-related eye movement performance (p < 0.001). A significant interaction was also demonstrated between these factors (p < 0.05), with the greatest decrement in performance observed when simulated hyperopia was combined with sustained near work. This combination resulted in performance reductions of between 5 and 24% across the range of academic-related measures. A significant moderate correlation was also found between the change in horizontal near heterophoria and the change in several of the academic-related outcome measures, after the addition of simulated hyperopia. A relatively low level of simulated bilateral hyperopia impaired children's performance on a range of academic-related outcome measures, with sustained near work further exacerbating this effect. Further investigations are required to determine the impact of correcting low levels of hyperopia on academic performance in children.


Active site of the solvated thiosulfate ion characterized by hydration structures and dynamics.
The reactivity of the terminated sulfur atom within the thiosulfate ion (S2O3(2-)) when it is involved in chemical reactions was investigated through the properties of the molecular hydration shell, obtained from the ab initio quantum mechanical change field molecular dynamics (QMCF MD) simulation. The average geometry indicated the significant effect of explicit water on the reduction of the S-S length, which was reflected in the splitting peaks of the spectrum for the stretching mode of this bond (ν(SS)). A further investigation on a simple model with various theoretical levels exhibited the hydrophobicity of the S-S bond. The evaluation of the molecular coordination number was sensitive to the radii of the atomic hydration spheres, which were obtained from the vague boundaries of the first peak in the atomic radial distribution functions. The number of actual contacts specified 6.8 water molecules interacting with the thiosulfate ion, and 2.4 extra waters located in the molecular hydration shell, forming a H-bonding network with the bulk water. The mean residence times for the water ligands distinguished the asymmetric strength of the hydration shell into a weaker sulfur and three stronger oxygen sites, instigating the terminated sulfur atom as the active site that is involved in chemical reactions.


Caveolin-2-deficient mice show increased sensitivity to endotoxemia.
Caveolin proteins are structural components of caveolae and are involved in the regulation of many biological processes. Recent studies have shown that caveolin-1 modulates inflammatory responses and is important for sepsis development. In the present study, we show that caveolin-1 and caveolin-2 have opposite roles in lipopolysaccharide (LPS)-induced sepsis using caveolin-deficient (Cav-1 (-/-) and Cav-2 (-/-) ) mice for each of these proteins. While Cav-1 (-/-) mice displayed delayed mortality following challenge with LPS, Cav-2 (-/-) mice were more sensitive to LPS compared to wild-type (WT). With Cav-2 (-/-) mice, this effect was associated with increased intestinal injury and increased intestinal permeability. This negative outcome was also correlated with enhanced expression of iNOS in epithelial intestinal cells, and enhanced production of nitric oxide (NO). By contrast, Cav-1 (-/-) mice demonstrated a decrease in iNOS expression with decreased NO production, but no alteration in intestinal permeability. The differential expression of iNOS was associated with a significant increase of STAT-1 activation in these mice. Intestinal cells of Cav-2 (-/-) mice showed increased phosphorylation of STAT-1 at tyrosine 701 compared to wild-type. However, Cav-1 (-/-) mice-derived intestinal cells showed decreased levels of phosphorylation of STAT-1 at tyrosine 701. Since caveolin-2 is almost completely absent in Cav-1 (-/-) mice, we conclude that it is not just the absence of caveolin-2 that is responsible for the observed effects, but that the balance between caveolin-1 and caveolin-2 is important for iNOS expression and ultimately for sepsis outcome.


Effect of conjunctival structure and inflammatory cell counts on intraocular pressure after trabeculectomy.
To evaluate the conjunctival structure and inflammatory cell counts and to determine the predictive value of these histological parameters for postoperative intraocular pressure (IOP) levels after trabeculectomy. A clinical and histological study was performed on consecutive patients. Postoperative (mean 32. 8 +/- 18.4 months; range 6-60 months) conjunctival biopsies of 36 eyes of 28 primary open-angle glaucoma patients who had trabeculectomy between 1992 and 1995 were included in the study. According to postoperative pressure control, patients with < or = 16 mm Hg and those with >16 mm Hg were taken as groups 1 and 2, respectively. The control group (group 3) consisted of 15 age-matched patients without glaucoma, who had received no topical therapy. We compared the conjunctival structure and cell counts within these groups. Goblet cells, acute inflammatory cells, chronic inflammatory cells, fibroblasts, epithelial thickness, vascular density, mucopolysaccharides and collagen compositions were determined in groups 1, 2 and 3. The number of goblet cells was significantly higher in group 1 (6.74 +/- 7.23) than group 2 (3. 09 +/- 2.77; p = 0.017). No statistical difference was observed in the number of acute inflammatory cells, chronic inflammatory cells, fibroblasts, epithelial thickness, vascular density, mucopolysaccharide or collagen compositions between groups 1 and 2 (p > 0.05). In addition, when groups 1 and 2 were compared with the control group, there was a significant decrease in goblet cells (p < 0.001), an increase in acute inflammatory cells, chronic inflammatory cells, fibroblasts, epithelial thickness and vascular density (p < 0.001), but there was no significant difference in mucopolysaccharide and collagen compositions (p > 0.05). This study suggests that at the time of surgery a high number of goblet cells may be a predictor of lowered IOP (< or = 16 mm Hg) following trabeculectomy without antimetabolite.


Hydrogen peroxide is a common signal for darkness- and ABA-induced stomatal closure in Pisum sativum.
The requirement for hydrogen peroxide (H2O2) generation and action during stomatal closure induced by darkness and abscisic acid (ABA) was investigated in pea (Pisum sativum L.). Stomatal closure induced by darkness or ABA was inhibited by the H2O2-scavenging enzyme catalase or the antioxidant N-acetyl cysteine (NAC), or by diphenylene iodonium (DPI), an inhibitor of the H2O2-generating enzyme NADPH oxidase. Exogenous H2O2 induced stomatal closure in a dose- and time-dependent manner, and H2O2 was also required for ABA-inhibition of stomatal opening in the light. H2O2 accumulation in guard cells was increased by darkness or ABA, as assessed with the fluorescent dye dichlorodihydrofluorescein diacetate (H2-DCFDA) and confocal microscopy. Such increases were inhibited by catalase, NAC or DPI, consistent with the effects of these compounds on stomatal apertures. Employing polymerase chain reaction (PCR) with degenerate oligonucleotide primers, several NADPH oxidase homologues were identified from pea genomic DNA that had substantial identity to the Arabidopsis thaliana (L.) Heynh. rboh (respiratory burst oxidase homologue) genes. Furthermore, an antibody raised against the tomato rboh identified immunoreactive proteins in epidermal, mesophyll and guard cells.


