Technical note: rapid prototyping of 3D grid arrays for image guided therapy quality assurance.
Three dimensional grid phantoms offer a number of advantages for measuring imaging related spatial inaccuracies for image guided surgery and radiotherapy. The authors examined the use of rapid prototyping technology for directly fabricating 3D grid phantoms from CAD drawings. We tested three different fabrication process materials, photopolymer jet with acrylic resin (PJ/AR), selective laser sintering with polyamide (SLS/P), and fused deposition modeling with acrylonitrile butadiene styrene (FDM/ABS). The test objects consisted of rectangular arrays of control points formed by the intersections of posts and struts (2 mm rectangular cross section) and spaced 8 mm apart in the x, y, and z directions. The PJ/AR phantom expanded after immersion in water which resulted in permanent warping of the structure. The surface of the FDM/ABS grid exhibited a regular pattern of depressions and ridges from the extrusion process. SLS/P showed the best combination of build accuracy, surface finish, and stability. Based on these findings, a grid phantom for assessing machine-dependent and frame-induced MR spatial distortions was fabricated to be used for quality assurance in stereotactic neurosurgical and radiotherapy procedures. The spatial uniformity of the SLS/P grid control point array was determined by CT imaging (0.6 x 0.6 x 0.625 mm3 resolution) and found suitable for the application, with over 97.5% of the control points located within 0.3 mm of the position specified in CAD drawing and none of the points off by more than 0.4 mm. Rapid prototyping is a flexible and cost effective alternative for development of customized grid phantoms for medical physics quality assurance.


Increased flanker task and forward digit span performance in caudate-nucleus-dependent response strategies.
One of two memory systems can be used to navigate in a new environment. Hippocampus-dependent spatial strategy consists of creating a cognitive map of an environment and caudate nucleus-dependent response strategy consists of memorizing a rigid sequence of turns. Spontaneous use of the response strategy is associated with greater activity and grey matter within the caudate nucleus while the spatial strategy is associated with greater activity and grey matter in the hippocampus. The caudate nucleus is involved in executive functions such as working memory, cognitive control and certain aspects of attention such as attentional disengaging. This study therefore aimed to investigate whether response learners would display better performance on tests of executive and attention functioning compared to spatial learners. Fifty participants completed the 4/8 virtual maze to assess navigational strategy, the forward and backward visual digit span and the Attention Network Test - Revised to assess both attention disengagement and cognitive control. Results revealed that response learners showed significantly higher working memory capacity, more efficient attention disengagement and better cognitive control. Results suggest that response learners, who putatively display more grey matter and activity in the caudate nucleus, are associated with better working memory span, cognitive control and attentional disengagement.


The role of the lateral prefrontal cortex and anterior cingulate in stimulus-response association reversals.
Many complex tasks require us to flexibly switch between behavioral rules, associations, and strategies. The prefrontal cerebral cortex is thought to be critical to the performance of such behaviors, although the relative contribution of different components of this structure and associated subcortical regions are not fully understood. We used functional magnetic resonance imaging to measure brain activity during a simple task which required repeated reversals of a rule linking a colored cue and a left/right motor response. Each trial comprised three discrete events separated by variable delay periods. A colored cue instructed which response was to be executed, followed by a go signal which told the subject to execute the response and a feedback instruction which indicated whether to "hold" or "flip" the rule linking the colored cue and response. The design allowed us to determine which brain regions were recruited by the specific demands of preparing a rule contingent motor response, executing such a response, evaluating the significance of the feedback, and reconfiguring stimulus-response (SR) associations. The results indicate that an increase in neural activity occurs within the anterior cingulate gyrus under conditions in which SR associations are labile. In contrast, lateral frontal regions are activated by unlikely/unexpected perceptual events regardless of their significance for behavior. A network of subcortical structures, including the mediodorsal nucleus of the thalamus and striatum were the only regions showing activity that was exclusively correlated with the neurocognitive demands of reversing SR associations. We conclude that lateral frontal regions act to evaluate the behavioral significance of perceptual events, whereas medial frontal-thalamic circuits are involved in monitoring and reconfiguring SR associations when necessary.


Correlates of crack abuse among drug-using incarcerated women: psychological trauma, social support, and coping behavior.
This investigation examines the relationship between psychological trauma and crack abuse among 158 women with a recent history of drug use who were incarcerated in a New York City jail facility. Interviewers obtained data on demographics, drug use, psychological trauma history, criminal history, social support, and coping behavior variables. Three-fourths of the total sample had used crack three or more times a week for a month in the past; a quarter had used other drugs, predominantly heroin, three or more times a week for a month in the past. Multiple logistic regression analysis was used to assess the association between adult psychological trauma variables (loss of custody of youngest child and lived in streets prior to arrest) and regular crack use in three sequential models. After adjusting for social support, coping behavior, demographics, and criminal history variables, women who had lost custody of their youngest child were 3.3 times more likely to be regular crack uses. Women who demonstrated more negative coping behavior and perceived themselves as having less emotional support were also more likely to be regular crack users. The association between childhood traumas (i.e., childhood sexual abuse, childhood physical abuse, parental alcohol abuse) and regular crack use was also assessed using multiple logistic regression; however, no significant associations were found between these childhood psychological traumas and regular crack use in both the unadjusted and adjusted models. Study findings underscore the importance of assessing environmental, interpersonal, and intrapersonal factors in tailoring treatment strategies for users of crack and other drugs.


Density-difference estimation.
We address the problem of estimating the difference between two probability densities. A naive approach is a two-step procedure of first estimating two densities separately and then computing their difference. However, this procedure does not necessarily work well because the first step is performed without regard to the second step, and thus a small estimation error incurred in the first stage can cause a big error in the second stage. In this letter, we propose a single-shot procedure for directly estimating the density difference without separately estimating two densities. We derive a nonparametric finite-sample error bound for the proposed single-shot density-difference estimator and show that it achieves the optimal convergence rate. We then show how the proposed density-difference estimator can be used in L²-distance approximation. Finally, we experimentally demonstrate the usefulness of the proposed method in robust distribution comparison such as class-prior estimation and change-point detection.


Astragaloside Alleviates Hepatic Fibrosis Function via PAR2 Signaling Pathway in Diabetic Rats.
Astragaloside (AGS) extracted from radix astragalin (Huangqi) has been considered to be beneficial to liver diseases. In this study, we examined the role played by AGS in alleviating hepatic fibrosis function via protease-activated receptor-2 (PAR2) mechanisms. We hypothesized that AGS affects PAR2 signaling pathway thereby improving hepatic function in rats with hepatic fibrosis induced by carbon tetrachloride (CCl4). We further hypothesized that AGS attenuates impaired hepatic function evoked by CCl4 to a greater degree in diabetic animals. ELISA and Western Blot analysis were used to examine PAR2 signaling pathway in diabetic CCl4-rats and non-diabetic CCl4-rats. AGS inhibited the protein expression of PAR2 and its downstream pathway PKA and PKCɛ in CCl4-rats. Notably, the effects of AGS were greater in CCl4-rats with diabetes. AGS also significantly attenuated the CCl4-induced upregulations of pro-inflammatory cytokines, namely interleukin-1β, interleukin-6 and tumor necrosis factor-α accompanied with decreases of collagenic parameters such as hexadecenoic acid, laminin and hydroxyproline. Additionally, AGS improved the CCl4-induced exaggerations of liver index and functions including alanine aminotransferase, aspartate aminotransferase. Moreover, TGF-β1, a marker of hepatic fibrosis, was increased in CCl4-rats and AGS inhibited increases in TGF-β1 induced by CCl4. AGS alleviates hepatic fibrosis by inhibiting PAR2 signaling expression and its effects are largely enhanced in diabetic animals. Targeting one or more of these signaling molecules may present new opportunities for treatment and management of hepatic fibrosis; and results of our study are likely to shed light on strategies for application of AGS because it has potentially greater therapeutic effectiveness for hepatic fibrosis in diabetes.


Investigating Intraindividual Variability of Psychological Needs Satisfaction and Relations With Subsequent Physical Activity.
Heeding recent calls to capture dynamic variability of physical activity (PA) motivation within a self-determination theory framework, this study examined the extent to which psychological needs satisfaction in PA predicted subsequent PA, disaggregating within-person and between-persons data. University students (N = 89) wore an ActiGraph GT3X accelerometer for 6 days and reported basic psychological needs satisfaction daily. Multilevel models examined whether competence, autonomy, and relatedness for the previous day's PA (>2,020 counts per minute) predicted the following day's minutes of PA (>2,020 counts per minute), controlling for previous-day PA. Participants who, on average, reported greater feelings of autonomy and competence tended to engage in more minutes of PA the following day. When participants reported feeling greater relatedness than what was typical for them, they tended to engage in more PA the following day. Psychological needs vary day to day, but how and to what extent they predict PA depends on the specific need.


Inhibition of tetrahydrobiopterin biosynthesis impairs endothelium-dependent relaxations in canine basilar artery.
Tetrahydrobiopterin is an essential cofactor in biosynthesis of nitric oxide. The present study was designed to determine the effect of decreased intracellular tetrahydrobiopterin levels on endothelial function of isolated cerebral arteries. Blood vessels were incubated for 6 h in minimum essential medium (MEM) in the presence or absence of a GTP cyclohydrolase I inhibitor, 2,4-diamino-6-hydroxypyrimidine (DAHP, 10(-2) M). Rings with and without endothelium were suspended for isometric force recording in the presence of a cyclooxygenase inhibitor, indomethacin (10(-5) M). In arteries with endothelium, DAHP significantly reduced intracellular levels of tetrahydrobiopterin. DAHP in combination with a precursor of the salvage pathway of tetrahydrobiopterin biosynthesis, sepiapterin (10(-4) M), not only restored but increased levels of tetrahydrobiopterin above control values. In DAHP-treated arteries, endothelium-dependent relaxations to bradykinin (10(-10)-10(-6) M) or calcium ionophore A23187 (10(-9)-10(-6) M) were significantly reduced, whereas endothelium-independent relaxations to a nitric oxide donor, 3-morpholinosydnonimine (10(-9)-10(-4) M), were not affected. When DAHP-treated arteries with endothelium were incubated with sepiapterin (10(-4) M) or superoxide dismutase (150 U/ml), relaxations to bradykinin and A23187 were restored to control levels. In contrast, superoxide dismutase did not affect endothelium-dependent relaxations in arteries incubated in MEM. A nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester (10(-4) M), abolished relaxations to bradykinin or A23187 in control arteries and in DAHP-treated arteries. These studies demonstrate that in cerebral arteries, decreased intracellular levels of tetrahydrobiopterin can reduce endothelium-dependent relaxations. Production of superoxide anions during activation of dysfunctional endothelial nitric oxide synthase appears to be responsible for the impairment of endothelial function.


