A new myocyte-specific enhancer-binding factor that recognizes a conserved element associated with multiple muscle-specific genes.
Exposure of skeletal myoblasts to growth factor-deficient medium results in transcriptional activation of muscle-specific genes, including the muscle creatine kinase gene (mck). Tissue specificity, developmental regulation, and high-level expression of mck are conferred primarily by a muscle-specific enhancer located between base pairs (bp) -1350 and -1048 relative to the transcription initiation site (E. A. Sternberg, G. Spizz, W. M. Perry, D. Vizard, T. Weil, and E. N. Olson, Mol. Cell. Biol. 8:2896-2909, 1988). To begin to define the regulatory mechanisms that mediate the selective activation of the mck enhancer in differentiating muscle cells, we have further delimited the boundaries of this enhancer and analyzed its interactions with nuclear factors from a variety of myogenic and nonmyogenic cell types. Deletion mutagenesis showed that the region between 1,204 and 1,095 bp upstream of mck functions as a weak muscle-specific enhancer that is dependent on an adjacent enhancer element for strong activity. This adjacent activating element does not exhibit enhancer activity in single copy but acts as a strong enhancer when multimerized. Gel retardation assays combined with DNase I footprinting and diethyl pyrocarbonate interference showed that a nuclear factor from differentiated C2 myotubes and BC3H1 myocytes recognized a conserved A + T-rich sequence within the peripheral activating region. This myocyte-specific enhancer-binding factor, designated MEF-2, was undetectable in nuclear extracts from C2 or BC3H1 myoblasts or several nonmyogenic cell lines. MEF-2 was first detectable within 2 h after exposure of myoblasts to mitogen-deficient medium and increased in abundance for 24 to 48 h thereafter. The appearance of MEF-2 required ongoing protein synthesis and was prevented by fibroblast growth factor and type beta transforming growth factor, which block the induction of muscle-specific genes. A myoblast-specific factor that is down regulated within 4 h after removal of growth factors was also found to bind to the MEF-2 recognition site. A 10-bp sequence, which was shown by DNase I footprinting and diethyl pyrocarbonate interference to interact directly with MEF-2, was identified within the rat and human mck enhancers, the rat myosin light-chain (mlc)-1/3 enhancer, and the chicken cardiac mlc-2A promoter. Oligomers corresponding to the region of the mlc-1/3 enhancer, which encompasses this conserved sequence, bound MEF-2 and competed for its binding to the mck enhancer. These results thus provide evidence for a novel myocyte-specific enhancer-binding factor, MEF-2, that is expressed early in the differentiation program and is suppressed by specific polypeptide growth factors. The ability of MEF-2 to recognize conserved activating elements associated with multiple-specific genes suggests that this factor may participate in the coordinate regulation of genes during myogenesis.


Oxidosqualene cyclases involved in the biosynthesis of triterpenoids in Quercus suber cork.
Cork is a water-impermeable, suberin-based material harboring lignin, (hemi)cellulose, and extractable small molecules (primarily triterpenoids). Extractables strongly influence the properties of suberin-based materials. Though these previous findings suggest a key role for triterpenoids in cork material quality, directly testing this idea is hindered in part because it is not known which genes control cork triterpenoid biosynthesis. Here, we used gas chromatography and mass spectrometry to determine that the majority (>85%) of non-polar extractables from cork were pentacyclic triterpenoids, primarily betulinic acid, friedelin, and hydroxy-friedelin. In other plants, triterpenoids are generated by oxidosqualene cyclases (OSCs). Accordingly, we mined Quercus suber EST libraries for OSC fragments to use in a RACE PCR-based approach and cloned three full-length OSC transcripts from cork (QsOSC1-3). Heterologous expression in Saccharomyces cerevisiae revealed that QsOSC1-3 respectively encoded enzymes with lupeol synthase, mixed α- and β-amyrin synthase, and mixed β-amyrin and friedelin synthase activities. These activities together account for the backbone structures of the major cork triterpenoids. Finally, we analyzed the sequences of QsOSC1-3 and other plant OSCs to identify residues associated with specific OSC activities, then combined this with analyses of Q. suber transcriptomic and genomic data to evaluate potential redundancies in cork triterpenoid biosynthesis.


Ultraviolet filters in stomatopod crustaceans: diversity, ecology and evolution.
Stomatopod crustaceans employ unique ultraviolet (UV) optical filters in order to tune the spectral sensitivities of their UV-sensitive photoreceptors. In the stomatopod species Neogonodactylus oerstedii, we previously found four filter types, produced by five distinct mycosporine-like amino acid pigments in the crystalline cones of their specialized midband ommatidial facets. This UV-spectral tuning array produces receptors with at least six distinct spectral sensitivities, despite expressing only two visual pigments. Here, we present a broad survey of these UV filters across the stomatopod order, examining their spectral absorption properties in 21 species from seven families in four superfamilies. We found that UV filters are present in three of the four superfamilies, and evolutionary character reconstruction implies that at least one class of UV filter was present in the ancestor of all modern stomatopods. Additionally, postlarval stomatopods were observed to produce the UV filters simultaneously alongside development of the adult eye. The absorbance properties of the filters are consistent within a species; however, between species we found a great deal of diversity, both in the number of filters and in their spectral absorbance characteristics. This diversity correlates with the habitat depth ranges of these species, suggesting that species living in shallow, UV-rich environments may tune their UV spectral sensitivities more aggressively. We also found additional, previously unrecognized UV filter types in the crystalline cones of the peripheral eye regions of some species, indicating the possibility for even greater stomatopod visual complexity than previously thought.


Gallstone Disease and the Risk of Ischemic Heart Disease.
Gallstone disease (GSD) is related to multiple cardiovascular risk factors; the present study was to prospectively examine the association between GSD and ischemic heart disease (IHD). We examined the association of GSD with IHD among 199 292 men and 288 081 women aged 30-79 years in the China Kadoorie Biobank study. Participants with cancer, heart disease, and stroke at baseline were excluded. Cox proportional hazards regression model was used to estimate the association of GSD with IHD. The prevalence of self-reported GSD was 3.7% in men and 7.3% in women at baseline. During 3 431 124 person-years of follow-up between 2004 and 2013 (median, 7.2 years), we documented 10 245 incident IHD cases in men and 14 714 in women. As compared with men without GSD at baseline, the multivariate-adjusted hazard ratio for IHD was 1.11 (95% confidence interval, 1.02-1.22) for men with GSD; the respective hazard ratio was 1.27 (95% confidence interval, 1.20-1.34) in women and 1.23 (95% confidence interval, 1.17-1.28) in the whole cohort. The sex difference in IHD risk associated with GSD was statistically significant (P=0.009 for interaction with sex). In addition, we found that the association between GSD and IHD was stronger in nonhypertensive than in hypertensive women (P<0.001 for interaction). In this large prospective study, the presence of GSD was associated with an increased risk of incident IHD, independent of other risk factors of cardiovascular disease. Our findings suggest novel prevention strategy to mitigate heart disease through improvement of gastrointestinal health.


"I see what you're saying": intrusive images from listening to a traumatic verbal report.
We tested the hypothesis that intrusive visual images could develop from listening to a traumatic verbal report. Eighty-six participants listened to a traumatic verbal report under one of three conditions: while shaping plasticine (visuospatial condition), while performing articulatory suppression (verbal condition), or with no extra task (control condition). Results showed that intrusive visual images developed from listening to the traumatic report. In line with the idea that central executive processes guide encoding of information, intrusion frequency was reduced in both the visuospatial and the verbal condition compared to the no task control condition. Overall, this pattern is similar to intrusive images from a traumatic film as found in earlier studies. This study provides a valuable addition to models of posttraumatic stress disorder and autobiographical memory. Additionally, the results have potential implications for therapists working with traumatized individuals.


Failure to clear persistent vaccine-derived neurovirulent poliovirus infection in an immunodeficient man.
Individuals who chronically excrete neurovirulent poliovirus of vaccine-origin are of considerable concern to the Global Polio Eradication programme. Chronic infection with such polioviruses is a recognised complication of hypogammaglobulinaemia. We did a series of in-vitro and in-vivo therapeutic studies, with a view to clearing persistent neurovirulent poliovirus infection in an individual with common variable immunodeficiency, using oral immunoglobulin, breast milk (as a source of secretory IgA), ribavirin, and the anti-picornaviral agent pleconaril. We undertook viral quantitation, antibody neutralisation and drug susceptibility assays, and viral gene sequencing. Long-term asymptomatic excretion of vaccine-derived neurovirulent poliovirus 2 was identified in this hypogammaglobulinaemic man, and was estimated to have persisted for up to 22 years. Despite demonstrable in-vitro neutralising activity of immunoglobulin and breast milk, and in-vitro antiviral activity of ribavirin, no treatment was successful at clearing the virus, although in one trial breast milk significantly reduced excretion levels temporarily. During the course of study, the virus developed reduced susceptibility to pleconaril, precluding the in-vivo use of this drug. Sequence analysis revealed the emergence of a methionine to leucine mutation adjacent to the likely binding site of pleconaril in these isolates. Chronic vaccine-associated poliovirus infection in hypogammaglobulinaemia is a difficult condition to treat. It represents a risk to the strategy to discontinue polio vaccination once global eradication has been achieved.


Weekly docetaxel in elderly patients with prostate cancer: efficacy and toxicity in patients at least 70 years of age compared with patients younger than 70 years.
We sought to determine whether age was significantly associated with efficacy and toxicity of weekly docetaxel in patients with metastatic androgen-independent prostate cancer (AIPC). Individual patient data were pooled from 2 phase II clinical trials of weekly docetaxel 36 mg/m(2) for 6 of every 8 weeks in men with metastatic AIPC. Baseline characteristics and outcome measures of men > 70 years of age (n = 52) were compared with patients < 70 of age (n = 34) using Pearson c2 test for categoric variables, Mann-Whitney U test for continuous variables, and log-rank test of Kaplan-Meier estimates for time-dependent variable. Multivariate analysis was used to adjust for any imbalances in baseline characteristics. At baseline, older patients had a lower hemoglobin level (P = 0.05) and a higher serum prostate-specific antigen (PSA; P = 0.04). The PSA response rate was 47% (95% CI, 33%-62%) in older patients and 40% (95% CI, 23%-59%) in younger patients (P = 0.75). Similarly, measurable disease response rate (P = 0.43), time to progression (P = 0.28), and survival (P = 0.52) were not affected by age in both univariate and multivariate analyses. There was also no difference in overall hematologic and nonhematologic toxicity > grade 2. This comparison of pooled individual patient data from 2 phase II studies of weekly docetaxel in AIPC did not reveal significant differences in efficacy or toxicity in men aged > 70 years compared with younger patients. These findings are consistent with the hypothesis that docetaxel chemotherapy in patients with AIPC is equally well tolerated and effective across a wide range of ages.