Formulation development and optimization: Encapsulated system of Atenolol and Glyburide.
Objective of this study is to develop; tablet-in-a capsule system, to deliver Atenolol 25mg and Glyburide 5mg in the hard gelatin capsule. In order to improve patient compliance and reduce problems associated with complex therapeutic regimen Atenolol (cardio-selective beta-blocker) and Glyburide (anti-diabetic; sulfonylurea) are commonly, prescribed to the diabetic hypertensive patient. Metgod: In present work six different formulations of Atenolol (AF1-AF6) and Glyburide (GF1-GF6) were prepared by direct compression method using Avicel, Lactose DC, Crospovidone and Magnesium Stearate in different proportions and encapsulated in hard gelatin shells. Post compression parameters i.e. weight variation, diameter variation, thickness variation, hardness variation, % friability, disintegration, % drug release were determined at different pH 1.2, 4.5 and 6.8, and subjected to dissolution profile comparison through similarity factor (ƒ2). Stability studies were performed and shelf lives were calculated by R-Gui Stab R console 2.15.2 and determined to be 15 and 27 months for Atenolol and Glyburide respectively. The percentage drug contents of Atenolol and Glyburide were estimated spectrophotometerically at 286 nm and 314.7 nm respectively. Formulations CF1-CF6 (encapsulated) were subjected to weight variation, disintegration and dissolution tests and subjected to model dependant analysis for dissolution studies. The simultaneous quantitation of Atenolol and Glyburide for content assay was done by HPLC method of analysis. formulation CF6 is showing highest coefficient of correlation values for all models applied. So we can conclude that the proposed system can improve patient compliance by increasing the ease of administration of two drugs together.


Human breast cancer cell cycle synchronization by estrogens and antiestrogens in culture.
The mechanisms by which estrogens and antiestrogens modulate breast cancer growth have not been totally defined. We have examined the cell cycle kinetic effects of estrogens and antiestrogens in cultured human breast cancer cell lines. In a previous study, we showed that tamoxifen induces a transition delay in early to mid-G1 phase of the cell cycle. In the present study, we show that this cell kinetic alteration by tamoxifen is dose dependent and that other antiestrogens have identical effects. As little as 0.01 to 0.1 microM tamoxifen reduces the S and G2 + M fractions and increases the G1 fraction of MCF-7 cells growing in medium with 5% charcoal-stripped bovine serum. More than 90% of cells are in G1 72 to 96 hr after the addition of 1 microM tamoxifen, a concentration achieved in patients treated with the drug. Nafoxidine and trioxifene have identical activity. Partial reversal of tamoxifen growth inhibition is observed with a simple change to tamoxifen-free medium. Complete reversal of the tamoxifen effect is observed with the addition of 17 beta-estradiol. By 24 hr after the addition of estrogen, 60 to 70% of tamoxifen-inhibited cells have progressed through G1 and into S phase, indicating that tamoxifen-treated cells remain viable. This estrogen "rescue" effect is observed even in the absence of a change to tamoxifen-free medium. A 100-fold-lower concentration of estradiol can totally reverse the inhibitory effects of 1.0 microM tamoxifen. Stimulation of the progression of G1 cells to enter S phase is also observed when estradiol is added to cells maintained for four days in medium with stripped serum, even in the absence of tamoxifen. Similar effects are observed in the estrogen receptor-positive ZR75-1 breast cancer cells. No effects of antiestrogens or estrogens are observed in the receptor-negative MDA-231 cells, suggesting that these effects are mediated through the estrogen receptor. In summary, antiestrogens and estrogens have prominent effects on the cell cycle kinetics of endocrine-dependent human breast cancer cells. Antiestrogens cause an accumulation of cells in G1 phase. Estrogen reverses this block with a synchronous cohort of cells progressing through the cell cycle. These data have important implications for the design of rational clinical trials of combined chemoendocrine therapy.


Ontogeny of head and caudal fin shape of an apex marine predator: The tiger shark (Galeocerdo cuvier).
How morphology changes with size can have profound effects on the life history and ecology of an animal. For apex predators that can impact higher level ecosystem processes, such changes may have consequences for other species. Tiger sharks (Galeocerdo cuvier) are an apex predator in tropical seas, and, as adults, are highly migratory. However, little is known about ontogenetic changes in their body form, especially in relation to two aspects of shape that influence locomotion (caudal fin) and feeding (head shape). We captured digital images of the heads and caudal fins of live tiger sharks from Southern Florida and the Bahamas ranging in body size (hence age), and quantified shape of each using elliptical Fourier analysis. This revealed changes in the shape of the head and caudal fin of tiger sharks across ontogeny. Smaller juvenile tiger sharks show an asymmetrical tail with the dorsal (upper) lobe being substantially larger than the ventral (lower) lobe, and transition to more symmetrical tail in larger adults, although the upper lobe remains relatively larger in adults. The heads of juvenile tiger sharks are more conical, which transition to relatively broader heads over ontogeny. We interpret these changes as a result of two ecological transitions. First, adult tiger sharks can undertake extensive migrations and a more symmetrical tail could be more efficient for swimming longer distances, although we did not test this possibility. Second, adult tiger sharks expand their diet to consume larger and more diverse prey with age (turtles, mammals, and elasmobranchs), which requires substantially greater bite area and force to process. In contrast, juvenile tiger sharks consume smaller prey, such as fishes, crustaceans, and invertebrates. Our data reveal significant morphological shifts in an apex predator, which could have effects for other species that tiger sharks consume and interact with.


