Functional tissue engineering: the role of biomechanics.
"Tissue engineering" uses implanted cells, scaffolds, DNA, protein, and/or protein fragments to replace or repair injured or diseased tissues and organs. Despite its early success, tissue engineers have faced challenges in repairing or replacing tissues that serve a predominantly biomechanical function. An evolving discipline called "functional tissue engineering" (FTE) seeks to address these challenges. In this paper, the authors present principles of functional tissue engineering that should be addressed when engineering repairs and replacements for load-bearing structures. First, in vivo stress/strain histories need to be measured for a variety of activities. These in vivo data provide mechanical thresholds that tissue repairs/replacements will likely encounter after surgery. Second, the mechanical properties of the native tissues must be established for subfailure and failure conditions. These "baseline data" provide parameters within the expected thresholds for different in vivo activities and beyond these levels if safety factors are to be incorporated. Third, a subset of these mechanical properties must be selected and prioritized. This subset is important, given that the mechanical properties of the designs are not expected to completely duplicate the properties of the native tissues. Fourth, standards must be set when evaluating the repairs/replacements after surgery so as to determine, "how good is good enough?" Some aspects of the repair outcome may be inferior, but other mechanical characteristics of the repairs and replacements might be suitable. New and improved methods must also be developed for assessing the function of engineered tissues. Fifth, the effects of physical factors on cellular activity must be determined in engineered tissues. Knowing these signals may shorten the iterations required to replace a tissue successfully and direct cellular activity and phenotype toward a desired end goal. Finally, to effect a better repair outcome, cell-matrix implants may benefit from being mechanically stimulated using in vitro "bioreactors" prior to implantation. Increasing evidence suggests that mechanical stress, as well as other physical factors, may significantly increase the biosynthetic activity of cells in bioartificial matrices. Incorporating each of these principles of functional tissue engineering should result in safer and more efficacious repairs and replacements for the surgeon and patient.


Estrogen inhibits lysyl oxidase and decreases mechanical function in engineered ligaments.
Women are more likely to suffer an anterior cruciate ligament (ACL) rupture than men, and the incidence of ACL rupture in women rises with increasing estrogen levels. We used an engineered ligament model to determine how an acute rise in estrogen decreases the mechanical properties of ligaments. Using fibroblasts isolated from human ACLs from male or female donors, we engineered ligaments and determined that ligaments made from female ACL cells had more collagen and were equal in strength to those made from male ACL cells. We then treated engineered ligaments for 14 days with low (5 pg/ml), medium (50 pg/ml), or high (500 pg/ml) estrogen, corresponding to the range of in vivo serum estrogen concentrations and found that collagen within the grafts increased without a commensurate increase in mechanical strength. Mimicking the menstrual cycle, with 12 days of low estrogen followed by 2 days of physiologically high estrogen, resulted in a decrease in engineered ligament mechanical function with no change in the amount of collagen in the graft. The decrease in mechanical stiffness corresponded with a 61.7 and 76.9% decrease in the activity of collagen cross-linker lysyl oxidase with 24 and 48 h of high estrogen, respectively. Similarly, grafts treated with the lysyl oxidase inhibitor β-aminoproprionitrile (BAPN) for 24 h showed a significant decrease in ligament mechanical strength [control (CON) = 1.58 ± 0.06 N; BAPN = 1.06 ± 0.13 N] and stiffness (CON = 7.7 ± 0.46 MPa; BAPN = 6.1 ± 0.71 MPa) without changing overall collagen levels (CON = 396 ± 11.5 μg; BAPN = 382 ± 11.6 μg). Together, these data suggest that the rise in estrogen during the follicular phase decreases lysyl oxidase activity in our engineered ligament model and if this occurs in vivo may decrease the stiffness of ligaments and contribute to the elevated rate of ACL rupture in women.


Assessment of progression of asbestosis in the sheep model by bronchoalveolar lavage and pulmonary function tests.
To study the relationship between the results of bronchoalveolar lavage and pulmonary function tests during induction and progression of asbestosis, three groups of six sheep were exposed repeatedly by intratracheal injection to either saline (controls), low doses of Canadian chrysotile UICC asbestos (cumulative exposure 328 mg) (low-dose group), or high doses of the same fibres (cumulative dose 2282 mg) (high-dose group) until there was clear evidence of alveolitis from the lung biopsy specimens of all sheep of the high-dose group. During the course of this induction and for the following eight months lung biopsies, bronchoalveolar lavage and pulmonary function tests were performed at two-month intervals. At the time of initial alveolitis in the high-dose group there was no significant change in the cellularity of the bronchoalveolar lavage fluid, but static lung compliance (Cst), vital capacity (VC), arterial oxygen tension (Pao2), and diffusion capacity (DL/VA) were significantly lower than in the other groups. In the following months, as the alveolitis evolved into a fibrosing process, macrophages and neutrophils from the bronchoalveolar lavage fluid increased significantly and pulmonary function deteriorated. Proteins and enzymes in the bronchoalveolar lavage fluid also increased significantly in the high-dose group. These data show that in the sheep model of asbestosis simple tests of pulmonary function correlate well with histological changes and changes in the bronchoalveolar lavage fluid in the course of the disease and can be used to assess progression of asbestosis.


Health-related quality of life trajectory of treatment-naive patients with Merkel cell carcinoma receiving avelumab.
Aim: To evaluate changes in health-related quality of life (HRQoL) in a Phase II trial (NCT02155647) of treatment-naive patients with metastatic Merkel cell carcinoma treated with avelumab (15-month follow-up). Materials & methods: Mixed-effect Models for Repeated Measures were applied to HRQoL data (FACT-M; EQ-5D-5L) to assess changes over time. Clinically derived progression-free survival was compared with HRQoL deterioration-free survival. Results: Overall, we saw relative stability in HRQoL among 116 included patients, with nonprogression associated with statistically and clinically meaningful better HRQoL compared with progressive disease. Deterioration-free survival rates (49-72% at 6 months, 40-58% at 12 months) were consistently higher/better compared with progression-free survival rates (41/31% at 6/12 months). Conclusion: These findings show unique longitudinal HRQoL data for treatment-naive metastatic Merkel cell carcinoma patients treated with avelumab. Clinical trial registration: NCT02155647 (ClinicalTrials.gov).


Liraglutide for type 2 diabetes mellitus.
Prevalence of type 2 diabetes mellitus (T2DM) is increasing. Management of this condition and minimizing the cardiovascular risks associated with it poses a significant burden on healthcare resources across the world. Currently available therapeutic agents are effective in glycemic management; however, the majority of these are associated with undesirable effects such as hypoglycemia and weight gain. Incretin-based therapies have been introduced over the last few years and are associated with less risk of hypoglycemia and weight gain. This review includes current challenges in the management of T2DM, and an overview of glucagon-like peptide-1 (GLP-1)-based therapies, in particular the results of Phase III clinical studies of recently approved liraglutide. Apart from glycemic control, multifactorial interventions are needed to minimize the cardiovascular risks associated with T2DM. Liraglutide is effective in improving glycemic control measured by HbA1c and it is also shown to improve weight. Recently, the National Institute of Health and Clinical Excellence in the UK has approved liraglutide 1.2 mg dose in dual and triple therapy for T2DM. Liraglutide, a once-daily GLP-1 analog, has a definite role in selected patients with T2DM and the long-term cardiovascular safety is currently being ascertained in ongoing trials.


The effect of a crunchy pseudo-chewing sound on perceived texture of softened foods.
Elderly individuals whose ability to chew and swallow has declined are often restricted to unpleasant diets of very soft food, leading to a poor appetite. To address this problem, we aimed to investigate the influence of altered auditory input of chewing sounds on the perception of food texture. The modified chewing sound was reported to influence the perception of food texture in normal foods. We investigated whether the perceived sensations of nursing care foods could be altered by providing altered auditory feedback of chewing sounds, even if the actual food texture is dull. Chewing sounds were generated using electromyogram (EMG) of the masseter. When the frequency properties of the EMG signal are modified and it is heard as a sound, it resembles a "crunchy" sound, much like that emitted by chewing, for example, root vegetables (EMG chewing sound). Thirty healthy adults took part in the experiment. In two conditions (with/without the EMG chewing sound), participants rated the taste, texture and evoked feelings of five kinds of nursing care foods using two questionnaires. When the "crunchy" EMG chewing sound was present, participants were more likely to evaluate food as having the property of stiffness. Moreover, foods were perceived as rougher and to have a greater number of ingredients in the condition with the EMG chewing sound, and satisfaction and pleasantness were also greater. In conclusion, the "crunchy" pseudo-chewing sound could influence the perception of food texture, even if the actual "crunchy" oral sensation is lacking. Considering the effect of altered auditory feedback while chewing, we can suppose that such a tool would be a useful technique to help people on texture-modified diets to enjoy their food.