Metagenomes obtained by 'deep sequencing' - what do they tell about the enhanced biological phosphorus removal communities?
Metagenomics enables studies of the genomic potential of complex microbial communities by sequencing bulk genomic DNA directly from the environment. Knowledge of the genetic potential of a community can be used to formulate and test ecological hypotheses about stability and performance. In this study deep metagenomics and fluorescence in situ hybridization (FISH) were used to study a full-scale wastewater treatment plant with enhanced biological phosphorus removal (EBPR), and the results were compared to an existing EBPR metagenome. EBPR is a widely used process that relies on a complex community of microorganisms to function properly. Insight into community and species level stability and dynamics is valuable for knowledge-driven optimization of the EBPR process. The metagenomes of the EBPR communities were distinct compared to metagenomes of communities from a wide range of other environments, which could be attributed to selection pressures of the EBPR process. The metabolic potential of one of the key microorganisms in the EPBR process, Accumulibacter, was investigated in more detail in the two plants, revealing a potential importance of phage predation on the dynamics of Accumulibacter populations. The results demonstrate that metagenomics can be used as a powerful tool for system wide characterization of the EBPR community as well as for a deeper understanding of the function of specific community members. Furthermore, we discuss and illustrate some of the general pitfalls in metagenomics and stress the need of additional DNA extraction independent information in metagenome studies.


Interactions between rotavirus and Suwannee River organic matter: aggregation, deposition, and adhesion force measurement.
Interactions between rotavirus and Suwannee River natural organic matter (NOM) were studied by time-resolved dynamic light scattering, quartz crystal microbalance, and atomic force microscopy. In NOM-containing NaCl solutions of up to 600 mM, rotavirus suspension remained stable for over 4 h. Atomic force microscopy (AFM) measurement for interaction force decay length at different ionic strengths showed that nonelectrostatic repulsive forces were mainly responsible for eliminating aggregation in NaCl solutions. Aggregation rates of rotavirus in solutions containing 20 mg C/L increased with divalent cation concentration until reaching a critical coagulation concentration of 30 mM CaCl(2) or 70 mM MgCl(2). Deposition kinetics of rotavirus on NOM-coated silica surface was studied using quartz crystal microbalance. Experimental attachment efficiencies for rotavirus adsorption to NOM-coated surface in MgCl(2) solution were lower than in CaCl(2) solution at a given divalent cation concentration. Stronger adhesion force was measured for virus-virus and virus-NOM interactions in CaCl(2) solution compared to those in MgCl(2) or NaCl solutions at the same ionic strength. This study suggested that divalent cation complexation with carboxylate groups in NOM and on virus surface was an important mechanism in the deposition and aggregation kinetics of rotavirus.


Variability in responses and temporal coding of tastants of similar quality in the nucleus of the solitary tract of the rat.
In the nucleus of the solitary tract (NTS), electrophysiological responses to taste stimuli representing four basic taste qualities (sweet, sour, salty, or bitter) can often be discriminated by spike count, although in units for which the number of spikes is variable across identical stimulus presentations, spike timing (i.e., temporal coding) can also support reliable discrimination. The present study examined the contribution of spike count and spike timing to the discrimination of stimuli that evoke the same taste quality but are of different chemical composition. Responses to between 3 and 21 repeated presentations of two pairs of quality-matched tastants were recorded from 38 single cells in the NTS of urethane-anesthetized rats. Temporal coding was assessed in 24 cells, most of which were tested with salty and sour tastants, using an information-theoretic approach. Within a given cell, responses to tastants of similar quality were generally closer in magnitude than responses to dissimilar tastants; however, tastants of similar quality often reversed their order of effectiveness across replicate sets of trials. Typically, discrimination between tastants of dissimilar qualities could be made by spike count. Responses to tastants of similar quality typically evoked more similar response magnitudes but were more frequently, and to a proportionally greater degree, distinguishable based on temporal information. Results showed that nearly every taste-responsive NTS cell has the capacity to generate temporal features in evoked spike trains that can be used to distinguish between stimuli of different qualities and chemical compositions.


Pelvic exenteration for locally advanced and recurrent rectal cancer-how much more?
There have been significant advances in the surgical management of locally advanced and recurrent rectal cancer in recent decades. Patient with advanced pelvic tumours involving adjacent organs and neurovascular structures, beyond the traditional mesorectal planes, who would have traditionally been considered irresectable at many centres, now undergo surgery routinely at specialised units. While high rates of morbidity and mortality were reported by the pioneers of pelvic exenteration (PE) in early literature, this is now considered historical data. In 2019, patients who undergo PE for advanced or recurrent rectal cancer can expect reasonable rates of long-term survival (up to 60% at 5 years) and acceptable morbidity and quality of life. This article describes the surgical techniques that have been developed for radical multivisceral pelvic resections and reviews contemporary outcomes.


Anemia, micronutrient deficiencies, malaria, hemoglobinopathies and malnutrition in young children and non-pregnant women in Ghana: Findings from a national survey.
Nationally representative data on the micronutrient status of Ghanaian women and children are very scarce. We aimed to document the current national prevalence of micronutrient deficiencies, anemia, malaria, inflammation, α-thalassemia, sickle cell disease and trait, and under- and over-nutrition in Ghana. In 2017, a two-stage cross-sectional design was applied to enroll pre-school children (6-59 months) and non-pregnant women (15-49 years) from three strata in Ghana: Northern, Middle and Southern Belt. Household and individual questionnaire data were collected along with blood samples. In total, 2123 households completed the household interviews, 1165 children and 973 women provided blood samples. Nationally, 35.6% (95%CI: 31.7,39.6) of children had anemia, 21.5% (18.4,25.0) had iron deficiency, 12.2% (10.1,14.7) had iron deficiency anemia, and 20.8% (18.1,23.9) had vitamin A deficiency; 20.3%(15.2,26.6) tested positive for malaria, 13.9% (11.1,17.3) for sickle trait plus disease, and 30.7% (27.5,34.2) for α-thalassemia. Anemia and micronutrient deficiencies were more prevalent in rural areas, poor households and in the Northern Belt. Stunting and wasting affected 21.4% (18.0,25.2) and 7.0% (5.1,9.5) of children, respectively. Stunting was more common in rural areas and in poor households. Among non-pregnant women, 21.7% (18.7,25.1) were anemic, 13.7% (11.2,16.6) iron deficient, 8.9% (6.7,11.7) had iron deficiency anemia, and 1.5% (0.8,2.9) were vitamin A deficient, 53.8% (47.6,60.0) were folate deficient, and 6.9% (4.8,9.8) were vitamin B12 deficient. Malaria parasitemia in women [8.4% (5.7,12.2)] was lower than in children, but the prevalence of sickle cell disease or trait and α-thalassemia were similar. Overweight [24.7% (21.0,28.8)] and obesity [14.3% (11.5,17.7)] were more common in wealthier, older, and urban women. Our findings demonstrate that anemia and several micronutrient deficiencies are highly present in Ghana calling for the strengthening of Ghana's food fortification program while overweight and obesity in women are constantly increasing and need to be addressed urgently through governmental policies and programs.


SU-E-I-36: A Study to Compare Operator Dosimeter Exposure versus Fluoroscopic Time and Air Kerma in Fluoroscopically Guided Interventional Procedures.
The total effective dose equivalent limit for occupationally radiation exposed persons has remained at 50 millisieverts per year since the 1960s. There is ongoing discussion whether this limit should be lowered. Whether or not it is lowered, all facilities have to adhere to the principle of ALARA (as low as reasonably achievable). In a hospital environment, the workers who typically have exposures which Result in ALARA investigations having to be performed include interventional radiologists, cardiologists, and mobile C-arm operators. At our institution there are a limited number of interventional radiologists with one person being responsible for performing the majority of the procedures and sometimes exceeding the ALARA investigational levels. A limited study was performed to compare the number, type of studies, cumulative air kerma, and fluoroscopic time versus the exposure measured by the personnel dosimeters worn by the radiologists. This was also related to the type of angiographic equipment in use. The length of fluoroscopic time and cumulative air kerma and cumulative DAP appeared to be related to the higher exposures received by the operator. The number of procedures performed did not correlate with the personnel dosimeter exposure; rather it was the type and complexity of a study and the length of fluoroscopic time that was related. The number of ALARA investigations required has decreased gradually. This could be attributed to several factors. These include increased use of available safety items. With an increase in the number of interventionalists the work load is distributed more equitably. The purchase of new angiographic equipment has resulted in better image quality and more shielding options for the operators. The operators are periodically reminded to keep fluoroscopic time to a minimum and use appropriate settings on the equipment. The equipment is calibrated to provide adequate image quality at mid-dose settings.


Mechanisms of transcription in eosinophils: GATA-1, but not GATA-2, transactivates the promoter of the eosinophil granule major basic protein gene.
Granule major basic protein (MBP) is expressed exclusively in eosinophils, basophils, and placental trophoblasts. To identify the cis-elements and transcription factors involved in regulating MBP expression, we subcloned 3.2 kb of sequence upstream of the exon 9 transcriptional start site (P2 promoter) and serial 5' deletions into the pXP2 luciferase reporter vector. An 80% decrement in promoter activity was obtained when MBP sequences between bp -117 to -67 were deleted. To identify transcription factors that bind to and transactivate through the bp -117 to -67 region, we first compared the upstream genomic sequences of human and murine MBP; a potential GATA binding consensus site was conserved in the 50-bp region between the two genes. To determine which GATA proteins bind this consensus site, we performed electrophoretic mobility shift assays (EMSAs), which showed that both GATA-1 and GATA-2 can bind to this consensus site. To determine the functionality of this site, we tested whether GATA-1 and GATA-2, either individually or in combination, can transactivate the MBP promoter in the Jurkat T cell line. Cotransfection with a GATA-1 expression vector produced 20-fold augmentation of MBP promoter activity, whereas GATA-2 had no activity. In contrast, combined cotransfection of GATA-1 and GATA-2 decreased the ability of GATA-1 to transactivate the MBP promoter by approximately 50%. Our results provide the first evidence for a GATA-1 target gene in eosinophils, a negative regulatory role for GATA-2 in MBP expression, and possibly eosinophil gene transcription in general during myelopoiesis.