When do response-related episodic retrieval effects co-occur with inhibition of return?
At some point, spatial priming effects more faithfully reflect response selection processes than they do attentional orienting or sensory processes. Findings from the spatial cueing literature suggest that two factors may be critical: (1) the amount of identity processing that is required in order to respond correctly (feature-based response hypothesis), and (2) the amount of spatial processing that is required in order to respond correctly (space-based response hypothesis). To test the first hypothesis, we manipulated whether observers made single keypress detection or two-choice localization responses to serially presented stimuli in peripheral vision and whether stimulus identity information processing was necessary before responding. Responses were always slowest when the target location repeated, consistent with an attentional orienting bias independent of keypress responding (i.e., inhibition of return; IOR). The localization procedure revealed a subtle additional cost for changing the target location and repeating a response, consistent with a response-related episodic retrieval effect predicted by the Theory of Event Coding (TEC). Neither effect was modulated by the need to discriminate features. To test the second hypothesis, we made spatial processing indispensable to response selection by requiring a decision between a detection and localization response, depending on where the target appeared. IOR was eliminated for detection, but not localization, responses, consistent with the TEC. Collectively, the findings suggest that the amount of space-based, but not feature-based, processing that is required to determine a response is responsible for the response retrieval effects that can co-occur with IOR.


Inhibition of SREBP by a small molecule, betulin, improves hyperlipidemia and insulin resistance and reduces atherosclerotic plaques.
Sterol regulatory element-binding proteins (SREBPs) are major transcription factors activating the expression of genes involved in biosynthesis of cholesterol, fatty acid and triglyceride. In this study, we identified a small molecule, betulin, that specifically inhibited the maturation of SREBP by inducing interaction of SREBP cleavage activating protein (SCAP) and Insig. Inhibition of SREBP by betulin decreased the biosynthesis of cholesterol and fatty acid. In vivo, betulin ameliorated diet-induced obesity, decreased the lipid contents in serum and tissues, and increased insulin sensitivity. Furthermore, betulin reduced the size and improved the stability of atherosclerotic plaques. Our study demonstrates that inhibition SREBP pathway can be employed as a therapeutic strategy to treat metabolic diseases including type II diabetes and atherosclerosis. Betulin, which is abundant in birch bark, could be a leading compound for development of drugs for hyperlipidemia.


Electromyographic and gait analysis of forty-three patients after rotationplasty.
Rotationplasty is considered to be a treatment option for patients who have had a primary malignant bone tumor of the distal part of the femur or the proximal part of the tibia. The present study was performed to evaluate the muscle activity, the kinetics (range of motion of the hip and knee joints), and the kinematics (joint moments) after rotationplasty and to determine whether there was an association between these parameters and the functional outcome. Forty-three patients who had been managed with rotationplasty for the treatment of a femoral or tibial bone tumor were evaluated clinically and functionally. The mean age (and standard deviation) at the time of follow-up was 24.4 +/- 10.7 years (range, eight to sixty-eight years), the mean age at the time of the procedure was 17.8 +/- 10.2 years (range, seven to sixty-three years), and the mean duration of follow-up was 6.7 +/- 4.9 years (range, 0.7 to eighteen years). Instrumented gait and electromyographic analyses were performed. The qualitative data were compared with the functional outcome, which was determined with the functional evaluation score of the Musculoskeletal Tumor Society. Gait analysis revealed a fairly normal walking pattern with a slight limp and a lateral lean of the trunk over the ipsilateral limb that led to a reduced joint moment in the hip (moment on involved side, 68 percent [compared with a control group]; moment on uninvolved side, 81 percent). The ranges of motion of the hips (uninvolved side, 42.0 +/- 8.2 degrees; involved side, 42.4 +/- 8.0 degrees) and the knees (uninvolved side, 59.7 +/- 5.0 degrees; involved side [former ankle joint], 58.1 +/- 11.6 degrees) were symmetrical even though the knee-motion pattern of the involved limb indicated a slightly reduced extensor mechanism in 51 percent (twenty-two) and a markedly reduced extensor mechanism in 35 percent (fifteen) of the forty-three patients. Electromyography revealed function of the muscles of the involved limb, with comparable amplitudes in the involved and uninvolved limbs. The leg muscles of the involved limb were active in the stance phase (the soleus and the lateral and medial heads of the gastrocnemius) and the swing phase (the peroneus longus and the tibialis anterior) according to their function in relation to the new knee joint. The patients had a good functional result, with a mean score of 23.9 +/- 2.7 of 30 points. With the numbers available for study, we could not show the duration of follow-up to be related to the overall outcome, but the age at the time of the operation was related to the total functional score as well as to gait and walking ability (p < 0.05). The results of the electromyographic and gait analyses demonstrated good functional restoration of gait following rotationplasty.


Urothelial or oral mucosa cells for tissue-engineered urethroplasty: A critical revision of the clinical outcome.
To report the clinical outcome of urethral reconstruction by cultured urothelial or oral mucosa cells for tissue-engineered urethroplasty. We systematically searched for studies reporting the use of tissue-engineered techniques for hypospadias and urethral stricture repair in humans in PubMed and Embase (OvidSP) through January, 1990 to June, 2018. We excluded studies based on titles that clearly were not related to the subject, studies in which tissue-engineered biomaterial were used only in laboratory or experimental animals, and in the absence of autologous cultured epithelial cells. Studies were also excluded if they were not published in English, had no disease background and adequate follow-up. Finally, we search all relevant abstract presented at two of the main urological meetings in the last 10 years: European Association of Urology (EAU) and American Urological Association (AUA). A total of six articles, reporting the clinical use of tissue-engineered techniques in humans, were fully reviewed in our review. The epithelial cells were harvested from the urethra (10 patients), the bladder (11 patients) and the mouth (104 patients). The tissue-engineered grafts were used in children for primary hypospadias repair in 16 cases, and in adults for posterior and anterior urethral strictures repair in 109 cases. Tissue-engineered grafts were showed working better in children for primary hypospadias repair than in adults for urethral strictures repair. One hundred and twenty-five patients received tissue-engineered urethroplasty using cultured epithelial cells for primary hypospadias or urethral strictures repair. The studies demonstrate a high degree of heterogeneity respect to epithelial cells (from urethra, bladder, and mouth), type of scaffold, etiology, site of urethral stricture, number of patients, follow-up and outcomes.


Proteogenomic studies in epithelial ovarian cancer: established knowledge and future needs.
There has been a concerted effort over the last decade to improve our understanding of the complex biology of ovarian cancer. A linear growth in published proteogenomic studies has addressed a variety of questions regarding its molecular pathogenesis. A number of genes have been identified by transcriptomic approaches, some of which are being investigated as putative tumor markers (HE4, OPN, Ep-CAM and Mesothelin), whilst others are potential targets for molecular therapeutic approaches (VEGF, IO4, EGFR, MUC1, CLDN4 and SLPI). Proteogenomics has the potential to further change our current characterization and treatment of ovarian cancer. Additional advances will depend on integrated study designs, interdisciplinary collaborations, use of robust high-throughput platforms, as well as uniform guidelines for bioinformatic analyses.


In Vivo Force Application Reveals a Fast Tissue Softening and External Friction Increase during Early Embryogenesis.
During development, cell-generated forces induce tissue-scale deformations to shape the organism [1,2]. The pattern and extent of these deformations depend not solely on the temporal and spatial profile of the generated force fields but also on the mechanical properties of the tissues that the forces act on. It is thus conceivable that, much like the cell-generated forces, the mechanical properties of tissues are modulated during development in order to drive morphogenesis toward specific developmental endpoints. Although many approaches have recently emerged to assess effective mechanical parameters of tissues [3-8], they could not quantitatively relate spatially localized force induction to tissue-scale deformations in vivo. Here, we present a method that overcomes this limitation. Our approach is based on the application of controlled forces on a single microparticle embedded in an individual cell of an embryo. Combining measurements of bead displacement with the analysis of induced deformation fields in a continuum mechanics framework, we quantify material properties of the tissue and follow their changes over time. In particular, we uncover a rapid change in tissue response occurring during Drosophila cellularization, resulting from a softening of the blastoderm and an increase of external friction. We find that the microtubule cytoskeleton is a major contributor to epithelial mechanics at this stage. We identify developmentally controlled modulations in perivitelline spacing that can account for the changes in friction. Overall, our method allows for the measurement of key mechanical parameters governing tissue-scale deformations and flows occurring during morphogenesis.


Negative refraction and planar focusing based on parity-time symmetric metasurfaces.
We introduce a new mechanism to realize negative refraction and planar focusing using a pair of parity-time symmetric metasurfaces. In contrast to existing solutions that achieve these effects with negative-index metamaterials or phase conjugating surfaces, the proposed parity-time symmetric lens enables loss-free, all-angle negative refraction and planar focusing in free space, without relying on bulk metamaterials or nonlinear effects. This concept may represent a pivotal step towards loss-free negative refraction and highly efficient planar focusing by exploiting the largely uncharted scattering properties of parity-time symmetric systems.