Impact microindentation in men with impaired fasting glucose and type 2 diabetes.
Individuals with type 2 diabetes (T2DM) are at increased fracture risk, with bone mineral density (BMD) measurements underestimating risk. Impact microindentation (IMI), a technique that measures bone microindentation distances, expressed as bone material strength index (BMSi), may improve fracture risk estimation in individuals with T2DM. This study describes the relationship between BMSi and glycaemia status in men and makes a comparison with bone measures from dual energy X-ray absorptiometry (DXA). Participants were 340 men aged 33-96 yr from the Geelong Osteoporosis Study. Impaired fasting glucose (IFG) was defined using fasting plasma glucose (FPG) between 5.5 and 6.9 mmol/L. Diabetes was defined as FPG ≥ 7.0 mmol/L, use of antihyperglycemic medication and/or self-report. Two participants with type 1 diabetes were excluded. BMSi was measured using an OsteoProbe. Femoral neck (FNBMD) and lumbar spine (LSBMD) were measured using DXA (Lunar Prodigy) and trabecular bone score (TBS) was calculated (TBS iNsight Version 2.2). Using linear regression techniques, the relationship between glycaemia status and BMSi was evaluated, adjusting for other potential confounders (including lifestyle factors, clinical measurements and FNBMD). Glycaemia status was also considered as a binary variable (T2DM vs normoglycaemia and IFG). There were 234 (68.8%) men with normoglycaemia, 59 (17.4%) with IFG and 47 (13.8%) with diabetes. When considering glycaemia status as a binary variable, men with T2DM had lower mean BMSi compared to those without T2DM (normoglycaemia and IFG combined) (79.8; 95%CI 77.0-82.6 vs 83.0; 82.2-83.8 p = 0.043) and this difference in BMSi was independent of FNBMD. No differences were observed for either FNBMD or LSBMD; however, TBS was lower (1.177; 1.121-1.233 vs 1.256; 1.240-1.272, p = 0.015, independent of FNBMD). For glycaemia status considered in three groups, there were no differences in mean BMSi values between men with normoglycaemia, IFG and T2DM (82.9 (95%CI 82.0-83.8), 83.5 (81.8-85.2) and 79.8 (77.0-82.6), respectively; ANCOVA, p = 0.104). Measures reflecting bone material properties and microarchitecture (BMSi and TBS) might be better than measures of bone mass (BMD) in identifying individuals with T2DM at risk of fracture.


The contribution of non-insulin-dependent diabetes to lower-extremity amputation in the community.
Despite the significant public health burden of lower-extremity amputations in diabetes mellitus, few data are available on the epidemiology of lower-extremity amputations in diabetes mellitus in the community setting. A retrospective incidence cohort study based in Rochester, Minn, was conducted. Among the 2015 diabetic individuals free of lower-extremity amputation at the diagnosis of diabetes mellitus, 57 individuals underwent 79 lower-extremity amputations (incidence, 375 per 100,000 person-years; 95% confidence interval, 297 to 467). Among the 1826 patients with non-insulin-dependent diabetes mellitus, 52 underwent 73 lower-extremity amputations, and the subsequent incidence of lower-extremity amputation among these residents was 388 per 100,000 person-years (95% confidence interval, 304 to 487). Of the 137 insulin-dependent diabetic patients, four subsequently underwent five lower-extremity amputations (incidence, 283 per 100,000 person-years; 95% confidence interval, 92 to 659). Twenty-five years after the diagnosis of diabetes mellitus, the cumulative risk of one lower-extremity amputation was 11.2% in insulin-dependent diabetes mellitus and 11.0% in non-insulin-dependent diabetes mellitus. When compared with lower-extremity amputation rates for Rochester residents without diabetes, patients with non-insulin-dependent diabetes mellitus were nearly 400 times more likely to undergo an initial transphalangeal amputation (rate ratio, 378.8) and had almost a 12-fold increased risk of a below-knee amputation (rate ratio, 11.8). In this community, more than 60% of lower-extremity amputations were attributable to non-insulin-dependent diabetes mellitus. These population-based data document the magnitude of the elevated risk of lower-extremity amputation among diabetic individuals. Efforts should be made to identify more precisely risk factors for amputation in diabetes and to intervene in the processes leading to amputation.


Mechanical stimuli on C2C12 myoblasts affect myoblast differentiation, focal adhesion kinase phosphorylation and galectin-1 expression: a proteomic approach.
Mechanical forces are crucial in the regulation of cell morphology and function. At the cellular level, these forces influence myoblast differentiation and fusion. In this study, we applied mechanical stimuli to embryonic muscle cells using magnetic microbeads, a method shown to apply stress to specific receptors on the cell surface. We showed that mechanical stimuli promote an increase in FAK (focal adhesion kinase) phosphorylation. In order to further shed light in the process of myoblast-induced differentiation by mechanical stimuli, we performed a proteomic analysis. Thirteen proteins were found to be affected by mechanical stimulation including galectin-1, annexin III and RhoGDI (Rho guanine-nucleotide-dissociation inhibitor). In this study, we demonstrate how the combination of this method of mechanical stimuli and proteomic analysis can be a powerful tool to detect proteins that are potentially interacting in biochemical pathways or complex cellular mechanisms during the process of myoblast differentiation. We determined an increase in expression and changes in cellular localization of galectin-1 in mechanically stimulated myoblasts. A potential involvement of galectin-1 in myoblast differentiation is presented.


Rivaroxaban or Aspirin for Extended Treatment of Venous Thromboembolism.
Although many patients with venous thromboembolism require extended treatment, it is uncertain whether it is better to use full- or lower-intensity anticoagulation therapy or aspirin. In this randomized, double-blind, phase 3 study, we assigned 3396 patients with venous thromboembolism to receive either once-daily rivaroxaban (at doses of 20 mg or 10 mg) or 100 mg of aspirin. All the study patients had completed 6 to 12 months of anticoagulation therapy and were in equipoise regarding the need for continued anticoagulation. Study drugs were administered for up to 12 months. The primary efficacy outcome was symptomatic recurrent fatal or nonfatal venous thromboembolism, and the principal safety outcome was major bleeding. A total of 3365 patients were included in the intention-to-treat analyses (median treatment duration, 351 days). The primary efficacy outcome occurred in 17 of 1107 patients (1.5%) receiving 20 mg of rivaroxaban and in 13 of 1127 patients (1.2%) receiving 10 mg of rivaroxaban, as compared with 50 of 1131 patients (4.4%) receiving aspirin (hazard ratio for 20 mg of rivaroxaban vs. aspirin, 0.34; 95% confidence interval [CI], 0.20 to 0.59; hazard ratio for 10 mg of rivaroxaban vs. aspirin, 0.26; 95% CI, 0.14 to 0.47; P<0.001 for both comparisons). Rates of major bleeding were 0.5% in the group receiving 20 mg of rivaroxaban, 0.4% in the group receiving 10 mg of rivaroxaban, and 0.3% in the aspirin group; the rates of clinically relevant nonmajor bleeding were 2.7%, 2.0%, and 1.8%, respectively. The incidence of adverse events was similar in all three groups. Among patients with venous thromboembolism in equipoise for continued anticoagulation, the risk of a recurrent event was significantly lower with rivaroxaban at either a treatment dose (20 mg) or a prophylactic dose (10 mg) than with aspirin, without a significant increase in bleeding rates. (Funded by Bayer Pharmaceuticals; EINSTEIN CHOICE ClinicalTrials.gov number, NCT02064439 .).


3D tensor factorization approach to single-frame model-free blind-image deconvolution.
By applying a bank of 2D Gabor filters to a blurred image, single-frame blind-image deconvolution (SF BID) is formulated as a 3D tensor factorization (TF) problem, with the key contribution that neither origin nor size of the spatially invariant blurring kernel is required to be known or estimated. Mixing matrix, the original image, and its spatial derivatives are identified from the factors in the Tucker3 model of the multichannel version of the blurred image. Previous approaches to 2D Gabor-filter-bank-based SF BID relied on 2D representation of the multichannel version of the blurred image and matrix factorization methods such as nonnegative matrix factorization (NMF) and independent component analysis (ICA). Unlike matrix factorization-based methods 3D TF preserves local structure in the image. Moreover, 3D TF based on the PARAFAC model is unique up to permutation and scales under very mild conditions. To achieve this, NMF and ICA respectively require enforcement of sparseness and statistical independence constraints on the original image and its spatial derivatives. These constraints are generally not satisfied. The 3D TF-based SF BID method is demonstrated on an experimental defocused red-green-blue image.


Investigation of foot-and-mouth disease outbreaks in the Mbala and Kazungula districts of Zambia.
Foot-and-mouth disease (FMD) is an acute, highly contagious viral infection of domestic and wild cloven-hoofed animals. It is known to be endemic in Zambia, with periodic outbreaks occurring in different geographical areas of the country. This study was conducted to investigate the presence of FMD virus (FMDV) in reported FMD-suspected cases in cattle from the Kazungula and Mbala districts of Zambia. Sixty epithelial tissues or oesophageal-pharyngeal (OP) scrapings (probang samples) were collected from Mbala (n = 51) and Kazungula (n = 9) and examined for FMDV. The FMDV viral RNA and serotypes were examined by realtime reverse transcription polymerase chain reaction (qRT-PCR) and antigen Enzyme- linked immunosorbent assay (ELISA), respectively. Twenty-two samples (36.7%) were positive for the FMDV genome by qRT-PCR with Cycle threshold (Ct) values ranging from 13 to 31. The FMDV-positive samples from epithelial tissues showed relatively higher Ct values compared to those obtained from OP scrapings, irrespective of geographical location. Forty percent (40%; n = 4) of epithelial tissues from Mbala were serotyped into SAT 2 serotype by antigen ELISA. Kazungula samples were serotyped into SAT 1. These findings indicated that Mbala and Kazungula districts had FMD outbreaks in 2012 that were ascribed to at least FMDV serotype SAT 2 and SAT 1 field strains. Furthermore, regular interaction between buffalos from the Mosi-o Tunya Park and domestic animals from surrounding areas could contribute to the occurrence of regular FMD outbreaks in Kazungula, whilst the uncontrolled animal movements across borders between Mbala and Nsumbawanga could be responsible for disease outbreaks in Mbala. In-depth molecular biological studies, including sequencing and phylogeny of the viruses, should be conducted to elucidate the complex epidemiology of FMD in Zambia, thereby providing valuable information needed for the rational control strategy of FMD in Zambia and neighbouring countries.