Phytocomponent p-hydroxycinnamic acid stimulates bone formation and inhibits bone resorption in rat femoral tissues in vitro.
The effect of cinnamic acid or its related compounds, which is present in many plants, on bone metabolism has not been clarified yet. The effect of cinnamic acid, p-hydroxycinnamic acid (HCA), ferulic acid, caffeic acid, or 3,4-dimethoxycinnamic acid (DCA) on bone calcium content in vitro was investigated. Rat femoral-diaphyseal (cortical bone) and -metaphyseal (trabecular bone) tissues were cultured for 48,h in Dulbecco's modified Eagle's medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. The presence of HCA (10(-5) or 10(-4),M) caused a significant increase in calcium content in the diaphyseal or metaphyseal tissues. Such an effect was not observed in the presence of cinnamic acid or other compounds at the concentration of 10(-5) or 10(-4),M. Alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the diaphyseal or metaphyseal tissues was significantly increased in the presence of HCA (10(-5) or 10(-4),M). The effect of HCA (10(-4),M) in increasing calcium content, alkaline phosphatase activity, and DNA content in the diaphyseal or metaphyseal tissues was completely prevented in the presence of cycloheximide (10(-6),M), an inhibitor of protein synthesis. Thus HCA had anabolic effects on bone components. The presence of parathyroid hormone (PTH; 10(-7),M), a bone-resorbing factor, caused a significant decrease in calcium content and a corresponding elevation in medium glucose consumption, lactic acid production or tartrate-resistant acid phosphatase (TRACP) activity in the diaphyseal or metaphyseal tissues. These alterations were completely prevented in the presence of HCA (10(-5) or 10(-4),M). This study demonstrates that p-hydroxycinnamic acid (HCA) has stimulatory effects on bone formation and inhibitory effects on bone resorption in tissue culture in vitro.


Piperine potentiates the hypocholesterolemic effect of curcumin in rats fed on a high fat diet.
It has previously been demonstrated that curcumin possesses a hypocholesterolemic effect and potentiates numerous pharmacological effects of curcumin, however, the mechanisms underlying this hypocholesterolemic effect and the interaction between curcumin and piperine remain to be elucidated. In the present study, male Sprague-Dawley rats were fed on a high-fat diet (HFD) to establish a hyperlipidemia (HLP) model. Co-administration of curcumin plus piperine was found to decrease the levels of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol in the serum and liver, as well as increase the levels of fecal TC, TG and total bile acid, compared with administration of curcumin alone. Curcumin plus piperine also markedly increased the levels of high-density lipoprotein cholesterol. Furthermore, compared with administration of curcumin alone, administration of curcumin plus piperine resulted in a significant upregulation of the activity and gene expression of apolipoprotein AI (ApoAI), lecithin cholesterol acyltransferase (LCAT), cholesterol 7α-hydroxylase (CYP7A1) and low-density lipoprotein receptor (LDLR). In conclusion, these results indicated that co-administration of curcumin plus piperine potentiates the hypocholesterolemic effects of curcumin by increasing the activity and gene expression of ApoAI, CYP7A1, LCAT and LDLR, providing a promising combination for the treatment of HLP.


Association of Thyroid Function with Severity of Coronary Artery Disease in Euthyroid Patients.
Thyroid hormone exerts multiple effects on the heart and vascular system. Variations of free T3 have been linked to coronary artery disease. We conducted a study to observe whether there is a relationship between the variation of the serum thyroid hormone levels (TSH, FT3 and FT4) and the presence and severity of CAD in the euthyroid patients. To study association of serum TSH, FT4 and FT3 levels within the normal range with presence and severity of coronary artery disease. A total of 100 euthyroid patients with stable angina, who underwent coronary angiography were enrolled in the study. Coronary artery disease was defined as >50% stenosis in the luminal diameter in at least one major epicardial coronary artery. The Gensini scoring system was used to define the severity of the CAD and serum TSH, FT3 and FT4 levels were measured by the chemiluminescence method. Single vessel disease was found in 23%, double vessel disease in 15% and triple vessel disease in 17% of patients. TSH and FT4 levels were also comparable between the groups. Normal coronary group had significantly higher mean FT3 values than triple vessel disease (p=0.004) and FT3 levels showed an inverse relation with Gensini score (Pearson's correlation =- 0.30) (p =0.002). A level of FT3 ≤ 2.7 predicted the severity of CAD with a 70% sensitivity and 60% specificity (area under curve (AUC): 0.755, p=0.001). In the absence of primary thyroid disease and acute coronary syndrome, the occurrence of CAD is associated with lower serum levels of FT3. FT3 and not the FT4 and TSH levels may be used as an indicator of increased risk for severe CAD. The present study clearly shows the existence of a strong association between the reduction of biologically active T3 and severity of coronary artery disease. However, low T3 state could be at first interpreted as just a biological risk factor of severe coronary artery disease; only the demonstration of beneficial effects on cardiovascular, end points of long term T3 replacement in CAD patients with low T3 state can answer this fundamental issue.


Magnetic field-induced switching of the radical-pair intersystem crossing mechanism in a donor-bridge-acceptor molecule for artificial photosynthesis.
A covalent, fixed-distance donor-bridge-acceptor (D-B-A) molecule was synthesized that upon photoexcitation undergoes ultrafast charge separation to yield a radical ion pair (RP) in which the spin-spin exchange interaction (2J) between the two radicals is sufficiently large to result in preferential RP intersystem crossing to the highest-energy RP eigenstate (T(+1)) at the 350 mT magnetic field characteristic of X-band (9.5 GHz) EPR spectroscopy. This behavior is unprecedented in covalent D-B-A molecules, and is evidenced by the time-resolved EPR (TREPR) spectrum at X-band of (3*)D-B-A derived from RP recombination, which shows all six canonical EPR transitions polarized in emission (e,e,e,e,e,e). In contrast, when the RP is photogenerated in a 3400 mT magnetic field, the TREPR triplet spectrum at W-band (94 GHz) of (3*)D-B-A displays the (a,e,e,a,a,e) polarization pattern characteristic of a weakly coupled RP precursor, similar to that observed in photosynthetic reaction center proteins, and indicates a switch to selective population of the lower-energy T(0) eigenstate.


Epidemiology of sudden death in young, competitive athletes due to blunt trauma.
Sudden deaths of young competitive athletes are highly visible events that have a substantial effect on families and communities. Recent attention has focused predominantly on cardiovascular causes, and less on traumatic organ damage. To define the clinical profile, epidemiology, and frequency of trauma-related deaths in young US athletes. We analyzed the 30-year US National Registry of Sudden Death in Young Athletes (1980-2009) by using systematic identification and tracking strategies. Of 1827 deaths of athletes aged 21 years or younger, 261 (14%) were caused by trauma-related injuries, usually involving the head and/or neck (mean: 16 ± 2 years; 90% male) in 22 sports. The highest number of events in a single year was 16 (1986), with an average of 9 per year throughout 30 years. The mortality rate was 0.11 in 100 000 participations (95% confidence interval: 0.08-0.15). The largest number of deaths was in football (148 [57%]), including 17 high school athletes who sustained concussions shortly before fatal head trauma ("second-impact syndrome"). Football deaths were more frequent in defensive players, although the single most common position involved was running back (61% of offensive players). In a large community-based national registry, sudden deaths caused by blunt trauma in young athletes aged 21 years or younger were relatively uncommon with 16 or fewer per year, about fourfold less than cardiovascular deaths. These fatalities were most frequent in football, and an important proportion of deaths after head blows in high school football were associated with a recent history of symptomatic concussion.


Explaining loss caused by tomato spotted wilt virus on tobacco with boreal winter weather: a Bayesian approach.
In North Carolina, Tomato spotted wilt virus (TSWV) has regularly been reported since 1997, with incidence being the highest in 2002. At the end of each season, a questionnaire is sent to the county agents to report disease losses. TSWV reported losses in 1993 to 2007 from 58 counties were available. A county-year combination was considered a case and, in total, 494 cases were analyzed. The winter months' temperature and precipitation significantly explained the reported TSWV loss (R(2) = 0.82). Specifically, the monthly average air temperature for December to February had a positive association with TSWV loss (P < 0.0001) whereas the total precipitation for the same months had a negative effect (P < 0.0001). Bayesian hierarchical models were implemented to include spatial and nonspatial random effects to investigate if there were significant spatial correlations or unexplained variability, respectively, and, thus, other significant variables that were ignored in the model development. The spatial random effects were not significant but the nonspatial random effects were significant in 36 cases. The importance of spring weather to dispersal of thrips and TSWV has been previously identified. Winter weather also may be a good indicator of potential available TSWV inoculum for the upcoming season.


Synthetic tolerance: three noncoding small RNAs, DsrA, ArcZ and RprA, acting supra-additively against acid stress.
Synthetic acid tolerance, especially during active cell growth, is a desirable phenotype for many biotechnological applications. Natively, acid resistance in Escherichia coli is largely a stationary-phase phenotype attributable to mechanisms mostly under the control of the stationary-phase sigma factor RpoS. We show that simultaneous overexpression of noncoding small RNAs (sRNAs), DsrA, RprA and ArcZ, which are translational RpoS activators, increased acid tolerance (based on a low-pH survival assay) supra-additively up to 8500-fold during active cell growth, and provided protection against carboxylic acid and oxidative stress. Overexpression of rpoS without its regulatory 5'-UTR resulted in inferior acid tolerance. The supra-additive effect of overexpressing the three sRNAs results from the impact their expression has on RpoS-protein levels, and the beneficial perturbation of the interconnected RpoS and H-NS networks, thus leading to superior tolerance during active growth. Unlike the overexpression of proteins, overexpression of sRNAs imposes hardly any metabolic burden on cells, and constitutes a more effective strain engineering strategy.


