The ability of calcaneal and multisite quantitative ultrasound variables in the identification of osteoporosis in women and men.
The aim of this study was to assess the ability of calcaneal and multisite quantitative ultrasound (QUS) parameters in the identification of osteoporosis in women and men. A total of 131 women (mean age 53.7±11.9 years; range, 21 to 79 years) and 109 men (mean age 57.8±13.7 years; range, 24 to 85 years) whose bone mineral density (BMD) at the spine and proximal femur was measured between January 2010 and January 2012, using dual-energy X-ray absorptiometry (DXA) were included. Acoustic bone properties were also examined using both a calcaneal and a multisite QUS. The receiver operating characteristic analysis with the calculation of areas under the curve (AUCs) to evaluate the ability of both QUS devices for the identification of osteoporosis. We also calculated a lower and an upper threshold at a specificity of 90% and at a sensitivity of 90%, respectively, for the identification of osteoporosis along with a threshold/cut-off value with the best compromise between sensitivity and specificity. All calcaneal QUS parameters showed significant AUCs within the range of 0.712 (for Broadband Ultrasound Attenuation [BUA]) and 0.764 (for Speed of Sound [SOS]) in women and ranging from 0.661 (for BUA) to 0.735 (for SOS) in men, while only radial SOS of the multisite QUS demonstrated a significant AUC value of 0.661 for identifying osteoporosis in women. A Quantitative Ultrasound Index T-score of -1.53 for women and -1.68 for men showed sensitivity and specificities around 70%. Based on the results of this study, all calcaneal QUS parameters in both women and men and possibly radial SOS measurements of the multisite QUS in women may be helpful for the identification of osteoporosis.


Inducible myocardial ischemia and outcomes in patients with coronary artery disease and left ventricular dysfunction.
The study objectives were to test the hypotheses that ischemia during stress testing has prognostic value and identifies those patients with coronary artery disease (CAD) with left ventricular (LV) dysfunction who derive the greatest benefit from coronary artery bypass grafting (CABG) compared with medical therapy. The clinical significance of stress-induced ischemia in patients with CAD and moderately to severely reduced LV ejection fraction (EF) is largely unknown. The STICH (Surgical Treatment for IsChemic Heart Failure) trial randomized patients with CAD and EF ≤35% to CABG or medical therapy. In the current study, we assessed the outcomes of those STICH patients who underwent a radionuclide (RN) stress test or a dobutamine stress echocardiogram (DSE). A test was considered positive for ischemia by RN testing if the summed difference score (difference in tracer activity between stress and rest) was ≥4 or if ≥2 of 16 segments were ischemic during DSE. Clinical endpoints were assessed by intention to treat during a median follow-up of 56 months. Of the 399 study patients (51 women, mean EF 26 ± 8%), 197 were randomized to CABG and 202 were randomized to medical therapy. Myocardial ischemia was induced during stress testing in 256 patients (64% of the study population). Patients with and without ischemia were similar in age, multivessel CAD, previous myocardial infarction, LV EF, LV volumes, and treatment allocation (all p = NS). There was no difference between patients with and without ischemia in all-cause mortality (hazard ratio: 1.08; 95% confidence interval: 0.77 to 1.50; p = 0.66), cardiovascular mortality, or all-cause mortality plus cardiovascular hospitalization. There was no interaction between ischemia and treatment for any clinical endpoint. In CAD with severe LV dysfunction, inducible myocardial ischemia does not identify patients with worse prognosis or those with greater benefit from CABG over optimal medical therapy. (Comparison of Surgical and Medical Treatment for Congestive Heart Failure and Coronary Artery Disease [STICH]; NCT00023595).


Piezoelectric harvest of osteo-odonto-lamina in modified osteo-odonto keratoprosthesis: A maxillofacial perspective.
We describe the piezo-osteotomy feasibility in rehabilitation and harvesting of osteo-odonto lamina in modified osteo-odonto keratoprosthesis (MOOKP) surgery. Surgery was evaluated regarding operative technique and success of the keratoprosthesis (KPros) in terms of perception to light (+ve) to finger counting (+ve). This retrospective cohort study included 12 patients undergone MOOKP surgery procedures. Harvesting of osteo-odonto-lamina was performed using piezosurgical osteotomy during 2007-2012. The mean follow up was 34 months (range 24-48 months). Of the 12 patients six patients had vision ≥6/12, four patients had vision <6/12, but >6/60 and one patients had vision ≤6/60. KPros was retained and functional in all the eyes after a minimum follow up of 24 months. There was mucosal overgrowth over the optical cylinder occurred in two cases over 4 years follow up which was corrected with trimming. Postoperative complication at donor site was seen in three cases, two cases with exposure of root of adjacent teeth and oro antral fistula in one. This study suggests that piezoelectric harvest of osteo-odonto-lamina is a valuable surgical option in patients undergoing MOOKP surgery, resulting in high success rate with less complication.


Microphysical particle parameters from extinction and backscatter lidar data by inversion with regularization: experiment.
We present effective radius, volume, surface-area, and number concentrations as well as mean complex refractive index of tropospheric particle size distributions based on lidar measurements at six wavelengths. The parameters are derived by means of an inversion algorithm that has been specifically designed for the inversion of available optical data sets. The data were taken on 20 June and on 20 July 1997 during the Aerosol Characterization Experiment ACE 2 (North Atlantic/Portugal) and on 9 August 1998 during the Lindenberg Aerosol Characterization Experiment LACE 98 (Lindenberg/Germany). Measurements on 20 June 1997 were taken in a clean-marine boundary layer, and a large value of 0.64 microm for the effective radius, a low value of 1.45 for the real part, and a negligible imaginary part of the complex refractive index were found. The single-scatter albedo was 0.98 at 532 nm. It was derived from the particle parameters with Mie-scattering calculations. In contrast, the particles were less than 0.2 microm in effective radius in a continental-polluted aerosol layer on 20 July 1997. The real part of the complex refractive index was approximately 1.6; the imaginary part showed values near 0.03i. The single-scatter albedo was 0.84. On 9 August 1998 an elevated particle layer located from 3000 to 6000 m was observed, which had originated from an area of biomass burning in northwestern Canada. Here the effective radius was approximately 0.24 mum, the real part of the complex refractive index was above 1.6, the imaginary part was approximately 0.04i, and the single-scatter albedo was 0.81. Excellent agreement has been found with results based on sunphotometer and in situ measurements that were performed during the field campaigns.


Management of Loose, Frequent Stools and Fecal Incontinence in a Chronic Mesenteric Ischemia Patient with Oral Serum-derived Bovine Immunoglobulin.
Chronic diarrhea with fecal incontinence (FI) is a severe, underreported, and intractable problem in many patients for which limited pharmaceutical options exist. A retrospective case history was collected after the administration of a prescription medical food composed of serum-derived bovine immunoglobulin/protein isolate (SBI) at 5 g once daily in a patient with chronic mesenteric ischemia (CMI) for chronic loose, frequent, and urgent stools. The patient was an 84-year-old white male with a 20-year history of progressively worsening chronic diarrhea with six to eight watery stools per day (Bristol Stool Form Scale, Type 7), urgency, nocturnal diarrhea, FI, and postprandial abdominal discomfort before administration of SBI. After four weeks of SBI administration, the patient had two to three soft, semi-formed stools (Bristol Stool Form Scale, Types 4 and 5) per day with no nocturnal diarrhea, urgency, or FI, as well as full resolution of abdominal discomfort. In addition, the patient expressed an enhanced quality of life (QoL): able to travel, attend social events, and perform tasks not possible before therapy. This case underscores how a safe, nutritional therapy may offer a new modality for physicians to address chronic loose, frequent stools with FI in patients with CMI in this difficult to manage gastrointestinal population.


Characterisation of taeniid cestodes by DNA analysis.
This review describes how the application of molecular technology is providing improved methods for characterising and identifying taeniid cestodes. Emphasis has been placed on the use of DNA approaches for specific diagnosis of the various taeniid life-cycle stages, unambiguous species and strain discrimination, determination of evolutionary and phylogenetic relationships and the assignment of function to key macromolecules based on inferred amino acid sequences corresponding to cloned cDNA sequences. In addition, the review describes methods for isolating DNA from taeniids, approaches for cloning genomic and mitochondrial DNA from these organisms and the use of DNA methods for strain characterisation and egg detection. Lastly, the review focuses on the polymerase chain reaction (PCR) which can provide the basis for exquisitely sensitive assays diagnostic for individual taeniid species. Coupled with direct sequencing techniques, PCR can also provide a valuable new method for examining genetic variation between and within the various taeniid species.


Mangosteen pericarp extract inhibits the formation of pentosidine and ameliorates skin elasticity.
The inhibition of advanced glycation end-products (AGEs) by daily meals is believed to become an effective prevention for lifestyle-related diseases. In the present study, the inhibitory effect of hot water extracts of mangosteen (Garcinia mangostana L.) pericarp (WEM) on the formation of pentosidine, one of AGEs, in vitro and in vivo and the remedial effect on skin conditions were measured. WEM significantly inhibited pentosidine formation during gelatin incubation with ribose. Several compounds purified from WEM, such as garcimangosone D and rhodanthenone B, were identified as inhibitors of pentosidine formation. Oral administration of WEM at 100 mg/day to volunteer subjects for 3 months reduced the serum pentosidine contents. Because obtaining skin biopsies from healthy volunteers is ethically difficult, AGE accumulation in the skin was estimated by a fluorescence detector. The oral administration of WEM significantly reduced the skin autofluorescence intensity, demonstrating that WEM also reduced AGE accumulation in the skin. Furthermore, the elasticity and moisture content of the skin was also improved by WEM. These results demonstrate that intakes of WEM reduces the glycation stress and results in the improvement of skin conditions.


Effects of acupuncture to treat fibromyalgia: a preliminary randomised controlled trial.
Acupuncture is often used to treat fibromyalgia (FM), but it remains unclear whether acupuncture is effective. This study aims to evaluate the effects of acupuncture on pain and quality of life (QoL) in FM patients. Sixteen patients (13 women and 3 men aged 25-63 years) suffering from FM were randomised into two groups: group A (n = 8) received five acupuncture treatments after the fifth week and group B received ten acupuncture treatments. Outcome measures used in this study were pain intensity (visual analogue scale, VAS) and the fibromyalgia impact questionnaire (FIQ). After the fifth week, pain intensity (U = 25.0; P = 0.022) in group B decreased and QoL (U = 24.5; P = 0.026) improved compared to group A. The present study suggests that acupuncture treatment is effective to relieve pain for FM patients in terms of QoL and FIQ.