Understanding the Nature of Weak Interactions between Functionalized Boranes and N2/O2, Promising Functional Groups for Gas Separations.
The separation of nitrogen and oxygen gases is considered as a very challenging process, since both O2 and N2 are nonpolar molecules with similar kinetic diameters. Electronic structure theory can provide a fundamental understanding of effects that can lead to selective binding of nitrogen or oxygen gas for the development of novel separation processes. Boranes can bind dinitrogen through a dative bond, where the boron acts as a σ acceptor and back-donates through π orbitals. To better understand these interactions, we have performed highly accurate CCSD(F12)(T) and CCSDT(Q) computations for the BH3-N2 and BH3-O2 complexes. The coupled-cluster binding energies were used as reference for benchmarking different density functionals, and larger functionalized boranes were examined at the M05/def2-TZVPPD level. Symmetry adapted perturbation theory (SAPT) calculations were performed for the elucidation of the nature of the interaction between nitrogen and substituted boranes and how direct or distal functionalizations affect the strength of the weak dative bonds. By use of these methods, several boranes were found to bind N2 over O2. These molecular species are promising functional groups for incorporation into the next generation of advanced materials for efficient N2/O2 separations.


Variation in pediatric traumatic brain injury outcomes in the United States.
To ascertain the degree of variation, by state of hospitalization, in outcomes associated with traumatic brain injury (TBI) in a pediatric population. A retrospective cohort study of pediatric patients admitted to a hospital with a TBI. Hospitals from states in the United States that voluntarily participate in the Agency for Healthcare Research and Quality's Healthcare Cost and Utilization Project. Pediatric (age ≤ 19 y) patients hospitalized for TBI (N=71,476) in the United States during 2001, 2004, 2007, and 2010. None. Primary outcome was proportion of patients discharged to rehabilitation after an acute care hospitalization among alive discharges. The secondary outcome was inpatient mortality. The relative risk of discharge to inpatient rehabilitation varied by as much as 3-fold among the states, and the relative risk of inpatient mortality varied by as much as nearly 2-fold. In the United States, approximately 1981 patients could be discharged to inpatient rehabilitation care if the observed variation in outcomes was eliminated. There was significant variation between states in both rehabilitation discharge and inpatient mortality after adjusting for variables known to affect each outcome. Future efforts should be focused on identifying the cause of this state-to-state variation, its relationship to patient outcome, and standardizing treatment across the United States.


Activation of protein kinase C (PKC) by 3,4-methylenedioxymethamphetamine (MDMA) occurs through the stimulation of serotonin receptors and transporter.
This report further characterizes the intermediate metabolic effects of the psychotropic amphetamine derivative, 3,4-methylenedioxymethamphetamine (MDMA or "ecstasy"), on the activity of second messenger-dependent kinases. Previous work has demonstrated that two injections of MDMA (20 mg/kg) elicits a prolonged translocation of the calcium and phospholipid-dependent enzyme, protein kinase C (PKC) in rats. However, because MDMA has actions at the 5-HT transporter and 5-HT2A/2C receptors, our experiments were directed at uncovering which of these many sites may be involved in this second messenger dependent response. A single injection of MDMA produced a time- and dose-dependent increase in the density of cortical and hippocampal PKC (as measured by 3H-phorbol 12,13-dibutyrate (PDBu) binding sites. MDMA-mediated PKC translocation was long-lasting and remained above control (saline-treated rats) for up to 24 h after injection. This effect was mimicked by another substituted amphetamine, p-chloroamphetamine (pCA), but with a temporal-response curve that was to the left of MDMA's. However, pure uptake inhibitors like fluoxetine, cocaine, and the selective 5-HT2A/2C agonist, DOB, were unable to produce a long-lasting translocation of PKC binding sites in rat cortex. Fluoxetine, a selective serotonin uptake inhibitor (SSRI) and ketanserin a 5-HT2A antagonist, attenuated PKC translocation by MDMA with differing efficacies; however, both compounds completely prevented the loss of 5-HT uptake sties after multiple doses of MDMA. These results suggest that MDMA increases PKC translocation by two interrelated mechanisms that involve 5-HT2A/2C receptors and the 5-HT transporter. This pathway appears to include: (1) the drug binding to the 5-HT transporter, (2) the release of cytosolic 5-HT stores into the extracellular space, and (3) the activation of post-synaptic 5-HT2A/2C receptors linked to G-protein-mediated phospholipid hydrolysis.


The Potential of Hyperspectral Patterns of Winter Wheat to Detect Changes in Soil Microbial Community Composition.
Reliable information on soil status and crop health is crucial for detecting and mitigating disasters like pollution or minimizing impact from soil-borne diseases. While infestation with an aggressive soil pathogen can be detected via reflected light spectra, it is unknown to what extent hyperspectral reflectance could be used to detect overall changes in soil biodiversity. We tested the hypotheses that spectra can be used to (1) separate plants growing with microbial communities from different farms; (2) to separate plants growing in different microbial communities due to different land use; and (3) separate plants according to microbial species loss. We measured hyperspectral reflectance patterns of winter wheat plants growing in sterilized soils inoculated with microbial suspensions under controlled conditions. Microbial communities varied due to geographical distance, land use and microbial species loss caused by serial dilution. After 3 months of growth in the presence of microbes from the two different farms plant hyperspectral reflectance patterns differed significantly from each other, while within farms the effects of land use via microbes on plant reflectance spectra were weak. Species loss via dilution on the other hand affected a number of spectral indices for some of the soils. Spectral reflectance can be indicative of differences in microbial communities, with the Renormalized Difference Vegetation Index the most common responding index. Also, a positive correlation was found between the Normalized Difference Vegetation Index and the bacterial species richness, which suggests that plants perform better with higher microbial diversity. There is considerable variation between the soil origins and currently it is not possible yet to make sufficient reliable predictions about the soil microbial community based on the spectral reflectance. We conclude that measuring plant hyperspectral reflectance has potential for detecting changes in microbial communities yet due to its sensitivity high replication is necessary and a strict sampling design to exclude other 'noise' factors.


Malignant extra-adrenal pelvic paraganglioma in a paediatric patient.
The extra-adrenal paraganglioma is a neoplasm originating in regional structures, uncommon in paediatrics. We report on a case of a 13-year-old patient who began with severe arterial hypertension, tachycardia, dilated cardiomyopathy and elevated levels of catecholamines in the blood and urine. The presence of a retrovesical pelvic mass in contact with the right vaginal dome was determined by imaging studies. A diagnosis of malignant extra-adrenal pelvic paraganglioma with lymph node metastases was reached through biopsy and the surgical resection of subsequent local recurrences. Paragangliomas are usually located in the paravertebral zones from the base of the skull to the retroperitoneum and are benign in 90% of cases. This kind of neoplasia is uncommon in paediatrics, especially those located in the pelvis. In cases of masses of a gynaecological origin, a differential diagnosis should be considered, and a histological and immunohistochemical study is essential in certifying the diagnosis.


Anti-inflammatory and antioxidant effects of Apium graveolens L. extracts mitigate against fatal acetaminophen-induced acute liver toxicity.
In the present work, antioxidant activity, total phenolics (TP), and total flavonoids (TF) contents of aqueous and methanol extracts of celery were determined, in addition to untargeted metabolites profiling its methanol celery root extract (MCRE) via UPLC-MS. Although MCRE exhibited the lowest TPC and TFC levels, it presented the most potential hydroxyl radical quenching effect using electron paramagnetic resonance spin trapping technique. Treatment of Acetaminophen-induced hepatotoxicity (AAH) rats with MCRE lowered serum levels of AST, ALT, ALP, TNF-α, and IL-1β significantly. Additionally, MCRE significantly increased total antioxidant capacity (TAC) and glutathione (GSH) levels relative to AAH rats. Strikingly, Kaplan-Meier survival analysis of all groups revealed a 100% prevention of acetaminophen-induced mortality of rats by MCRE pretreatment (100 mg/kg/day). MCRE prevented AAH-associated severe weight loss and elicited normal behavior in the rescued rats. Our results suggest that pretreatment with MCRE can mitigate against overdosed acetaminophen-induced acute liver failure and warrant further investigations on the potential of postinjury intervention. PRACTICAL APPLICATIONS: Acetaminophen-induced hepatotoxicity (AAH) accounts for alerting numbers of overdose-related acute liver failure and liver transplant cases with increased morbidity and mortality rates. Currently proposed mechanisms implicate mitochondria-mediated oxidative stress and inflammation in the pathogenesis of AAH, which underline current interventions employing antioxidants to combat liver damage by over-dosed acetaminophen. The present work uncovers potent protective effects of some celery extracts (and their fractions) against acetaminophen-induced oxidative stress and inflammation. Treatment of rats with fatal liver injury with methanol extract of celery root significantly reduced secretion of liver enzymes and markedly decreased inflammatory as well as oxidative stress markers in these animals. This, in turn, rescued challenged rats exposed to fatal doses of acetaminophen completely, which establishes methanol extracts of celery roots as effective therapeutic intervention against AAH. The antioxidant capacity of the extracts was determined using EPR technique, and the secondary metabolites related to antioxidant activity were characterized via UPLC-MS.