Mediterranean grassland soil C-N compound turnover is dependent on rainfall and depth, and is mediated by genomically divergent microorganisms.
Soil microbial activity drives the carbon and nitrogen cycles and is an important determinant of atmospheric trace gas turnover, yet most soils are dominated by microorganisms with unknown metabolic capacities. Even Acidobacteria, among the most abundant bacteria in soil, remain poorly characterized, and functions across groups such as Verrucomicrobia, Gemmatimonadetes, Chloroflexi and Rokubacteria are understudied. Here, we have resolved 60 metagenomic and 20 proteomic data sets from a Mediterranean grassland soil ecosystem and recovered 793 near-complete microbial genomes from 18 phyla, representing around one-third of all microorganisms detected. Importantly, this enabled extensive genomics-based metabolic predictions for these communities. Acidobacteria from multiple previously unstudied classes have genomes that encode large enzyme complements for complex carbohydrate degradation. Alternatively, most microorganisms encode carbohydrate esterases that strip readily accessible methyl and acetyl groups from polymers like pectin and xylan, forming methanol and acetate, the availability of which could explain the high prevalence of C1 metabolism and acetate utilization in genomes. Microorganism abundances among samples collected at three soil depths and under natural and amended rainfall regimes indicate statistically higher associations of inorganic nitrogen metabolism and carbon degradation in deep and shallow soils, respectively. This partitioning decreased in samples under extended spring rainfall, indicating that long-term climate alteration can affect both carbon and nitrogen cycling. Overall, by leveraging natural and experimental gradients with genome-resolved metabolic profiles, we link microorganisms lacking prior genomic characterization to specific roles in complex carbon, C1, nitrate and ammonia transformations, and constrain factors that impact their distributions in soil.


Nutrition and Exercise Interventions to Improve Body Composition for Persons with Overweight or Obesity Near Retirement Age: A Systematic Review and Network Meta-Analysis of Randomized Controlled Trials.
The retirement phase is an opportunity to integrate healthy (nutrition/exercise) habits into daily life. We conducted this systematic review to assess which nutrition and exercise interventions most effectively improve body composition (fat/muscle mass), body mass index (BMI), and waist circumference (WC) in persons with obesity/overweight near retirement age (ages 55-70 y). We conducted a systematic review and network meta-analysis (NMA) of randomized controlled trials, searching 4 databases from their inception up to July 12, 2022. The NMA was based on a random effects model, pooled mean differences, standardized mean differences, their 95% confidence intervals, and correlations with multi-arm studies. Subgroup and sensitivity analyses were also conducted. Ninety-two studies were included, 66 of which with 4957 participants could be used for the NMA. Identified interventions were clustered into 12 groups: no intervention, energy restriction (i.e., 500-1000 kcal), energy restriction plus high-protein intake (1.1-1.7 g/kg/body weight), intermittent fasting, mixed exercise (aerobic and resistance), resistance training, aerobic training, high protein plus resistance training, energy restriction plus high protein plus exercise, energy restriction plus resistance training, energy restriction plus aerobic training, and energy restriction plus mixed exercise. Intervention durations ranged from 8 wk to 6 mo. Body fat was reduced with energy restriction plus any exercise or plus high-protein intake. Energy restriction alone was less effective and tended to decrease muscle mass. Muscle mass was only significantly increased with mixed exercise. All other interventions including exercise effectively preserved muscle mass. A BMI and/or WC decrease was achieved with all interventions except aerobic training/resistance training alone or resistance training plus high protein. Overall, the most effective strategy for nearly all outcomes was combining energy restriction with resistance training or mixed exercise and high protein. Health care professionals involved in the management of persons with obesity need to be aware that an energy-restricted diet alone may contribute to sarcopenic obesity in persons near retirement age. This network meta-analysis is registered at https://www.crd.york.ac.uk/prospero/ as CRD42021276465.


Acetate and CO2 assimilation by Methanothrix concilii.
Biosynthetic pathways in Methanothrix concilii, a recently isolated aceticlastic methanogen, were studied by 13C-nuclear magnetic resonance spectroscopy. Labeling patterns of amino acids, lipids, and carbohydrates were determined. Similar to other methanogens, acetate was carboxylated to pyruvate, which was further converted to amino acids by various biosynthetic pathways. The origin of carbon atoms in glutamate, proline, and arginine clearly showed that an incomplete tricarboxylic acid cycle operating in the oxidative direction was used for their biosynthesis. Isoleucine was synthesized via citramalate, which is a typical route for methanogens. As with Methanosarcina barkeri, an extensive exchange of the label between the carboxyl group of acetate and CO2 was observed. Lipids predominantly contained diphytanyl chains, the labeling of which indicated that biosynthesis proceeded through mevalonic acid. Labeling of the C-1,6 of glucose from [2-13C]acetate is consistent with a glucogenic route for carbohydrate biosynthesis. Except for the different origins of the methyl group of methionine, the metabolic properties of Methanothrix concilii are closely related to those of Methanosarcina barkeri.


Sequencing crop genomes: approaches and applications.
Many challenges face plant scientists, in particular those working on crop production, such as a projected increase in population, decrease in water and arable land, changes in weather patterns and predictability. Advances in genome sequencing and resequencing can and should play a role in our response to meeting these challenges. However, several barriers prevent rapid and effective deployment of these tools to a wide variety of crops. Because of the complexity of crop genomes, de novo sequencing with next-generation sequencing technologies is a process fraught with difficulties that then create roadblocks to the utilization of these genome sequences for crop improvement. Collecting rapid and accurate phenotypes in crop plants is a hindrance to integrating genomics with crop improvement, and advances in informatics are needed to put these tools in the hands of the scientists on the ground.


Longitudinal evaluation of physician payment reform and team-based care for chronic disease management and prevention.
We evaluated a large-scale transition of primary care physicians to blended capitation models and team-based care in Ontario, Canada, to understand the effect of each type of reform on the management and prevention of chronic disease. We used population-based administrative data to assess monitoring of diabetes mellitus and screening for cervical, breast and colorectal cancer among patients belonging to team-based capitation, non-team-based capitation or enhanced fee-for-service medical homes as of Mar. 31, 2011 (n = 10 675 480). We used Poisson regression models to examine these associations for 2011. We then used a fitted nonlinear model to compare changes in outcomes between 2001 and 2011 by type of medical home. In 2011, patients in a team-based capitation setting were more likely than those in an enhanced fee-for-service setting to receive diabetes monitoring (39.7% v. 31.6%, adjusted relative risk [RR] 1.22, 95% confidence interval [CI] 1.18 to 1.25), mammography (76.6% v. 71.5%, adjusted RR 1.06, 95% CI 1.06 to 1.07) and colorectal cancer screening (63.0% v. 60.9%, adjusted RR 1.03, 95% CI 1.02 to 1.04). Over time, patients in medical homes with team-based capitation experienced the greatest improvement in diabetes monitoring (absolute difference in improvement 10.6% [95% CI 7.9% to 13.2%] compared with enhanced fee for service; 6.4% [95% CI 3.8% to 9.1%] compared with non-team-based capitation) and cervical cancer screening (absolute difference in improvement 7.0% [95% CI 5.5% to 8.5%] compared with enhanced fee for service; 5.3% [95% CI 3.8% to 6.8%] compared with non-team-based capitation). For breast and colorectal cancer screening, there were no significant differences in change over time between different types of medical homes. The shift to capitation payment and the addition of team-based care in Ontario were associated with moderate improvements in processes related to diabetes care, but the effects on cancer screening were less clear.


Barriers to mental health care and predictors of treatment dropout in the South African Stress and Health Study.
This study used data from the South African Stress and Health Study (SASH) to examine both structural and attitudinal barriers to treatment initiation among South Africans with mental disorders and to investigate predictors of treatment dropout. Face-to-face interviews were conducted with 4,315 adult South Africans living in households or hostel quarters. The interview included a core diagnostic assessment of past-12-month mental disorders and assessments of disorder severity, service use, and barriers to treatment. Multivariate logistic regression models were used to determine predictors of not seeking treatment in relation to disorder severity and sociodemographic characteristics, as well as factors that were predictive of premature treatment discontinuation by participants who had received mental health treatment in the previous 12 months. Predictors of dropout were identified by cross-tabulation and discrete-time survival analysis. Of the 4,315 adults, 729 (16.9% weighted) met criteria for a mental disorder in the past 12 months. Across all levels of severity, the most frequently cited reason for not seeking professional treatment was a low perceived need for treatment. Among those who recognized the need but did not access treatment during the past 12 months (7.2%), attitudinal barriers to treatment seeking were reported more commonly than structural barriers (100% and 34%, respectively). Of the 182 respondents who received treatment (25% weighted), 20% discontinued prematurely. Various factors, such as substance use disorders and absence of health insurance, increased the odds of treatment dropout. Low rates of treatment seeking and high treatment dropout rates for common mental disorders among South Africans are a major concern. Public health efforts to improve treatment of mental disorders should consider the multiple influences on treatment initiation and discontinuation.


Identification and characterisation of short chain rhamnolipid production in a previously uninvestigated, non-pathogenic marine pseudomonad.
This study aimed to identify and characterise biosurfactant compounds produced by bacteria associated with a marine eukaryotic phytoplankton bloom. One strain, designated MCTG214(3b1), was isolated by enrichment with polycyclic aromatic hydrocarbons and based on 16S rDNA, and gyrB sequencing was found to belong to the genus Pseudomonas, however not related to P. aeruginosa. Cell-free supernatant samples of strain MCTG214(3b1) at stationary phase showed significant reductions in surface tension. HPLC-MS and NMR analysis of these samples indicated the presence of five different rhamnolipid (RL) congeners. Di-rhamnolipids accounted for 87% relative abundance and all congeners possessed fatty acid moieties consisting of 8-12 carbons. PCR screening of strain MCTG214(3b1) DNA revealed homologues to the P. aeruginosa RL synthesis genes rhlA and rhlB; however, no rhlC homologue was identified. Using the Galleria mellonella larvae model, strain MCTG214(3b1) was demonstrated to be far less pathogenic than P. aeruginosa. This study identifies for the first time a significantly high level of synthesis of short chain di-rhamnolipids by a non-pathogenic marine Pseudomonas species. We postulate that RL synthesis in Pseudomonas sp. MCTG214(3b1) is carried out by enzymes expressed from rhlA/B homologues similar to those of P. aeruginosa; however, a lack of rhlC potentially indicates the presence of a second novel rhamnosyltransferase responsible for the di-rhamnolipid congeners identified by HPLC-MS.