Self-perceived oral function in elderly residents in a suburban area of Stockholm, Sweden.
The primary aim was to describe self-perceived oral health and function in a group of adults aged 75 to 84 years. The secondary aim was to study the agreement between self-reported oral function and clinical findings. A 5% sample (150 subjects) was selected at random from the total population (2,910) of 75 to 84-year-old residents in suburban Stockholm, Sweden. The inclusion criterion was that the individual lived independently, without any community assistance. A questionnaire covering self-perceived chewing ability, mouth dryness and bleeding gums was sent by mail. The total response rate was 86%. Of these 129 respondents, a 25.0% subsample was randomly selected for clinical examination by a dental hygienist. No radiographs were used. The average age of the individuals was 78.7 years (SD 2.71). Most were satisfied with their oral status and function: > 75.0% reported good chewing ability, correlating with the number of remaining teeth and dentures. Mouth dryness was reported by 41.5% of respondents to the questionnaire; one case was diagnosed in the subsample which underwent clinical examination. Bleeding gums were reported by 11.8% of respondents; clinical examination disclosed bleeding on probing in all participants. Mouth dryness and bleeding gums were not associated with chewing ability. Most subjects reported satisfactory chewing ability. Deterioration in oral function correlated with fewer remaining teeth and removable dentures. In relation to the clinical findings, mouth dryness was 'over-reported' and bleeding gums were 'under-reported'.


Expression of antisense SnRK1 protein kinase sequence causes abnormal pollen development and male sterility in transgenic barley.
A chimaeric gene was constructed comprising a wheat high molecular weight glutenin subunit gene promoter, a 304-bp sucrose non-fermenting-1-related (SnRK1) protein kinase sequence in the antisense orientation, and the cauliflower mosaic virus 35S RNA gene terminator. Transgenic barley plants containing the antisense SnRK1 chimaeric gene were produced by particle bombardment of barley immature embryos with the aim of obtaining plants expressing the antisense SnRK1 sequence in the seeds. Despite the fact that the promoter was expected to be active only in seeds, two independent transgenic lines were found to fail to transmit the transgene to the T1 generation. These T0 plants had matured and died before this was discovered, but subsequently four other independent transgenic lines were found to be affected in the same way. Cytological analysis of the pollen grains in these lines showed that about 50% were normal but the rest had arrested at the binucleate stage of development, were small, pear-shaped, contained little or no starch and were non-functional. The presence of antisense SnRK1 transcripts was detected in the anthers of the four lines analyzed and a ubiquitin promoter/UidA (Gus) gene, one of the marker genes codelivered with the antisense gene, was found to be expressed only in the abnormal pollen. Expression analyses confirmed that SnRK1 is expressed in barley anthers and that expression of one class of SnRK1 transcripts (SnRK1b) was reduced in the abnormal lines. All of the abnormal lines showed approximately 50% seed set, and none of the transgenes were detected in the T1 generation.


CXCL16 and oxLDL are induced in the onset of diabetic nephropathy.
Diabetic nephropathy (DN) is a major cause of end-stage renal failure worldwide. Oxidative stress has been reported to be a major culprit of the disease and increased oxidized low density lipoprotein (oxLDL) immune complexes were found in patients with DN. In this study we present evidence, that CXCL16 is the main receptor in human podocytes mediating the uptake of oxLDL. In contrast, in primary tubular cells CD36 was mainly involved in the uptake of oxLDL. We further demonstrate that oxLDL down-regulated alpha(3)-integrin expression and increased the production of fibronectin in human podocytes. In addition, oxLDL uptake induced the production of reactive oxygen species (ROS) in human podocytes. Inhibition of oxLDL uptake by CXCL16 blocking antibodies abrogated the fibronectin and ROS production and restored alpha(3) integrin expression in human podocytes. Furthermore we present evidence that hyperglycaemic conditions increased CXCL16 and reduced ADAM10 expression in podocytes. Importantly, in streptozotocin-induced diabetic mice an early induction of CXCL16 was accompanied by higher levels of oxLDL. Finally immunofluorescence analysis in biopsies of patients with DN revealed increased glomerular CXCL16 expression, which was paralleled by high levels of oxLDL. In summary, regulation of CXCL16, ADAM10 and oxLDL expression may be an early event in the onset of DN and therefore all three proteins may represent potential new targets for diagnosis and therapeutic intervention in DN.


Serum Antibodies to HPV16 Early Proteins Warrant Investigation as Potential Biomarkers for Risk Stratification and Recurrence of HPV-Associated Oropharyngeal Cancer.
Human papillomavirus (HPV) is responsible for increasing incidence of oropharyngeal cancer. At present, there are no biomarkers in the surveillance algorithm for HPV-positive oropharyngeal cancer (HPV-OPC). HPV16 E6 antibody precedes oropharyngeal cancer diagnosis. If HPV16 E6 indeed precedes primary diagnosis, it is similarly expected to precede disease recurrence and may have a potential role as a biomarker for surveillance of HPV-OPC. To determine whether HPV antibody titers have a potential role as early markers of disease recurrence or prognosis, a retrospective pilot study was designed to determine whether HPV16 early antibody titers E6, E7, E1, and E2 decrease after treatment of HPV16-positive OPC. Trends in pretreatment, early (≤6 months after treatment), and late posttreatment (>6 months after treatment) HPV16 antibody titers were examined. There were 43, 34, and 52 subjects with serum samples available for pretreatment, early, and late posttreatment intervals. Mean pretreatment antibody levels were higher than posttreatment antibody levels. Average antibody levels decreased significantly over time for E6 (Ptrend = 0.001) and E7 (Ptrend < 0.001). Six disease recurrences were observed during the follow-up period (median, 4.4 years). In univariate analysis, a log-unit increase in pretreatment E6 titer was significantly associated with increased risk of disease recurrence (HR, 5.42; 95% CI, 1.1-25.7; P = 0.03). Therefore, levels of antibodies to HPV16 early oncoproteins decline after therapy. Higher E6 titers at diagnosis are associated with significant increases in the risk of recurrence. These data support the prospective evaluation of HPV16 antibodies as markers of surveillance and for risk stratification at diagnosis.


Identification of research hypotheses and new knowledge from scientific literature.
Text mining (TM) methods have been used extensively to extract relations and events from the literature. In addition, TM techniques have been used to extract various types or dimensions of interpretative information, known as Meta-Knowledge (MK), from the context of relations and events, e.g. negation, speculation, certainty and knowledge type. However, most existing methods have focussed on the extraction of individual dimensions of MK, without investigating how they can be combined to obtain even richer contextual information. In this paper, we describe a novel, supervised method to extract new MK dimensions that encode Research Hypotheses (an author's intended knowledge gain) and New Knowledge (an author's findings). The method incorporates various features, including a combination of simple MK dimensions. We identify previously explored dimensions and then use a random forest to combine these with linguistic features into a classification model. To facilitate evaluation of the model, we have enriched two existing corpora annotated with relations and events, i.e., a subset of the GENIA-MK corpus and the EU-ADR corpus, by adding attributes to encode whether each relation or event corresponds to Research Hypothesis or New Knowledge. In the GENIA-MK corpus, these new attributes complement simpler MK dimensions that had previously been annotated. We show that our approach is able to assign different types of MK dimensions to relations and events with a high degree of accuracy. Firstly, our method is able to improve upon the previously reported state of the art performance for an existing dimension, i.e., Knowledge Type. Secondly, we also demonstrate high F1-score in predicting the new dimensions of Research Hypothesis (GENIA: 0.914, EU-ADR 0.802) and New Knowledge (GENIA: 0.829, EU-ADR 0.836). We have presented a novel approach for predicting New Knowledge and Research Hypothesis, which combines simple MK dimensions to achieve high F1-scores. The extraction of such information is valuable for a number of practical TM applications.


Adaptation to reef habitats through selection on the coral animal and its associated microbiome.
Spatially adjacent habitats on coral reefs can represent highly distinct environments, often harbouring different coral communities. Yet, certain coral species thrive across divergent environments. It is unknown whether the forces of selection are sufficiently strong to overcome the counteracting effects of the typically high gene flow over short distances, and for local adaptation to occur. We screened the coral genome (using restriction site-associated sequencing) and characterized both the dinoflagellate photosymbiont- and tissue-associated prokaryote microbiomes (using metabarcoding) of a reef flat and slope population of the reef-building coral, Pocillopora damicornis, at two locations on Heron Island in the southern Great Barrier Reef. Reef flat and slope populations were separated by <100 m horizontally and ~5 m vertically, and the two study locations were separated by ~1 km. For the coral host, genetic divergence between habitats was much greater than between locations, suggesting limited gene flow between the flat and slope populations. Consistent with environmental selection, outlier loci primarily belonged to the conserved, minimal cellular stress response, likely reflecting adaptation to the different temperature and irradiance regimes on the reef flat and slope. The prokaryote community differed across both habitat and, to a lesser extent, location, whereas the dinoflagellate photosymbionts differed by habitat but not location. The observed intraspecific diversity associated with divergent habitats supports that environmental adaptation involves multiple members of the coral holobiont. Adaptive alleles or microbial associations present in coral populations from the environmentally variable reef flat may provide a source of adaptive variation for assisted evolution approaches, through assisted gene flow, artificial cross-breeding or probiotic inoculations, with the aim to increase climate resilience in the slope populations.


Synaptic dysfunction and abnormal behaviors in mice lacking major isoforms of Shank3.
SHANK3 is a synaptic scaffolding protein enriched in the postsynaptic density (PSD) of excitatory synapses. Small microdeletions and point mutations in SHANK3 have been identified in a small subgroup of individuals with autism spectrum disorder (ASD) and intellectual disability. SHANK3 also plays a key role in the chromosome 22q13.3 microdeletion syndrome (Phelan-McDermid syndrome), which includes ASD and cognitive dysfunction as major clinical features. To evaluate the role of Shank3 in vivo, we disrupted major isoforms of the gene in mice by deleting exons 4-9. Isoform-specific Shank3(e4-9) homozygous mutant mice display abnormal social behaviors, communication patterns, repetitive behaviors and learning and memory. Shank3(e4-9) male mice display more severe impairments than females in motor coordination. Shank3(e4-9) mice have reduced levels of Homer1b/c, GKAP and GluA1 at the PSD, and show attenuated activity-dependent redistribution of GluA1-containing AMPA receptors. Subtle morphological alterations in dendritic spines are also observed. Although synaptic transmission is normal in CA1 hippocampus, long-term potentiation is deficient in Shank3(e4-9) mice. We conclude that loss of major Shank3 species produces biochemical, cellular and morphological changes, leading to behavioral abnormalities in mice that bear similarities to human ASD patients with SHANK3 mutations.