Combination of temsirolimus (CCI-779) with chemoradiation in newly diagnosed glioblastoma multiforme (GBM) (NCCTG trial N027D) is associated with increased infectious risks.
The mammalian target of rapamycin (mTOR) functions within the phosphoinositide 3-kinase/Akt signaling pathway as a critical modulator of cell survival. The mTOR inhibitor temsirolimus (CCI-779) was combined with chemoradiotherapy in glioblastoma multiforme (GBM) patients in a dose-escalation phase I trial. The first 12 patients were treated with CCI-779 combined with radiation/temozolomide and adjuvant temozolomide. A second cohort of 13 patients was treated with concurrent CCI-779/radiation/temozolomide followed by adjuvant temozolomide monotherapy. Concomitant and adjuvant CCI-779 was associated with a high rate (3 of 12 patients) of grade 4/5 infections. By limiting CCI-779 treatment to the radiation/temozolomide phase and using antibiotic prophylaxis, the rate of infections was reduced, although 2 of 13 patients developed exacerbation of pre-existing fungal or viral infections. Dose-limiting toxicities were observed in 2 of 13 patients with this modified schedule. Weekly CCI-779 (50 mg/week) combined with radiation/temozolomide is the recommended phase II dose and schedule. The immune profile of patients in the second cohort was assessed before, during, and after CCI-779 therapy. There was robust suppression of helper and cytotoxic T cells, B cells, natural killer, cells and elevation of regulatory T cells during CCI-779/radiation/temozolomide therapy with recovery to baseline levels during adjuvant temozolomide of cytotoxic T cells, natural killer cells, and regulatory T cells. The increased infection rate observed with CCI-779 combined with chemoradiotherapy in GBM was reduced with antibiotic prophylaxis and by limiting the duration of CCI-779 therapy. The combined suppressive effects of CCI-779 and temozolomide therapy on discrete immune compartments likely contributed to the increased infectious risks observed.


Radiation hardness of high-Q silicon nitride microresonators for space compatible integrated optics.
Integrated optics has distinct advantages for applications in space because it integrates many elements onto a monolithic, robust chip. As the development of different building blocks for integrated optics advances, it is of interest to answer the important question of their resistance with respect to ionizing radiation. Here we investigate effects of proton radiation on high-Q (θ(10⁶)) silicon nitride microresonators formed by a waveguide ring. We show that the irradiation with high-energy protons has no lasting effect on the linear optical losses of the microresonators.


The Central Subfoveal Bouquet in Idiopathic Epiretinal Membranes.
To report both the unoperated clinical course and the surgical outcomes of eyes with a central foveal bouquet (CB) secondary to idiopathic epiretinal membranes (iERMs). Retrospective, consecutive, and observational case series. All patients examined between January 1, 2014, and December 31, 2019, for evaluation of epiretinal membrane with a CB lesion identified on spectral domain optical coherence tomography (SD-OCT) were included. Exclusion criteria included vitreoretinal comorbidities associated with secondary ERMs and an absence of CB lesions on SD-OCT. Patients were divided into two groups: those who were followed with observation (Group I) and those who received surgery (Group II). Each group had 3 different types of mechanical abnormalities of the CB previously described as cotton ball sign, subfoveal detachment, or acquired vitelliform lesion, without a subanalysis discrimination. Best-corrected visual acuity (BCVA) at baseline and last follow-up, subjective metamorphopsia, central retinal thickness (CMT), mechanical stress lesions of the CB, and resolution or evolution of the CB lesions during the follow-up interval. Two hundred seventy-six eyes with iERM were reviewed, and 46 eyes met the inclusion criteria. Among these, 21 of 46 (46%) were observed, and 25 of 46 (54%) underwent surgery. Metamorphopsia was identified in 61.9% of patients in Group I and 81.2% of patients in Group II, at baseline. The mean BCVA was 0.19 ± 0.17 (20/30) in Group I and 0.31 ± 0.33 (20/40) in Group II at presentation. At the final exam, patients in Group I achieved a mean BCVA of 0.24 ± 0.18 (20/30), while patients in Group II obtained a mean BCVA of 0.15 ± 0.21 (20/30). Spontaneous resolution of the CB sign occurred in 5 of 21 eyes (23.8%) that were observed, whereas, after surgery, the CB sign resolved in 16 of 25 eyes (61.5%). Mean CMT was 422 ± 84.2μm in Group I and 531 ± 143.9μm in Group II, at baseline, while at the latest follow-up, the mean CMT was 400 ± 40.8μm in the cases followed with observation and 454 ± 148.7μm in the surgical cases. The clinical course and surgical outcomes of CB findings in iERM are favorable in terms of visual acuity. However, those receiving surgery had an increase in visual acuity and resolution of the CB abnormality.


Influence of military contraceptive policy changes on contraception use and childbirth rates among new recruits.
Unplanned pregnancy is a common problem among United States servicewomen. Variation among service branches in contraceptive education and access during initial training is associated with differences in contraceptive use and childbirth rates despite access to a uniform health benefit including no-cost reproductive healthcare and contraception. However, it is unclear whether changes in branch-specific contraceptive policies can influence reproductive outcomes among junior enlisted women in that service branch. To assess the longitudinal effect of contraceptive policy changes on contraception use and childbirth rates among military recruits. Secondary analysis was performed of insurance records from 70,852 servicewomen who started basic training between October 2013 and December 2016, assessing the longitudinal impact of a Navy policy change expanding contraceptive access during basic training implemented in January 2015, and a Marine Corps policy change restricting contraceptive access during basic training implemented in January 2016 on the following: contraception use (pills, patches, rings, injectable, implantable, and intrauterine) at 6 months, long-acting reversible contraception use at 6 months, and childbirth prior to 24 months after service entry. We used logistic and Cox regression models, adjusted for age group, to compare outcomes of women in the Navy and Marine Corps who started basic training before and after their service branch's policy change with outcomes among women in the Army and Air Force. Compared to the longitudinal difference observed among women attending Army or Air Force basic training, changing policies to increase contraceptive access during Navy basic training in January 2015 increased contraception use from 33.1% of sailors to 39.2% of sailors before and after the policy change (interaction term odds ratio, 1.31; 95% confidence interval, 1.22-1.41) and long-acting reversible contraception use 11.0% to 22.7% (odds ratio, 1.78; 95% confidence interval, 1.50-2.08). However, this policy change was not associated with a decline in childbirth rates among sailors (7.5% versus 6.1%) relative to the change among women in the Army and Air Force over the same time period (interaction term hazard ratio, 0.90; 95% confidence interval, 0.79-1.03). The January 2016 Marine Corps policy change decreased contraception use (29.6% to 24.4%; odds ratio, 0.78; 95% confidence interval, 0.70-0.88), long-acting reversible contraception use 14.6% to 7.3% (odds ratio, 0.39; 95% confidence interval, 0.31-0.48), and increased childbirth rates (8.0% to 9.6%; hazard ratio, 1.26; 95% confidence interval, 1.03-1.55) among Marines compared to outcomes in the Army and Air Force over the same time period. Basic training contraceptive policy influences contraception use among junior enlisted servicewomen. Implementing best practices across the military may increase contraception use and decrease childbirth rates among junior enlisted servicewomen.


The independent associations of protein consumption with body fat and glycaemic control in adult Chinese.
High-protein diets were popular in weight control. However, the role of protein intake in adiposity and related metabolic conditions among general populations is not clear. We aimed to evaluate the associations of protein intake with adiposity and glycaemic control among adult Chinese in a nationwide population-based survey. The data were from China Health and Nutrition Survey 2009. A total of 9360 men and women aged 18 years or older were included. Body fat percentage was calculated using validated Chinese-specific equations. Dietary intake levels of macronutrients were evaluated by food-weighing approach combined with a 3-day food intake recall. Averagely, our participants have 12.5% energy intake from dietary protein. With multivariate adjustment including total energy intake, the odds ratios (95% CIs) of excessive adiposity (body fat percentage ≥ 20/30% for men/women), and central obesity (waist circumference ≥ 90/80 cm for men/women) were 1.51 (1.30, 1.75) and 1.40 (1.21, 1.62), respectively, comparing extreme quintiles of relative protein intake, while fat and carbohydrate were not associated with adiposity indices. Moreover, higher relative protein intake was associated with elevated concentration of fasting glucose (β ± SE: 1.233 ± 0.583), fasting insulin (23.211 ± 9.191), glycated hemoglobin (1.057 ± 0.369), and insulin resistance indicated by homeostasis model assessment of insulin resistance (7.558 ± 2.928) (all P < 0.05). Further adjusting for body mass index attenuated the associations. In Chinese adults, higher habitual protein consumption may be associated with higher adiposity and worse glycaemic control, independent of total energy intake.


Anti-inflammatory effects of octadecylamine-functionalized nanodiamond on primary human macrophages.
Chronic inflammatory disorders such as rheumatoid arthritis are characterized by excessive pro-inflammatory or "M1" activation of macrophages, the primary cells of the innate immune system. Current treatments include delivery of glucocorticoids (e.g. dexamethasone - Dex), which reduce pro-inflammatory M1 behaviour in macrophages. However, these treatments have many off-target effects on cells other than macrophages, resulting in broad immunosuppression. To limit such side effects, drug-incorporated nano- and microparticles may be used to selectively target macrophages via phagocytosis, because of their roles as highly effective phagocytes in the body. In this study, surface-modified nanodiamond (ND) was explored as a platform for the delivery of dexamethasone to macrophages because of ND's rich surface chemistry, which contributes to ND's high potential as a versatile drug delivery platform. After finding that octadecylamine-functionalized nanodiamond (ND-ODA) enhanced adsorption of Dex compared to carboxylated ND, the effects of Dex, ND-ODA, and Dex-adsorbed ND-ODA on primary human macrophage gene expression were characterized. Surprisingly, even in the absence of Dex, ND-ODA had strong anti-inflammatory effects, as determined by multiplex gene expression via NanoString and by protein secretion analysis via ELISA. ND-ODA also inhibited expression of M2a markers yet increased the expression of M2c markers and phagocytic receptors. Interestingly, the adsorption of Dex to ND-ODA further increased some anti-inflammatory effects, but abrogated the effect on phagocytic receptors, compared to its individual components. Overall, the ability of ND-ODA to promote anti-inflammatory and pro-phagocytic behaviour in macrophages, even in the absence of loaded drugs, suggests its potential for use as an anti-inflammatory therapeutic to directly target macrophages through phagocytosis.