Atrial natriuretic peptide in humans. Production and clearance by various tissues.
Although it is assumed that the human heart secretes atrial peptides, direct proof is not available. We therefore measured immunoreactive atrial natriuretic peptide levels in coronary sinus blood and simultaneously in femoral arterial and venous blood from patients before and during stepwise incremental atrial pacing of up to 200 beats per minute. Since the fate of circulating atrial peptides is unknown, we also measured immunoreactive atrial natriuretic peptide concentrations in arterial and venous blood across the liver, kidney, lower limb, and lung in patients undergoing cardiological investigation. Peptide levels in coronary sinus blood were higher than in samples from the femoral artery or vein. As the heart rate was accelerated by atrial pacing, peptide concentrations increased in coronary sinus blood and to a lesser extent in peripheral samples. Whereas the levels in venous blood draining the liver, kidney, and lower limb were approximately 50% of those in arterial blood, concentrations were similar in samples drawn simultaneously from the pulmonary artery and the aorta. These results show that the human heart produces immunoreactive atrial natriuretic peptide and that secretion increases with atrial tachycardia. The liver, kidney, and lower limb remove the peptide from arterial blood, but there is little change in its concentration during circulation of blood through the lungs.


Inappropriate use of antibiotic prophylaxis to prevent infective endocarditis in obstetric patients.
To evaluate infective endocarditis prophylaxis practices during the intrapartum period and to assess obstetric providers' adherence to the American Heart Association and American College of Obstetrics and Gynecology guidelines for infective endocarditis prophylaxis. We performed a chart review of pharmacy, electronic nursing, and physician records to report this case series of obstetric patients who received infective endocarditis prophylaxis during the intrapartum period at a single tertiary referral care center during a 1-year study period from August 1, 2004, to July 31, 2005. Fifty patients received antibiotics for infective endocarditis prophylaxis. Three of the 50 patients who received infective endocarditis prophylaxis had high-risk cardiac lesions and three other patients had moderate-risk cardiac lesions and evidence for intrapartum infection. Thus, only six patients (12.0%, 95% confidence interval 4.5%-24.3%) met the American Heart Association and American College of Obstetricians and Gynecologists criteria for an appropriate indication for infective endocarditis prophylaxis. Of these six patients who had an appropriate indication for infective endocarditis prophylaxis, only three (50.0%, 95% confidence interval 11.8%-88.2%) received appropriate antibiotic regimens. Antibiotics are frequently given to obstetric patients during pregnancy. Although many obstetric patients receive antibiotics for recommended indications, some patients, as our study shows, do not. A concerted effort by all practitioners and institutions to reduce the amount of inappropriate antibiotics given to obstetric patients will have positive public health effects in addition to benefiting individual mothers and neonates.


Assessment of the Cambodian National Biodigester Program.
The National Biodigester Program (NBP) was established in 2006 to build and maintain household biodigesters in Cambodia. In its first six years, the program installed almost 20,000 biodigesters and established an in-country network of local financiers, construction companies, skilled masons, bioslurry specialists and after-care technicians accessible to rural users in 14 of 24 Cambodian provinces. Since its initial success, the program's adoption rates have stalled despite increasing government support and high rates of user satisfaction. Building on an initial evaluation of the NBP in 2013, this updated assessment identifies multiple changes in its second implementation phase that have undermined the initial momentum of the program. Abrupt interruptions in institutional support, deteriorating supply side services (access to construction agents, masons, repair services) and reduced access to credit for farmers have eroded the service network that the first implementation phase established. Structural changes in the rural economy may also contribute to declining demand. Government support to another biodigester program which offers a lower investment price, but does not provide after-sales services has also undercut the long-term implementation strategy of the NBP. The paper finds that despite these programmatic changes, the installed biodigesters continue to perform according to expectation and to be maintained and valued by their users, but the future viability of the program remains uncertain.


The secretory immunoglobulin A response to Mycobacterium tuberculosis in a childhood population.
We report on the measurement of saliva anti-Purified Protein Derivative sIgA and 38kDa antibodies from 127 children, of whom 31 were strong tuberculosis suspects and 96 were healthy contact children. The results concerning the percentage of children with antibody reactivity to PPD and 38kDa antigens showed that, of these 2 antigens, 38kDa induced higher reactivity in patients positive and negative for the Tuberculin Skin Test (28% and 16.6%, respectively) in comparison to controls positive and negative for the TST (11.7% and 7.1%, respectively). There was a statistically significant difference between patients positive and controls negative for the TST. In relation to the Purified Protein Derivative antigen, while 14.2% of patients positive for the TST showed antibody reactivity to the PPD antigen, no patients negative for the TST had reactivity to this antigen. The findings suggest that these two antigens seem be associated with a different development of the mucosal defence mechanisms mediated by sIgA against Mycobacterium tuberculosis.


Living with a venous leg ulcer: a descriptive study of patients' experiences.
This study aimed to describe the experience of living with a venous leg ulcer from the patient's perspective, to answer the question 'What is it like to live with a venous leg ulcer?' Investigating the experience of illness is important in understanding how disease processes affect people, how they understand and cope with these processes, and how care may be given most effectively. Little research was identified which examined these areas with reference to people with leg ulceration. A qualitative approach was taken in the following study, using phenomenological methods to describe the experience of venous leg ulceration. Informal unstructured interviews were conducted with 13 informants, guided by their descriptive priorities. Tape recordings were then transcribed and analysed for recurrent themes and their meanings. It was found that certain symptoms of ulceration, principally pain, were the overwhelming feature of the experience. These symptoms caused significant restrictions in people's lives, particularly in their ability to walk and go out. Treatment was not described as being efficacious in ameliorating these symptoms, but nevertheless great confidence and trust was placed in the expertise of nurses. People coped with the experience mainly by a process of normalizing its components. It is concluded that if treatment is to meet the needs of those suffering venous leg ulceration, then symptom control must be the highest priority.