Digital holographic interferometer using simultaneously three lasers and a single monochrome sensor for 3D displacement measurements.
The use of digital holographic interferometry for 3D measurements using simultaneously three illumination directions was demonstrated by Saucedo et al. (Optics Express 14(4) 2006). The technique records two consecutive images where each one contains three holograms in it, e.g., one before the deformation and one after the deformation. A short coherence length laser must be used to obtain the simultaneous 3D information from the same laser source. In this manuscript we present an extension of this technique now illuminating simultaneously with three different lasers at 458, 532 and 633 nm, and using only one high resolution monochrome CMOS sensor. This new configuration gives the opportunity to use long coherence length lasers allowing the measurement of large object areas. A series of digital holographic interferograms are recorded and the information corresponding to each laser is isolated in the Fourier spectral domain where the corresponding phase difference is calculated. Experimental results render the orthogonal displacement components u, v and w during a simple load deformation.


Brachial artery endothelial function is stable across a menstrual and oral contraceptive pill cycle but lower in premenopausal women than in age-matched men.
Sex hormone concentrations differ between men, premenopausal women with natural menstrual cycles (NAT), and premenopausal women using oral contraceptive pills (OCP), as well as across menstrual or OCP phases. This study sought to investigate how differences in sex hormones, particularly estradiol, between men and women and across cycle phases might influence brachial artery endothelial function. Fifty-three healthy adults (22 ± 3 yr, 20 men, 15 NAT women, and 18 second-, third-, or fourth-generation OCP women) underwent assessments of sex hormones and endothelial [flow-mediated dilation (FMD) test] and smooth muscle [nitroglycerin (NTG) test] function. Men were tested three times at 1-wk intervals, and women were tested three times throughout a single menstrual or OCP cycle (NAT: menstrual, midfollicular, and luteal phases and OCP: placebo/no pill, "early", and "late" active pill phases). Endogenous estradiol concentration was comparable between men and women in their NAT menstrual or OCP placebo phase ( P = 0.36) but increased throughout a NAT cycle ( P < 0.001). Allometrically scaled FMD did not change across a NAT or OCP cycle but was lower in both groups of women than in men ( P = 0.005), whereas scaled NTG was lower only in NAT women ( P = 0.001). Changes in estradiol across a NAT cycle were not associated with changes in relative FMD ( r2 = 0.01, P = 0.62) or NTG ( r2 = 0.09, P = 0.13). Duration of OCP use was negatively associated with the average relative FMD for second-generation OCP users only ( r = -0.65, P = 0.04). Our findings suggest that brachial endothelial function is unaffected by cyclic hormonal changes in premenopausal women but may be negatively impacted by longer-term use of second-generation OCPs. NEW & NOTEWORTHY We demonstrate that brachial artery flow-mediated dilation does not change across a menstrual or oral contraceptive pill cycle in premenopausal women but is lower in women than in men. Although unaffected by within-cycle changes in estradiol, brachial flow-mediated dilation is negatively correlated with duration of oral contraceptive pill use for second-generation pills.


Clique size and network characteristics in hyperlink cinema. Constraints of evolved psychology.
Hyperlink cinema is an emergent film genre that seeks to push the boundaries of the medium in order to mirror contemporary life in the globalized community. Films in the genre thus create an interacting network across space and time in such a way as to suggest that people's lives can intersect on scales that would not have been possible without modern technologies of travel and communication. This allows us to test the hypothesis that new kinds of media might permit us to break through the natural cognitive constraints that limit the number and quality of social relationships we can manage in the conventional face-to-face world. We used network analysis to test this hypothesis with data from 12 hyperlink films, using 10 motion pictures from a more conventional film genre as a control. We found few differences between hyperlink cinema films and the control genre, and few differences between hyperlink cinema films and either the real world or classical drama (e.g., Shakespeare's plays). Conversation group size seems to be especially resilient to alteration. It seems that, despite many efficiency advantages, modern media are unable to circumvent the constraints imposed by our evolved psychology.


Detection of low-renin hypertension; evaluation of out-patient renin-stimulating methods.
1. Three-renin-stimulating methods for detection of low-renin hypertension have been compared. First, renin activity was measured in hospital patients after 5 days of sodium restriction and 3 h ambulation. Secondly, renin activity was measured after frusemide stimualtion [0.42 mmol (140 mg) in 18 h] and 3 h ambulation. Thirdly, renin activity was measured after 5 days of chlorthalidone treatment [0.3 mmol (100 mg/day]ured after 5 days of chlorthalidone treatment [0.3 mmol (100 mg)/day] and 3 h ambulation. The last two tests were done with the subjects as out-patients without any dietary regimen. 2. In eleven normotensive control subjects and twenty hypertensive patients the results after frusemide were not comparable with those after sodium restriction since the frusemide test did not identify the same renin-suppressed hypertensive subjects as the sodium-restriction procedure. 3. After 5 days of chlorthalidone treatment in the renin values in eleven control subjects as well as in thirty-eight hypertensive patients were significantly higher than after sodium restriction. The values obtained after each procedure were closely correlated. 4. Thus the out-patinet chlorthalidone procedure identified similar sub-groups of patients as having low- or normal-renin hypertension as did the inpatient sodium-restriction test.


Dynamic changes in Bach1 expression in the kidney of rhabdomyolysis-associated acute kidney injury.
Free heme, a pro-oxidant released from myoglobin, is thought to contribute to the pathogenesis of rhabdomyolysis-associated acute kidney injury (RM-AKI), because renal overexpression of heme oxygenase-1 (HO-1), the rate-limiting enzyme in heme catabolism, confers protection against RM-AKI. BTB and CNC homology 1 (Bach1) is a heme-responsive transcription factor that represses HO-1. Here, we examined the changes with time in the gene expression of Bach1, HO-1, and δ-aminolevulinate synthase (ALAS1, a heme biosynthetic enzyme) in the rat kidney using an RM-AKI model induced by the injection of 50% glycerol (10 mL/kg body weight) into bilateral limbs. We also examined the protein expression of Bach1 in the nucleus and cytosol, and HO-1 in the rat kidney. Glycerol treatment induced significant elevation of serum creatinine kinase and aspartate aminotransferase levels followed by the marked elevation of serum blood urea nitrogen and creatinine levels, which caused serious damage to renal tubules. Following glycerol treatment, HO-1 mRNA and protein levels were significantly up-regulated, while ALAS1 mRNA expression was down-regulated, suggesting an increase in the free renal heme concentration. The Bach1 mRNA level was drastically increased 3 h after glycerol treatment, and the increased level was maintained for 12 h. Nuclear Bach1 protein levels were significantly decreased 3 h after treatment. Conversely, cytosolic Bach1 protein levels abruptly increased after 6 h. In conclusion, we demonstrate the dynamic changes in Bach1 expression in a rat model of RM-AKI. Our findings suggest that the increase in Bach1 mRNA and cytosolic Bach1 protein expression may reflect de novo Bach1 protein synthesis to compensate for the depletion of nuclear Bach1 protein caused by the induction of HO-1 by free heme.


The impact of automatic enrollment in 401(k) plans on future retirement accumulations: a simulation study based on plan design modifications of large plan sponsors.
SIGNIFICANCE OF AUTO-ENROLLMENT: Automatic enrollment of participants in 401(k) plans, which was encouraged by provisions in the Pension Protect Act of 2006, is designed to overcome the drawbacks of voluntary enrollment by getting more workers to save in their work place retirement plan. Auto-enrollment for 401(k) plans has been demonstrated by previous EBRI research to have substantial potential benefits for some employees. NEW EBRI RESEARCH: This EBRI study analyzes plan-specific data of 1,000 large defined contribution plans for salaried employees from Benefit SpecSelect (Hewitt Associates LLC) in 2005 and 2009 to compare a subsample of plan sponsors that did not have auto-enrollment in 2005 but that had adopted it in 2009. Actual plan information on both actual auto-enrollment and actual match rate information were coded both before and after adoption of auto-enrollment from 225 large 401(k) plan sponsors and found that the average change was positive under auto-enrollment in each of the following three categories: The first-tier match rate, the effective match rate, the average total employer contribution rate. MODELING ANALYSIS: This analysis created a series of simulation programs using these data. The analysis indicates that the adoption of automatic enrollment in 401(k) plans is likely to have a very significant positive impact (even greater than EBRI projected in 2008) in generating additional retirement savings for many workers, especially for young and low-income workers: Under baseline assumptions, the median 401(k) accumulations for the lowest-income quartile of workers currently age 25-29 (assuming all 401(k) plans were voluntary enrollment plans as typified by the 225 large plan sponsors described above) would only be 0.08 times final earnings at age 65. However, if all 401(k) plans are assumed to be using the large plan sponsor auto-enrollment provisions, the median 401(k) accumulations for the lowest-income quartile jumps to 4.96 times final earnings (if 401(k) participants revert back to the default contribution when they change jobs) and 5.33 times final earnings (if they retain their previous contribution level when they change jobs). There are also large increases even for high-income workers: The multiple under a voluntary enrollment scenario is 2.41 times final earnings compared with 9.15 or 9.81 under auto-enrollment, depending on the assumptions for employee reversion to default contribution rates upon job change. Future EBRI research will examine the extent to which the increased 401(k) generosity resulted from modifications to defined benefit plans as pension plans were closed or frozen.