The alchemy of immune privilege explored from a neuroimmunological perspective.
The term 'immune privilege' (IP) generally describes the protection of vital structures, such as the brain, the eye, or the pregnant uterus, from the potentially damaging effects of an inflammatory immune response. Initially, barriers physically camouflaging such organs were thought to shield autoantigens from immune recognition and inflammation. This simplistic concept gave way to a much more complex understanding of IP, which reflects an entire network of interacting immunoregulatory processes and immunosuppressive microenvironments. Also, the number of organs and tissues that enjoy relative IP has grown considerably. This is not surprising since many different organs are constantly exposed to major, potentially damaging inflammatory events, for example, skin, gut, or lung-without evidence for excessive inflammation under physiological conditions. Focusing on fetotrophoblast IP as well as on hair-follicle-associated IP (an underappreciated, yet biologically fascinating, clinically important IP model), we summarize here key regulatory cues that operate in immunoprivileged tissues. Proposing novel concepts of how IP may collapse, for example, by exposure to psychosocial, stress-associated inflammation, we develop related strategies for how IP may be manipulated clinically so as to achieve IP protection and restoration.


Fine structure observations of phagotrophic activity by plasmodia of Physarum polycephalum.
Scanning electron microscopic observations of feeding plasmodia show three characteristic features: 1) extension of multilobed pseudopodia protruding from the leading edge of the plasmodium as it advances onto the surface of a food particle, 2) confluence of the lobes to form a sheath-like pseudopodium attached to the surface of the food particle, and 3) protrusion of small nodules with thin lamellar projections from the leading edge of the plasmodium. Sections through freeze-dried preparations of the feeding plasmodium exhibit a highly convoluted under surface in contact with loosened starch grains that appear to be released by extracellular digestion. The cytoplasm, viewed by transmission electron microscopy, contains branched, internally penetrating canals (ca. 2 microns wide) enclosing engulfed starch grains. Starch grains in the deeper part of the canals are more electron dense and appear to be digested. Micropseudopodia (70-80 nm dia.), projecting from the surface of the canals, protrude toward and into the ingested starch grains. Digestive marker enzyme (acid phosphatase) activity was detected cytochemically in food particles penetrated by micropseudopodia indicating a digestive role for these structures not reported previously.


Enhanced sensitivity to cytochrome c-induced apoptosis mediated by PHAPI in breast cancer cells.
Apoptotic signaling defects both promote tumorigenesis and confound chemotherapy. Typically, chemotherapeutics stimulate cytochrome c release to the cytoplasm, thereby activating the apoptosome. Although cancer cells can be refractory to cytochrome c release, many malignant cells also exhibit defects in cytochrome c-induced apoptosome activation, further promoting chemotherapeutic resistance. We have found that breast cancer cells display an unusual sensitivity to cytochrome c-induced apoptosis when compared with their normal counterparts. This sensitivity, not observed in other cancers, resulted from enhanced recruitment of caspase-9 to the Apaf-1 caspase recruitment domain. Augmented caspase activation was mediated by PHAPI, which is overexpressed in breast cancers. Furthermore, cytochrome c microinjection into mammary epithelial cells preferentially killed malignant cells, suggesting that this phenomenon might be exploited for chemotherapeutic purposes.


Psychosocial risks and work-related stress in developing countries: health impact, priorities, barriers and solutions.
The current research explores experts' perceptions of psychosocial risks and work-related stress in emerging economies and developing countries. This paper focuses on knowledge of potential health impact of psychosocial risks and preliminary priorities for action, and discusses potential barriers and solutions to addressing psychosocial risks and work-related stress in developing countries. This research applied a mixed methodology including semi-structured interviews, two rounds of an online Delphi survey, and four focus groups. Twenty nine experts with expertise in occupational health were interviewed. Seventy four experts responded to the first round of an online Delphi survey and 53 responded to the second round. Four groups of experts with a total of 37 active participants with specific or broader knowledge about developing country contexts participated in focus group discussions. High concern was expressed for the need to address psychosocial risks and work-related stress and their health impact. Developing country experts' knowledge about these issues was comparable to knowledge from industrialized countries, however, application of expert knowledge was reported to be weak in developing countries. Socio-economic conditions were regarded as important considerations. Priorities to be addressed were identified, and barriers to implementing possible solutions were proposed. The future research and action paradigms in relation to psychosocial risk management will need to be broadened to include the larger social, political and economic contexts in developing countries beyond issues focusing solely on the working environment. Work-related psychosocial risks and the emerging priority of work-related stress should urgently be included in the research and political agendas and action frameworks of developing countries.


Revealing covariance structures in fourier transform infrared and Raman microspectroscopy spectra: a study on pork muscle fiber tissue subjected to different processing parameters.
The aim of this study was to investigate the correlation patterns between Fourier transform infrared (FT-IR) and Raman microspectroscopic data obtained from pork muscle tissue, which helped to improve the interpretation and band assignment of the observed spectral features. The pork muscle tissue was subjected to different processing factors, including aging, salting, and heat treatment, in order to induce the necessary degree of variation of the spectra. For comparing the information gained from the two spectroscopic techniques with respect to the experimental design, multiblock principal component analysis (MPCA) was utilized for data analysis. The results showed that both FT-IR and Raman spectra were mostly affected by heat treatment, followed by the variation in salt content. Furthermore, it could be observed that IR amide I, II, and III band components appear to be effected to a different degree by brine-salting and heating. FT-IR bands assigned to specific protein secondary structures could be related to different Raman C-C stretching bands. The Raman C-C skeletal stretching bands at 1,031, 1,061, and 1,081 cm(-1) are related to the IR bands indicative of aggregated beta-structures, while the Raman bands at 901 cm(-1) and 934 cm(-1) showed a strong correlation with IR bands assigned to a alpha-helical structures. At the same time, the IR band at 1,610 cm(-1), which formerly was assigned to tyrosine in spectra originating from pork muscle, did not show a correlation to the strong tyrosine doublet at 827 and 852 cm(-1) found in Raman spectra, leading to the conclusion that the IR band at 1,610 cm(-1) found in pork muscle tissue is not originating from tyrosine.


Sex differences in mortality after acute myocardial infarction: changes from 1994 to 2006.
Previous studies have shown that women younger than 55 years have higher hospital mortality rates after acute myocardial infarction (MI) than age-matched men. We examined whether such mortality differences have decreased in recent years. We investigated temporal trends in the hospital case-fatality rates of MI by sex and age from June 1, 1994, through December 31, 2006. The study population included 916,380 patients from the National Registry of Myocardial Infarction with a confirmed diagnosis of MI. In-hospital mortality decreased markedly between 1994 and 2006 in all patients but more so in women than men. The mortality reduction in 2006 relative to 1994 was largest in women younger than 55 years (52.9%) and lowest in men younger than 55 years (33.3%). In patients younger than 55 years, the absolute decrease in mortality was 3 times larger in women than men (2.7% vs 0.9%). As a result, the excess mortality in younger women (<55 years) compared with men was less pronounced in 2004-2006 (unadjusted odds ratio, 1.32; 95% confidence interval, 1.07-1.67) than it was in 1994-1995 (unadjusted odds ratio, 1.93; 95% confidence interval, 1.67-2.24). The sex difference in mortality decrease was lower in older patients (P = .004 for the interaction among sex, age, and year). Changes in comorbidity and clinical severity features at admission accounted for more than 90% of these mortality trends. In recent years, women, particularly younger ones, experienced larger improvements in hospital mortality after MI than men. The narrowing of the mortality gap between younger women and men is largely attributable to temporal changes in risk profiles.


Competence-impeding electronic games and players' aggressive feelings, thoughts, and behaviors.
[Correction Notice: An Erratum for this article was reported in Vol 106(3) of Journal of Personality and Social Psychology (see record 2014-07574-006). In the article, the name of author Edward Deci was missing his middle name initial and should have read as Edward L. Deci. In addition, an incorrect version of figure 1 was published.] Recent studies have examined whether electronic games foster aggression. At present, the extent to which games contribute to aggression and the mechanisms through which such links may exist are hotly debated points. In current research we tested a motivational hypothesis derived from self-determination theory-that gaming would be associated with indicators of human aggression to the degree that the interactive elements of games serve to impede players' fundamental psychological need for competence. Seven studies, using multiple methods to manipulate player competence and a range of approaches for evaluating aggression, indicated that competence-impeding play led to higher levels of aggressive feelings, easier access to aggressive thoughts, and a greater likelihood of enacting aggressive behavior. Results indicated that player perceived competence was positively related to gaming motivation, a factor that was, in turn, negatively associated with player aggression. Overall, this pattern of effects was found to be independent of the presence or absence of violent game contents. We discuss the results in respect to research focused on psychological need frustration and satisfaction and as they regard gaming-related aggression literature.


Hypercholesterolemia and atherosclerosis change vascular reactivity in rabbits by different mechanisms.
Vasomotor reactivity was assessed in vitro in arterial segments obtained from rabbits with different stages of atherosclerosis. Rabbits were fed a standard chow diet (controls) or a cholesterol-enriched diet to induce hypercholesterolemia and atherosclerosis. A third group received the hydroxymethylglutaryl coenzyme A reductase inhibitor, lovastatin, simultaneously with the cholesterol diet. Contractile responses of thoracic aortas to norepinephrine, serotonin, and potassium-rich solution, as well as endothelium-dependent dilations to acetylcholine, were compared after 2 and 4 months on the respective diet. Additionally, plasma cholesterol levels and the amount of plaques covering the intimal surface (as a percentage of the intimal surface) were determined; transmission electron microscopy of atherosclerotic arteries was also performed. After 2 months, the only difference was an enhancement of contractile responses to serotonin in the cholesterol-fed versus the control group. After 4 months on the diet, contractile responses to serotonin were further enhanced, and norepinephrine- and potassium-induced vasoconstrictions were now also significantly enhanced in cholesterol-fed animals versus controls. Endothelium-dependent vasodilations were simultaneously reduced in cholesterol-fed animals. These alterations were partly prevented in cholesterol-fed and lovastatin-treated animals. Suppression of nitric oxide synthesis in control aortas by NG-nitro-L-arginine did not reveal any significant increases in contractile responses. Contractile responses to serotonin were enhanced after 2 months on the diet but before the appearance of intimal plaques, whereas attenuation of endothelium-dependent dilations, as well as the further enhancement of contractile responses to serotonin and to other agonists, were closely correlated with the degree of intimal plaques after 4 months on the diet.(ABSTRACT TRUNCATED AT 250 WORDS)


Optimal management of complications associated with achondroplasia.
Achondroplasia is the most common form of skeletal dysplasia, resulting in disproportionate short stature, and affects over 250,000 people worldwide. Individuals with achondroplasia demonstrate a number of well-recognized anatomical features that impact on growth and development, with a complex array of medical issues that are best managed through a multidisciplinary team approach. The complexity of this presentation, whereby individual impairments may impact upon multiple activity and participation areas, requires consideration and discussion under a broad framework to gain a more thorough understanding of the experience of this condition for individuals with achondroplasia. This paper examines the general literature and research evidence on the medical and health aspects of individuals with achondroplasia and presents a pictorial model of achondroplasia based on The International Classification of Functioning, Disability, and Health (ICF). An expanded model of the ICF will be used to review and present the current literature pertaining to the musculoskeletal, neurological, cardiorespiratory, and ear, nose, and throat impairments and complications across the lifespan, with discussion on the impact of these impairments upon activity and participation performance. Further research is required to fully identify factors influencing participation and to help develop strategies to address these factors.