The optimal screening interval for gastric cancer using esophago-gastro-duodenoscopy in Japan.
Gastric cancer is one of the most significant diseases, and esophago-gastro-duodenoscopy (EGD) is one of screening methods for gastric cancer. This study was conducted to identify the optimal screening interval for gastric cancer using EGD in healthy adults. A retrospective cohort study was conducted on 3,723 healthy participants without a known diagnosis of gastric cancer at baseline from January 2005 to December 2010. Participants underwent annual health screenings, including EGD, at the Center for Preventive Medicine at St Luke's International Hospital, a community teaching hospital in Japan. Participants with cytological abnormalities underwent further examination. A generalized estimating equation (GEE) was used to analyze the longitudinal data. We decided 0.5% of incidence of gastric cancer as a cutoff point for interval. The mean age (SD) of the participants was 55 (11) years, and 1,879 (50.5%) were male. During the study period, gastric cancer was detected in 35 participants. However, the incidence varied based on their ages. In the age groups <40, 40-49, 50-59, 60-69 and ≥70 years old, the 5-year cumulative incidences (95%CI) of gastric cancer were 0% (0-0%), 0.3% (0.1-1.0%), 1.0% (0.5-1.8%), 1.4% (0.8-2.4%) and 1.9% (0.8-3.8%), respectively. The odds ratios of the incidence of gastric cancer per year, which were evaluated using GEE models for the age groups 40-49, 50-59, 60-69 and ≥70 years old, were 1.51 (95%CI: 0.91-2.49), 1.94 (95%CI: 1.31-2.86), 1.59 (95%CI: 1.23-2.06) and 1.46 (95%CI: 1.06-2.02), respectively. A screening for gastric cancer using EGD may be appropriate annually for healthy people over 70 years old, every two or three years for people 60-69 years old and every four years for people 50-59 years old. People younger than 50 years old may only need repeat screenings every five years or more.


What happens to intraocular pressure at high altitude?
To investigate changes of intraocular pressure on ascent to high altitude. The Apex 2 medical research expedition provided the opportunity to measure intraocular pressure (IOP) and central corneal thickness (CCT) in 76 healthy lowlanders. They all arrived in La Paz, Bolivia (altitude, 3700 m), where they spent 4 days before being driven more than 2 hours to the Cosmic Physics Laboratory at Chacaltaya (5200 m) where they stayed for 7 days. IOP and CCT were measured with a hand-held tonometer and ultrasound pachymetry on the first, third, and seventh days at 5200 m. Pre- and postexpedition CCT and postexpedition IOP readings at sea-level were also measured. IOP increased significantly from baseline after acute exposure to altitude before returning to baseline with time. IOP at baseline, change in IOP from baseline, and IOP at altitude did not predict symptoms of acute mountain sickness (AMS) or development of high-altitude retinopathy (HAR). Acute exposure to altitude caused a statistically significant but clinically insignificant increase in IOP. This finding may be partially explained by the change in CCT. IOP returned to baseline levels and possibly lower with prolonged exposure to altitude. Changes in IOP at altitude are not predictive of symptoms of acute mountain sickness (AMS) or development of high-altitude retinopathy (HAR).


Molecular organization of 5S rDNA in sharks of the genus Rhizoprionodon: insights into the evolutionary dynamics of 5S rDNA in vertebrate genomes.
In this study, we attempted a molecular characterization of the 5S rDNA in two closely related species of carcharhiniform sharks, Rhizoprionodon lalandii and Rhizoprionodon porosus, as well as a further comparative analysis of available data on lampreys, several fish groups and other vertebrates. Our data show that Rhizoprionodon sharks carry two 5S rDNA classes in their genomes: a short repeat class (termed class I) composed of approximately 185 bp repeats, and a large repeat class (termed class II) arrayed in approximately 465 bp units. These classes were differentiated by several base substitutions in the 5S coding region and by completely distinct non-transcribed spacers (NTS). In class II, both species showed a similar composition for both the gene coding region and the NTS region. In contrast, class I varied extensively both within and between the two shark species. A comparative analysis of 5S rRNA gene sequences of elasmobranchs and other vertebrates showed that class I is closely related to the bony fishes, whereas the class II gene formed a separate cartilaginous clade. The presence of two variant classes of 5S rDNA in sharks likely maintains the tendency for dual ribosomal classes observed in other fish species. The present data regarding the 5S rDNA organization provide insights into the dynamics and evolution of this multigene family in the fish genome, and they may also be useful in clarifying aspects of vertebrate genome evolution.


Imaging-assisted anticancer nanotherapy.
Cancer nanomedicines are submicrometer-sized formulations designed to improve the biodistribution of anticancer drugs, resulting in less off-target localization, altered toxicity profiles, improved target site accumulation and enhanced efficacy. Together, these beneficial features have resulted in the regulatory approval of about a dozen nanomedicines for the treatment of solid and hematological malignancies. In recent years, significant progress has been made in combining nanomedicines with imaging, to better understand key aspects of the tumor-targeted drug delivery process, and to address the high inter- and intra-individual heterogeneity in the Enhanced Permeability and Retention (EPR) effect. Strategies explored in this regard have included the use of traditional imaging techniques, companion diagnostics and nanotheranostics. Preclinically, integrating imaging in nanomedicine and drug delivery research has enabled the non-invasive and quantitative assessment of nanocarrier biodistribution, target site accumulation and (triggered) drug release. Clinically, imaging has been emerging as a promising tool for patient stratification, which is urgently needed to improve the translation of cancer nanomedicines. We here summarize recent progress in imaging-assisted anticancer nanotherapy and we discuss future strategies to improve the performance of cancer nanomedicines in patients.


Reorganization of cortical blood flow and transcranial magnetic stimulation maps in human subjects after upper limb amputation.
1. Two complimentary techniques were used to study cortical function in six human upper limb amputees: positron emission tomographic (PET) measurements of regional cerebral blood flow (rCBF) were made in subjects during limb movements to study activation of the primary motor (M1), primary somatosensory (S1), and association cortices; and electromyographic responses to transcranial magnetic stimulation (TMS) were measured in proximal upper limb muscles to assess the excitability of corticospinal neurons in subjects at rest. 2. To explore possible cortical mechanisms governing the phantom limb phenomenon, PET and TMS findings were compared between subjects with acquired, traumatic upper limb amputations (n = 3), in whom phantom limb symptoms were prominent, and congenital upper limb amputees (n = 3) without phantom limbs. 3. Paced shoulder movements were associated with significant blood flow increases in the contralateral M1/S1 cortex of both groups of amputees. In traumatic amputees, these increases were present over a wider area and were of significantly greater magnitude in the partially deafferented cortex contralateral to the amputation. In congenital amputees blood flow increases were also present over a wider area in the partially deafferented M1/S1 cortex, but their magnitude was not significantly different from that in the normally afferented M1/S1 cortex. 4. Abnormal blood flow increases also were present in the partially deafferented M1/S1 cortex of traumatic amputees during movement of the ipsilateral, intact arm. Abnormal ipsilateral M1/S1 responses were not present during movement of the intact arm in the congenital group. 5. TMS studies showed that the abnormal blood flow increases in the partially deafferented M1 cortex of traumatic amputees were associated with increased corticospinal excitability.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparing Propensity Score Methods for Creating Comparable Cohorts of Chiropractic Users and Nonusers in Older, Multiply Comorbid Medicare Patients With Chronic Low Back Pain.
Patients who use complementary and integrative health services like chiropractic manipulative treatment (CMT) often have different characteristics than do patients who do not, and these differences can confound attempts to compare outcomes across treatment groups, particularly in observational studies when selection bias may occur. The purposes of this study were to provide an overview on how propensity scoring methods can be used to address selection bias by balancing treatment groups on key variables and to use Medicare data to compare different methods for doing so. We described 2 propensity score methods (matching and weighting). Then we used Medicare data from 2006 to 2012 on older, multiply comorbid patients who had a chronic low back pain episode to demonstrate the impact of applying methods on the balance of demographics of patients between 2 treatment groups (those who received only CMT and those who received no CMT during their episodes). Before application of propensity score methods, patients who used only CMT had different characteristics from those who did not. Propensity score matching diminished observed differences across the treatment groups at the expense of reduced sample size. However, propensity score weighting achieved balance in patient characteristics between the groups and allowed us to keep the entire sample. Although propensity score matching and weighting have similar effects in terms of balancing covariates, weighting has the advantage of maintaining sample size, preserving external validity, and generalizing more naturally to comparisons of 3 or more treatment groups. Researchers should carefully consider which propensity score method to use, as using different methods can generate different results.


Sedentary behaviour and health: mapping environmental and social contexts to underpin chronic disease prevention.
The time that children and adults spend sedentary-put simply, doing too much sitting as distinct from doing too little physical activity-has recently been proposed as a population-wide, ubiquitous influence on health outcomes. It has been argued that sedentary time is likely to be additional to the risks associated with insufficient moderate-to-vigorous physical activity. New evidence identifies relationships of too much sitting with overweight and obesity, type 2 diabetes, cardiovascular disease, some cancers and other adverse health outcomes. There is a need for a broader base of evidence on the likely health benefits of changing the relevant sedentary behaviours, particularly gathering evidence on underlying mechanisms and dose-response relationships. However, as remains the case for physical activity, there is a research agenda to be pursued in order to identify the potentially modifiable environmental and social determinants of sedentary behaviour. Such evidence is required so as to understand what might need to be changed in order to influence sedentary behaviours and to work towards population-wide impacts on prolonged sitting time. In this context, the research agenda needs to focus particularly on what can inform broad, evidence-based environmental and policy initiatives. We consider what has been learned from research on relationships of environmental and social attributes and physical activity; provide an overview of recent-emerging evidence on relationships of environmental attributes with sedentary behaviour; argue for the importance of conducting international comparative studies and addressing life-stage issues and socioeconomic inequalities and we propose a conceptual model within which this research agenda may be addressed.