Discriminative-stimulus effects of adenosine analogs: mediation by adenosine A2 receptors.
Squirrel monkeys were trained to discriminate either the nonselective adenosine analog 5'-N-ethylcarboxamide adenosine (NECA) or the A1-selective analog N6-cyclopentyladenosine (CPA) from saline. After i.v. injections of 0.03 mumol/kg of NECA or 1.0 mumol/kg of CPA, 10 consecutive responses on one lever produced food, whereas after i.v. injections of saline, 10 consecutive responses on the alternate lever produced food. The discriminative-stimulus effects of the adenosine analogs NECA, CPA, 2-chloroadenosine and the R- and S-isomers of N6-phenylisopropyladenosine (PIA), of the adenosine antagonists caffeine and 8-cyclopentyltheophylline (CPT), and of selected drugs from other classes (haloperidol, pentobarbital, diazepam and morphine) were determined by administering cumulative doses i.v. before sequential components of the experimental session. All adenosine analogs engendered dose-related increases in the percentage of responses on the NECA- or CPA-associated levers reaching a maximum of greater than or equal to 90%. The rank order of potency was similar in both NECA-trained and CPA-trained monkeys: NECA greater than 2-chloroadenosine greater than or equal to CPA greater than or equal to R-PIA greater than S-PIA. None of the other drugs had discriminative-stimulus effects comparable to those of the adenosine analogs. In additional studies, the nonselective adenosine antagonist caffeine or the A1-selective antagonist CPT were administered before cumulative doses of NECA or CPA. Both adenosine antagonists blocked the discriminative-stimulus effects of NECA and CPA, with CPT being about one order of magnitude more potent than caffeine.(ABSTRACT TRUNCATED AT 250 WORDS)


Tor1a+/- mice develop dystonia-like movements via a striatal dopaminergic dysregulation triggered by peripheral nerve injury.
Isolated generalized dystonia is a central motor network disorder characterized by twisted movements or postures. The most frequent genetic cause is a GAG deletion in the Tor1a (DYT1) gene encoding torsinA with a reduced penetrance of 30-40 % suggesting additional genetic or environmental modifiers. Development of dystonia-like movements after a standardized peripheral nerve crush lesion in wild type (wt) and Tor1a+/- mice, that express 50 % torsinA only, was assessed by scoring of hindlimb movements during tail suspension, by rotarod testing and by computer-assisted gait analysis. Western blot analysis was performed for dopamine transporter (DAT), D1 and D2 receptors from striatal and quantitative RT-PCR analysis for DAT from midbrain dissections. Autoradiography was used to assess the functional DAT binding in striatum. Striatal dopamine and its metabolites were analyzed by high performance liquid chromatography. After nerve crush injury, we found abnormal posturing in the lesioned hindlimb of both mutant and wt mice indicating the profound influence of the nerve lesion (15x vs. 12x relative to control) resembling human peripheral pseudodystonia. In mutant mice the phenotypic abnormalities were increased by about 40 % (p < 0.05). This was accompanied by complex alterations of striatal dopamine homeostasis. Pharmacological blockade of dopamine synthesis reduced severity of dystonia-like movements, whereas treatment with L-Dopa aggravated these but only in mutant mice suggesting a DYT1 related central component relevant to the development of abnormal involuntary movements. Our findings suggest that upon peripheral nerve injury reduced torsinA concentration and environmental stressors may act in concert in causing the central motor network dysfunction of DYT1 dystonia.


Recruitment of CREB binding protein is sufficient for CREB-mediated gene activation.
Phosphorylation of the transcription factor CREB leads to the recruitment of the coactivator, CREB binding protein (CBP). Recent studies have suggested that CBP recruitment is not sufficient for CREB function, however. We have identified a conserved protein-protein interaction motif within the CBP-binding domains of CREB and another transcription factor, SREBP (sterol-responsive element binding protein). In contrast to CREB, SREBP interacts with CBP in the absence of phosphorylation. We have exploited the conservation of this interaction motif to test whether CBP recruitment to CREB is sufficient for transcriptional activation. Substitution of six nonconserved amino acids from SREBP into the activation domain of CREB confers high-affinity, phosphorylation-independent CBP binding. The mutated CREB molecule, CREB(DIEDML), activates transcription in F9 teratocarcinoma and PC12 cells even in the absence of protein kinase A (PKA). Addition of exogenous CBP augments the level of transcription mediated by CREB(DIEDML), and adenovirus 12S E1A blocks transcription, implicating CBP in the activation process. Thus, recruitment of CBP to CREB is sufficient for transcriptional activation. Addition of PKA stimulates transcription induced by CREB(DIEDML) further, suggesting that a phosphorylation event downstream from CBP recruitment augments CREB signaling.


Surface properties of polyetheretherketone after different laboratory and chairside polishing protocols.
Polyetheretherketone (PEEK) can be used as a framework material for fixed dental prostheses. However, information about laboratory and chairside polishing methods is still scarce. The purpose of this in vitro study was to determine the effects of laboratory and chairside polishing methods on the surface roughness (SR) and surface free energy (SFE) of PEEK, an autopolymerizing poly(methyl methacrylate), and a veneering composite resin. For each of the 3 materials, 80 specimens were prepared (N=240) and divided into 7 polishing groups and 1 control group (n=10). The 7 groups were split into 4 laboratory protocols: polishing paste (Abraso), a second polishing paste (Opal L), silicone polisher (Ceragum), and diamond grinder (Diagen-Turbo grinder). The other 3 groups were chairside protocols: rainbow technique (Super-Snap kit), polishing paste (Prisma gloss), and a polishing system (Enhance finishing). Machine polishing with SiC P4000 served as the control treatment. The protocols' average SRs and SFEs were measured, and their surface topographies were evaluated with scanning electron microscopy (SEM). The logarithmically transformed data were analyzed using covariance analysis, 2-way and 1-way ANOVA, and partial correlation (α=.05). The polishing protocol exerted the highest influence on SR and SFE values (P<.001; SR: partial eta squared ηP2=.970; SFE: ηP2=.450), followed by material group (P<.001, SR: ηP2=.319; SFE: ηP2=.429). The interaction effect of the binary combinations of the 2 independent parameters (polishing protocol and material group) was also significant (P<.001, SR: ηP2=.681; SFE: ηP2=.365). Chairside methods presented lower SR values than laboratory methods, and specimens polished using the 2-body mode showed higher SR than did specimens polished using the 3-body mode.


Methodological quality of English-language genetic guidelines on hereditary breast-cancer screening and management: an evaluation using the AGREE instrument.
We examined the methodological quality of guidelines on syndromes conferring genetic susceptibility to breast cancer. PubMed, EMBASE, and Google were searched for guidelines published up to October 2010. All guidelines in English were included. The Appraisal of Guidelines, Research and Evaluation (AGREE) instrument was used to assess the quality of the guidelines, and their reported evidence base was evaluated. Thirteen guidelines were deemed eligible: seven had been developed by independent associations, and the other six had national/state endorsements. Four guidelines performed satisfactorily, achieving a score of greater than 50% in all six AGREE domains. Mean ± SD standardized scores for the six AGREE domains were: 90 ± 9% for 'scope and purpose', 51 ± 18% for 'stakeholder involvement', 55 ± 27% for 'rigour of development', 80 ± 11% for 'clarity and presentation', 37 ± 32% for 'applicability', and 47 ± 38% for 'editorial independence'. Ten of the thirteen guidelines were found to be based on research evidence. Given the ethical implications and the high costs of genetic testing for hereditary breast cancer, guidelines on this topic should provide clear and evidence-based recommendations. Our analysis shows that there is scope for improving many aspects of the methodological quality of current guidelines. The AGREE instrument is a useful tool, and could be used profitably by guidelines developers to improve the quality of recommendations.


Effects of intermittent pressure-induced strain on the electrophysiology of cultured human chondrocytes: evidence for the presence of stretch-activated membrane ion channels.
1. Cyclical pressurization of cultured chondrocytes results in increases in cyclic AMP and in the rate of proteoglycan synthesis. Intermittent increases in hydrostatic pressure are also associated with hyperpolarization of chondrocyte cell membranes and activation of Ca(2+)-dependent K(+)-ion channels but the physiological basis for this response to mechanical stimulation is unclear. 2. Experiments have been undertaken to better define the types of ion channels involved and to explore the possibility that the hyperpolarization response associated with cyclical pressurization of chondrocytes follows activation of stretch-activated ion channels. 3. The mean membrane potential of chondrocytes in non-confluent monolayer cell culture rose from -15.3 +/- 0.24 mV to -21.1 +/- 0.28 mV (n = 60, P < 0.0001) after intermittent pressurization (0.33 Hz, 16 kPa, 20 min). 4. Strain gauge measurements showed that cyclical pressurization was associated with strain on the base of the culture plate. The amplitude of the hyperpolarization response was proportional to the microstrain to which cells were subjected. 5. Membrane hyperpolarization did not occur when chondrocytes were subjected to cyclical pressurization in rigid glass culture dishes or plastic dishes positioned in the pressurization chamber so as to avoid bending of the base of the culture dish. 6. Indirect evidence that the hyperpolarization response after intermittent pressure-induced strain was associated with stimulation of stretch-activated ion channels was obtained from experiments with gadolinium, amiloride and hexamethylene amiloride, each of which abolished hyperpolarization. 7. Experiments with apamin, charybdotoxin and iberiotoxin showed that the Ca(2+)-activated K+ channels involved in the hyperpolarization response are apamin-sensitive, charybdotoxin- and iberiotoxin-resistant, low-conductance channels. 8. Somatostatin and cadmium chloride, which block L-type calcium channels, abolished strain-induced chondrocyte hyperpolarization. EGTA, which chelates extracellular Ca2+, reduced the response to 48% of control values, and thapsigargin, which raises intracellular Ca2+ by inhibition of Ca(2+)-ATPase in endoplasmic reticulum, caused hyperpolarization independently with further hyperpolarization after pressure-induced strain. These data indicate that chondrocyte hyperpolarization was dependent on intracellular Ca2+ concentrations. 9. Further work is required to determine whether stretch-activated ion channels shown to be associated with chondrocyte hyperpolarization after cyclical pressure-induced strain are also involved in the signal transduction process that leads to increases in proteoglycan synthesis.


Nme gene family evolutionary history reveals pre-metazoan origins and high conservation between humans and the sea anemone, Nematostella vectensis.
The Nme gene family is involved in multiple physiological and pathological processes such as cellular differentiation, development, metastatic dissemination, and cilia functions. Despite the known importance of Nme genes and their use as clinical markers of tumor aggressiveness, the associated cellular mechanisms remain poorly understood. Over the last 20 years, several non-vertebrate model species have been used to investigate Nme functions. However, the evolutionary history of the family remains poorly understood outside the vertebrate lineage. The aim of the study was thus to elucidate the evolutionary history of the Nme gene family in Metazoans. Using a total of 21 eukaryote species including 14 metazoans, the evolutionary history of Nme genes was reconstructed in the metazoan lineage. We demonstrated that the complexity of the Nme gene family, initially thought to be restricted to chordates, was also shared by the metazoan ancestor. We also provide evidence suggesting that the complexity of the family is mainly a eukaryotic innovation, with the exception of Nme8 that is likely to be a choanoflagellate/metazoan innovation. Highly conserved gene structure, genomic linkage, and protein domains were identified among metazoans, some features being also conserved in eukaryotes. When considering the entire Nme family, the starlet sea anemone is the studied metazoan species exhibiting the most conserved gene and protein sequence features with humans. In addition, we were able to show that most of the proteins known to interact with human NME proteins were also found in starlet sea anemone. Together, our observations further support the association of Nme genes with key cellular functions that have been conserved throughout metazoan evolution. Future investigations of evolutionarily conserved Nme gene functions using the starlet sea anemone could shed new light on a wide variety of key developmental and cellular processes.