Topical buparvaquone formulations for the treatment of cutaneous leishmaniasis.
As the part of a study to develop buparvaquone (BPQ) formulations for the treatment of cutaneous leishmaniasis, the topical delivery of BPQ and one of its prodrugs from a range of formulations was evaluated. In previous studies, BPQ and its prodrugs were shown to be potent antileishmanials in-vitro, with ED50 values in the nanomolar range. 3-Phosphono-oxymethyl-buparvaquone (3-POM-BPQ) was the most potent antileishmanial and was chosen, together with the parent drug, for further investigation. The ability of the parent and prodrug formulations to cross human and murine skin was tested in-vitro using the Franz diffusion cells. Formulations intended for topical application containing either BPQ or 3-POM-BPQ were developed using excipients that were either acceptable for topical use (GRAS or FDA inactive ingredients) or currently going through the regulatory process. BPQ was shown to penetrate both human epidermal membranes and full thickness BALB/c skin from a range of formulations (gels, emulsions). Similarly, 3-POM-BPQ penetrated full-thickness BALB/c skin from several gel formulations. In-vitro binding studies showed that BPQ bound melanin in a dose-dependent manner and preferably bound to delipidized skin over untreated BALB/c skin (on a weight to weight basis). The results confirm that BPQ and its prodrug 3-POM-BPQ can penetrate the skin from several formulations, making them potentially interesting candidates for further investigation of topical formulations using in-vivo models of cutaneous leishmaniasis.


SU-E-T-426: Comparison of HDR Brachytherapy for Cervix Cancer Using an Adaptive Simulated Annealing Program and Oncentra- for Simultaneously Integrated Boost.
High dose rate (HDR) volumetric brachytherapy is an effective method of treating advanced cervix carcinoma. Local failure is associated with multiple factors including higher maximal standardized uptake value (SUV) values in fluorodeoxyglucose positron emission tomography (FDG- PET) scans. The purpose of this study is to evaluate the ability to simultaneously boost regions of high SUV values using an in-house adaptive simulated annealing (ASA) algorithm and the Oncentra® (Nucletron V.B., Veenendaal, The Netherlands) treatment planning system, thereby potentially improving local control. Five cervix cancers were evaluated for brachytherapy treatment (tandem/ring and/or interstitial needles). MRI and PET images were obtained post-implant and fused with treatment planning CTs to define a high-risk (HR) CTV (cervix and tumor on MRI) and GTV (volume with >50% of the maximum SUV on PET). The prescribed dose was 5-6 Gy to the HR CTV and 7-9 Gy to the GTV. Treatment plans were first generated in Oncentra® with IPSA followed by manual graphic optimization by the physician. Plans were also independently optimized using the ASA program. The two plans were compared side by side and one was chosen for treatment. Dose-volume parameters including D90, V100 of targets, D2cc to the critical organs, and generalized equivalent uniform dose (gEUD) of all structures were compared between the ASA and the Oncentra plans. Both ASA and Oncentra plans were considered acceptable by the physician in four of five cases. Two ASA plans were chosen due to better critical organ sparing and tumor coverage. Two Oncentra plans were preferred because of lower doses to critical organs. One ASA plan was not accepted because of a higher bowel dose. Both ASA and Oncentra® planning methods produce acceptable treatment plans for optimized brachytherapy of cervix carcinoma. Continued studies are warranted to further determine the relative strength of each method. This study was supported in part by a research grant from Varian Medical System Inc.


The frequency and consequences of head injury in epileptic seizures.
The frequency and sequelae of head injury in epileptic seizures were assessed. Two hundred and fifty five resident patients with chronic long term epilepsy (165M, 90F) of average age 54 years were studied for a year and 43 patients in a short term assessment unit (22M, 21F) of average age 26 years were studied for a month. A total of 27,934 seizures were recorded, of which 12,626 (45.2%) were associated with falls. There were 766 significant head injuries. Four hundred and twenty two required simple dressing and observation, and 341 required sutures (average number of sutures 4.5). There was one confirmed skull fracture, one confirmed extradural haemorrhage and one confirmed subdural haemorrhage. Thus 2.7% of all seizures resulted in a head injury (6.1% of seizures associated with falling). Simple dressing was required in 1.5%; 1.2% required sutures. Only one in every 9311 seizures (one in every 4208 seizures associated with falls) resulted in skull fracture, extradural or subdural haemorrhage. Minor seizure related head injury is therefore relatively common, while severe head injury is rare.


Team performance in networked supervisory control of unmanned air vehicles: effects of automation, working memory, and communication content.
Assess team performance within a net-worked supervisory control setting while manipulating automated decision aids and monitoring team communication and working memory ability. Networked systems such as multi-unmanned air vehicle (UAV) supervision have complex properties that make prediction of human-system performance difficult. Automated decision aid can provide valuable information to operators, individual abilities can limit or facilitate team performance, and team communication patterns can alter how effectively individuals work together. We hypothesized that reliable automation, higher working memory capacity, and increased communication rates of task-relevant information would offset performance decrements attributed to high task load. Two-person teams performed a simulated air defense task with two levels of task load and three levels of automated aid reliability. Teams communicated and received decision aid messages via chat window text messages. Task Load x Automation effects were significant across all performance measures. Reliable automation limited the decline in team performance with increasing task load. Average team spatial working memory was a stronger predictor than other measures of team working memory. Frequency of team rapport and enemy location communications positively related to team performance, and word count was negatively related to team performance. Reliable decision aiding mitigated team performance decline during increased task load during multi-UAV supervisory control. Team spatial working memory, communication of spatial information, and team rapport predicted team success. An automated decision aid can improve team performance under high task load. Assessment of spatial working memory and the communication of task-relevant information can help in operator and team selection in supervisory control systems.


Phase transitions of multivalent proteins can promote clustering of membrane receptors.
Clustering of proteins into micrometer-sized structures at membranes is observed in many signaling pathways. Most models of clustering are specific to particular systems, and relationships between physical properties of the clusters and their molecular components are not well understood. We report biochemical reconstitution on supported lipid bilayers of protein clusters containing the adhesion receptor Nephrin and its cytoplasmic partners, Nck and N-WASP. With Nephrin attached to the bilayer, multivalent interactions enable these proteins to polymerize on the membrane surface and undergo two-dimensional phase separation, producing micrometer-sized clusters. Dynamics and thermodynamics of the clusters are modulated by the valencies and affinities of the interacting species. In the presence of the Arp2/3 complex, the clusters assemble actin filaments, suggesting that clustering of regulatory factors could promote local actin assembly at membranes. Interactions between multivalent proteins could be a general mechanism for cytoplasmic adaptor proteins to organize membrane receptors into micrometer-scale signaling zones.


Transcriptome and excretory-secretory proteome of infective-stage larvae of the nematode Gnathostoma spinigerum reveal potential immunodiagnostic targets for development.
Gnathostoma spinigerum is a harmful parasitic nematode that causes severe morbidity and mortality in humans and animals. Effective drugs and vaccines and reliable diagnostic methods are needed to prevent and control the associated diseases; however, the lack of genome, transcriptome, and proteome databases remains a major limitation. In this study, transcriptomic and secretomic analyses of advanced third-stage larvae of G. spinigerum (aL3Gs) were performed using next-generation sequencing, bioinformatics, and proteomics. An analysis that incorporated transcriptome and bioinformatics data to predict excretory-secretory proteins (ESPs) classified 171 and 292 proteins into classical and non-classical secretory groups, respectively. Proteins with proteolytic (metalloprotease), cell signaling regulatory (i.e., kinases and phosphatase), and metabolic regulatory function (i.e., glucose and lipid metabolism) were significantly upregulated in the transcriptome and secretome. A two-dimensional (2D) immunomic analysis of aL3Gs-ESPs with G. spinigerum-infected human sera and related helminthiases suggested that the serine protease inhibitor (serpin) was a promising antigenic target for the further development of gnathostomiasis immunodiagnostic methods. The transcriptome and excretory-secretory proteome of aL3Gs can facilitate an understanding of the basic molecular biology of the parasite and identifying multiple associated factors, possibly promoting the discovery of novel drugs and vaccines. The 2D-immunomic analysis identified serpin, a protein secreted from aL3Gs, as an interesting candidate for immunodiagnosis that warrants immediate evaluation and validation.