A novel member of the family Hepeviridae from cutthroat trout (Oncorhynchus clarkii).
Beginning in 1988, the Chinook salmon embryo (CHSE-214) cell line was used to isolate a novel virus from spawning adult trout in the state of California, USA. Termed the cutthroat trout (Oncorhynchus clarkii) virus (CTV), the small, round virus was not associated with disease, but was subsequently found to be present in an increasing number of trout populations in the western USA, likely by a combination of improved surveillance activities and the shipment of infected eggs to new locations. Here, we report that the full length genome of the 1988 Heenan Lake isolate of CTV consisted of 7269 nucleotides of positive-sense, single-stranded RNA beginning with a 5' untranslated region (UTR), followed by three open reading frames (ORFs), a 3' UTR and ending in a polyA tail. The genome of CTV was similar in size and organization to that of Hepatitis E virus (HEV) with which it shared the highest nucleotide and amino acid sequence identities. Similar to the genomes of human, rodent or avian hepeviruses, ORF 1 encoded a large, non-structural polyprotein that included conserved methyltransferase, protease, helicase and polymerase domains, while ORF 2 encoded the structural capsid protein and ORF 3 the phosphoprotein. Together, our data indicated that CTV was clearly a member of the family Hepeviridae, although the level of amino acid sequence identity with the ORFs of mammalian or avian hepeviruses (13-27%) may be sufficiently low to warrant the creation of a novel genus. We also performed a phylogenetic analysis using a 262nt region within ORF 1 for 63 isolates of CTV obtained from seven species of trout reared in various geographic locations in the western USA. While the sequences fell into two genetic clades, the overall nucleotide diversity was low (less than 8.4%) and many isolates differed by only 1-2 nucleotides, suggesting an epidemiological link. Finally, we showed that CTV was able to form persistently infected cultures of the CHSE-214 cell line that may have use in research on the biology or treatment of hepevirus infections of humans or other animals.


Agreement between anthropometric and dual-energy X-ray absorptiometry assessments of lower-limb volumes and composition estimates in youth-club rugby athletes.
The purpose of this study was to assess the agreement of lower-limb volume estimates based on anthropometry and dual-energy X-ray absorptiometry (DXA) as a reference method in male rugby athletes. Predictive models using body mass and skinfolds were tested to improve the relative agreement between protocols (anthropometry, DXA). Rugby players (n = 41; 19.9 ± 2.2 years) volunteered for the study. Lower-limb total and fat-free volumes were estimated by anthropometry and also derived using DXA. Cross-validation between the anthropometry technique and DXA was then performed. Lower-limb volume estimates by anthropometry overestimated reference values and tended to be further from the reference values with the increase of scale. For the total sample, standard errors of measurement for volume estimates by anthropometry were 1.99 L and 1.34 L for total and fat-free volumes, respectively. Correlations with reference values were 0.81 for lower-limb volume and 0.90 for lower-limb fat-free volume. Correlations between estimated prediction equations and reference values showed higher correlations (r = 0.96 for lower-limb volume and r = 0.93 for lower-limb fat-free volume) compared with anthropometric estimates. Overall, the agreement of anthropometry method to quantify lower-limb volumes with DXA as a reference in young adult rugby players is acceptable and is a practical method when more expensive and complex techniques are not available. The consideration of body mass and lower-limb skinfolds increases the precision of lower-limb volume estimates using anthropometry in the young adult rugby players.


The investigation of adrenal involvement in carbapenem resistant Acinetobacter baumannii sepsis: experimental mouse model
In the last years, incidence of carbapenem resistant Acinetobacter baumannii sepsis is increasing with high mortality. However, it is not clear whether this is due to inadequate antimicrobial choice or a more severe clinical course. We aimed to evaluate the inflammation and adrenal involvement in the carbapenem resistant A. baumannii by using experimental mouse model sepsis. Balb/c female mice were randomly put into control and three sepsis groups ( A. baumannii susceptible to carbapenem-CSAB-, A. baumannii resistant to carbapenem-CRAB-, Escherichia coli). A total of sixty mice were included in this study with each group having 15 mice. Mice were sacrificed 72 h after bacterial inoculation, and blood was taken from each mouse for the assessment of cytokines and corticosterone. Both adrenal glands were dissected; one was used for culture and the other was used for histopathological examination. Bacterial loads of organs were calculated as CFU/g. The histopathological changes, bacterial levels in adrenal and cytokine and corticosterone levels were assessed and compared among the groups. The bacterial level was higher in E. coli (108, 45 ±30, 55 log10 CFU/g) (mean±SD) than other sepsis groups. The lowest level of corticosterone was observed in the E. coli group (p < 0.001). TNF alpha level was highest in the CRAB and E. coli group and this difference was statistically significant than control group (p < 0.05). The IL-6 level in CRAB was significantly higher than the control group (10, 20 pg/mL). The adrenal gland congestion was significantly severe in all the sepsis groups compared to the control. In the group comparison, congestion was significantly more severe in the E. coli group than in CSAB and CRAB groups. Adrenal involvement and inflammatory reactions are seen in E. coli sepsis and in CRAB sepsis. These findings will be focused on in future clinical trials.


Nothing to sneeze at: a dynamic and integrative computational model of an influenza A virion.
The influenza virus is surrounded by an envelope composed of a lipid bilayer and integral membrane proteins. Understanding the structural dynamics of the membrane envelope provides biophysical insights into aspects of viral function, such as the wide-ranging survival times of the virion in different environments. We have combined experimental data from X-ray crystallography, nuclear magnetic resonance spectroscopy, cryo-electron microscopy, and lipidomics to build a model of the intact influenza A virion. This is the basis of microsecond-scale coarse-grained molecular dynamics simulations of the virion, providing simulations at different temperatures and with varying lipid compositions. The presence of the Forssman glycolipid alters a number of biophysical properties of the virion, resulting in reduced mobility of bilayer lipid and protein species. Reduced mobility in the virion membrane may confer physical robustness to changes in environmental conditions. Our simulations indicate that viral spike proteins do not aggregate and thus are competent for multivalent immunoglobulin G interactions.


The local and referred pain patterns of the longus colli muscle.
The longus colli muscle is a neck flexor believed to play an important role in pain originating in the neck region, including pain resulting from whiplash injuries. Despite the clinical importance attributed to it, the pain referral pattern of the longus colli has previously been described only in a small cohort of subjects. Here, we aim to delineate the pain referral pattern of the longus colli muscle. Thirty-five healthy volunteers underwent deep massage of the longus colli followed by dry needling of the muscle. The subjects depicted the distribution of the pain they experienced on a blank manikin. Their drawings were digitized and used to produce pain pattern histogram maps. The pain referral pattern during deep massage and needling of the longus colli was primarily local, with referral to the ipsilateral ear and lateral to the ipsilateral eye. Some subjects reported pain on the contralateral side of the neck.


War, space, and the evolution of Old World complex societies.
How did human societies evolve from small groups, integrated by face-to-face cooperation, to huge anonymous societies of today, typically organized as states? Why is there so much variation in the ability of different human populations to construct viable states? Existing theories are usually formulated as verbal models and, as a result, do not yield sharply defined, quantitative predictions that could be unambiguously tested with data. Here we develop a cultural evolutionary model that predicts where and when the largest-scale complex societies arose in human history. The central premise of the model, which we test, is that costly institutions that enabled large human groups to function without splitting up evolved as a result of intense competition between societies-primarily warfare. Warfare intensity, in turn, depended on the spread of historically attested military technologies (e.g., chariots and cavalry) and on geographic factors (e.g., rugged landscape). The model was simulated within a realistic landscape of the Afroeurasian landmass and its predictions were tested against a large dataset documenting the spatiotemporal distribution of historical large-scale societies in Afroeurasia between 1,500 BCE and 1,500 CE. The model-predicted pattern of spread of large-scale societies was very similar to the observed one. Overall, the model explained 65% of variance in the data. An alternative model, omitting the effect of diffusing military technologies, explained only 16% of variance. Our results support theories that emphasize the role of institutions in state-building and suggest a possible explanation why a long history of statehood is positively correlated with political stability, institutional quality, and income per capita.


The multidrug resistance P-glycoprotein. Oligomeric state and intramolecular interactions.
The human multidrug resistance P-glycoprotein (P-gp), a member of the ATP-binding cassette (ABC) superfamily of transporters, is frequently responsible for the failure of chemotherapy by virtue of its ability to export hydrophobic cytotoxic drugs from cells. Elucidating the inter- and intramolecular interactions of this protein is critical to understanding its cellular function and mechanism of action. Toward this end, we have used both biochemical and genetic techniques to probe potential oligomerization interactions of P-gp. Differentially epitope-tagged P-gp molecules did not co-immunoprecipitate when co-expressed in HEK293 cells or when co-translated in vitro, demonstrating that P-gp is monomeric in both the presence and absence of detergents. The two cytoplasmic domains of P-gp did not interact with each other in vivo when co-expressed as gene fusions in yeast. In contrast, the homologous domains of the transporter associated with antigen processing (TAP), which reside on separate polypeptides and must form a heterodimeric transporter (TAP1/TAP2), did interact in this system, suggesting a role for these domains in TAP dimerization. Implications for understanding the subunit organization of ABC transporters are discussed.


Absence of the RNF213 p.R4810K variant may indicate a severe form of pediatric moyamoya disease in Japanese patients.
The authors' objective was to investigate the influence of the RNF213 p.R4810K variant on the clinical presentation and outcomes of Japanese pediatric patients with moyamoya disease. A total of 129 Japanese patients with pediatric-onset moyamoya disease (onset age ≤ 15 years) who visited the authors' department from 2012 to 2020 participated in this study. After RNF213 p.R4810K genotyping of each patient was performed, the relationship between genotype and clinical presentation or outcomes, including onset age, initial presentation, surgical outcomes, and subsequent cerebrovascular events, was evaluated. Patients without the p.R4810K variant were tested for RNF213 variants other than p.R4810K. The authors especially focused on the results of patients who presented with moyamoya disease at younger than 1 year of age (infantile onset). Compared with the patients with heterozygous variants, patients without the p.R4810K variant were younger at onset (7.1 ± 3.7 vs 4.4 ± 0.9 years), and all 4 patients with infantile onset lacked the p.R4810K variant. A greater proportion of patients without the p.R4810K variant presented with infarction than patients with the heterozygous variant (24.0% vs 7.6%) and a decreased proportion presented with transient ischemic attack (36.0% vs 71.7%). No significant correlation was observed between p.R4810K genotype and clinical outcomes, including surgical outcomes and subsequent cerebrovascular events; however, a decreased proportion of patients without the p.R4810K variant had good surgical outcomes compared with that of patients with the heterozygous variant (76.5% vs 92.2%). Among the 25 patients without the p.R4810K variant, 8 rare variants other than p.R4810K were identified. Three of 4 patients with infantile onset had RNF213 variants other than p.R4810K, which had a more severe functional effect on this gene than p.R4810K. Absence of the RNF213 p.R4810K variant may be a novel biomarker for identification of a severe form of pediatric moyamoya disease.