Brain MRI white matter hyperintensities and one-carbon cycle metabolism in non-geriatric outpatients with major depressive disorder (Part I).
The objective of the present work was to study the interrelationship between white matter hyperintensities (WMHs), cardiovascular risk factors and elements of the one-carbon cycle including serum folate, vitamin B12, and homocysteine levels in a relatively young sample of outpatients with major depressive disorder (MDD), and to compare the severity of white matter hyperintensities in MDD patients and healthy volunteers. Fifty MDD outpatients (34% women, age 40.6+/-10.3 years), free of psychotropic medications for at least 2 weeks before enrollment, underwent magnetic resonance imaging (MRI) scans of the brain to detect T2 WMHs and also had (1) serum folate, vitamin B12, homocysteine and cholesterol levels measured, and (2) cardiovascular risk factors assessed during the same study visit. Thirty-five healthy comparison subjects (40% women, age 39.2+/-9.8 years) also underwent brain MRI scans. Hypofolatemia, hypertension and age independently predicted a greater severity of total brain WMHs. Separately, the same factors also predicted a greater severity of subcortical WMHs. Hypofolatemic and hypertensive patients had more severe WMHs than normal controls. In light of the adverse impact of WMHs on a number of health-related outcomes later in life, hypofolatemia and hypertension may represent modifiable risk factors to prevent the occurrence of such adverse outcomes.


Infliximab in recalcitrant generalized pustular arthropatic psoriasis.
Generalized pustular psoriasis is an unstable inflammatory type of psoriasis, with widespread areas of erythema and sterile pustules, associated with fever and systemic symptoms. Infliximab is a monoclonal antibody with anti-TNFalpha activity, approved for use in psoriasis. We describe a male patient with a long history of stable arthropathic psoriasis, hospitalized with a generalized pustular psoriasis and acute exacerbation of articular complaints. The disease was resistant to multiple therapies (acitretin, methotrexate and corticosteroids), so the patient was started on infliximab, with a very rapid response of both cutaneous and articular symptoms. He had complete clearing of lesions at week 12, and marked improvement of the articular symptoms. No recurrence occurred at 8 months of follow-up with infliximab every 8 weeks. Infliximab had an extremely rapid therapeutic action response on a recalcitrant generalized pustular psoriasis. The articular response was also excellent, with significant improvement of quality of life.


Measurement of single-diffractive dijet production in proton-proton collisions at s = 8 Te with the CMS and TOTEM experiments.
Measurements are presented of the single-diffractive dijet cross section and the diffractive cross section as a function of the proton fractional momentum loss ξ and the four-momentum transfer squared t. Both processes p p → p X and p p → X p , i.e. with the proton scattering to either side of the interaction point, are measured, where X includes at least two jets; the results of the two processes are averaged. The analyses are based on data collected simultaneously with the CMS and TOTEM detectors at the LHC in proton-proton collisions at s = 8 Te during a dedicated run with β ∗ = 90 m at low instantaneous luminosity and correspond to an integrated luminosity of 37.5 nb - 1 . The single-diffractive dijet cross section σ jj p X , in the kinematic region ξ < 0.1 , 0.03 < | t | < 1 Ge 2 , with at least two jets with transverse momentum p T > 40 Ge , and pseudorapidity | η | < 4.4 , is 21.7 ± 0.9 (stat) - 3.3 + 3.0 (syst) ± 0.9 (lumi) nb . The ratio of the single-diffractive to inclusive dijet yields, normalised per unit of ξ , is presented as a function of x, the longitudinal momentum fraction of the proton carried by the struck parton. The ratio in the kinematic region defined above, for x values in the range - 2.9 ≤ log 10 x ≤ - 1.6 , is R = ( σ jj p X / Δ ξ ) / σ jj = 0.025 ± 0.001 (stat) ± 0.003 (syst) , where σ jj p X and σ jj are the single-diffractive and inclusive dijet cross sections, respectively. The results are compared with predictions from models of diffractive and nondiffractive interactions. Monte Carlo predictions based on the HERA diffractive parton distribution functions agree well with the data when corrected for the effect of soft rescattering between the spectator partons.


Minimally Invasive Percutaneous Spinopelvic Fixation for Unstable Pelvic Ring Fracture Performed With the Patient in a Lateral Position.
Spinopelvic fixation provides a strong fixation for unstable pelvic ring fractures. However, the technique is usually performed with the patient in the prone position, with the applied weight on the anterior superior iliac crests aggravating fracture displacement. We developed a novel approach for minimally invasive percutaneous spinopelvic fixation that is performed with the patient in a lateral (side lying) position. We describe the application of our technique for the treatment of a bilateral pelvic ring and acetabulum fracture in a 79-year-old woman injured in a traffic accident. Initial posterior fixation was performed with the patient in the left-side lying position, using bilateral pedicle screws at L3 and L4 and a left sacral-alar iliac screw and 2 right iliac screws inserted under navigation. The lateral and cranial displacement of the right pelvic ring was reduced percutaneously. One week after this initial surgery, we proceeded with an open anterior reduction and internal fixation of the left pelvic ring and acetabulum fracture. The postoperative course was uneventful and clinical outcomes were satisfactory. Reduction of a pelvic ring fracture in a lateral position, with subsequent spinopelvic fixation, is a reasonable option for the treatment of an unstable pelvic ring fracture.


CDC25A(Q110del): a novel cell division cycle 25A isoform aberrantly expressed in non-small cell lung cancer.
Lung cancer remains number one cause of cancer related deaths worldwide. Cell cycle deregulation plays a major role in the pathogenesis of Non-Small Cell Lung Cancer (NSCLC). CDC25A represents a critical cell cycle regulator that enhances cell cycle progression. In this study we aimed to investigate the role of a novel CDC25A transcriptional variant, CDC25A(Q110del), on the regulation of the CDC25A protein, and its impact on prognosis of NSCLC patients. Here we report a novel CDC25A transcript variant with codon 110 (Glutamine) deletion, that we termed CDC25A(Q110del) in NSCLC cells. In 9 (75%) of the 12 NSCLC cell lines, CDC25A(Q110del) expression accounted for more than 20% of the CDC25A transcripts. Biological effects of CDC25A(Q110del) were investigated in H1299 and HEK-293F cells using UV radiation, flowcytometry, cyclohexamide treatment, and confocal microscopy. Compared to CDC25A(wt), CDC25A(Q110del) protein had longer half-life; cells expressing CDC25A(Q110del) were more resistant to UV irradiation and showed more mitotic activity. Taqman-PCR was used to quantify CDC25A(Q110del) expression levels in 88 primary NSCLC tumor/normal tissue pairs. In patients with NSCLC, Kaplan Meier curves showed tumors expressing higher levels of CDC25A(Q110del) relative to the adjacent lung tissues to have significantly inferior overall survival (P = .0018). Here we identified CDC25A(Q110del) as a novel transcriptional variant of CDC25A in NSCLC. The sequence-specific nature of the abnormality could be a prognostic indicator in NSCLC patients as well as a candidate target for future therapeutic strategies.