Pemetrexed induced acute kidney injury in patients with non-small cell lung cancer: reversible and chronic renal damage.
Pemetrexed (Alimta(®)) (PEM) is an antifolate antineoplastic agent effective in several tumor types, such as non-small-cell lung cancer (NSCLC) and mesothelioma, among others. It is almost exclusively excreted by the kidney and an eGFR lower 45 mL/min is a contraindication for its use: above this level PEM administration is considered safe and dose adjustment is not required. Although there are some reported cases of PEM-induced renal injury, its incidence and the negative effects on patients' outcome has not been systematically evaluated. We report a retrospective evaluation on the incidence of PEM-induced renal injury in patients affected by NSCLC. Between June 2010 and March 2012 a total of 38 NSCLC patients were treated at our hospital. In 29 of them other possible cause of renal injury were excluded and thus they were eligible to be analysed. Although by protocol all of them had eGFR >45 mL/min at baseline, six patients (average eGFR 56.2 ± 11.5 mL/min/1.73 m(2)) developed AKI (21 %). In these six patients PEM-induced myelosuppression was more severe and hospitalization was longer. Kidney function completely recovered in four patients whereas in the other two deterioration of renal function was irreversible. The number of patients with baseline eGFR <60 mL/min/1.73 m(2) was higher (4/6) in the group that developed AKI as compared to those who did not (6/23) (p < 0.05). There is no clear cut eGFR above which PEM may be used without potential risks of renal toxicity. If PEM has to be used, all the coexisting risk factors for AKI should be possibly corrected.


A low-power programmable neural spike detection channel with embedded calibration and data compression.
This paper reports a programmable 400 μm pitch neural spike recording channel, fabricated in a 130 nm standard CMOS technology, which implements amplification, filtering, digitization, analog spike detection plus feature extraction, and self-calibration functionalities. It can operate in two different output modes: 1) signal tracking, in which the neural signal is sampled and transmitted as raw data; and 2) feature extraction, in which the spikes of the neural signal are detected and encoded by piece-wise linear curves. Additionally, the channel offers a foreground calibration procedure in which the amplification gain and the passband of the embedded filter can be self-adjusted. The amplification stage obtains a noise efficiency factor of 2.16 and an input referred noise of 2.84 μVrms over a nominal bandwidth of 167 Hz-6.9 kHz. The channel includes a reconfigurable 8-bit analog-to-digital converter combined with a 3-bit controlled programmable gain amplifier for adjusting the input signal to the full scale range of the converter. This combined block achieves an overall energy consumption per conversion of 102 fJ at 90 kS/s. The energy consumed by the circuit elements which are strictly related to the digitization process is 14.12 fJ at the same conversion rate. The complete channel consumes 2.8 μW at 1.2 V voltage supply when operated in the signal tracking mode, and 3.1 μW when the feature extraction mode is enabled.


Is chronic total coronary occlusion a risk factor for long-term outcome after minimally invasive bypass grafting of the left anterior descending artery?
Chronic total occlusion (CTO) of coronary vessels is still a challenge for percutaneous coronary intervention and recent data show unfavorable long-term results compared with medical therapy. It is unclear whether CTO is also a negative predictor for long-term outcome in minimally invasive bypass grafting. From 1996 to 2007 minimally invasive surgical revascularization of the left internal mammary artery to the left anterior descending artery (LAD) was performed in 1,800 patients. Demographic data, risk factors, perioperative outcome, and annual follow-up were obtained from all patients. Estimated survival and freedom from major adverse cardiac and cerebrovascular events or recurrence of angina with log-rank tests and Cox regression analysis for identification of independent risk factors were calculated for patients with (420 patients) and without (1,380 patients) CTO of the LAD. Revascularization of the LAD could be completed in all but one patient (99.8% success rate with CTO). At 5 years estimated overall survival was 90.5% (95% confidence interval [CI] 85.8 to 95.5) with CTO and 90.4% (95% CI 85.8 to 95.1) without CTO (p = 0.91). Freedom from major adverse cardiac and cerebrovascular events and angina with or without CTO at 5 years was 83.2% (95% CI 77.6 to 88.8) and 85.5% (95% CI 82.6 to 88.1), respectively (p = 0.64). Chronic occlusion of the target vessel and other preoperative factors were not identified as risk factors for major adverse cardiac and cerebrovascular events during follow-up. As opposed to percutaneous coronary intervention, minimally invasive bypass grafting of a totally occluded LAD is almost always possible and chronic occlusion is not a negative predictor for short and long-term outcome. Minimally invasive bypass grafting of the LAD should be considered the treatment of choice for chronically occluded left anterior descending arteries.


Cell guidance ligands, receptors and complexes - orchestrating signalling in time and space.
Members of four cell guidance molecule families (the netrins, slits, ephrins and semaphorins) interact with their cognate cell surface receptors to guide cells during development and maintain tissue homeostasis. Integrated structure and cell-based analyses are providing insight into the mechanisms by which these signalling systems can deliver myriad outcomes that require exquisite accuracy in timing and location. Here we review recent advances in our understanding of the roles of oligomeric states, auto-inhibition, signalling assembly size and composition in cell guidance cue function.


Safety and efficacy of drug-eluting stents compared with bare metal stents in ST-elevation myocardial infarction.
Drug-eluting stents (DES) reduce restenosis and the need for repeat revascularization, but patients with ST-segment elevation myocardial infarction (STEMI) were excluded from many of the trials that established the safety and efficacy of DES. Because of the unstable nature of lesions associated with STEMI, these patients are considered high risk, and often experience higher rates of adverse events. There is concern that DES may increase the risk of stent thrombosis, particularly late and very late stent thrombosis, in STEMI patients. Evidence also suggests that although DES reduce target vessel revascularization, this benefit may be lost after extended follow-up due to procedures necessitated by increased stent thrombosis. Several randomized trials, meta-analyses, and registry studies have been conducted to compare DES with bare metal stents in patients with STEMI, but many of the studies are not large scale and the length of follow-up has been limited in duration. This review summarizes the data comparing DES with bare metal stents in patients with STEMI.


Prefrontal activity during response inhibition decreases over time in the postpartum period.
The postpartum period is characterized by complex hormonal changes, but human imaging studies in the postpartum period have thus far predominantly focused on the neural correlates of maternal behavior or postpartum depression, whereas longitudinal studies on neural correlates of cognitive function across the postpartum period in healthy women are lacking. The aim of this study was to longitudinally examine response inhibition, as a measure of executive function, during the postpartum period and its neural correlates in healthy postpartum women and non-postpartum controls. Thirteen healthy postpartum women underwent event-related functional magnetic resonance imaging while performing a Go/NoGo task. The first assessment was made within 48 h of delivery, and the second at 4-7 weeks postpartum. In addition, 13 healthy women examined twice during the menstrual cycle were included as non-postpartum controls. In postpartum women region of interest analyses revealed task-related decreased activations in the right inferior frontal gyrus, right anterior cingulate, and bilateral precentral gyri at the late postpartum assessment. Generally, postpartum women displayed lower activity during response inhibition in the bilateral inferior frontal gyri and precentral gyri compared to non-postpartum controls. No differences in performance on the Go/NoGo task were found between time-points or between groups. In conclusion, this study has discovered that brain activity in prefrontal areas during a response inhibition task decreases throughout the course of the first postpartum weeks and is lower than in non-postpartum controls. Further studies on the normal adaptive brain activity changes that occur during the postpartum period are warranted.


Developmental trajectories of form perception: A story of attention.
The developmental trajectories of selective and divided attention were examined in relation to the processing of hierarchically integrated stimuli. The participants included children in 4 age groups (6, 8, 10, and 12 years) and a group of young adults (24 years) who completed 2 computer-based attention tasks. In the selective attention task, the participants were instructed to attend to only 1 level of analysis and ignore the other. In the divided attention task, participants were told that the target could appear at either level, and the probability that a target would appear at either the global or local level was manipulated. For both of the tasks, distinct and qualitative developmental shifts were evident both between 6 and 8 years of age and between 8 and 10 years of age. Attention to the global form developed prior to, and may have been a prerequisite of, attention to the local form. These gains in attentional control occurred in terms of selective attention, sensitivity to the probability of bias, and relative efficiency in processing global and local targets. The clear developmental trajectory is consistent with the emergent role of voluntary attention in the processing of these types of stimuli.


Bioaerosol Sampling for Respiratory Viruses in Singapore's Mass Rapid Transit Network.
As a leading global city with a high population density, Singapore is at risk for the introduction of novel biological threats. This risk has been recently reinforced by human epidemics in Singapore of SARS coronavirus, 2009 pandemic H1N1 influenza A virus, and enterovirus 71. Other major threats to Singapore include MERS-coronavirus and various avian and swine influenza viruses. The ability to quickly identify and robustly track such threats to initiate an early emergency response remains a significant challenge. In an effort to enhance respiratory virus surveillance in Singapore, our team conducted a pilot study employing a noninvasive bioaerosol sampling method to detect respiratory viruses in Singapore's Mass Rapid Transit (MRT) network. Over a period of 52 weeks, 89 aerosol samples were collected during peak MRT ridership hours. Nine (10%) tested positive for adenovirus, four (4.5%) tested positive for respiratory syncytial virus type A, and one (1%) tested positive for influenza A virus using real-time RT-PCR/PCR. To our knowledge, this is the first time molecular evidence for any infectious respiratory agent has been collected from Singapore's MRT. Our pilot study data support the possibility of employing bioaerosol samplers in crowded public spaces to noninvasively monitor for respiratory viruses circulating in communities.


Co-ingesting milk fat with micellar casein does not affect postprandial protein handling in healthy older men.
Dietary protein digestion and absorption plays an important role in modulating postprandial muscle protein synthesis. The impact of co-ingesting other macronutrients with dietary protein on protein digestion and absorption and the subsequent muscle protein synthetic response remains largely unexplored. This study investigated the impact of co-ingesting milk fat with micellar casein on dietary protein-derived amino acid appearance in the circulation and the subsequent postprandial muscle protein synthetic response in healthy older men. Twenty-four healthy, older males (age: 65 ± 1 y, BMI: 25.7 ± 0.5 kg/m2) received a primed continuous infusion of L-[ring-2H5]-phenylalanine and L-[1-13C]-leucine and ingested 20 g intrinsically L-[1-13C]-phenylalanine and L-[1-13C]-leucine-labeled casein with (PRO + FAT; n = 12) or without (PRO; n = 12) 26.7 g milk fat. Plasma samples and muscle biopsies were collected in both the postabsorptive and postprandial state. Release of dietary protein-derived phenylalanine into the circulation increased following protein ingestion (P < 0.001) and tended to be higher in PRO compared with PRO + FAT (Time × Treatment P = 0.076). No differences were observed in dietary protein-derived plasma phenylalanine availability (52 ± 2 vs 52 ± 3% in PRO vs PRO + FAT, respectively; P = 0.868). Myofibrillar protein synthesis rates did not differ between treatments, calculated using either the L-[ring-2H5]-phenylalanine (0.036 ± 0.003 vs 0.036 ± 0.004 %/h after PRO vs PRO + FAT, respectively; P = 0.933) or L-[1-13C]-leucine (0.051 ± 0.004 vs 0.046 ± 0.004 %/h, respectively; P = 0.480) tracer. In accordance, no differences were observed in myofibrillar protein-bound L-[1-13C]-phenylalanine enrichments between treatments (0.018 ± 0.002 vs 0.014 ± 0.001 MPE, respectively; P = 0.173). Co-ingesting milk fat with micellar casein does not impair protein-derived phenylalanine appearance in the circulation and does not modulate postprandial myofibrillar protein synthesis rates. NCT01680146 (http://www.clinicaltrials.gov/).