Clinical outcomes and radiological instability following decompressive lumbar laminectomy for degenerative spinal stenosis: a comparison of patients undergoing concomitant arthrodesis versus decompression alone.
One hundred twenty-four patients with degenerative lumbar stenosis underwent decompression with fusion (32 patients) and without fusion (92 patients) during a 30-month period between 1986 and 1988. Patient-reported satisfaction at a mean follow-up period of 5.8 years (range 4.6-6.8 years) revealed a 79% good or fair outcome and a 21% poor outcome (26 patients). Seven patients (6%) developed lumbar instability, three patients (2%) developed new stenosis at an adjacent unoperated level, and three patients (2%) developed a new disc herniation between 2 and 5 years after surgery. Progressive postoperative spondylolisthesis occurred in 31% of patients with normal preoperative alignment (mean 7.8 mm, range 2-20 mm) and in 73% of patients with preoperative subluxation (mean 5.1 mm, range 2-13 mm) in whom fusion was not attained. Radiological progression did not correlate well with patient-reported outcome. The major conclusions from this study are the following: 1) the majority of patients respond well to this surgery, but complication (22%) and late deterioration (10%) rates are not insignificant; 2) radiological instability is common after decompression for degenerative lumbar spinal stenosis, but this correlates poorly with clinical outcome; 3) there are no definitive clinical or radiological factors that preoperatively predict patients at risk for a poor outcome; 4) post-operative radiological instability is more likely to occur when the following criteria are present: preoperative spondy-degenerated L-4 or a markedly degenerated L-3 disc; and when a radical and extensive decompression greater than one level is planned; and 5) the group at greatest risk for a poor outcome consists of those patients with normal preoperative alignment who do not suffer slippage following surgery.


[Health counseling in primary care doctors' offices: a new wind! The Health Coaching Program of the Swiss College of Primary Care Medicine].
The Health Coaching Program facilitates health behavior counseling in all areas of primary medical care: prevention, therapy and rehabilitation, i.e. wherever the patient is the decisive agent of change. Health Coaching gives the patient the main role. The physician becomes his coach. Health Coaching offers skills training and simple algorithms with a colour-coded visual tool to assist patient and physician through the 4 steps of developing awareness, building motivation, preparing a personal health project and implementing it. Health Coaching was tested successfully by 20 family doctors during 12 months: of 1045 patients invited 91% enrolled; 37% completed all four steps; one half achieved a positive behavior change. Acceptance and feasibility were high in physicians and patients. Nationwide dissemination is now in preparation.


EAACI Biologicals Guidelines-dupilumab for children and adults with moderate-to-severe atopic dermatitis.
Atopic dermatitis imposes a significant burden on patients, families and healthcare systems. Management is difficult, due to disease heterogeneity, co-morbidities, complexity in care pathways and differences between national or regional healthcare systems. Better understanding of the mechanisms has enabled a stratified approach to the management of atopic dermatitis, supporting the use of targeted treatments with biologicals. However, there are still many issues that require further clarification. These include the definition of response, strategies to enhance the responder rate, the duration of treatment and its regimen (in the clinic or home-based), its cost-effectiveness and long-term safety. The EAACI Guidelines on the use of dupilumab in atopic dermatitis follow the GRADE approach in formulating recommendations for each outcome and age group. In addition, future approaches and research priorities are discussed.


Lanosterol Disrupts Aggregation of Human γD-Crystallin by Binding to the Hydrophobic Dimerization Interface.
Cataracts are a leading cause of vision impairment, which stem from the misfolding and aggregation of crystallins in the eye lens. Despite its prevalence and severity, the detailed mechanism by which misfolded crystallins aggregate into cataracts remains elusive. Recently, in vitro and in vivo experiments demonstrated that lanosterol, a steroid-type compound found in human and animal eyes, can not only prevent cataract formation but also reverse the formation. Inspired by these experimental observations, we investigate the preventive activity of lanosterol in the aggregate formation of human γD-crystallins (HγD-Crys) using all atom molecular dynamics (MD) simulation and free energy perturbation (FEP) techniques. Our results reveal that lanosterol preferentially binds to the HγD-Crys hydrophobic dimerization interface, in particular, to the structured C-terminus (near residues 135-165) with a stronger binding affinity than the unfolded N-terminus. Furthermore, we observe that the C-terminal binding is more favorable than lanosterol self-aggregation, further attesting to lanosterol's efficacy. Finally, we compare the binding free energy of lanosterol with cholesterol using alchemical transformation and discuss the possible correlation of the molecular geometry of steroids with binding affinity.


Suppression of subthalamic nucleus activity by micromagnetic stimulation.
Magnetic stimulation delivered via 0.5-mm diameter coils was recently shown to activate retinal neurons; the small coil size raises the possibility that micromagnetic stimulation ( μMS) could underlie a new generation of implanted neural prosthetics. Such an approach has several inherent advantages over conventional electric stimulation, including the potential for selective activation of neuronal targets as well as less susceptibility to inflammatory responses. The viability of μMS for some applications, e.g., deep brain stimulation (DBS), may require suppression (rather than creation) of neuronal activity, however, and therefore we explore here whether (μMS) could, in fact, suppress activity. While single pulses elicited weak and inconsistent spiking in neurons of the mouse subthalamic nucleus (in vitro), repetitive stimulation effectively suppressed activity in ∼ 70% of targeted neurons. This is the same percentage suppressed by conventional electric stimulation; with both modalities, suppression occurred only after an initial increase in spiking. The latency to the onset of suppression was inversely correlated to the energy of the stimulus waveform: larger amplitudes and lower frequencies had the fastest onset of suppression. These findings continue to support the viability of μMS as a next-generation implantable neural prosthetic.


Change in salivary physiological stress markers by spa bathing.
We assessed the stress relief effect of spa bathing by measuring sensitive salivary stress markers, cortisol and chromogranin A (CgA). From 12 healthy males, saliva samples were collected immediately before and after spa bathing, and 30 min after that. Salivary cortisol and CgA levels were determined by ELISA. Salivary cortisol levels decreased after spa bathing. This tendency was more pronounced in individuals with higher levels of stress. The high-stress group showed lower salivary CgA levels after spa bathing, while the low-stress group higher salivary CgA levels in the same condition. These findings suggest that the spa bathing has a moderate affect on the stress relief.


The Aging Males' Symptoms (AMS) scale: review of its methodological characteristics.
The current paper reviews data from different sources to get a closer impression on the psychometric and other methodological characteristics of the Aging Males' Symptoms (AMS) scale gathered recently. The scale was designed and standardized as self-administered scale to (a) to assess symptoms of aging (independent from those which are disease-related) between groups of males under different conditions, (b) to evaluate the severity of symptoms over time, and (c) to measure changes pre- and post androgen replacement therapy. The scale is in widespread use (14 languages). Original data from different studies in many countries were centrally analysed to evaluate reliability and validity of the AMS. Reliability measures (consistency and test-retest stability) were found to be good across countries, although the sample size was sometimes small. The internal structure of the AMS in healthy and androgen deficient males, and across countries was sufficiently similar to conclude that the scale really measures the same phenomenon. The sub-scores and total score correlations were high (0.8-0.9) but lower among the sub-scales (0.5-0.7). This however suggests that the subscales are not fully independent. The comparison with other scales for aging males or screening instruments for androgen deficiency showed sufficiently good correlations, illustrating a good criterion-oriented validity. The same is true for the comparison with the generic quality-of-life scale SF36 where also high correlation coefficients have been shown. Methodological analyses of a treatment study of symptomatic males with testosterone demonstrated the ability of the AMS scale to measure treatment effect, irrespective of the severity of complaints before therapy. It was also shown that the AMS result can predict the independently generated (physician's) opinion about the individual treatment effect. The currently available methodological evidence points towards a high quality of the AMS scale to measure and to compare HRQoL of aging males over time or before/after treatment, it suggests a high reliability and high validity as far as the process of construct validation could be pressed ahead yet. But certainly more data will become available, particularly from ongoing clinical studies.


T cell receptor signal strength in Treg and iNKT cell development demonstrated by a novel fluorescent reporter mouse.
The ability of antigen receptors to engage self-ligands with varying affinity is crucial for lymphocyte development. To further explore this concept, we generated transgenic mice expressing GFP from the immediate early gene Nr4a1 (Nur77) locus. GFP was up-regulated in lymphocytes by antigen receptor stimulation but not by inflammatory stimuli. In T cells, GFP was induced during positive selection, required major histocompatibility complex for maintenance, and directly correlated with the strength of T cell receptor (TCR) stimulus. Thus, our results define a novel tool for studying antigen receptor activation in vivo. Using this model, we show that regulatory T cells (T(reg) cells) and invariant NKT cells (iNKT cells) perceived stronger TCR signals than conventional T cells during development. However, although T(reg) cells continued to perceive strong TCR signals in the periphery, iNKT cells did not. Finally, we show that T(reg) cell progenitors compete for recognition of rare stimulatory TCR self-ligands.


Titanium oral implants: surface characteristics, interface biology and clinical outcome.
Bone-anchored titanium implants have revolutionized oral healthcare. Surface properties of oral titanium implants play decisive roles for molecular interactions, cellular response and bone regeneration. Nevertheless, the role of specific surface properties, such as chemical and phase composition and nanoscale features, for the biological in vivo performance remains to be established. Partly, this is due to limited transfer of state-of-the-art preparation techniques to complex three-dimensional geometries, analytical tools and access to minute, intact interfacial layers. As judged by the available results of a few randomized clinical trials, there is no evidence that any particular type of oral implant has superior long-term success. Important insights into the recruitment of mesenchymal stem cells, cell-cell communication at the interface and high-resolution imaging of the interface between the surface oxide and the biological host are prerequisites for the understanding of the mechanisms of osseointegration. Strategies for development of the next generation of material surface modifications for compromised tissue are likely to include time and functionally programmed properties, pharmacological modulation and incorporation of cellular components.


Evidence-based practice is a time of opportunity for nursing.
The debate regarding evidence-based practice (EBP) continues to rage within nursing. This article reviews the existing situation regarding policy and the ability of nurses to engage with EBP. Recommendations are made regarding moving the debate away from a focus on barriers to EBP towards seizing it as an opportunity. The article suggests that a fundamental change in attitude is required by nurses in order for the nursing profession to progress. There is a need to recognize that the nature of health services are changing and that nurses need to grasp the opportunities this makes available, but in order to do this nurses need support.