Towards defining nutrient conditions encountered by the rice blast fungus during host infection.
Fungal diseases cause enormous crop losses, but defining the nutrient conditions encountered by the pathogen remains elusive. Here, we generated a mutant strain of the devastating rice pathogen Magnaporthe oryzae impaired for de novo methionine biosynthesis. The resulting methionine-requiring strain grew strongly on synthetic minimal media supplemented with methionine, aspartate or complex mixtures of partially digested proteins, but could not establish disease in rice leaves. Live-cell-imaging showed the mutant could produce normal appressoria and enter host cells but failed to develop, indicating the availability or accessibility of aspartate and methionine is limited in the plant. This is the first report to demonstrate the utility of combining biochemical genetics, plate growth tests and live-cell-imaging to indicate what nutrients might not be readily available to the fungal pathogen in rice host cells.


Glucagon-like Peptide 1 Receptor Signaling in Acinar Cells Causes Growth-Dependent Release of Pancreatic Enzymes.
Incretin-based therapies are widely used for type 2 diabetes and now also for obesity, but they are associated with elevated plasma levels of pancreatic enzymes and perhaps a modestly increased risk of acute pancreatitis. However, little is known about the effects of the incretin hormone glucagon-like peptide 1 (GLP-1) on the exocrine pancreas. Here, we identify GLP-1 receptors on pancreatic acini and analyze the impact of receptor activation in humans, rodents, isolated acini, and cell lines from the exocrine pancreas. GLP-1 did not directly stimulate amylase or lipase release. However, we saw that GLP-1 induces phosphorylation of the epidermal growth factor receptor and activation of Foxo1, resulting in cell growth with concomitant enzyme release. Our work uncovers GLP-1-induced signaling pathways in the exocrine pancreas and suggests that increases in amylase and lipase levels in subjects treated with GLP-1 receptor agonists reflect adaptive growth rather than early-stage pancreatitis.


Current approaches for predicting a lack of response to anti-EGFR therapy in KRAS wild-type patients.
Targeting epidermal growth factor receptor (EGFR) has been one of the most effective colorectal cancer strategies. Anti-EGFR antibodies function by binding to the extracellular domain of EGFR, preventing its activation, and ultimately providing clinical benefit. KRAS mutations in codons 12 and 13 are recognized prognostic and predictive biomarkers that should be analyzed at the clinic prior to the administration of anti-EGFR therapy. However, still an important fraction of KRAS wild-type patients do not respond to the treatment. The identification of additional genetic determinants of primary or secondary resistance to EGFR targeted therapy for further improving the selection of patients is urgent. Herein, we review the latest published literature highlighting the most important genes that may predict resistance to anti-EGFR monoclonal antibodies in colorectal cancer patients. According to the available findings, the evaluation of BRAF, NRAS, PIK3CA, and PTEN status could be the right strategy to select patients who are likely to respond to anti-EGFR therapies. In the future, the combination of those biomarkers will help establish consensus that can be introduced into clinical practice.


K-Ras mutations and N-Ras mutations in childhood acute leukemias with or without mixed-lineage leukemia gene rearrangements.
It is believed that Ras mutations drive the proliferation of leukemic cells. The objective of this study was to investigate the association of Ras mutations with childhood acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) with special reference to the presence or absence of mixed-lineage leukemia gene (MLL) rearrangements. Bone marrow samples from 313 children with B-precursor ALL and 130 children with de novo AML were studied at diagnosis. Southern blot analysis was used to detect MLL rearrangements, and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was used to detect common MLL fusion transcripts. Complementary DNA panhandle PCR was used to identify the infrequent or unknown MLL partner genes. DNA PCR or RT-PCR followed by direct sequencing was performed to detect mutations at codons 12, 13, and 61 of the N-Ras and K-Ras genes. Twenty of 313 patients with B-precursor ALL and 17 of 130 patients with de novo AML had MLL rearrangements. N-Ras mutations were detected in 2 of 20 patients with MLL-positive ALL and in 27 of 293 patients with MLL-negative ALL (P = 1.000). N-Ras mutations were detected in 2 of 17 patients with MLL-positive AML and in 14 of 113 patients with MLL-negative AML (P = 1.000). K-Ras mutations were present in 8 of 20 patients with MLL-positive ALL compared with 32 of 293 patients with MLL-negative ALL (P = 0.001). K-Ras mutations were detected in 3 of 17 patients with MLL-positive AML compared with 5 of 113 patients with MLL-negative AML (P = 0.069). Ras mutations were detected in 20.8% of patients with childhood B-precursor ALL and in 17.7% of patients with childhood AML. MLL-positive B-precursor ALL was associated closely with Ras mutations (50%), especially with K-Ras mutations (40%), whereas MLL-positive AML was not associated with Ras mutations.


Professional perspectives about pharmacogenetic testing and managing ancillary findings.
Pharmacogenetic (PGx) tests, intended to inform therapeutic decision making through prediction of patient likelihood to respond to or experience an adverse effect from a specific treatment, may also generate ancillary, or incidental, disease information unrelated to the purpose for which the test was ordered. To assess attitudes toward PGx testing, ancillary disease risk information, and related clinical issues, we conducted a series of focus groups among health professionals. Twenty-one primary care and genetics professionals from Durham, NC, were recruited to participate in three focus groups (two of primary care professionals [PCPs] and one of geneticists). Overall, interest in PGx testing was positive, though enthusiasm was reserved among PCPs due to concerns about clinical utility, insurance coverage, delay of treatment, and ability to communicate and interpret ancillary disease risk information. Although many PCPs felt an obligation to disclose information about ancillary disease risk, geneticists did not believe that it was always necessary, noting the complexities of genetic risk results such as incomplete penetrance. This pilot study found that health professionals' interest in the use of PGx testing was limited by concerns about the lack of evidence of clinical utility and their ability to interpret and communicate ancillary disease risk information to patients. Additional educational resources, access to genetic specialists, and clear clinical guidelines about the use of PGx testing would greatly facilitate appropriate use of testing.


Elucidating selection processes for antibiotic resistance in sewage treatment plants using metagenomics.
Sewage treatment plants (STPs) have repeatedly been suggested as "hotspots" for the emergence and dissemination of antibiotic-resistant bacteria. A critical question still unanswered is if selection pressures within STPs, caused by residual antibiotics or other co-selective agents, are sufficient to specifically promote resistance. To address this, we employed shotgun metagenomic sequencing of samples from different steps of the treatment process in three Swedish STPs. In parallel, concentrations of selected antibiotics, biocides and metals were analyzed. We found that concentrations of tetracycline and ciprofloxacin in the influent were above predicted concentrations for resistance selection, however, there was no consistent enrichment of resistance genes to any particular class of antibiotics in the STPs, neither for biocide and metal resistance genes. The most substantial change of the bacterial communities compared to human feces occurred already in the sewage pipes, manifested by a strong shift from obligate to facultative anaerobes. Through the treatment process, resistance genes against antibiotics, biocides and metals were not reduced to the same extent as fecal bacteria. The OXA-48 gene was consistently enriched in surplus and digested sludge. We find this worrying as OXA-48, still rare in Swedish clinical isolates, provides resistance to carbapenems, one of our most critically important classes of antibiotics. Taken together, metagenomics analyses did not provide clear support for specific antibiotic resistance selection. However, stronger selective forces affecting gross taxonomic composition, and with that resistance gene abundances, limit interpretability. Comprehensive analyses of resistant/non-resistant strains within relevant species are therefore warranted.


Food antigen-induced immune responses in Crohn's disease patients and experimental colitis mice.
In Crohn's disease (CD), the involvement of food antigens in immune responses remains unclear. The objective of this study was to detect immune responses against food antigens in CD patients and examine the mechanism in a mouse model of colitis. We enrolled 98 CD patients, 50 ulcerative colitis patients, and 52 healthy controls (HCs) to compare the levels of serum immunoglobulin (Ig)Gs against 88 foods. The presence of serum IgGs against foods was also examined in interleukin (IL)-10 knockout (KO) mice in which CD4(+) T cell activation by antigenic food protein was assessed. Mice transferred with IL-10 KO cells received diets with or without food antigens, and the development of colitis was evaluated. The prevalence of IgGs against various foods, especially vegetables, grains, and nuts, was significantly higher in CD patients than in HCs. Similarly, the prevalence of IgGs against food proteins was higher in IL-10 KO mice than in BALB/c mice. Beta-conglycinin, identified as an antigenic food proteins in IL-10 KO mice, induced CD4(+) T cell production of interferon-γ and IL-17 through dendritic cell antigen presentation. Elimination of the food antigens ameliorated the development of colitis in mice without altering the composition of their intestinal microbiota. In CD colitis mice, intestinal inflammation via CD4(+) T cell hyperactivation was induced by food antigens associated with high serum IgG levels and was ameliorated by the elimination of food antigens. This disrupted immunological tolerance to food antigen, which might act as an exacerbating factor, remains to be elucidated in CD patients.


A systematic review of the use of telehealth in Asian countries.
We conducted a systematic review of the literature on telehealth in Asia. The Medline database was searched, together with three specialist journals, for peer-reviewed articles published in the ten years to June 2007 which were related to any telehealth application involving one or more Asian country. Out of the 1504 abstracts retrieved, 109 articles were selected by two independent reviewers for the final review. The number of published articles on telehealth in Asia increased during the review period. The largest number of studies were conducted in Japan (37%). Most telehealth applications were based on the store-and-forward modality (43%), with 35% using videoconferencing and 15% using a hybrid approach. Most of the studies were descriptive (75%) and only eight included a control group against which telehealth was compared. The most common means of telecommunication was ISDN lines, which were employed in 32% of the studies. Some 40% of the studies mentioned improved quality of health care; about 20% mentioned improved access to health care. Although most studies mentioned cost, only 13 of them assessed resource utilization and cost. The overall findings gave a generally optimistic picture of telehealth in Asia. However, there is a lack of good quality studies.