Molecules on rails: friction anisotropy and preferential sliding directions of organic nanocrystallites on two-dimensional materials.
Two-dimensional (2D) materials are envisaged as ultra-thin solid lubricants for nanomechanical systems. So far, their frictional properties at the nanoscale have been studied by standard friction force microscopy. However, lateral manipulation of nanoparticles is a more suitable method to study the dependence of friction on the crystallography of two contacting surfaces. Still, such experiments are lacking. In this study, we combine atomic force microscopy (AFM) based lateral manipulation and molecular dynamics simulations in order to investigate the movements of organic needle-like nanocrystallites grown by van der Waals epitaxy on graphene and hexagonal boron nitride. We observe that nanoneedle fragments - when pushed by an AFM tip - do not move along the original pushing directions. Instead, they slide on the 2D materials preferentially along the needles' growth directions, which act as invisible rails along commensurate directions. Further, when the nanocrystallites were rotated by applying a torque with the AFM tip across the preferential sliding directions, we find an increase of the torsional signal of the AFM cantilever. We demonstrate in conjunction with simulations that both, the significant friction anisotropy and preferential sliding directions are determined by the complex epitaxial relation and arise from the commensurate and incommensurate states between the organic nanocrystallites and the 2D materials.


Productive Aging in the Social Work Profession: A Comparison of Licensed Workers 60 Years and Older with Their Younger Counterparts.
This article reports the findings of an online survey in 13 U.S. states that compared the self-described demographics, wellness factors (mental health, physical health, and substance misuse), practice factors (fields of practice and work environment issues), and feelings about being a social worker (compassion satisfaction, workplace stress, being glad one chose social work and feeling valued as a professional in society) of 870 employed licensed social workers age 60 and older to 4076 licensed social workers under age 60. The results indicate that the older social workers were more likely to be male and white, less likely to report mental health problems, and more likely to work exclusively in private practice. Although older workers reported more serious physical health problems, they rated their physical health more favorably than their younger counterparts. In both bivariate and multivariate analyses, the older social workers scored significantly higher in compassion satisfaction, being glad they chose social work as a career, and feeling valued as a professional in society. The older social workers scored lower in workplace stress at levels that were statistically significant in a bivariate analysis, but not in a multivariate analysis.


New Cyt-like δ-endotoxins from Dickeya dadantii: structure and aphicidal activity.
In the track of new biopesticides, four genes namely cytA, cytB, cytC and cytD encoding proteins homologous to Bacillus thuringiensis (Bt) Cyt toxins have been identified in the plant pathogenic bacteria Dickeya dadantii genome. Here we show that three Cyt-like δ-endotoxins from D. dadantii (CytA, CytB and CytC) are toxic to the pathogen of the pea aphid Acyrthosiphon pisum in terms of both mortality and growth rate. The phylogenetic analysis of the comprehensive set of Cyt toxins available in genomic databases shows that the whole family is of limited taxonomic occurrence, though in quite diverse microbial taxa. From a structure-function perspective the 3D structure of CytC and its backbone dynamics in solution have been determined by NMR. CytC adopts a cytolysin fold, structurally classified as a Cyt2-like protein. Moreover, the identification of a putative lipid binding pocket in CytC structure, which has been probably maintained in most members of the Cyt-toxin family, could support the importance of this lipid binding cavity for the mechanism of action of the whole family. This integrative approach provided significant insights into the evolutionary and functional history of D. dadantii Cyt toxins, which appears to be interesting leads for biopesticides.


Tracheobronchial rupture due to blunt trauma in children: report of two cases.
Tracheobronchial tree injuries occur in a small number of patients after blunt chest trauma, and their occurrence is uncommon in the pediatric trauma population. The authors report two male children, one with a tracheal rupture, and the other with disruption of the main right bronchus. Mediastinal and subcutaneous emphysema resulting in airway obstruction were noted in Case 1 and soft-tissue emphysema, pneumomediastinum and tension pneumothorax were evident in Case 2 at the time of presentation. In the child with bronchial disruption, a major airway injury was suspected early on, because of a massive air leak despite two properly placed chest tubes. The definitive diagnosis was established bronchoscopically, and thoracotomy and primary repair were performed. The child with rupture of the posterior tracheal wall was diagnosed at an early stage by bronchoscopy and he was successfully managed without surgery.


C1q Binding Activity of De Novo Donor-specific HLA Antibodies in Renal Transplant Recipients With and Without Antibody-mediated Rejection.
Complement fixation by donor-specific HLA antibodies (DSA) is a primary mechanism for antibody-mediated damage of organ allografts. Using a recently developed kit that measures C1q binding to distinguish complement fixing and nonfixing antibodies, studies showed that C1q + DSAs have a higher risk of rejection and graft loss compared to C1q-DSA. The objective of this study was to assess the ability of the C1q-binding assay to identify clinically significant de novo DSA in renal transplant recipients and to define the properties of DSA that confer C1q binding ability. The DSA-positive sera from 34 kidney recipients, 19 with biopsy-proven antibody-mediated rejection (AMR) + and 15 who were AMR-, were assayed in C1q-binding assays (C1q Screen; One Lambda, Inc. Canoga Park, CA). The correlation between C1q-binding activity, presence of AMR, DSA mean fluorescence intensity (MFI) values, and immunoglobulin G isotype was determined. Fifty-three percent (10/19) of sera from AMR+ patients had C1q + DSA, whereas only 13% (2/15) of sera from AMR- patients contained C1q + DSA. C1q + DSA exhibited significantly higher MFI values regardless of whether they were from AMR+ or AMR- patients (16,118 ± 6698 vs 6429 ± 4003; P < 0.0001). C1q + DSA converted to C1q - when diluted to a comparable MFI level as the C1q - DSA from AMR- patients, and some C1q - antibodies converted to C1q + when concentrated to MFI levels comparable to those observed for AMR+/C1q + sera. The C1q binding activity by de novo DSA in patients with AMR largely reflects differences in antibody strength. The C1q assay does not appear to distinguish functionally distinct DSA with clinical significance.


Assessment of collagen-induced arthritis using cyanine 5.5 conjugated with hydrophobically modified glycol chitosan nanoparticles: correlation with 18F-fluorodeoxyglucose positron emission tomography data.
To evaluate the potential and correlation between near-infrared fluorescence (NIRF) imaging using cyanine 5.5 conjugated with hydrophobically modified glycol chitosan nanoparticles (HGC-Cy5.5) and (18)F-fluorodeoxyglucose-positron emission tomography ((18)F-FDG-PET) imaging of collagen-induced arthritis (CIA). We used 10 CIA and 3 normal mice. Nine days after the injecting collagen twice, microPET imaging was performed 40 minutes after the intravenous injection of 9.3 MBq (18)F-FDG in 200 µL PBS. One day later, NIRF imaging was performed two hours after the intravenous injection of HGC-cy5.5 (5 mg/kg). We assessed the correlation between these two modalities in the knees and ankles of CIA mice. The mean standardized uptake values of (18)F-FDG for knees and ankles were 1.68 ± 0.76 and 0.79 ± 0.71, respectively, for CIA mice; and 0.57 ± 0.17 and 0.54 ± 0.20 respectively for control mice. From the NIRF images, the total photon counts per 30 mm(2) for knees and ankles were 2.32 ± 1.54 × 10(5) and 2.75 ± 1.51 × 10(5), respectively, for CIA mice, and 1.22 ± 0.27 × 10(5) and 0.88 ± 0.24 × 10(5), respectively, for control mice. These two modalities showed a moderate correlation for knees (r = 0.604, p = 0.005) and ankles (r = 0.464, p = 0.039). Moreover, both HGC-Cy5.5 (p = 0.002) and (18)F-FDG-PET (p = 0.005) imaging also showed statistically significant differences between CIA and normal mice. NIRF imaging using HGC-Cy5.5 was moderately correlated with (18)F-FDG-PET imaging in the CIA model. As such, HGC-Cy5.5 imaging can be used for the early detection of rheumatoid arthritis.


Supra-optimal expression of the cold-regulated OsMyb4 transcription factor in transgenic rice changes the complexity of transcriptional network with major effects on stress tolerance and panicle development.
The R2R3-type OsMyb4 transcription factor of rice has been shown to play a role in the regulation of osmotic adjustment in heterologous overexpression studies. However, the exact composition and organization of its underlying transcriptional network has not been established to be a robust tool for stress tolerance enhancement by regulon engineering. OsMyb4 network was dissected based on commonalities between the global chilling stress transcriptome and the transcriptome configured by OsMyb4 overexpression. OsMyb4 controls a hierarchical network comprised of several regulatory sub-clusters associated with cellular defense and rescue, metabolism and development. It regulates target genes either directly or indirectly through intermediary MYB, ERF, bZIP, NAC, ARF and CCAAT-HAP transcription factors. Regulatory sub-clusters have different combinations of MYB-like, GCC-box-like, ERD1-box-like, ABRE-like, G-box-like, as1/ocs/TGA-like, AuxRE-like, gibberellic acid response element (GARE)-like and JAre-like cis-elements. Cold-dependent network activity enhanced cellular antioxidant capacity through radical scavenging mechanisms and increased activities of phenylpropanoid and isoprenoid metabolic processes involving various abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), ethylene and reactive oxygen species (ROS) responsive genes. OsMyb4 network is independent of drought response element binding protein/C-repeat binding factor (DREB/CBF) and its sub-regulons operate with possible co-regulators including nuclear factor-Y. Because of its upstream position in the network hierarchy, OsMyb4 functions quantitatively and pleiotrophically. Supra-optimal expression causes misexpression of alternative targets with costly trade-offs to panicle development.