Site-specific conjugation of an antibody on a gold nanoparticle surface for one-step diagnosis of prostate specific antigen with dynamic light scattering.
Small dimensions of gold nanoparticles (AuNPs) necessitate antibodies to be immobilized in an oriented fashion in order to conserve their antigen binding activity for proper function. In this study, we used the previously described UV-NBS method to site-specifically incorporate a thioctic acid (TA) functionality into antibodies at the conserved nucleotide-binding site (NBS). Modified antibodies were immobilized on the AuNP surface in an oriented manner utilizing the newly incorporated TA functionality while maintaining the antibody structure and activity. The resulting antibody functionalized AuNPs via the UV-NBS method demonstrated significantly enhanced antigen detection capabilities and improved antigen detection sensitivity with a high level of selectivity when compared to other commonly used AuNP functionalization methods. Our results demonstrate that the limit of detection (LOD) for AuNPs functionalized via the UV-NBS method was 55 pM PSA, which is 40, 851, and 5873-fold improved over the other immobilization methods: EDC-NHS, thiol reduction, and ionic interaction, respectively. Consequently, the UV-NBS method provides a universal, site-specific functionalization method that generates highly sensitive and more stable antibody functionalized AuNPs which are amenable to any available detection and treatment assay with potential significant implications.


Effect of Environmental Temperature and Humidity on Permethrin Biomarkers of Exposure in U.S. Soldiers Wearing Permethrin-Treated Uniforms.
Environmental factors, including high temperature and humidity, can influence dermal absorption of chemicals. Soldiers can be dermally exposed to permethrin while wearing permethrin-treated uniforms. This study aimed at examining the effects of high temperature and a combined high temperature and humid environment on permethrin absorption compared with ambient conditions when wearing a permethrin-treated uniform. Twenty-seven male enlisted soldiers wore study-issued permethrin-treated army uniforms for 33 consecutive hours in three different environments: 1) simulated high temperature (35°C, 40% relative humidity [rh]) (n = 10), 2) simulated high temperature and humidity (30°C, 70% rh) (n = 10), and 3) ambient conditions (13°C, 60% rh) (n = 7). Spot urine samples, collected at 21 scheduled time points before, during, and after wearing the study uniforms, were analyzed for permethrin exposure biomarkers (3-phenoxybenzoic acid, cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid) and creatinine. Biomarker concentrations were 60-90% higher in the heat and combined heat/humidity groups (P < 0.001-0.022) than the ambient group. Also, the average daily permethrin dose, calculated 12 hours after removing the treated uniforms, was significantly higher in the heat (P = 0.01) and the heat/humidity (P = 0.03) groups than the ambient group. There were no significant differences in biomarker concentrations or computed average daily dose between the heat and the heat/humidity groups. Both hot and combined hot and humid environmental conditions significantly increased permethrin absorption in soldiers wearing permethrin-treated uniforms.


Mechanism of inhibition of HIV-1 reverse transcriptase by nonnucleoside inhibitors.
The mechanism of inhibition of HIV-1 reverse transcriptase by three nonnucleoside inhibitors is described. Nevirapine, O-TIBO, and CI-TIBO each bind to a hydrophobic pocket in the enzyme-DNA complex close to the active site catalytic residues. Pre-steady-state kinetic analysis was used to establish the mechanism of inhibition by these noncompetitive inhibitors. Analysis of the pre-steady-state burst of DNA polymerization indicated that inhibitors blocked the chemical reaction, but did not interfere with nucleotide binding or the nucleotide-induced conformational change. Rather, in the presence of saturating concentrations of the inhibitors, the nucleoside triphosphate bound tightly (Kd, 100 nM), but nonproductively. The data suggest that an inhibitor combining the functionalities of a nonnucleoside inhibitor and a nucleotide analog could bind very tightly and specifically to reverse transcriptase and could be effective in the treatment of AIDS.


Facilitators and barriers to weight loss and weight loss maintenance: a qualitative exploration.
The present study aimed to explore facilitators and barriers to weight loss (WL) and weight loss maintenance (WLM) in women who participated in a primary, 18-week comparative trial that promoted WL with an energy-restricted diet. Twenty-three women participated in seven focus groups conducted by a moderator and co-facilitator using open-ended questions and probes. Focus groups were held in a private room and audio tape-recorded. Tapes were transcribed verbatim and thematic analysis was used to evaluate transcripts for common themes. Accountability to others, social support, planning ahead, awareness and mindfulness of food choices, basic nutrition education, portion control, exercise, and self-motivation were perceived as key facilitators for WL and WLM by women. Identified barriers included life transitions, health status changes, internal factors, environmental pressures, lack of accountability and an absence of social support. Future interventions should address these salient facilitators and barriers to promote sustainable changes in women across their WL and WLM journeys.


Activated eosinophils upregulate the metastasis suppressor molecule E-cadherin on prostate tumor cells.
Cell adhesion molecules (CAMs) play an important role in cancer metastasis by facilitating attachment to vascular endothelia, invasion and spread into secondary tissue sites. We have shown that activated eosinophils (EosA) inhibited the growth of prostate cancer (Pca) cells in vitro. In the present study, we examined the ability of EosA 24 hr conditioned supernatants (EosAcs) to modulate the expression of ICAM-1, VCAM-1, ELAM-1, E-cadherin and N-cadherin expression on human Pca cell lines, Du-145 and PC-3 by flow cytometry. TNF-alpha, IL-10 and IL-12 were also evaluated. ICAM-1, expressed on PC-3 and DU 145 cells, was enhanced by TNF-alpha and IL-10. ELAM-1 was present on DU 145 cells but absent on PC-3. TNF-alpha and IL-10 enhanced ELAM-1 on DU 145, but EosA 24 hr supematants failed to do so. All three cytokines, namely IL-10, IL-12 and TNF-alpha-induced ELAM-1 on PC-3 tumor cells. Although VCAM-1 was absent on DU 145 and PC-3 cells, it was expressed on DU-145 cells after exposure to EosA: tumor cell co-cultures, and was expressed on PC-3 following exposure to IL-10 and IL-12. N-cadherin and E-cadherin were both expressed on DU-145. While N-cadherin was expressed on PC-3 cells, E-cadherin was not. N-cadherin was enhanced on DU-145 and PC-3 cells following exposure to EosA co-culture and upregulated on PC-3 by IL-10 and EosA 24 hr supernatants, but decreased by IL-12. E-cadherin was up-regulated on DU 145 cells following co-culture with EosA and was induced on PC-3 by IL-10 and IL-12, but not by EosA co-culture and 24 hr supematants. In conclusion, inflammatory and non-inflammatory cytokines modulate CAM expression on Pca cells; EosA and EosA 24 hr supernatants also exerted modulatory activity of CAM expression. Most significantly, the metastasis suppressor molecule, E-cadherin was enhanced on DU 145 cells by EosA and induced on PC-3 by IL-10 and IL-12 both of which are produced by EosA. This suggests potential use of these cytokines in immunotherapeutic strategies for prostate cancer and its metastasis.


Altered architecture and functional consequences of the mesolimbic dopamine system in cannabis dependence.
Cannabinoid withdrawal produces a hypofunction of mesencephalic dopamine neurons that impinge upon medium spiny neurons (MSN) of the forebrain. After chronic treatment with two structurally different cannabinoid agonists, Delta(9)-tetrahydrocannabinol and CP55 940 (CP) rats were withdrawn spontaneously and pharmacologically with the CB1 antagonist SR141716A (SR). In these two conditions, evaluation of tyrosine hydroxylase (TH)-positive neurons revealed significant morphometrical reductions in the ventrotegmental area but not substantia nigra pars compacta of withdrawn rats. Similarly, confocal analysis of Golgi-Cox-stained sections of the nucleus accumbens revealed a decrease in the shell, but not the core, of the spines' density of withdrawn rats. Administration of the CB1 antagonist SR to control rats, provoked structural abnormalities reminiscent of those observed in withdrawal conditions and support the regulatory role of cannabinoids in neurogenesis, axonal growth and synaptogenesis by acting as eu-proliferative signals through the CB1 receptors. Further, these measures were incorporated into a realistic computational model that predicts a strong reduction in the excitability of morphologically altered MSN, yielding a significant reduction in action potential output. These pieces of evidence support the tenet that withdrawal from addictive compounds alters functioning of the mesolimbic system and provide direct morphological evidence for functional abnormalities associated with cannabinoid dependence at the level of dopaminergic neurons and their postsynaptic counterpart and are coherent with recent hypothesis underscoring a hypodopaminergic state as a distinctive feature of the 'addicted brain'.


Study protocol: E-freeze - freezing of embryos in assisted conception: a randomised controlled trial evaluating the clinical and cost effectiveness of a policy of freezing embryos followed by thawed frozen embryo transfer compared with a policy of fresh embryo transfer, in women undergoing in vitro fertilisation.
Infertility affects one in seven couples; many of these need in vitro fertilisation (IVF). IVF involves external hormones to stimulate a woman's ovaries to produce eggs which are harvested surgically. Embryos, created in the laboratory by mixing eggs with sperm, are grown in culture for a few days before being replaced within the uterus (fresh embryo transfer). Spare embryos are usually frozen with a view to transfer at a later point in time - especially if the initial fresh transfer does not result in a pregnancy. Despite improvements in technology, IVF success rates remain low with an overall live birth rate of 25-30% per treatment. Additionally, there are concerns about health outcomes for mothers and babies conceived through IVF, particularly after fresh embryo transfer, including maternal ovarian hyperstimulation syndrome (OHSS) and preterm delivery. It is believed that high levels of hormones during ovarian stimulation could create a relatively hostile environment for embryo implantation whilst increasing the risk of OHSS. It has been suggested that freezing all embryos with the intention of thawing and replacing them within the uterus at a later stage (thawed frozen embryo transfer) instead of fresh embryo transfer, may lead to improved pregnancy rates and fewer complications. We aim to compare the clinical and cost effectiveness of fresh and thawed frozen embryo transfer, with the primary aim of identifying any difference in the chance of having a healthy baby. E-Freeze is a pragmatic, multicentre two-arm parallel group randomised controlled trial where women aged ≥18 and < 42 years, with at least three good quality embryos are randomly allocated to receive either a fresh or thawed frozen embryo transfer. The primary outcome is a healthy baby, defined as a term, singleton, live birth with appropriate weight for gestation. Cost effectiveness will be calculated from a healthcare and societal perspective. E-Freeze will determine the relative benefits of fresh and thawed frozen embryo transfer in terms of improving the chance of having a healthy baby. The results of this pragmatic study have the potential to be directly transferred to clinical practice. ISRCTN registry: ISRCTN61225414 . Date assigned 29/12/2015.