How bulk and surface properties of Ti4SiC3, V4SiC3, Nb4SiC3 and Zr4SiC3 tune reactivity: a computational study.
We present several in silico insights into the MAX-phase of early transition metal silicon carbides and explore how these affect carbon dioxide hydrogenation. Periodic density functional methodology is applied to models of Ti4SiC3, V4SiC3, Nb4SiC3 and Zr4SiC3. We find that silicon and carbon terminations are unstable, with sintering occurring in vacuum and significant reconstruction taking place under an oxidising environment. In contrast, the metal terminated surfaces are highly stable and very active towards CO2 reduction. However, we predict that under reaction conditions these surfaces are likely to be oxidised. These results are compared to studies on comparable materials and we predict optimal values for hydrogen evolution and CO2 reduction.


Feasibility, acceptability and potential effectiveness of a mobile health (mHealth) weight management programme for New Zealand adults.
Mobile health (mHealth) behaviour change programmes use mobile phones and the internet to deliver health information and behaviour change support to participants. Such programmes offer a potentially cost-effective way to reach many individuals who do not currently access weight loss services. We developed a mHealth weight management programme using proven face-to-face behaviour change techniques and incorporating target population input. Our aim was to evaluate the feasibility, acceptability and potential effectiveness of this programme for ethnically diverse adults with a view to informing a larger trial. Fifty three adults who had a BMI of ≥25 kg/m(2) and wanted to lose weight (81% female, mean age 42 years, mean BMI 35.7 kg/m(2), 26% Maori, 34% Pacific) received the eight-week mHealth weight loss programme. Anthropometric measures were taken at two face-to-face assessments at baseline and 12-weeks (i.e. four weeks after cessation of intervention). Twelve-week follow-up measurements were available for 36/53 participants (68%). Non-completers were younger and more likely to be male and of Pacific ethnicity. Thirty five participants (66%) reported reading 'all or most' text messages sent and 96% responded to at least one text data collection question over the eight-week active intervention period. Eighty one per cent of participants logged in to the study website at least once during the eight-week study period. In the intention-to-treat analysis, mean weight change was -1.0 kg (SD 3.1) at 12 weeks (p = 0.024) and change in BMI was -0.34 kg/m(2) (SD 1.1) (p = 0.026). In the completers only analysis (n = 36), mean weight change was -1.4 kg (SD 3.6) (p = 0.023) and change in BMI was -0.50 kg/m(2) (SD 1.3) (p = 0.025). A mHealth weight management programme is feasible to deliver to an ethnically diverse population. Changes in body weight and BMI at 12 weeks indicate that the programme could be effective in supporting people with weight loss. However, the high dropout rate indicates a need for further improvements to the programme. ACTRN12612000850875.


Comprehensive behavioral analysis of the Cdkl5 knockout mice revealed significant enhancement in anxiety- and fear-related behaviors and impairment in both acquisition and long-term retention of spatial reference memory.
Mutations in the Cyclin-dependent kinase-like 5 (CDKL5) gene cause severe neurodevelopmental disorders. Recently we have generated Cdkl5 KO mice by targeting exon 2 on the C57BL/6N background, and demonstrated postsynaptic overaccumulation of GluN2B-containing N-methyl-D-aspartate (NMDA) receptors in the hippocampus. In the current study, we subjected the Cdkl5 KO mice to a battery of comprehensive behavioral tests, aiming to reveal the effects of loss of CDKL5 in a whole perspective of motor, emotional, social, and cognition/memory functions, and to identify its undetermined roles. The neurological screen, rotarod, hot plate, prepulse inhibition, light/dark transition, open field, elevated plus maze, Porsolt forced swim, tail suspension, one-chamber and three-chamber social interaction, 24-h home cage monitoring, contextual and cued fear conditioning, Barnes maze, and T-maze tests were applied on adult Cdkl5 -/Y and +/Y mice. Cdkl5 -/Y mice showed a mild alteration in the gait. Analyses of emotional behaviors revealed significantly enhanced anxiety-like behaviors of Cdkl5 -/Y mice. Depressive-like behaviors and social interaction of Cdkl5 -/Y mice were uniquely altered. The contextual and cued fear conditioning of Cdkl5 -/Y mice were comparable to control mice; however, Cdkl5 -/Y mice showed a significantly increased freezing time and a significantly decreased distance traveled during the pretone period in the altered context. Both acquisition and long-term retention of spatial reference memory were significantly impaired. The morphometric analysis of hippocampal CA1 pyramidal neurons revealed impaired dendritic arborization and immature spine development in Cdkl5 -/Y mice. These results indicate that CDKL5 plays significant roles in regulating emotional behaviors especially on anxiety- and fear-related responses, and in both acquisition and long-term retention of spatial reference memory, which suggests that focus and special attention should be paid to the specific mechanisms of these deficits in the CDKL5 deficiency disorder.


A literature review to identify factors that determine policies for influenza vaccination.
To conduct a literature review of influenza vaccination policy, describing roles and interactions between stakeholders and the factors influencing policy-making. Major databases were searched using keywords related to influenza vaccination, decision-making and health policy. Titles and abstracts were screened according to defined criteria using independent reviewers. Selected articles were analysed and compared against a checklist. 342 papers were identified, but only 111 included. A wide range of countries was represented in articles published in 1994-2012. We identified numerous stakeholders at the national and international level and found a variety of interactions between them. Using these data, we suggest a scheme for the most important stakeholders and their interactions. Determinants of policy-making were mainly related to the vaccine/disease, political-economic context, and stakeholders communication. The most relevant evidence was clinical/epidemiological studies. After the 2009 pandemic: the importance of mathematical modelling and ethical issues was greater; and the need for better communication between stakeholders was emphasised. The relevance of evidence and factors influencing policy-making varied between countries, according to complex interactions between the stakeholders involved at different levels of decision-making process. These interactions remain unclear, especially at national level, where the most important influenza policy decisions are made. To better define and understand the exact interactions and use of evidence, we recommend undertaking future qualitative studies at national level using small number of countries.


Correlation between antibodies and histology in celiac disease: incidence of celiac disease is higher than expected in the pediatric population.
The present study aims to report on the correlation between the degree of negativity of anti-endomysial antibodies and anti-tissue transglutaminase antibodies in the IgA and IgG classes with regard to histological grade, in 44 newly diagnosed children with celiac disease (CD). Samples with negative antibodies, but a positive histology from a 5-year program searching for CD in the pediatric population were collected. A total of 4247 biopsy samples were used in this study. We documented that certain pediatric patients are seronegative, while the disease is active and the incidence of CD is higher than expected in the pediatric population. This is an important finding, which demonstrates the lack of association between autoantibodies and lesions, and justifies the use of biopsies for an accurate CD diagnosis and the importance of revising the diagnostic criteria in a clinical, endoscopic and serological context. We recommend a more active search for incidences of the disease in the pediatric population. Serological markers are not the main method for the diagnosis of CD as they are considered to only have a supporting role clinically. Biopsies of the small intestine are always necessary for the diagnosis of CD in these patients.


The irrationality of categorical perception.
Perception is often categorical: the perceptual system selects one interpretation of a stimulus even when evidence in favor of other interpretations is appreciable. Such categorization is potentially in conflict with normative decision theory, which mandates that the utility of various courses of action should depend on the probabilities of all possible states of the world, not just that of the one perceived. If these probabilities are lost as a result of categorization, choice will be suboptimal. Here we test for such irrationality in a task that requires human observers to combine perceptual evidence with the uncertain consequences of action. Observers made rapid pointing movements to targets on a touch screen, with rewards determined by perceptual and motor uncertainty. Across both visual and auditory decision tasks, observers consistently placed too much weight on perceptual uncertainty relative to action uncertainty. We show that this suboptimality can be explained as a consequence of categorical perception. Our findings indicate that normative decision making may be fundamentally constrained by the architecture of the perceptual system.


Consumer holographic read-only memory reader with mastering and replication technology.
What is believed to be a novel holographic design for read-only memory systems allows a compact low-cost consumer drive within a 10 mm drive height, using a lensless phase conjugate readout and a combination of polytopic and angle multiplexing. A two-step mastering method enables production of high-efficiency holographic masters, and fast replication is possible by using only a series of plane-wave illuminations. Mastering and replication techniques are verified experimentally with an array of 125 holograms with no measured bit errors.