A quality-improvement approach to urgent-care antibiotic stewardship for respiratory tract infections during the COVID-19 pandemic: Lessons learned.
We investigated a decrease in antibiotic prescribing for respiratory illnesses in 2 academic urgent-care clinics during the coronavirus disease 2019 (COVID-19) pandemic using semistructured clinician interviews. We conducted a quality-improvement project from November 2020 to May 2021. We investigated provider antibiotic decision making using a mixed-methods explanatory design including interviews. We analyzed transcripts using a thematic framework approach to identify emergent themes. Our performance measure was antibiotic prescribing rate (APR) for encounters with respiratory diagnosis billing codes. We extracted billing and prescribing data from the electronic medical record and assessed differences using run charts, p charts and generalized linear regression. We observed significant reductions in the APR early during the COVID-19 pandemic (relative risk [RR], 0.20; 95% confidence interval [CI], 0.17-0.25), which was maintained over the study period (P < .001). The average APRs were 14% before the COVID-19 pandemic, 4% during the QI project, and 7% after the project. All providers prescribed less antibiotics for respiratory encounters during COVID-19, but only 25% felt their practice had changed. Themes from provider interviews included changing patient expectations and provider approach to respiratory encounters during COVID-19, the impact of increased telemedicine encounters, and the changing epidemiology of non-COVID-19 respiratory infections. Our findings suggest that the decrease in APR was likely multifactorial. The average APR decreased significantly during the pandemic. Although the APR was slightly higher after the QI project, it did not reach prepandemic levels. Future studies should explore how these factors, including changing patient expectations, can be leveraged to improve urgent-care antibiotic stewardship.


The antifibrotic drug pirfenidone promotes pulmonary cavitation and drug resistance in a mouse model of chronic tuberculosis.
Pirfenidone is a recently approved antifibrotic drug for the treatment of idiopathic pulmonary fibrosis (IPF). Because tuberculosis (TB) is characterized by granulomatous inflammation in conjunction with parenchymal destruction and replacement fibrosis, we sought to determine whether the addition of pirfenidone as an adjunctive, host-directed therapy provides a beneficial effect during antimicrobial treatment of TB. We hypothesized that pirfenidone's antiinflammatory and antifibrotic properties would reduce inflammatory lung damage and increase antimicrobial drug penetration in granulomas to accelerate treatment response. The effectiveness of adjunctive pirfenidone during TB drug therapy was evaluated using a murine model of chronic TB. Mice treated with standard therapy 2HRZ/4HR (H, isoniazid; R, rifampin; and Z, pyrazinamide) were compared with 2 alternative regimens containing pirfenidone (Pf) (2HRZPf/4HRPf and 2HRZPf/4HR). Contrary to our hypothesis, adjunctive pirfenidone use leads to reduced bacterial clearance and increased relapse rates. This treatment failure is closely associated with the emergence of isoniazid monoresistant bacilli, increased cavitation, and significant lung pathology. While antifibrotic agents may eventually be used as part of adjunctive host-directed therapy of TB, this study clearly demonstrates that caution must be exercised. Moreover, as pirfenidone becomes more widely used in clinical practice, increased patient monitoring would be required in endemic TB settings.


Inhibition of peptidoglycan transpeptidase by beta-lactam antibiotics: structure-activity relationships.
The inhibitory activities of representative beta-lactam compounds, such as penicillins G and N, cephalosporins C and G, clavulanic acid, nocardicin A and thienamycin against Escherichia coli KN-126 and Bacillus megaterium KM peptidoglycan transpeptidases were studied. Their modes of action against E. coli are discussed on the basis of the results and the published binding data for penicillin binding proteins. The effects of modifications at position 3 and 7 of the cephalosporin and those at alpha-carbon of the benzyl side-chain of cephalosporin G and penicillin G were studied. The introduction of an amino group at this position in cephalosporin G together with the removal of an acetoxy group from the acetoxymethyl group at position 3 reduced the inhibitory activity against E. coli transpeptidase considerably. The activity was restored by the replacement of the methyl group at position 3 of cephalexin with chlorine. The restoration was accompanied by about 15-fold increase in the lytic activity of cephachlor against E. coli.


The diesel exhaust in miners study: I. Overview of the exposure assessment process.
This report provides an overview of the exposure assessment process for an epidemiologic study that investigated mortality, with a special focus on lung cancer, associated with diesel exhaust (DE) exposure among miners. Details of several components are provided in four other reports. A major challenge for this study was the development of quantitative estimates of historical exposures to DE. There is no single standard method for assessing the totality of DE, so respirable elemental carbon (REC), a component of DE, was selected as the primary surrogate in this study. Air monitoring surveys at seven of the eight study mining facilities were conducted between 1998 and 2001 and provided reference personal REC exposure levels and measurements for other agents and DE components in the mining environment. (The eighth facility had closed permanently prior to the surveys.) Exposure estimates were developed for mining facility/department/job/year combinations. A hierarchical grouping strategy was developed for assigning exposure levels to underground jobs [based on job titles, on the amount of time spent in various areas of the underground mine, and on similar carbon monoxide (CO, another DE component) concentrations] and to surface jobs (based on the use of, or proximity to, diesel-powered equipment). Time trends in air concentrations for underground jobs were estimated from mining facility-specific prediction models using diesel equipment horsepower, total air flow rates exhausted from the underground mines, and, because there were no historical REC measurements, historical measurements of CO. Exposures to potentially confounding agents, i.e. respirable dust, silica, radon, asbestos, and non-diesel sources of polycyclic aromatic hydrocarbons, also were assessed. Accuracy and reliability of the estimated REC exposures levels were evaluated by comparison with several smaller datasets and by development of alternative time trend models. During 1998-2001, the average measured REC exposure level by facility ranged from 40 to 384 μg m⁻³ for the underground workers and from 2 to 6 μg m⁻³ for the surface workers. For one prevalent underground job, 'miner operator', the maximum annual REC exposure estimate by facility ranged up to 685% greater than the corresponding 1998-2001 value. A comparison of the historical CO estimates from the time trend models with 1976-1977 CO measurements not used in the modeling found an overall median relative difference of 29%. Other comparisons showed similar levels of agreement. The assessment process indicated large differences in REC exposure levels over time and across the underground operations. Method evaluations indicated that the final estimates were consistent with those from alternative time trend models and demonstrated moderate to high agreement with external data.