Corneal biomechanics, thickness and optic disc morphology in children with optic disc tilt.
To determine the associations between corneal biomechanical parameters as measured by the Reichert Ocular Response Analyser (ORA) and disc morphology and retinal nerve fibre layer thickness (RNFL) measured by the Heidelberg Retinal Tomograph (HRT) II in Singaporean children. This is a cross-sectional study conducted on a subset of children enrolled in the Singapore Cohort Study of the Risk Factors of Myopia (SCORM). Corneal hysteresis (CH), corneal resistance factor (CRF) and central corneal thickness (CCT) were measured with the ORA. Optic disc morphology and RNFL thickness were assessed by the HRT II. Cycloplegic refraction and ultrasound A-scans were also performed, and disc tilt was assayed from stereo photographs. 102 subjects (mean age 12.01 (SD 0.57) years; range 11-14 years) were included in the study. The mean CH was 12.00 (1.40) mm Hg, the mean CRF was 11.99 (1.65) mm Hg, and the mean CCT was 581.12 (33.53) mum. Eyes with tilted discs had significantly longer axial lengths and more myopic refraction than eyes without tilted discs. There were no significant correlations between CH, CRF or CCT and the HRT II parameters, after the application of the Bonferroni correction. When stratified for disc tilt, however, the global disc area was significantly correlated with CCT (r = -0.49, p = 0.001). Corneal biomechanical properties as measured with the ORA do not vary with optic disc parameters or RNFL. Central corneal thickness is correlated with disc area in Singaporean schoolchildren with tilted discs. This relationship may influence glaucoma risk in myopic subjects.


Skeletal Muscle Ratio: A Complete Mediator of Physical Activity and HbA1C in Type 2 Diabetes.
An increase in the physical activity level reduces body weight, decreases body fat, increases skeletal muscle mass, and improves serum glucose; however, the influence of body composition parameters on the relationship between physical activity and serum glucose remains unclear. This study investigated whether skeletal muscle and visceral fat affect the relationship between high physical activity and long-term serum glucose goals. This cross-sectional study recruited patients with type 2 diabetes. The Chinese version of the International Physical Activity Questionnaire was used for estimating the physical activity level, and a bioimpedance device was used to measure the skeletal muscle ratio (skeletal muscle mass/total body weight, %) and visceral fat area (cm2). Hierarchical logistic regression models and mediation tests were conducted according to Hayes' procedures. Of the total 543 Chinese individuals with type 2 diabetes enrolled, HbA1C levels of fewer than half (n = 243, 44.8%) met the target of ≤7.0%. The skeletal muscle ratio was found to be a complete mediator (OR = 0.920, 95% CI: 0.848 to 0.998; indirect effect: -0.238, 95% CI: -0.525 to -0.020) of the relationship between high physical activity and the target HbA1C level after controlling for visceral fat area (indirect effect: -0.013, 95% CI: -0.183 to 0.156), age, time since diabetes diagnosis, and rice intake. Nurses should include an increase in the skeletal muscle ratio as an objective in physical activity interventions for patients with type 2 diabetes to help them achieve their long-term serum glucose goals.


Reconstruction and modeling protein translocation and compartmentalization in Escherichia coli at the genome-scale.
Membranes play a crucial role in cellular functions. Membranes provide a physical barrier, control the trafficking of substances entering and leaving the cell, and are a major determinant of cellular ultra-structure. In addition, components embedded within the membrane participate in cell signaling, energy transduction, and other critical cellular functions. All these processes must share the limited space in the membrane; thus it represents a notable constraint on cellular functions. Membrane- and location-based processes have not yet been reconstructed and explicitly integrated into genome-scale models. The recent genome-scale model of metabolism and protein expression in Escherichia coli (called a ME-model) computes the complete composition of the proteome required to perform whole cell functions. Here we expand the ME-model to include (1) a reconstruction of protein translocation pathways, (2) assignment of all cellular proteins to one of four compartments (cytoplasm, inner membrane, periplasm, and outer membrane) and a translocation pathway, (3) experimentally determined translocase catalytic and porin diffusion rates, and (4) a novel membrane constraint that reflects cell morphology. Comparison of computations performed with this expanded ME-model, named iJL1678-ME, against available experimental data reveals that the model accurately describes translocation pathway expression and the functional proteome by compartmentalized mass. iJL1678-ME enables the computation of cellular phenotypes through an integrated computation of proteome composition, abundance, and activity in four cellular compartments (cytoplasm, periplasm, inner and outer membrane). Reconstruction and validation of the model has demonstrated that the iJL1678-ME is capable of capturing the functional content of membranes, cellular compartment-specific composition, and that it can be utilized to examine the effect of perturbing an expanded set of network components. iJL1678-ME takes a notable step towards the inclusion of cellular ultra-structure in genome-scale models.


Forecasting fire season severity in South America using sea surface temperature anomalies.
Fires in South America cause forest degradation and contribute to carbon emissions associated with land use change. We investigated the relationship between year-to-year changes in fire activity in South America and sea surface temperatures. We found that the Oceanic Niño Index was correlated with interannual fire activity in the eastern Amazon, whereas the Atlantic Multidecadal Oscillation index was more closely linked with fires in the southern and southwestern Amazon. Combining these two climate indices, we developed an empirical model to forecast regional fire season severity with lead times of 3 to 5 months. Our approach may contribute to the development of an early warning system for anticipating the vulnerability of Amazon forests to fires, thus enabling more effective management with benefits for climate and air quality.


PAK1 is a therapeutic target in acute myeloid leukemia and myelodysplastic syndrome.
Poor clinical outcome of acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) has been attributed to failure of current chemotherapeutic regimens to target leukemic stem cells. We recently identified p21-activated kinase (PAK1) as a downstream effector molecule of H2.0-like homeobox (HLX), a gene functionally relevant for AML pathogenesis. In this study, we find that inhibition of PAK1 activity by small molecule inhibitors or by RNA interference leads to profound leukemia inhibitory effects both in vitro and in vivo. Inhibition of PAK1 induces differentiation and apoptosis of AML cells through downregulation of the MYC oncogene and a core network of MYC target genes. Importantly, we find that inhibition of PAK1 inhibits primary human leukemic cells including immature leukemic stem cell-enriched populations. Moreover, we find that PAK1 upregulation occurs during disease progression and is relevant for patient survival in MDS. Our studies highlight PAK1 as a novel target in AML and MDS and support the use of PAK1 inhibitors as a therapeutic strategy in these diseases.


Class-specific features of neuronal wiring.
Brain function relies on specificity of synaptic connectivity patterns among different classes of neurons. Yet, the substrates of specificity in complex neuropil remain largely unknown. We search for imprints of specificity in the layout of axonal and dendritic arbors from the rat neocortex. An analysis of 3D reconstructions of pairs consisting of pyramidal cells (PCs) and GABAergic interneurons (GIs) revealed that the layout of GI axons is specific. This specificity is manifested in a relatively high tortuosity, small branch length of these axons, and correlations of their trajectories with the positions of postsynaptic neuron dendrites. Axons of PCs show no such specificity, usually taking a relatively straight course through neuropil. However, wiring patterns among PCs hold a large potential for circuit remodeling and specificity through growth and retraction of dendritic spines. Our results define distinct class-specific rules in establishing synaptic connectivity, which could be crucial in formulating a canonical cortical circuit.


Speech and language therapies to improve pragmatics and discourse skills in patients with schizophrenia.
Individuals with schizophrenia display speech and language impairments that greatly impact their integration to the society. The aim of this systematic review was to identify the importance of speech and language therapy (SLT) as part of rehabilitation curriculums for patients with schizophrenia emphasizing on the speech and language abilities assessed, the therapy setting and the therapeutic approach. This article reviewed 18 studies testing the effects of language therapy or training in 433 adults diagnosed with schizophrenia. Results showed that 14 studies out of 18 lead to improvements in language and/or speech abilities. Most of these studies comprised pragmatic or expressive discursive skills being the only aim of the therapy or part of it. The therapy settings vary widely ranging from twice daily individual therapy to once weekly group therapy. The therapeutic approach was mainly operant conditioning. Although the evidence tends to show that certain areas of language are treatable through therapy, it remains difficult to state the type of approach that should be favoured and implemented to treat language impairments in schizophrenia.


The importance of the microenvironment in breast cancer progression: recapitulation of mammary tumorigenesis using a unique human mammary epithelial cell model and a three-dimensional culture assay.
The extracellular matrix (ECM) is a dominant regulator of tissue development and homeostasis. "Designer microenvironments" in culture and in vivo model systems have shown that the ECM regulates growth, differentiation, and apoptosis in murine and human mammary epithelial cells (MEC) through a hierarchy of transcriptional events involving the intricate interplay between soluble and physical signaling pathways. Furthermore, these studies have shown that these pathways direct and in turn are influenced by the tissue structure. Tissue structure is directed by the cooperative interactions of the cell-cell and cell-ECM pathways and can be modified by stromal factors. Not surprisingly then, loss of tissue structure and alterations in ECM components are associated with the appearance and dissemination of breast tumors, and malignancy is associated with perturbations in cell adhesion, changes in adhesion molecules, and a stromal reaction. Several lines of evidence now support the contention that the pathogenesis of breast cancer is determined (at least in part) by the dynamic interplay between the ductal epithelial cells, the microenvironment, and the tissue structure (acini). Thus, to understand the mechanisms involved in carcinogenesis, the role of the microenvironment (ECM as well as the stromal cells) with respect to tissue structure should be considered and studied. Towards this goal, we have established a unique human MEC model of tumorigenesis, which in concert with a three-dimensional assay, recapitulates many of the genetic and morphological changes observed in breast in cancer in vivo. We are currently using this system to understand the role of the microenvironment and tissue structure in breast cancer progression.