Protease-Inhibitor Interaction Predictions: Lessons on the Complexity of Protein-Protein Interactions.
Protein interactions shape proteome function and thus biology. Identification of protein interactions is a major goal in molecular biology, but biochemical methods, although improving, remain limited in coverage and accuracy. Whereas computational predictions can guide biochemical experiments, low validation rates of predictions remain a major limitation. Here, we investigated computational methods in the prediction of a specific type of interaction, the inhibitory interactions between proteases and their inhibitors. Proteases generate thousands of proteoforms that dynamically shape the functional state of proteomes. Despite the important regulatory role of proteases, knowledge of their inhibitors remains largely incomplete with the vast majority of proteases lacking an annotated inhibitor. To link inhibitors to their target proteases on a large scale, we applied computational methods to predict inhibitory interactions between proteases and their inhibitors based on complementary data, including coexpression, phylogenetic similarity, structural information, co-annotation, and colocalization, and also surveyed general protein interaction networks for potential inhibitory interactions. In testing nine predicted interactions biochemically, we validated the inhibition of kallikrein 5 by serpin B12. Despite the use of a wide array of complementary data, we found a high false positive rate of computational predictions in biochemical follow-up. Based on a protease-specific definition of true negatives derived from the biochemical classification of proteases and inhibitors, we analyzed prediction accuracy of individual features, thereby we identified feature-specific limitations, which also affected general protein interaction prediction methods. Interestingly, proteases were often not coexpressed with most of their functional inhibitors, contrary to what is commonly assumed and extrapolated predominantly from cell culture experiments. Predictions of inhibitory interactions were indeed more challenging than predictions of nonproteolytic and noninhibitory interactions. In summary, we describe a novel and well-defined but difficult protein interaction prediction task and thereby highlight limitations of computational interaction prediction methods.


Contribution of NADPH oxidase to membrane CD38 internalization and activation in coronary arterial myocytes.
The CD38-ADP-ribosylcyclase-mediated Ca(2+) signaling pathway importantly contributes to the vasomotor response in different arteries. Although there is evidence indicating that the activation of CD38-ADP-ribosylcyclase is associated with CD38 internalization, the molecular mechanism mediating CD38 internalization and consequent activation in response to a variety of physiological and pathological stimuli remains poorly understood. Recent studies have shown that CD38 may sense redox signals and is thereby activated to produce cellular response and that the NADPH oxidase isoform, NOX1, is a major resource to produce superoxide (O2·-)) in coronary arterial myocytes (CAMs) in response to muscarinic receptor agonist, which uses CD38-ADP-ribosylcyclase signaling pathway to exert its action in these CAMs. These findings led us hypothesize that NOX1-derived O2·- serves in an autocrine fashion to enhance CD38 internalization, leading to redox activation of CD38-ADP-ribosylcyclase activity in mouse CAMs. To test this hypothesis, confocal microscopy, flow cytometry and a membrane protein biotinylation assay were used in the present study. We first demonstrated that CD38 internalization induced by endothelin-1 (ET-1) was inhibited by silencing of NOX1 gene, but not NOX4 gene. Correspondingly, NOX1 gene silencing abolished ET-1-induced O2·- production and increased CD38-ADP-ribosylcyclase activity in CAMs, while activation of NOX1 by overexpression of Rac1 or Vav2 or administration of exogenous O2·- significantly increased CD38 internalization in CAMs. Lastly, ET-1 was found to markedly increase membrane raft clustering as shown by increased colocalization of cholera toxin-B with CD38 and NOX1. Taken together, these results provide direct evidence that Rac1-NOX1-dependent O2·- production mediates CD38 internalization in CAMs, which may represent an important mechanism linking receptor activation with CD38 activity in these cells.


Effector-independent motor sequence representations exist in extrinsic and intrinsic reference frames.
Many daily activities rely on the ability to produce meaningful sequences of movements. Motor sequences can be learned in an effector-specific fashion (such that benefits of training are restricted to the trained hand) or an effector-independent manner (meaning that learning also facilitates performance with the untrained hand). Effector-independent knowledge can be represented in extrinsic/world-centered or in intrinsic/body-centered coordinates. Here, we used functional magnetic resonance imaging (fMRI) and multivoxel pattern analysis to determine the distribution of intrinsic and extrinsic finger sequence representations across the human neocortex. Participants practiced four sequences with one hand for 4 d, and then performed these sequences during fMRI with both left and right hand. Between hands, these sequences were equivalent in extrinsic or intrinsic space, or were unrelated. In dorsal premotor cortex (PMd), we found that sequence-specific activity patterns correlated higher for extrinsic than for unrelated pairs, providing evidence for an extrinsic sequence representation. In contrast, primary sensory and motor cortices showed effector-independent representations in intrinsic space, with considerable overlap of the two reference frames in caudal PMd. These results suggest that effector-independent representations exist not only in world-centered, but also in body-centered coordinates, and that PMd may be involved in transforming sequential knowledge between the two. Moreover, although effector-independent sequence representations were found bilaterally, they were stronger in the hemisphere contralateral to the trained hand. This indicates that intermanual transfer relies on motor memories that are laid down during training in both hemispheres, but preferentially draws upon sequential knowledge represented in the trained hemisphere.


Allogeneic marrow transplantation in patients with acute myeloid leukemia in first remission: a randomized trial of two irradiation regimens.
A randomized trial of 12.0 Gy versus 15.75 Gy of total body irradiation (TBI) was performed in patients with acute myeloid leukemia undergoing allogeneic marrow transplantation while in first complete remission. All patients received 120 mg/kg cyclophosphamide followed by TBI and marrow from HLA-identical siblings. Cyclosporine and methotrexate were used for prophylaxis against acute graft-versus-host disease (GVHD). Thirty-four patients received 2.0-Gy fractions of irradiation daily for 6 days and 37 received 2.25-Gy fractions daily for 7 days. The 3-year actuarial probabilities for relapse-free survival were 0.58 for the patients who received 12.0 Gy and 0.59 for those who received 15.75 Gy. The 3-year probabilities of relapse were 0.35 for the 12.0 Gy group and 0.12 for the 15.75 Gy group (P = .06). The 3-year probabilities of transplant-related mortality were 0.12 and 0.32, respectively (P = .04). The probability of moderate to severe acute GVHD was 0.21 for the 12.0 Gy group and 0.48 for the 15.75 Gy group (P = .02). Patients exposed to the higher irradiation dose received less immunoprophylaxis against, and had a higher incidence of, acute GVHD. The increased dose of TBI significantly reduced the probability of relapse but did not improve survival because of increased mortality from causes other than relapse.


Factors responsible for increased susceptibility of mice to intestinal colonization after treatment with streptomycin.
Streptomycin sulfate (5 mg/ml) was added to the drinking water of Swiss white mice. After treatment for 1 week, the mice were challenged orogastrically with 10(8) Pseudomonas aeruginosa cells. The organism failed to multiply in the intestinal tract of either treated or untreated animals, but could be recovered from contents and tissues after 48 h. In a previous study, Salmonella typhimurium was shown to multiply in the intestines of streptomycin-treated but not untreated mice when 10(3) organisms were used as inoculum. Streptomycin administration had little effect on Eh, protein or carbohydrate concentrations of cecal contents, or intestinal motility. However, it caused a statistically significant increase in water content and pH of contents and a decrease in the concentrations of acetic, propionic, butyric, and valeric acids. S. typhimurium multiplied in pooled cecal contents obtained from both streptomycin-treated and untreated animals, but its multiplication rate and total populations were significantly greater in contents from treated animals. P. aeruginosa did not multiply in contents from either treated or untreated mice. Similar results were obtained when the organisms were inoculated into nutrient broth adjusted to simulate the pH levels and volatile fatty acid (VFA) concentrations in cecal contents of treated and untreated mice. The addition of brain heart infusion broth to cecal contents from untreated animals, in concentrations that support multiplication of S. typhimurium and P. aeruginosa, did not reverse inhibition. The addition of VFA to cecal contents from treated animals to equal the concentration in cecal contents from untreated animals caused inhibition of a magnitude observed in cecal contents from untreated animals. The results indicate that VFA operating at the pH level of cecal contents of conventional mice inhibit the multiplication of both S. typhimurium and P. aeruginosa and restrict colonization of the intestine by these organisms. The decrease in VFA concentrations that occurs as a result of streptomycin administration adequately explains the increased susceptibility of treated mice to colonization with S. typhimurium. It does not explain the increased susceptibility of treated mice to P. aeruginosa colonization, however.


Parents' reflections on managing their children's diabetes with insulin pumps.
To describe the experiences of parents managing the type 1 diabetes of their young children using an insulin pump. Qualitative description, based on principles of naturalistic inquiry. Open-ended, in-depth interviews were conducted with 21 mothers and fathers of 16 children less than 12 years old in the eastern part of the United States. The children had been on pump treatment from 3 to 36 months. Data were managed and analyzed using qualitative content analysis. Parents reported becoming comfortable with pump management 10 days to 2-3 months after the child started using it. During this adjustment period, they had to "rethink" how they managed the disease. Although using the pump required vigilance and more frequent glucose checking than did multiple daily injections (MDI), the pump provided better glucose control, easier disease management, fewer variables to manipulate, and more flexible meal times. A few parents were tempted to more tightly control the child's glucose levels, which could potentially increase parental stress. Fathers were actively involved in the day-to-day management and benefited from opportunities to participate in the child's care. They emphasized the importance of practicing the tasks associated with diabetes management to improve their confidence when caring for the child alone. Parents reported that everyone in the family experienced more freedom, flexibility, and spontaneity in their daily lives. For this sample of parents, the health and management benefits of their children's use of the insulin pump outweighed the adjustment challenges involved in changing from MDI.


Evaluation of magnetic resonance safety of veterinary radiofrequency identification devices at 1 T.
Implants containing metallic components have the potential to become heated or move within the patient while in the magnetic resonance (MR) environment. Despite containing a ferromagnetic core and having been in use for over 20 years, no information is available on the safety of veterinary radiofrequency identification devices during MR examinations. These devices are the most commonly encountered metallic implants in dogs and cats undergoing MR imaging. Three commercial veterinary microchips were evaluated for safety in the MR environment at 1 T. Parameters tested were translational force, torque, heating, artifact production, and function. Translation and torque were larger than that expected from normal activity under normal gravity. No significant heating was observed. Signal void artifacts may affect diagnosis if they are too close to the area of clinical importance. Microchip function was unaffected by routine clinical MR imaging. Capsule formation around devices is a major factor in counteracting translation and torque. Our findings support that is acceptable for patients to undergo MR imaging with this 1 T system following an interval of 3 months postimplantation to allow capsule growth. Because of the complex interactions involved, these observations may not be translatable to MR scanners of different field strength and/or manufacturer. Further safety testing of these and other radiofrequency identification devices is therefore recommended at different field strengths and equipment specifications.