Diminished specific activity of cytosolic protein kinase C in sciatic nerve of streptozocin-induced diabetic rats and its correction by dietary myo-inositol.
The impaired Na(+)-K(+)-ATPase activity in peripheral nerve from diabetic rats is prevented by dietary myo-inositol (MI) supplementation in vivo and corrected by protein kinase C (PKC) agonists in vitro, suggesting that PKC may mediate the effects of nerve MI depletion on Na(+)-K(+)-ATPase activity. However, little is known about the effect of diabetes on PKC activity or peptide in rat peripheral nerve. Therefore, the effect of streptozocin-induced diabetes and dietary MI supplementation on the activity and distribution of PKC in rat sciatic nerve homogenates and cytosolic and particulate fractions was explored with histone phosphorylation assay and Western-blot analysis. PKC activity but not peptide was selectively decreased in the cytosolic fraction by streptozocin-induced diabetes, and this abnormality was partially corrected by dietary MI supplementation. These results suggest that altered MI metabolism may affect nerve PKC specific activity, and this alteration may play a role in reduced Na(+)-K(+)-ATPase activity and blunted regenerative response in diabetic nerve.


Histological evaluation of the effect of smoking on peripheral small adenocarcinomas of the lung.
As there is little information on the histologic characteristics of adenocarcinoma in smokers, we histologically examined the effect of smoking on the carcinogenesis and progression of peripheral small lung adenocarcinomas. Two hundred thirty-six consecutive patients with peripheral adenocarcinoma of the lung 30 mm or less in diameter were studied. Prognosis, histology, and location of the adenocarcinoma were compared among patients with a Brinkman index (B.I.) of 0, 1 to 500, and more than 500. The ratio of smokers to nonsmokers was 1.4:1. The rate of carcinogenesis in the upper region of the lung (S1-3) was 1.4 times as high that in the lower region (S4-10) in smokers, but almost equal in the two regions in nonsmokers. Outcome tended to be worse in patients with a B.I. of more than 500 than in those with a B.I. of less than or equal to 500 for adenocarcinomas 30 mm or less in diameter (p = 0.0855), and was significantly worse for adenocarcinomas 20 mm or less in diameter (p = 0.0359). Patients with a high B.I. tended to have invasive adenocarcinoma (IAC) without a bronchioloalveolar carcinoma (BAC) component (IAC - BAC) or IAC with a BAC component (IAC + BAC) rather than noninvasive adenocarcinoma. For adenocarcinomas as a whole, B.I. was correlated with several pathologic prognostic factors, including pathologic stage, lymphatic permeation, vascular invasion, presence of a solid component, necrosis, and modified scar grade, particularly in the upper region. Specifically, in IAC + BAC, B.I. was correlated with modified scar grade and the presence of a solid component. In IAC - BAC, B.I. was correlated with the presence of a solid component and necrosis. Small adenocarcinoma in smokers seems to occur frequently in the upper region of the lung, shows invasive features more frequently, and shows greater progression and dedifferentiation than that in nonsmokers. Tobacco-smoking may have an effect on the carcinogenesis and multistep progression of peripheral lung adenocarcinoma 30 mm or less in diameter.


Development and screening of brain-targeted lipid-based nanoparticles with enhanced cell penetration and gene delivery properties.
The potential of gene therapy for treatment of neurological disorders can be explored using designed lipid-based nanoparticles such as liposomes, which have demonstrated ability to deliver nucleic acid to brain cells. We synthesized liposomes conjugated to cell-penetrating peptides (CPPs) (vascular endothelial-cadherin-derived peptide [pVec], pentapeptide QLPVM and HIV-1 trans-activating protein [TAT]) and transferrin (Tf) ligand, and examined the influence of surface modifications on the liposome delivery capacity and transfection efficiency of encapsulated plasmid DNA. The design of liposomes was based on targeting molecular recognition of transferrin receptor overexpressed on the blood-brain barrier (BBB) with enhanced internalization ability of CPPs. CPP-Tf-liposomes were characterized by particle size distribution, zeta potential, protection of encapsulated plasmid DNA, uptake mechanisms and transfection efficiencies. An in vitro triple co-culture BBB model selected the liposomal formulations that were able to cross the in vitro BBB and subsequently, transfect primary neuronal cells. The in vivo biodistribution and biocompatibility of selected formulations were also investigated in mice. Liposomal formulations were able to protect the encapsulated plasmid DNA against enzymatic degradation and presented low hemolytic potential and low cytotoxicity at 100 nM phospholipid concentration. Cellular internalization of nanoparticles occurred via multiple endocytosis pathways. CPP-Tf-conjugated liposomes mediated robust transfection of brain endothelial (bEnd.3), primary glial and primary neuronal cells. Liposomes modified with Tf and TAT demonstrated superior ability to cross the barrier layer and subsequently, transfect neuronal cells compared to other formulations. Quantification of fluorescently labeled liposomes and in vivo imaging demonstrated that this system could efficiently overcome the BBB and penetrate the brain of mice (7.7% penetration of injected dose). In vitro screening platforms are important tools to enhance the success of brain-targeted gene delivery systems. The potential of TAT-Tf-liposomes as efficient brain-targeted gene carriers in vitro and in vivo was suggested to be related to the presence of selected moieties on the nanoparticle surface.


The efficacy and cost of aprotinin in children undergoing reoperative open heart surgery.
We performed a prospective, randomized, placebo-controlled, double-blind trial to assess the efficacy of aprotinin in 61 children (median age 3.7 yr) undergoing reoperative open heart surgery (OHS). Three demographically similar groups were studied: large-dose aprotinin (ALD), small-dose aprotinin (ASD), and placebo (P). Over the first 24 postoperative hours fewer patients in the aprotinin groups received packed red cells (ALD, 53%; ASD, 89%; and P, 95%; P = 0.001), platelets (ALD, 32%; ASD, 50%; and P, 65%; P = 0.04), and fresh frozen plasma (ALD, 16%; ASD, 17%; and P, 60%; P = 0.003) than placebo patients. Most importantly, aprotinin patients had fewer exposures to banked blood components (ALD, median 1 U; and ASD, median 2 U) than P (median 6 U; P = 0.001), with no difference in overall complication rate. Use of aprotinin was associated with a savings in the patient charges for blood components, operating room time, and duration of hospitalization. In conclusion, aprotinin decreased the number of units of banked blood components used during the first 24 postoperative hours in reoperative pediatric OHS. Aprotinin thus decreases the risks associated with exposure to banked blood components and reduces hospital charges.


From quiescence to proliferation: Cdk oscillations drive the mammalian cell cycle.
We recently proposed a detailed model describing the dynamics of the network of cyclin-dependent kinases (Cdks) driving the mammalian cell cycle (Gérard and Goldbeter, 2009). The model contains four modules, each centered around one cyclin/Cdk complex. Cyclin D/Cdk4-6 and cyclin E/Cdk2 promote progression in G1 and elicit the G1/S transition, respectively; cyclin A/Cdk2 ensures progression in S and the transition S/G2, while the activity of cyclin B/Cdk1 brings about the G2/M transition. This model shows that in the presence of sufficient amounts of growth factor the Cdk network is capable of temporal self-organization in the form of sustained oscillations, which correspond to the ordered, sequential activation of the various cyclin/Cdk complexes that control the successive phases of the cell cycle. The results suggest that the switch from cellular quiescence to cell proliferation corresponds to the transition from a stable steady state to sustained oscillations in the Cdk network. The transition depends on a finely tuned balance between factors that promote or hinder progression in the cell cycle. We show that the transition from quiescence to proliferation can occur in multiple ways that alter this balance. By resorting to bifurcation diagrams, we analyze the mechanism of oscillations in the Cdk network. Finally, we show that the complexity of the detailed model can be greatly reduced, without losing its key dynamical properties, by considering a skeleton model for the Cdk network. Using such a skeleton model for the mammalian cell cycle we show that positive feedback (PF) loops enhance the amplitude and the robustness of Cdk oscillations with respect to molecular noise. We compare the relative merits of the detailed and skeleton versions of the model for the Cdk network driving the mammalian cell cycle.


5-Methyl-N-(8-(5,6,7,8-tetrahydroacridin-9-ylamino)octyl)-5H-indolo[2,3-b]quinolin-11-amine: a highly potent human cholinesterase inhibitor.
The synthesis, cholinesterase inhibition, molecular modelling and ADME properties of novel tacrine-neocryptolepine heterodimers are described. Compound 3 [5-methyl-N-(8-(5,6,7,8-tetrahydroacridin-9-ylamino)octyl)-5H-indolo[2,3-b]quinolin-11-amine], showing a moderate inhibition of the Aβ1-42 self-aggregation (26.5% at a 1 : 5 ratio with Aβ1-42), and a calculated log BB value (0.27) indicating excellent potential BBB penetration, is a highly potent human cholinesterase inhibitor [IC50 (hAChE) = 0.95 ± 0.04 nM; IC50 (hBuChE) = 2.29 ± 0.14 nM] which can be listed among the most potent hAChE inhibitors so far identified, and is not hepatotoxic in vitro at the concentrations at which the ChEs are inhibited. A molecular modeling study was also undertaken in order to elucidate the AChE and the BuChE bind modes of all the new compounds. The docking results show that all of them bind to AChE in extended conformations and to BuChE in folded conformations. Moreover, these studies revealed that the length of the linker is crucial to binding both the catalytic anionic site and the peripheral anionic site.


Activation of the beta-catenin gene by interstitial deletions involving exon 3 in primary colorectal carcinomas without adenomatous polyposis coli mutations.
Among 222 primary colorectal cancers we examined, 58 showed no detectable APC mutations by the protein truncation test. We screened those 58 tumors for somatic mutations in the beta-catenin gene. Although amino acid substitutions in serine or threonine residues in exon 3 had been reported, we found no such mutations; however, in seven tumors, we detected somatic interstitial deletions of 234-760 bp, each of which included all or part of exon 3. Short nucleotide sequences at both ends of each deletion were either identical or complementary, indicating that repeated or inversely repeated sequences were involved in the somatic rearrangements. Reverse transcription-PCR experiments using RNAs isolated from three of these seven tumors detected transcripts that lacked exon 3, in addition to the normal transcript. In one of these cases, we confirmed accumulation of aberrant beta-catenin protein in cytoplasm and nuclei of cancer cells by Western and immunohistochemical analyses. This result suggested that, in the absence of a peptide encoded by exon 3, beta-catenin is stabilized and has a dominant oncogenic effect on colorectal tumorigenesis.