Bedside differentiation of vestibular neuritis from central "vestibular pseudoneuritis".
Acute unilateral peripheral and central vestibular lesions can cause similar signs and symptoms, but they require different diagnostics and management. We therefore correlated clinical signs to differentiate vestibular neuritis (40 patients) from central "vestibular pseudoneuritis" (43 patients) in the acute situation with the final diagnosis assessed by neuroimaging. Skew deviation was the only specific but non-sensitive (40%) sign for pseudoneuritis. None of the other isolated signs (head thrust test, saccadic pursuit, gaze evoked nystagmus, subjective visual vertical) were reliable; however, multivariate logistic regression increased their sensitivity and specificity to 92%.


Single adult kidney stem/progenitor cells reconstitute three-dimensional nephron structures in vitro.
The kidneys are formed during development from two distinct primordial tissues, the metanephric mesenchyme and the ureteric bud. The metanephric mesenchyme develops into the kidney nephron, the minimal functional unit of the kidney. A nephron consists of several segments and regulates water, electrolyte, and acid-base homeostasis in addition to secreting certain hormones. It has been predicted that the kidney will be among the last organs successfully regenerated in vitro due to its complex structure and multiple functions. Here, we show that adult kidney stem/progenitor cells (KS cells), derived from the S3 segment of adult rat kidney nephrons, can reconstitute a three-dimensional kidney-like structure in vitro. Kidney-like structures were formed when a cluster of KS cells was suspended in an extracellular matrix gel and cultured in the presence of several growth factors. Morphological analyses revealed that these kidney-like structures contained every substructure of the kidney, including glomeruli, proximal tubules, the loop of Henle, distal tubules, and collecting ducts, but no vasculature. Our results demonstrate that a cluster of tissue stem/progenitor cells has the ability to reconstitute the minimum unit of its organ of origin by differentiating into specialized cells in the correct location. This process differs from embryonic kidney development, which requires the mutual induction of two different populations of progenitors, metanephric mesenchymal cells and ureteric bud cells.


A mutation update on the LDS-associated genes TGFB2/3 and SMAD2/3.
The Loeys-Dietz syndrome (LDS) is a connective tissue disorder affecting the cardiovascular, skeletal, and ocular system. Most typically, LDS patients present with aortic aneurysms and arterial tortuosity, hypertelorism, and bifid/broad uvula or cleft palate. Initially, mutations in transforming growth factor-β (TGF-β) receptors (TGFBR1 and TGFBR2) were described to cause LDS, hereby leading to impaired TGF-β signaling. More recently, TGF-β ligands, TGFB2 and TGFB3, as well as intracellular downstream effectors of the TGF-β pathway, SMAD2 and SMAD3, were shown to be involved in LDS. This emphasizes the role of disturbed TGF-β signaling in LDS pathogenesis. Since most literature so far has focused on TGFBR1/2, we provide a comprehensive review on the known and some novel TGFB2/3 and SMAD2/3 mutations. For TGFB2 and SMAD3, the clinical manifestations, both of the patients previously described in the literature and our newly reported patients, are summarized in detail. This clearly indicates that LDS concerns a disorder with a broad phenotypical spectrum that is still emerging as more patients will be identified. All mutations described here are present in the corresponding Leiden Open Variant Database.


The "not so simple" ankle fracture: avoiding problems and pitfalls to improve patient outcomes.
Ankle fractures are among the most common injuries managed by orthopaedic surgeons. Many ankle fractures are simple, with straightforward management leading to successful outcomes. Some fractures, however, are challenging, and debate arises regarding the best treatment to achieve an optimal outcome. Some patients have medical comorbidities that increase the risk for complications or may require modifications to standard surgical techniques and fixation methods. Several recent investigations have highlighted the pitfalls in accurately reducing syndesmotic injuries. Controversy remains regarding the number and diameter of screws, the duration of weight-bearing limitations, and the need or timing of screw removal. Open reduction may allow more accurate reduction than standard closed methods. Direct fixation of associated posterior malleolus fractures may provide improved syndesmotic stability. Posterior malleolus fractures vary in size and can be classified based on the orientation of the fracture line. As the size of the posterior malleolus fracture fragment increases, the load pattern in the ankle is altered. Direct or indirect reduction and surgical fixation may be required to prevent posterior talar subluxation and restore articular congruency. The supination-adduction fracture pattern is also important to recognize. Articular depression of the medial tibial plafond may require reduction and bone grafting. Optimal fixation requires directing screws parallel to the ankle joint or using a buttress plate. Identifying ankle fractures that may present additional treatment challenges is essential to achieving a successful outcome. A careful review of radiographs and CT scans, a thorough patient assessment, and detailed preoperative planning are needed to improve patient outcomes.