Kremen1 restricts Dkk activity during posterior lateral line development in zebrafish.
Canonical Wnt signaling plays crucial roles during development and disease. How Wnt signaling is modulated in different in vivo contexts is currently not well understood. Here, we investigate the modulation of Wnt signaling in the posterior lateral line primordium (pLLP), a cohort of ~100 cells that collectively migrate along the trunk of the zebrafish embryo. The pLLP comprises proliferative progenitor cells and organized epithelial cells that will form the mechanosensory organs of the posterior lateral line. Wnt signaling is active in the leading progenitor zone of the pLLP and restricted from the trailing zone through expression of the secreted Wnt inhibitors dkk1b and dkk2. We have identified a zebrafish strain, krm1(nl10), which carries a mutation in the kremen1 gene, a non-obligate co-receptor for the Dkk family of proteins. Previous studies have shown that Kremen1 inhibits Wnt signaling by facilitating internalization of the Kremen1-Dkk-Lrp5/6 complex. Surprisingly, we found that disruption of Kremen1 in the pLLP exhibited molecular and cellular phenotypes associated with a decrease rather than overactivation of Wnt signaling. Transplantation of wild-type cells into the mutant primordia failed to rescue the krm1(nl10) phenotype, thus revealing that the effects of Kremen1 loss are non-cell-autonomous. Finally, ectopic expression of Dkk1b-mTangerine protein revealed larger spread of the fusion protein in the mutant primordia compared with the wild type. Based on our data, we propose a novel mechanism in which Kremen1 modulates Wnt activity by restricting the range of secreted Dkk proteins during collective cell migration in the pLLP.


Behavior of macrosomic and appropriate-for-gestational-age newborns.
To compare the behavior of macrosomic newborns who were vaginally delivered of healthy mothers without diabetes with that of non-macrosomic, appropriate-for-gestational-age (AGA) newborns. Newborns were recruited conveniently from a tertiary hospital. Newborns were examined at 12-24 and 36-48 hours of age, using the Brazelton Neonatal Behavioral Assessment Scale (NBAS). Thirty macrosomic newborns who were delivered vaginally were matched with AGA newborns for ethnicity, maternal education, parity, and obstetric medications. Dimensions scores derived from the individual NBAS items measured reflex functioning, response decrement, orientation, motor processes, range of state, autonomic stability, and regulation of state. Macrosomic newborns performed weaker than AGA newborns on the reflex and motor dimensions. Both groups displayed improved motor scores on Day 2, but regulation of state scores were weaker. For orientation, AGA newborns scored higher on Day 1, and macrosomic newborns scored higher on Day 2. Increased head, limb, and body mass of macrosomic newborns, compared with adjacent and overall muscle strength, might have interfered with the execution of coordinated movements. Nurses can inform mothers of changes they can expect in their newborns' behavior.


Inpatient child mortality by travel time to hospital in a rural area of Tanzania.
To investigate the association, if any, between child mortality and distance to the nearest hospital. The study was based on data from a 1-year study of the cause of illness in febrile paediatric admissions to a district hospital in north-east Tanzania. All villages in the catchment population were geolocated, and travel times were estimated from availability of local transport. Using bands of travel time to hospital, we compared admission rates, inpatient case fatality rates and child mortality rates in the catchment population using inpatient deaths as the numerator. Three thousand hundred and eleven children under the age of 5 years were included of whom 4.6% died; 2307 were admitted from <3 h away of whom 3.4% died and 804 were admitted from ≥ 3 h away of whom 8.0% died. The admission rate declined from 125/1000 catchment population at <3 h away to 25/1000 at ≥ 3 h away, and the corresponding hospital deaths/catchment population were 4.3/1000 and 2.0/1000, respectively. Children admitted from more than 3 h away were more likely to be male, had a longer pre-admission duration of illness and a shorter time between admission and death. Assuming uniform mortality in the catchment population, the predicted number of deaths not benefiting from hospital admission prior to death increased by 21.4% per hour of travel time to hospital. If the same admission and death rates that were found at <3 h from the hospital applied to the whole catchment population and if hospital care conferred a 30% survival benefit compared to home care, then 10.3% of childhood deaths due to febrile illness in the catchment population would have been averted. The mortality impact of poor access to hospital care in areas of high paediatric mortality is likely to be substantial although uncertainty over the mortality benefit of inpatient care is the largest constraint in making an accurate estimate.


Recognition and Management of Nonaccidental Esophageal Perforation.
Nonaccidental trauma is a common pediatric concern that often goes unrecognized. Although most patients present with bruising, burns, fractures, and head trauma, it is critical that physicians be able to diagnose and treat atypical presentations such as pharyngeal and esophageal trauma. In this report, we describe the presentation and management of a 5-week-old girl with an inflicted esophageal perforation.


Lysozyme-stabilized gold nanoclusters as a novel fluorescence probe for cyanide recognition.
Lysozyme-stabilized gold nanoclusters (Lys-AuNCs) have been synthesized and utilized as a fluorescent probe for selective detection of cyanide (CN(-)). Lys-AuNCs had an average size of 4 nm and showed a red emission at 650 nm (λex=370 nm). The fluorescence of Lys-AuNCs could be quenched by CN(-). An excellent sensitivity and selectivity toward the detection of CN(-) in aqueous solution was observed. The fluorescence intensity was linear with the CN(-) concentration in the range of 5.00×10(-6)M-1.20×10(-4) M with a detection limit as low as 1.9×10(-7) M. Also, the addition of CN(-) to Lys-AuNCs could induce an obvious color change from light yellow to colorless. Correspondingly, a bright red fluorescence disappeared and a blue fluorescence appeared. The results indicated that Lys-AuNCs could be applied in detection of cyanide on environmental aspects.


RENEB intercomparison exercises analyzing micronuclei (Cytokinesis-block Micronucleus Assay).
In the framework of the 'Realizing the European Network of Biodosimetry' (RENEB) project, two intercomparison exercises were conducted to assess the suitability of an optimized version of the cytokinesis-block micronucleus assay, and to evaluate the capacity of a large laboratory network performing biodosimetry for radiation emergency triages. Twelve European institutions participated in the first exercise, and four non-RENEB labs were added in the second one. Irradiated blood samples were shipped to participating labs, whose task was to culture these samples and provide a blind dose estimate. Micronucleus analysis was performed by automated, semi-automated and manual procedures. The dose estimates provided by network laboratories were in good agreement with true administered doses. The most accurate estimates were reported for low dose points (≤ 0.94 Gy). For higher dose points (≥ 2.7 Gy) a larger variation in estimates was observed, though in the second exercise the number of acceptable estimates increased satisfactorily. Higher accuracy was achieved with the semi-automated method. The results of the two exercises performed by our network demonstrate that the micronucleus assay is a useful tool for large-scale radiation emergencies, and can be successfully implemented within a large network of laboratories.


Dosing and monitoring of low-molecular-weight heparin in high-risk pregnancy: single-center experience.
To evaluate dosing requirements and monitoring patterns of low-molecular-weight heparin (LMWH) when used in high-risk pregnancy. Retrospective, observational, cohort study. University-affiliated medical center. Forty-nine women treated with LMWH between 2001 and 2005 for either prophylaxis or treatment of venous thromboembolism during pregnancy and monitored with antifactor Xa activity. Data were obtained on 53 pregnancies in the 49 women. The primary outcome was change in dosing requirements of LMWH throughout pregnancy as determined by the corresponding antifactor Xa activity peak levels. Mean starting doses of twice-daily enoxaparin and doses most proximate to delivery were 39.2 mg (range 30-60 mg) and 55.0 mg (range 30-100 mg, p=0.06), respectively, for the prophylaxis group and 83.0 mg (range 30-180 mg) and 85.7 mg (range 30-160 mg, p=0.41), respectively, for the therapeutic group. Weight-based mean starting doses and doses most proximate to delivery were 0.46 and 0.62 mg/kg (p=0.03), respectively, for the prophylaxis group and 0.90 and 0.87 mg/kg (p=0.29), respectively, for the therapeutic group. Dose changes were required in 9 (69%) of 13 pregnancies and 21 (55%) of 38 pregnancies (data from two of the 40 pregnancies were excluded-one in a patient receiving dalteparin, and one in a patient with mitral valve replacement who had higher antifactor Xa goals) in the prophylaxis and therapeutic groups, respectively, to achieve target antifactor Xa activity. The weight-based prophylactic dose was consistently 0.6 mg/kg in all three trimesters, achieving a mean ± SD target antifactor Xa activity of 0.39 ± 0.18 units/ml, whereas the therapeutic dose was 0.9 mg/kg to maintain antifactor Xa activity of 0.71 ± 0.22 units/ml. Dose changes for LMWH throughout pregnancy as guided by antifactor Xa activity were common. A significant increase in the LMWH dose requirements in the prophylactic group suggests that more frequent monitoring of antifactor Xa activity may be appropriate in pregnant patients to maintain target anticoagulant levels.