Dietary practices, dining out behavior, and physical activity correlates of weight loss maintenance.
Loss of excess weight can improve blood lipids, insulin sensitivity, and blood pressure. However, data are scant on behavioral strategies related to maintenance of weight loss. We examined dietary practices, physical activity, and self-efficacy among adults self-reported to be successful at maintaining weight loss. Using the 2004 Styles survey, a mailed survey of U.S. adults aged 18 years or older, we examined behaviors associated with weight loss maintenance among people who reported trying to lose weight. We analyzed data on number of daily fruit and vegetable servings, minutes per week of physical activity, dining out behavior, and confidence in one's ability to engage in behavioral strategies. We conducted frequency and multivariable logistic regression analyses. More men (35.5%) than women (27.7%) were classified as successful weight loss maintainers. Compared with adults who reported eating at a fast-food restaurant two or more times per week, adults who reported not eating at fast-food restaurants were more successful at weight loss maintenance (odds ratio, 1.62; 95% confidence interval, 1.09-2.42). Compared with adults who consumed fewer than five fruit and vegetable servings per day and were sedentary, adults who consumed fewer than five fruit and vegetable servings per day and accrued 420 minutes or more per week of physical activity or consumed five or more fruit and vegetable servings and accrued 150 minutes or more per week of activity were more successful at weight loss maintenance. The behavioral strategy of reducing consumption of fast foods could assist people in keeping weight off. The combined approach of consuming five or more fruit and vegetable servings per day and attaining 150 minutes or more per week of physical activity was a common strategy among adults successful at weight loss maintenance.


Characterizing export of land-based microplastics to the estuary - Part I: Application of integrated geospatial microplastic transport models to assess tire and road wear particles in the Seine watershed.
Human and ecological exposure to micro- and nanoplastic materials (abbreviated as MP, < 5 mm) occurs in both aquatic and terrestrial environments. Recent reviews prioritize the need for assessments linking spatially distributed MP releases with terrestrial and freshwater transport processes, thereby providing a better understanding of the factors affecting MP distribution to the sea. Tire and road wear particles (TRWP) have an estimated generation rate of 1 kg tread inhabitant-1 year-1 in Europe, but the fate of this MP source in watersheds has not been systematically assessed. An integrated temporally and geospatially resolved watershed-scale MP modeling methodology was applied to TRWP fate and transport in the Seine (France) watershed. The mass balance considers TRWP generation and terrestrial transport to soil, air, and roadways, as well as freshwater transport processes including particle heteroaggregation, degradation and sedimentation within subcatchments. The per capita TRWP mass release estimate in the Seine watershed was 1.8 kg inhabitant-1 yr-1. The model estimates indicated that 18% of this release was transported to freshwater and 2% was exported to the estuary, which demonstrated the potential for appreciable capture, degradation, and retention of TRWP prior to export. The modeled pseudo-steady state sediment concentrations were consistent with measurements from the Seine watershed supporting the plausibility of the predicted trapping efficiency of approximately 90%. The approach supported the efficient completion of local and global sensitivity analyses presented in Part II of this study, and can be adapted to the assessment of other MPs.


The Time-Course of Sentence Meaning Composition. N400 Effects of the Interaction between Context-Induced and Lexically Stored Affordances.
Contemporary semantic theories can be classified along two dimensions: (i) the way and time-course in which contextual factors influence sentence truth-conditions; and (ii) whether and to what extent comprehension involves sensory, motor and emotional processes. In order to explore this theoretical space, our ERP study investigates the time-course of the interaction between the lexically specified telic component of a noun (the function of the object to which the noun refers to, e.g., a funnel is generally used to pour liquids into containers) and an ad-hoc affordance contextually induced by the situation described in the discourse. We found that, if preceded by a neutral discourse context, a verb incongruent with the noun's telic component as in "She uses the funnel to hang her coat" elicited an enhanced N400 compared to a congruent verb as in "She uses the funnel to pour water into a container." However, if the situation introduced in the preceding discourse induced a new function for the object as an ad-hoc affordance (e.g., the funnel is glued to the wall and the agent wants to hang the coat), we observed a crossing-over regarding the direction of the N400 effect: comparing the ad-hoc affordance-inducing context with the neutral context, the N400 for the incongruent verb was significantly reduced, whereas the N400 for the congruent verb was significantly enhanced. We explain these results as a consequence of the incorporation of the contextually triggered ad-hoc affordance into the meaning of the noun. Combining these results with an analysis of semantic similarity values between test sentences and contexts, we argue that one possibility is that the incorporation of an ad-hoc affordance may be explained on the basis of the mental simulation of concurrent motor information.


Tissue distribution and pharmacokinetics of centchroman. A new nonsteroidal postcoital contraceptive agent and its 7-desmethyl metabolite in female rats after a single oral dose.
This study reports assay methodology, tissue distribution, and the basic pharmacokinetic behavior of centchroman and its 7-desmethyl metabolite [7-desmethyl centchroman (DMC)] after a single 12.5 mg/kg po dose in young female rats. Plasma, liver, lung, spleen, uterus, and adipose tissue were collected at various time intervals up to 14 days after dose. Reversed-phase HPLC, coupled with fluorescence detector, was used for simultaneous determination of centchroman and DMC in biosamples. The drug and metabolite were quantitated up to 2 and 5 ng/ml in plasma and 10 and 20 ng/g in tissues, respectively. The assay method was validated in terms of accuracy, precision, interassay, and intraassay variability, and was found to be reliable and reproducible. Peak centchroman levels in all of the tissues were found between 8-12 hr, whereas DMC peaks appeared between 8 and 24 hr, except that in liver the first peak of 1.2 micrograms/g appeared in the 1-hr sample. Tissue-to-plasma concentration ratios of centchroman were > 200 times in the lung; > 100 times in the spleen, liver, and adipose tissue; and > 40 times in the uterus at maxima in each tissue. Similarly, tissue concentrations of DMC were > 350 times in the lung, > 100 times in the liver and spleen, and > 25 times in the uterus and adipose tissue than in the plasma. High tissue-to-plasma concentration ratios of metabolites than the parent drug are indicative of its greater affinity for tissues. Terminal half-life of the centchroman and DMC in plasma were 24.1 and 36.6 hr, respectively. The mean residence time of centchroman was highest in the liver (78.4 hr), followed by the uterus (72.7 hr), adipose tissue (47.5 hr), lung (46 hr), spleen (44.1 hr), and plasma (37.7 hr). The mean residence time of DMC was also highest in the liver (133.7 hr), followed by the uterus (122 hr), adipose tissue (85.2 hr), lung (62.6 hr), spleen (62.6 hr), and plasma (48.2 hr).


Temperature-dependent development, cold tolerance, and potential distribution of Cricotopus lebetis (Diptera: Chironomidae), a tip miner of Hydrilla verticillata (Hydrocharitaceae).
A chironomid midge, Cricotopus lebetis (Sublette) (Diptera: Chironomidae), was discovered attacking the apical meristems of Hydrilla verticillata (L.f. Royle) in Crystal River, Citrus Co., Florida in 1992. The larvae mine the stems of H. verticillata and cause basal branching and stunting of the plant. Temperature-dependent development, cold tolerance, and the potential distribution of the midge were investigated. The results of the temperature-dependent development study showed that optimal temperatures for larval development were between 20 and 30°C, and these data were used to construct a map of the potential number of generations per year of C. lebetis in Florida. Data from the cold tolerance study, in conjunction with historical weather data, were used to generate a predicted distribution of C. lebetis in the United States. A distribution was also predicted using an ecological niche modeling approach by characterizing the climate at locations where C. lebetis is known to occur and then finding other locations with similar climate. The distributions predicted using the two modeling approaches were not significantly different and suggested that much of the southeastern United States was climatically suitable for C. lebetis.


Tracking lexical consolidation with ERPs: Lexical and semantic-priming effects on N400 and LPC responses to newly-learned words.
Novel words can be recalled immediately and after little exposure, but require a post-learning consolidation period to show word-like behaviour such as lexical competition. This pattern is thought to reflect a qualitative shift from episodic to lexical representations. However, several studies have reported immediate effects of meaningful novel words on semantic processing, suggesting that integration of novel word meanings may not require consolidation. The current study synthesises and extends these findings by showing a dissociation between lexical and semantic effects on the electrophysiological (N400, LPC) response to novel words. The difference in N400 amplitude between novel and existing words (a lexical effect) decreased significantly after a 24-h consolidation period, providing novel support for the hypothesis that offline consolidation aids lexicalisation. In contrast, novel words preceded by semantically related primes elicited a more positive LPC response (a semantic-priming effect) both before and after consolidation, indicating that certain semantic effects can be observed even when words have not been fully lexicalised. We propose that novel meanings immediately start to contribute to semantic processing, but that the underlying neural processes may shift from strategic to more automatic with consolidation.


How Structural Violence, Prohibition, and Stigma Have Paralyzed North American Responses to Opioid Overdose.
As of 2020, North America is now into the fifth year of an unprecedented increase in drug overdose deaths driven by a toxic, unpredictable, and unregulated drug supply. While the genesis and drivers of and response to the opioid overdose crisis have wide regional variations, structural violence, prohibitions against illicit drug use, and stigma consistently play a central role. The criminalization of users of illicit drugs has led directly not only to users' incarceration, but also to their marginalization and isolation and to violence, entrenched poverty, and a vicious cycle of trauma. This policy has created an environment wherein any initiatives to prevent and reverse overdoses have been severely restricted. While a war on drugs and the people who use them has been widely criticized as destructive and unwinnable, the criminal policies that support the war on drugs have not changed even in response to this unprecedented crisis.