First Report of Diaporthe australafricana Associated with Stem Canker on Blueberry in Chile.
Stem cankers of blueberry (Vaccinium corymbosum L.) have been observed on as much as 15% of the plants in plantations in central and southern Chile since 2006. Symptoms consisted of apical necrosis of the shoots and brown-to-reddish necrotic lesions on the stems. Internally, a brown-to-reddish discoloration of the vascular tissue can be observed. Twenty, single-plant samples were collected in 12 blueberry plantings (approximately 33°27' to 40°53'S). Isolations from the margins of the necrotic lesions on the stems were made by plating small pieces (5 mm) on potato dextrose agar acidified with 0.5 μl/ml of 92% lactic acid (APDA). The plates were incubated at 20°C for 5 to 7 days, and hyphal tips of white colonies with septate and hyaline mycelium were transferred to APDA. Colonies were then transferred to autoclaved Pinus radiata needles on 2% water agar and incubated for 20 days at 20°C. Twelve isolates producing black pycnidia and alpha conidia were tentatively identified as a Phomopsis sp. (teleomoph Diaporthe Nitschke). Other fungi, including Botryosphaeriaceae spp. and Pestalotiopsis spp., were also isolated. Alpha conidia were smooth, unicellular, hyaline, fusoid, biguttulate, and 6.4 to 7.9 × 2.3 to 3.3 μm (n = 20). Beta conidia were not observed. The internal transcribed spacer (ITS) region of the rDNA was amplified using primers ITS1 and ITS2 (4) and sequenced. BLASTn analysis of the 473-bp fragment (GenBank Accession No. JQ045712) showed 100% identity to Diaporthe australafricana Crous & J.M. van Niekerk from Vitis vinifera (3). The pathogenicity of D. australafricana was studied on blueberry cv. O'Neal using detached stems (n = 4) in the laboratory, on 2-year-old potted plants (n = 4) in a greenhouse, and on attached stems of mature plants (n = 4) established in the ground. Inoculations were done by placing mycelial plugs taken from 7-day-old APDA cultures in a 7-mm long incision made on the stems. Inoculations with sterile mycelium plugs served as negative controls. Inoculation sites were wrapped with Parafilm to avoid rapid dehydration. Dark brown, necrotic lesions on the internal tissues were obtained on all inoculated stems 15 days after inoculation. Mean lesion lengths were 18.0 ± 7.4 mm on detached stems, 7.8 ± 6.9 mm on stems of 2-year-old plants, and 7.3 ± 2.5 mm on mature plants in the field. No symptoms developed on control stems. Reisolations were successful in 100% of the inoculated stems and D. australafricana was confirmed by the presence of pycnidia and alpha conidia. To our knowledge, this is the first report of D. australafricana causing stem canker in V. corymbosum. Previously, this pathogen has been reported to be affecting Vitis vinifera in Australia and South Africa (3). These results do not exclude that other plant-pathogenic fungi may be involved in this syndrome (1,2). References: (1) J. G. Espinoza et al. Plant Dis 92:1407, 2008. (2) J. G. Espinoza et al. Plant Dis. 93:1187, 2009. (3) J. M. van Niekerk et al. Australas. Plant Pathol. 34:27, 2005. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, NY, 1990.


Identification and characterization of microRNAs from Phaeodactylum tricornutum by high-throughput sequencing and bioinformatics analysis.
Diatoms, which are important planktons widespread in various aquatic environments, are believed to play a vital role in primary production as well as silica cycling. The genomes of the pennate diatom Phaeodactylum tricornutum and the centric diatom Thalassiosira pseudonana have been sequenced, revealing some characteristics of the diatoms' mosaic genome as well as some features of their fatty acid metabolism and urea cycle, and indicating their unusual properties. To identify microRNAs (miRNAs) from P. tricornutum and to study their probable roles in nitrogen and silicon metabolism, we constructed and sequenced small RNA (sRNA) libraries from P. tricornutum under normal (PT1), nitrogen-limited (PT2) and silicon-limited (PT3) conditions. A total of 13 miRNAs were identified. They were probable P. tricornutum-specific novel miRNAs. These miRNAs were sequenced from P. tricornutum under normal, nitrogen-limited and/or silicon-limited conditions, and their potential targets were involved in various processes, such as signal transduction, protein amino acid phosphorylation, fatty acid biosynthetic process, regulation of transcription and so on. Our results indicated that P. tricornutum contained novel miRNAs that have no identifiable homologs in other organisms and that they might play important regulator roles in P. tricornutum metabolism.


Communication: Evaluating non-empirical double hybrid functionals for spin-state energetics in transition-metal complexes.
The computationally assisted, accelerated design of inorganic functional materials often relies on the ability of a given electronic structure method to return the correct electronic ground state of the material in question. Outlining difficulties with current density functionals and wave function-based approaches, we highlight why double hybrid density functionals represent promising candidates for this purpose. In turn, we show that PBE0-DH (and PBE-QIDH) offers a significant improvement over its hybrid parent functional PBE0 [as well as B3LYP* and coupled cluster singles and doubles with perturbative triples (CCSD(T))] when computing spin-state splitting energies, using high-level diffusion Monte Carlo calculations as a reference. We refer to the opposing influence of Hartree-Fock (HF) exchange and MP2, which permits higher levels of HF exchange and a concomitant reduction in electronic density error, as the reason for the improved performance of double-hybrid functionals relative to hybrid functionals. Additionally, using 16 transition metal (Fe and Co) complexes, we show that low-spin states are stabilised by increasing contributions from MP2 within the double hybrid formulation. Furthermore, this stabilisation effect is more prominent for high field strength ligands than low field strength ligands.


Structure and function of the conserved core of histone deposition protein Asf1.
Asf1 is a ubiquitous eukaryotic histone binding and deposition protein that mediates nucleosome formation in vitro and is required for genome stability in vivo. Studies in a variety of organisms have defined Asf1's role as a histone chaperone during DNA replication through specific interactions with histones H3/H4 and the histone deposition factor CAF-I. In addition to its role in replication, conserved interactions with proteins involved in chromatin silencing, transcription, chromatin remodeling, and DNA repair have also established Asf1 as an important component of a number of chromatin assembly and modulation complexes. We demonstrate that the highly conserved N-terminal domain of S. cerevisiae Asf1 (Asf1N) is the core region that mediates all tested functions of the full-length protein. The crystal structure of this core domain, determined to 1.5 A resolution, reveals a compact immunoglobulin-like beta sandwich fold topped by three helical linkers. The surface of Asf1 displays a conserved hydrophobic groove flanked on one side by an area of strong electronegative surface potential. These regions represent potential binding sites for histones and other interacting proteins. The structural model also allowed us to interpret mutagenesis studies of the human Asf1a/HIRA interaction and to functionally define the region of Asf1 responsible for Hir1-dependent telomeric silencing in budding yeast. The evolutionarily conserved, N-terminal 155 amino acids of histone deposition protein Asf1 are functional in vitro and in vivo. This core region of Asf1 adopts a compact immunoglobulin-fold structure with distinct surface characteristics, including a Hir protein binding region required for gene silencing.