Biological affinities and regional microevolution among pre-Hispanic communities of Colombia's Northern Andes.
Dental non-metric data were used to examine the biological continuity of pre-Hispanic peoples of Colombia's Northern Andes, including highland, lowland and coastal peoples. This report contributes to studies regarding the peopling of South America by establishing a benchmark comparison that includes pre-Hispanic populations of the Northern Andes. The sample consisted of a total of 583 individuals from 56 cemeteries ranging in time from the Early Holocene (10,000 BP) to the Final Late Holocene (500 BP). Permanent dentitions from individuals between 5 and 40 years of age were scored for 87 dental traits based on the ASUDAS. A divergence matrix was programmed using the Smith's Mean Measure of Divergence equation (MMD). Bartlett's adjustment and Ascombe transformation were considered into MMD calculations. Principal Coordenate analysis was applied based on MMD matrix scores. A clear group was found that associated Initial Late Holocene samples with Final Late Holocene samples. Early Holocene samples are very different to that, and Middle Holocene samples show as morphologically intermediate series. A comparison of the frequencies by time and period showed that a limited biological continuity existed. Interbreeding among initial populations of the same regions is expressed in similar frequencies of dental traits within Early Holocene and Middle Holocene samples. Early Holocene samples did not match with Sinodont pattern according to discriminant function analysis. These findings help us to better understand the settlement process of human groups in the Northern Andes and its relationship with migratory movements in South America.


Strategies for the enrichment and identification of basic proteins in proteome projects.
Two-dimensional gel electrophoresis (2-DE) is currently the method of choice for separating complex mixtures of proteins for visual comparison in proteome analysis. This technology, however, is biased against certain classes of proteins including low abundance and hydrophobic proteins. Proteins with extremely alkaline isoelectric points (pI) are often very poorly represented using 2-DE technology, even when complex mixtures are separated using commercially available pH 6-11 or pH 7-10 immobilized pH gradients. The genome of the human gut pathogen, Helicobacter pylori, is dominated by genes encoding basic proteins, and is therefore a useful model for examining methodology suitable for separating such proteins. H. pylori proteins were separated on pH 6-11 and novel pH 9-12 immobilized pH gradients and 65 protein spots were subjected to matrix-assisted laser desorption/ionization-time of flight mass spectrometry, leading to the identification of 49 unique proteins. No proteins were characterized with a theoretical pI of greater than 10.23. A second approach to examine extremely alkaline proteins (pI > 9.0) utilized a prefractionation isoelectric focusing. Proteins were separated into two fractions using Gradiflow technology, and the extremely basic fraction subjected to both sodium dodecyl sulphate-polyacrylamide gel electrophoresis and liquid chromatography (LC) - tandem mass spectrometry post-tryptic digest, allowing the identification of 17 and 13 proteins, respectively. Gradiflow separations were highly specific for proteins with pI > 9.0, however, a single LC separation only allowed the identification of peptides from highly abundant proteins. These methods and those encompassing multiple LC 'dimensions' may be a useful complement to 2-DE for 'near-to-total' proteome coverage in the alkaline pH range.


Comparative transcriptomes profiling of photoperiod-sensitive male sterile rice Nongken 58S during the male sterility transition between short-day and long-day.
Photoperiod-sensitive genic male sterile (PGMS) rice, Nongken 58S, was discovered in 1973. It has been widely used for the production of hybrid rice, and great achievements have been made in improving rice yields. However, the mechanism of the male sterility transition in PGMS rice remains to be determined. To investigate the transcriptome during the male sterility transition in PGMS rice, the transcriptome of Nongken 58S under short-day (SD) and long-day (LD) at the glume primordium differentiation and pistil/stamen primordium forming stages was compared. Seventy-three and 128 differentially expressed genes (DEGs) were identified at the glume primordium differentiation and pistil/stamen primordium forming stages, respectively. Five and 22 genes were markedly up-regulated (≥ 5-fold), and two and five genes were considerably down-regulated (≥ 5-fold) under SD during the male sterility transition. Gene ontology annotation and pathway analysis revealed that four biological processes and the circadian rhythms and the flowering pathways coordinately regulated the male sterility transition. Further quantitative PCR analysis demonstrated that the circadian rhythms of OsPRR1, OsPRR37, OsGI, Hd1, OsLHY and OsDof in leaves were obviously different between Nongken 58S and Nongken 58 under LD conditions. Moreover, both OsPRR37 and Hd1 in the inflorescence displayed differences between Nongken 58S and Nongken 58 under both LD and SD conditions. The results presented here indicate that the transcriptome in Nongken 58S was significantly suppressed under LD conditions. Among these DEGs, the circadian rhythm and the flowering pathway were involved in the male sterility transition. Furthermore, these pathways were coordinately involved in the male sterility transition in PGMS rice.


Inhibition of choroidal neovascularization with an anti-inflammatory carotenoid astaxanthin.
Astaxanthin (AST) is a carotenoid found in marine animals and vegetables. The purpose of the present study was to investigate the effect of AST on the development of experimental choroidal neovascularization (CNV) with underlying cellular and molecular mechanisms. Laser photocoagulation was used to induce CNV in C57BL/6J mice. Mice were pretreated with intraperitoneal injections of AST daily for 3 days before photocoagulation, and treatments were continued daily until the end of the study. CNV response was analyzed by volumetric measurements 1 week after laser injury. Retinal pigment epithelium-choroid levels of IkappaB-alpha, intercellular adhesion molecule (ICAM)-1, monocyte chemotactic protein (MCP)-1, interleukin (IL)-6, vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR)-1, and VEGFR-2 were examined by Western blotting or ELISA. AST was applied to capillary endothelial (b-End3) cells, macrophages, and RPE cells to analyze the activation of NF-kappaB and the expression of inflammatory molecules. The index of CNV volume was significantly suppressed by treatment with AST compared with that in vehicle-treated animals. AST treatment led to significant inhibition of macrophage infiltration into CNV and of the in vivo and in vitro expression of inflammation-related molecules, including VEGF, IL-6, ICAM-1, MCP-1, VEGFR-1, and VEGFR-2. Importantly, AST suppressed the activation of the NF-kappaB pathway, including IkappaB-alpha degradation and p65 nuclear translocation. AST treatment, together with inflammatory processes including NF-kappaB activation, subsequent upregulation of inflammatory molecules, and macrophage infiltration, led to significant suppression of CNV development. The present study suggests the possibility of AST supplementation as a therapeutic strategy to suppress CNV associated with AMD.


Use of Ball Blanket in attention-deficit/hyperactivity disorder sleeping problems.
Based on actigraphic surveillance, attention-deficit/hyperactivity disorder (ADHD) symptom rating and sleep diary, this study will evaluate the effect of Ball Blanket on sleep for a sample of 8-13-year-old children with ADHD. Case-control study. A child and adolescent psychiatric department of a teaching hospital. 21 children aged 8-13 years with a diagnosis of ADHD and 21 healthy control subjects. Sleep was monitored by parent-completed sleep diaries and 28 nights of actigraphy. For 14 of those days, the child slept with a Ball Blanket. The sleep latency, number of awakenings and total length of sleep was measured, as was the possible influence on parent- and teacher-rated ADHD symptom load. The results of this study will show that the time it takes for a child to fall asleep is shortened when using a Ball Blanket. The time it takes to fall asleep when using the Ball Blanket is found to be at the same level as the healthy control subjects. Teacher rating of symptoms show an improvement in both activity levels and attention span of approximately 10% after using the Ball Blankets. The results of this study show that the use of Ball Blankets is a relevant and effective treatment method with regard to minimizing sleep onset latency. We find that the use of Ball Blankets for 14-days improves the time it takes to fall asleep, individual day-to-day variation and the number of awakenings to a level that compares with those found in the healthy control group. Furthermore, we find that the use of Ball Blankets significantly reduces the number of nights that the ADHD child spends more than 30 min falling asleep from 19% to 0%.


Pleiomorphism in Mycobacterium.
Morphological variants in mycobacterial cultures under different growth conditions, including aging of the culture, have been shown to include fibrous aggregates, biofilms, coccoids, and spores. Here we discuss the diversity in shape and size changes demonstrated by bacterial cells with special reference to pleiomorphism observed in Mycobacterium spp. in response to nutritional and other environmental stresses. Inherent asymmetry in cell division and compartmentalization of cell interior under different growth conditions might contribute toward the observed pleiomorphism in mycobacteria. The regulatory genes comprising the bacterial signaling pathway responsible for initiating morphogenesis are speculated upon from bioinformatic identifications of genes for known sensors, kinases, and phosphatases existing in mycobacterial genomes as well as on the basis of what is known in other bacteria.


Solid-state study of the structure and host-guest chemistry of cucurbituril-ferrocene inclusion complexes.
Inclusion complexes of ferrocene (Fc) with cucurbit[n]urils (n = 7, 8) have been prepared via a rapid microwave-assisted hydrothermal approach. Solids were isolated and characterised by elemental analysis, powder X-ray diffraction (PXRD), spectroscopic, and thermoanalytical methods. The UV-Vis spectra support the presence of Fc in Fc@CB7 and a mixture of Fc and ferrocenium ions in Fc@CB8. Partial oxidation of Fc to Fc+ takes place in situ mainly due to the presence of acid of crystallisation in CB8. On the basis of PXRD, the complex Fc@CB8 is classified into an isostructural series that is formed by several CB8-containing compounds that crystallise in the space group I41/a and have similar unit cell dimensions and CB8 packing motifs. The FT-IR and Raman spectra of Fc@CB7 are compared with those of the CB7 host and the Fc guest starting materials, revealing significant frequency shifts of some Fc-centered vibrational modes upon complexation. Blueshifts of the Fe-Cp stretching and ring tilt bands are attributed to encapsulation of Fc monomers in a constrained environment, leading to restricted motion effects and/or a change in the Fc conformation from staggered to eclipsed. The absence of comparable shifts for Fc@CB8 point to a weaker host-guest interaction as a consequence of the larger cavity size. The different host-guest interactions are also evident through a comparison of the 13C{1H} CP MAS NMR spectra. Thermogravimetric analysis for the inclusion compounds reveals that sublimation of Fc is inhibited by molecular encapsulation to the extent that oxidative decomposition of the organoiron species takes place concurrently with cucurbituril decomposition, leading to the formation of hematite, α-Fe2O3.


Inter-laboratory comparison of channelized hotelling observer computation.
The task-based assessment of image quality using model observers is increasingly used for the assessment of different imaging modalities. However, the performance computation of model observers needs standardization as well as a well-established trust in its implementation methodology and uncertainty estimation. The purpose of this work was to determine the degree of equivalence of the channelized Hotelling observer performance and uncertainty estimation using an intercomparison exercise. Image samples to estimate model observer performance for detection tasks were generated from two-dimensional CT image slices of a uniform water phantom. A common set of images was sent to participating laboratories to perform and document the following tasks: (a) estimate the detectability index of a well-defined CHO and its uncertainty in three conditions involving different sized targets all at the same dose, and (b) apply this CHO to an image set where ground truth was unknown to participants (lower image dose). In addition, and on an optional basis, we asked the participating laboratories to (c) estimate the performance of real human observers from a psychophysical experiment of their choice. Each of the 13 participating laboratories was confidentially assigned a participant number and image sets could be downloaded through a secure server. Results were distributed with each participant recognizable by its number and then each laboratory was able to modify their results with justification as model observer calculation are not yet a routine and potentially error prone. Detectability index increased with signal size for all participants and was very consistent for 6 mm sized target while showing higher variability for 8 and 10 mm sized target. There was one order of magnitude between the lowest and the largest uncertainty estimation. This intercomparison helped define the state of the art of model observer performance computation and with thirteen participants, reflects openness and trust within the medical imaging community. The performance of a CHO with explicitly defined channels and a relatively large number of test images was consistently estimated by all participants. In contrast, the paper demonstrates that there is no agreement on estimating the variance of detectability in the training and testing setting.


Effects of common mutations in the SARS-CoV-2 Spike RBD and its ligand, the human ACE2 receptor on binding affinity and kinetics.
The interaction between the SARS-CoV-2 virus Spike protein receptor binding domain (RBD) and the ACE2 cell surface protein is required for viral infection of cells. Mutations in the RBD are present in SARS-CoV-2 variants of concern that have emerged independently worldwide. For example, the B.1.1.7 lineage has a mutation (N501Y) in its Spike RBD that enhances binding to ACE2. There are also ACE2 alleles in humans with mutations in the RBD binding site. Here we perform a detailed affinity and kinetics analysis of the effect of five common RBD mutations (K417N, K417T, N501Y, E484K, and S477N) and two common ACE2 mutations (S19P and K26R) on the RBD/ACE2 interaction. We analysed the effects of individual RBD mutations and combinations found in new SARS-CoV-2 Alpha (B.1.1.7), Beta (B.1.351), and Gamma (P1) variants. Most of these mutations increased the affinity of the RBD/ACE2 interaction. The exceptions were mutations K417N/T, which decreased the affinity. Taken together with other studies, our results suggest that the N501Y and S477N mutations enhance transmission primarily by enhancing binding, the K417N/T mutations facilitate immune escape, and the E484K mutation enhances binding and immune escape.


Two enzymes bound to one transfer RNA assume alternative conformations for consecutive reactions.
In most bacteria and all archaea, glutamyl-tRNA synthetase (GluRS) glutamylates both tRNA(Glu) and tRNA(Gln), and then Glu-tRNA(Gln) is selectively converted to Gln-tRNA(Gln) by a tRNA-dependent amidotransferase. The mechanisms by which the two enzymes recognize their substrate tRNA(s), and how they cooperate with each other in Gln-tRNA(Gln) synthesis, remain to be determined. Here we report the formation of the 'glutamine transamidosome' from the bacterium Thermotoga maritima, consisting of tRNA(Gln), GluRS and the heterotrimeric amidotransferase GatCAB, and its crystal structure at 3.35 A resolution. The anticodon-binding body of GluRS recognizes the common features of tRNA(Gln) and tRNA(Glu), whereas the tail body of GatCAB recognizes the outer corner of the L-shaped tRNA(Gln) in a tRNA(Gln)-specific manner. GluRS is in the productive form, as its catalytic body binds to the amino-acid-acceptor arm of tRNA(Gln). In contrast, GatCAB is in the non-productive form: the catalytic body of GatCAB contacts that of GluRS and is located near the acceptor stem of tRNA(Gln), in an appropriate site to wait for the completion of Glu-tRNA(Gln) formation by GluRS. We identified the hinges between the catalytic and anticodon-binding bodies of GluRS and between the catalytic and tail bodies of GatCAB, which allow both GluRS and GatCAB to adopt the productive and non-productive forms. The catalytic bodies of the two enzymes compete for the acceptor arm of tRNA(Gln) and therefore cannot assume their productive forms simultaneously. The transition from the present glutamylation state, with the productive GluRS and the non-productive GatCAB, to the putative amidation state, with the non-productive GluRS and the productive GatCAB, requires an intermediate state with the two enzymes in their non-productive forms, for steric reasons. The proposed mechanism explains how the transamidosome efficiently performs the two consecutive steps of Gln-tRNA(Gln) formation, with a low risk of releasing the unstable intermediate Glu-tRNA(Gln).


The Therapeutic Potential of Piceatannol, a Natural Stilbene, in Metabolic Diseases: A Review.
Metabolic disease comprises a set of risk factors highly associated with obesity and insulin resistance and is a consequence of central adiposity, hyperglycemia, and dyslipidemia. Furthermore, obesity increases the risk of the development of metabolic disease due to ectopic fat deposition, low-grade inflammation, and systemic energy disorders caused by dysregulated adipose tissue function. Piceatannol is a naturally occurring polyphenolic stilbene found in various fruits and vegetables and has been reported to exhibit anticancer and anti-inflammatory properties. In addition, recently reported beneficial effects of piceatannol on hypercholesterolemia, atherosclerosis, and angiogenesis underscore its therapeutic potential in cardiovascular disease. However, investigation of its role in metabolic disease is still in its infancy. This review intensively summarizes in vitro and in vivo studies supporting the potential therapeutic effects of piceatannol in metabolic disease, including inhibition of adipogenesis and lipid metabolism in adipocytes, and regulation of hyperlipidemia, hyperglycemia, insulin resistance, and fatty acid-induced inflammation and oxidative stress.


Dynamic response-by-response models of matching behavior in rhesus monkeys.
We studied the choice behavior of 2 monkeys in a discrete-trial task with reinforcement contingencies similar to those Herrnstein (1961) used when he described the matching law. In each session, the monkeys experienced blocks of discrete trials at different relative-reinforcer frequencies or magnitudes with unsignalled transitions between the blocks. Steady-state data following adjustment to each transition were well characterized by the generalized matching law; response ratios undermatched reinforcer frequency ratios but matched reinforcer magnitude ratios. We modelled response-by-response behavior with linear models that used past reinforcers as well as past choices to predict the monkeys' choices on each trial. We found that more recently obtained reinforcers more strongly influenced choice behavior. Perhaps surprisingly, we also found that the monkeys' actions were influenced by the pattern of their own past choices. It was necessary to incorporate both past reinforcers and past choices in order to accurately capture steady-state behavior as well as the fluctuations during block transitions and the response-by-response patterns of behavior. Our results suggest that simple reinforcement learning models must account for the effects of past choices to accurately characterize behavior in this task, and that models with these properties provide a conceptual tool for studying how both past reinforcers and past choices are integrated by the neural systems that generate behavior.


Analysis of uncoupling protein 2-deficient mice upon anaesthesia and sedation revealed a role for UCP2 in locomotion.
General anaesthesia is associated with hypothermia, oxidative stress, and immune depression. Uncoupling Protein (UCP2) is a member of the mitochondrial carrier family present in many organs including the spleen, the lung and the brain. A role of UCP2 in the activation of the inflammatory/immune cells, in the secretion of hormones, and in the excitability of neurons by regulating the production of reactive oxygen species has been discussed. Because of the side effects of anaesthesia listed above, we aimed to question the expression and the function of UCP2 during anaesthesia. Induction of anaesthesia with ketamine (20 mg/kg) or isoflurane (3.6%) and induction of sedation with the α2 adrenergic receptor agonist medetomidine (0.2 mg/kg) stimulated infiltration of immune cells in the lung and increased UCP2 protein content in the lung, in both immune and non-immune cells. UCP2 content in the lung inversely correlated with body temperature decrease induced by medetomidine treatment. Challenge of the Ucp2(-/-) mice with isoflurane and medetomidine revealed an earlier behavioral recovery phenotype. Transponder analysis of body temperature and activity showed no difference between Ucp2(-/-) and control mice in basal conditions. However, upon an acute decrease of body temperature induced by medetomidine, Ucp2(-/-) mice exhibited increased locomotion activity. Together, these results show that UCP2 is rapidly mobilized during anaesthesia and sedation in immune cells, and suggest a role of UCP2 in locomotion.


ERK1/2 and p38α/β signaling in tumor cell quiescence: opportunities to control dormant residual disease.
Systemic minimal residual disease after primary tumor treatment can remain asymptomatic for decades. This is thought to be due to the presence of dormant disseminated tumor cells (DTC) or micrometastases in different organs. DTCs lodged in brain, lungs, livers, and/or bone are a major clinical problem because they are the founders of metastasis, which ultimately kill cancer patients. The problem is further aggravated by our lack of understanding of DTC biology. In consequence, there are almost no rational therapies to prevent dormant DTCs from surviving and expanding. Several cancers, including melanoma as well as breast, prostate, and colorectal carcinomas, undergo dormant periods before metastatic recurrences develop. Here we review our experience in studying the cross-talk between ERK1/2 and p38α/β signaling in models of early cancer progression, dissemination, and DTC dormancy. We also provide some potential translational and clinical applications of these findings and describe how some currently used therapies might be useful to control dormant disease. Finally, we draw caution on the use of p38 inhibitors currently in clinical trials for different diseases as these may accelerate metastasis development.


Dental caries: prospects for prevention.
Combined utilization of measures now available or imminent could reduce caries of the crowns of the teeth to the point of negligibility as a public health problem, if public desire were great enough to motivate changes in some of our habits (64). Universal optimum application of fluoride and substitution of starchy foods for sugary ones (or even simply judicious consumption of sugar) would alone do most of the job. Sealing of susceptible occlusal areas with adhesive polymers promises to protect the sites where fluoride evidently cannot be maximally effective. It seems unlikely that any single measure will be found sufficient to control this multifactorial disease. Consequently, we must continue the search for new means to increase the caries resistance of teeth, to reduce the cariogenicity of foodstuffs, and to check the deleterious activities of cariogenic bacteria. Anticaries food additives and antibacterial agents for intraoral use seem to be approaching practicability. Past performance warrants expectation that ongoing fundamental investigations will produce leads for future development and application.


A method for compression of intra-cortically-recorded neural signals dedicated to implantable brain-machine interfaces.
This paper proposes an efficient data compression technique dedicated to implantable intra-cortical neural recording devices. The proposed technique benefits from processing neural signals in the Discrete Haar Wavelet Transform space, a new spike extraction approach, and a novel data framing scheme to telemeter the recorded neural information to the outside world. Based on the proposed technique, a 64-channel neural signal processor was designed and prototyped as a part of a wireless implantable extra-cellular neural recording microsystem. Designed in a 0.13- μ m standard CMOS process, the 64-channel neural signal processor reported in this paper occupies ∼ 0.206 mm(2) of silicon area, and consumes 94.18 μW when operating under a 1.2-V supply voltage at a master clock frequency of 1.28 MHz.