Point Prevalence Survey of Antibiotic Use across 13 Hospitals in Uganda.
Standardized monitoring of antibiotic use underpins the effective implementation of antimicrobial stewardship interventions in combatting antimicrobial resistance (AMR). To date, few studies have assessed antibiotic use in hospitals in Uganda to identify gaps that require intervention. This study applied the World Health Organization's standardized point prevalence survey methodology to assess antibiotic use in 13 public and private not-for-profit hospitals across the country. Data for 1077 patients and 1387 prescriptions were collected between December 2020 and April 2021 and analyzed to understand the characteristics of antibiotic use and the prevalence of the types of antibiotics to assess compliance with Uganda Clinical Guidelines; and classify antibiotics according to the WHO Access, Watch, and Reserve classification. This study found that 74% of patients were on one or more antibiotics. Compliance with Uganda Clinical Guidelines was low (30%); Watch-classified antibiotics were used to a high degree (44% of prescriptions), mainly driven by the wide use of ceftriaxone, which was the most frequently used antibiotic (37% of prescriptions). The results of this study identify key areas for the improvement of antimicrobial stewardship in Uganda and are important benchmarks for future evaluations.


Precision control of an invasive ant on an ecologically sensitive tropical island: a principle with wide applicability.
Effective management of invasive ants is an important priority for many conservation programs but can be difficult to achieve, especially within ecologically sensitive habitats. This study assesses the efficacy and nontarget risk of a precision ant baiting method aiming to reduce a population of the invasive big-headed ant Pheidole megacephala on a tropical island of great conservation value. Area-wide application of a formicidal bait, delivered in bait stations, resulted in the rapid decline of 8 ha of P. megacephala. Effective suppression remained throughout the succeeding 11-month monitoring period. We detected no negative effects of baiting on nontarget arthropods. Indeed, species richness of nontarget ants and abundance of other soil-surface arthropods increased significantly after P. megacephala suppression. This bait station method minimized bait exposure to nontarget organisms and was cost effective and adaptable to target species density. However, it was only effective over short distances and required thorough bait placement. This method would therefore be most appropriate for localized P. megacephala infestations where the prevention of nontarget impacts is essential. The methodology used here would be applicable to other sensitive tropical environments.


Single-route CNS prophylaxis for aggressive non-Hodgkin lymphomas: real-world outcomes from 21 US academic institutions.
Prophylaxis is commonly used to prevent central nervous sy stem (CNS) relapse in diffuse large B-cell lymphoma (DLBCL), with no clear standard of care. We retrospectively evaluated 1162 adult patients across 21 US academic centers with DLBCL or similar histologies who received single-route CNS prophylaxis as part of frontline therapy between 2013 and 2019. Prophylaxis was administered intrathecally(IT) in 894 (77%) and using systemic high-dose methotrexate (HD-MTX) in 236 (20%); 32 patients (3%) switched route due to toxicity and were assessed separately. By CNS-International Prognostic Index (IPI), 18% were considered low-risk, 51% moderate, and 30% high. Double-hit lymphoma (DHL) was confirmed in 243 of 866 evaluable patients (21%). Sixty-four patients (5.7%) had CNS relapse after median 7.1 months from diagnosis, including 15 of 64 (23%) within the first 6 months. There was no significant difference in CNS relapse between IT and HD-MTX recipients (5.4% vs 6.8%, P = .4), including after propensity score matching to account for differences between respective recipient groups. Weighting by CNS-IPI, expected vs observed CNS relapse rates were nearly identical (5.8% vs 5.7%). Testicular involvement was associated with high risk of CNS relapse (11.3%) despite most having lower CNS-IPI scores. DHL did not significantly predict for CNS relapse after single-route prophylaxis, including with adjustment for treatment regimen and other factors. This large study of CNS prophylaxis recipients with DLBCL found no significant difference in CNS relapse rates between routes of administration. Relapse rates among high-risk subgroups remain elevated, and reconsideration of prophylaxis strategies in DLBCL is of critical need.


Dynamics of G6PD activity in patients receiving weekly primaquine for therapy of Plasmodium vivax malaria.
Acute Plasmodium vivax malaria is associated with haemolysis, bone marrow suppression, reticulocytopenia, and post-treatment reticulocytosis leading to haemoglobin recovery. Little is known how malaria affects glucose-6-phosphate dehydrogenase (G6PD) activity and whether changes in activity when patients present may lead qualitative tests, like the fluorescent spot test (FST), to misdiagnose G6PD deficient (G6PDd) patients as G6PD normal (G6PDn). Giving primaquine or tafenoquine to such patients could result in severe haemolysis. We investigated the G6PD genotype, G6PD enzyme activity over time and the baseline FST phenotype in Cambodians with acute P. vivax malaria treated with 3-day dihydroartemisinin piperaquine and weekly primaquine, 0·75 mg/kg x8 doses. Of 75 recruited patients (males 63), aged 5-63 years (median 24), 15 were G6PDd males (14 Viangchan, 1 Canton), 3 were G6PD Viangchan heterozygous females, and 57 were G6PDn; 6 patients had α/β-thalassaemia and 26 had HbE. Median (range) Day0 G6PD activities were 0·85 U/g Hb (0·10-1·36) and 11·4 U/g Hb (6·67-16·78) in G6PDd and G6PDn patients, respectively, rising significantly to 1·45 (0·36-5·54, p<0.01) and 12·0 (8·1-17·4, p = 0.04) U/g Hb on Day7, then falling to ~Day0 values by Day56. Day0 G6PD activity did not correlate (p = 0.28) with the Day0 reticulocyte counts but both correlated over time. The FST diagnosed correctly 17/18 G6PDd patients, misclassifying one heterozygous female as G6PDn. In Cambodia, acute P. vivax malaria did not elevate G6PD activities in our small sample of G6PDd patients to levels that would result in a false normal qualitative test. Low G6PDd enzyme activity at disease presentation increases upon parasite clearance, parallel to reticulocytosis. More work is needed in G6PDd heterozygous females to ascertain the effect of P. vivax on their G6PD activities. The trial was registered (ACTRN12613000003774) with the Australia New Zealand Clinical trials (https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=363399&isReview=true).