Y1 receptor ligand-based nanomicelle as a novel nanoprobe for glioma-targeted imaging and therapy.
Due to the molecular and cellular heterogeneity of glioma, discovery of novel targeted sites and ligands for glioma imaging and therapy remains challenging. Neuropeptide Y (NPY) Y1 receptors (Y1Rs) are highly over expressed in various brain tumors including glioma, and can serve as potential targeting sites for glioma imaging and therapy. Here, we show by in vivo fluorescent imaging that a highly selective Y1R ligand, [Asn6, Pro34] NPY (AP-NPY), facilitated circumvention of the blood brain barrier (BBB) by nanomicelles specifically targeting glioma. Modification with AP-NPY stabilized doxorubicin-loaded nanomicelles in the normal physiological state and promoted drug release in the acidic tumor microenvironment. Furthermore, targeted delivery of AP-NPY nanomicelles improved the therapeutic efficacy of doxorubicin for glioma, producing a prolonged survival rate. These results suggest that Y1R is a novel targeted receptor, and its selective ligand AP-NPY improves BBB permeability and glioma targeting. Our study paves the way for developing a novel delivery system for diagnosis and treatment of glioma in which Y1Rs are over expressed.


Mitochondrial survivin inhibits apoptosis and promotes tumorigenesis.
Evasion of apoptosis is a hallmark of cancer, but the molecular circuitries of this process are not understood. Here we show that survivin, a member of the inhibitor of apoptosis gene family that is overexpressed in cancer, exists in a novel mitochondrial pool in tumor cells. In response to cell death stimulation, mitochondrial survivin is rapidly discharged in the cytosol, where it prevents caspase activation and inhibits apoptosis. Selective targeting of survivin to mitochondria enhances colony formation in soft agar, accelerates tumor growth in immunocompromised animals, and abolishes tumor cell apoptosis in vivo. Therefore, mitochondrial survivin orchestrates a novel pathway of apoptosis inhibition, which contributes to tumor progression.


Adolescent boys' physical fighting and adult life outcomes: Examining the interplay with intelligence.
Although it is well known that adolescent delinquent behavior is related to poor outcomes in adulthood, longitudinal research on specific acts of delinquency and their interplay with important individual characteristics in predicting future outcomes is scarce. We aimed to examine how physical fighting-one of the most common acts of violent delinquency among adolescent boys-is related to adult life success in several domains, and how intelligence influences these associations. The study used data from 1,083 boys that participated in the population-based longitudinal Young in Norway Study, following adolescents from 1992 to 2015, by combining self-reports at four time points with comprehensive information from registers. Results showed that adolescent boys' physical fighting was associated with poor adult outcomes in the domains of employment, education, and criminal behavior. Associations remained significant even after controlling for conduct problems in general-which isolated the effects of fighting from other delinquent acts-as well as from a variety of other potential confounders. Detailed analyses on the interplay of physical fighting and intelligence showed that some parts of the associations between adolescent boys' fighting and several adverse adult outcomes could be ascribed to lower intelligence among the fighters. Moreover, intelligence moderated the relationship between physical fighting and adult education. Adolescent fighting was not related to educational attainment among boys with high intelligence, whereas boys with lower intelligence experienced detrimental effects of adolescent fighting. The analyses show the importance of considering adolescent boys' physical fighting as a potential risk factor for future social marginalization.


Attitudes towards and beliefs about colorectal cancer and screening using the faecal occult blood test within the Italian-Australian community.
Studies with minority ethnic communities worldwide reveal important differences in the content of beliefs about cancer and attitudes towards screening. Current initiatives in colorectal cancer (CRC) screening highlight the importance of identifying any illness-specific beliefs that might influence participation rates within the targeted age-range. We conducted semi-structured interviews with 20 Italian-Australians aged between 50 and 78 years, living in Adelaide, South Australia. Qualitative data from the interviews were analysed using framework analysis. Participants articulated specific beliefs about the nature of cancer, risk factors, prevention possibilities, and variety of potential barriers and benefits to faecal occult blood testing (FOBT). Although participants'beliefs overlapped with conventional medical models of cancer, the results also demonstrated the presence of specific cultural perceptions that might influence FOBT participation. Our results suggest that models used to inform communication about cancer need to be sensitive to culture specific concerns. Within the context of the older Italian-Australian community, there is a suggestion that self and response efficacy may be serious barriers to screening behavior and that bi-lingual, verbal delivery of information may be the most effective mode of communication to increase screening participation.


Changes in the prevalence of breast feeding in preterm infants discharged from neonatal units: a register study over 10 years.
There are indications that the prevalence of exclusively breastfed preterm infants is decreasing in Sweden. The objective was to investigate trends in exclusive breast feeding at discharge from Swedish neonatal units and associated factors in preterm infants. This is a register study with data from the Swedish Neonatal Quality Register. Data from 29 445 preterm infants (gestational age (GA) <37 weeks) who were born during the period 2004-2013 were retrieved. Data included maternal, perinatal and neonatal characteristics. Data were analysed for the whole population as well as for 3 GA groups. From 2004 to 2013, the prevalence of exclusive breast feeding decreased, in extremely preterm (GA 22-27 weeks) from 55% to 16%, in very preterm (GA 28-31 weeks) from 41% to 34% and in moderately preterm infants (GA 32-36 weeks) from 64% to 49%. The decline was statistically significant (p<0.001) in all 3 GA groups. This decline remained significant when adjustments were made for factors negatively associated with exclusive breast feeding and which became more prevalent during the study period, that is, small for GA (all groups) and maternal mental illness (very preterm and moderately preterm infants). In the past 10 years, Sweden has experienced a lower rate of exclusive breast feeding in preterm infants, especially in extremely preterm infants. The factors analysed in this study explain only a small proportion of this decline. The decline in exclusive breast feeding at discharge from neonatal units raises concern and present challenges to the units to support and promote breast feeding.


Modeling and Analysis of Micro-Spacecraft Attitude Sensing with Gyrowheel.
This paper proposes two kinds of approaches of angular rate sensing for micro-spacecraft with a gyrowheel (GW), which can combine attitude sensing with attitude control into one single device to achieve a compact micro-spacecraft design. In this implementation, during the three-dimensional attitude control torques being produced, two-dimensional spacecraft angular rates can be sensed from the signals of the GW sensors, such as the currents of the torque coils, the tilt angles of the rotor, the motor rotation, etc. This paper focuses on the problems of the angular rate sensing with the GW at large tilt angles of the rotor. For this purpose, a novel real-time linearization approach based on Lyapunov's linearization theory is proposed, and a GW linearized measurement model at arbitrary tilt angles of the rotor is derived. Furthermore, by representing the two-dimensional rotor tilt angles and tilt control torques as complex quantities and separating the twice periodic terms about the motor spin speed, the linearized measurement model at smaller tilt angles of the rotor is given and simplified. According to the respective characteristics, the application schemes of the two measurement models are analyzed from the engineering perspective. Finally, the simulation results are presented to demonstrate the effectiveness of the proposed strategy.


SMO expression in colorectal cancer: associations with clinical, pathological, and molecular features.
Smoothened, frizzled family receptor (SMO) is an important component of the hedgehog signaling pathway, which has been implicated in various human carcinomas. However, clinical, molecular, and prognostic associations of SMO expression in colorectal cancer remain unclear. Using a database of 735 colon and rectal cancers in the Nurse's Health Study and the Health Professionals Follow-up Study, we examined the relationship of tumor SMO expression (assessed by immunohistochemistry) to prognosis, and to clinical, pathological, and tumor molecular features, including mutations of KRAS, BRAF, and PIK3CA, microsatellite instability, CpG island methylator phenotype (CIMP), LINE-1 methylation, and expression of phosphorylated AKT and CTNNB1. SMO expression was detected in 370 tumors (50 %). In multivariate logistic regression analysis, SMO expression was independently inversely associated with phosphorylated AKT expression [odds ratio (OR) 0.48; 95 % confidence interval (CI) 0.34-0.67] and CTNNB1 nuclear localization (OR 0.48; 95 % CI 0.35-0.67). SMO expression was not significantly associated with colorectal cancer-specific or overall survival. However, in CIMP-high tumors, but not CIMP-low/0 tumors, SMO expression was significantly associated with better colorectal cancer-specific survival (log-rank P = 0.012; multivariate hazard ratio, 0.36; 95 % CI 0.13-0.95; P interaction = 0.035, for SMO and CIMP status). Our data reveal novel potential associations between the hedgehog, the WNT/CTNNB1, and the PI3K (phosphatidylinositol-4,5-bisphosphonate 3-kinase)/AKT pathways, supporting pivotal roles of SMO and hedgehog signaling in pathway networking. SMO expression in colorectal cancer may interact with tumor CIMP status to affect patient prognosis, although confirmation by future studies is needed.


A role for the G12 family of heterotrimeric G proteins in prostate cancer invasion.
Many studies have suggested a role for the members of the G12 family of heterotrimeric G proteins (Galpha12 and Galpha13) in oncogenesis and tumor cell growth. However, few studies have examined G12 signaling in actual human cancers. In this study, we examined the role of G12 signaling in prostate cancer. We found that expression of the G12 proteins is significantly elevated in prostate cancer. Interestingly, expression of the activated forms of Galpha12 or Galpha13 in the PC3 and DU145 prostate cancer cell lines did not promote cancer cell growth. Instead, expression of the activated forms of Galpha12 or Galpha13 in these cell lines induced cell invasion through the activation of the RhoA family of G proteins. Furthermore, inhibition of G12 signaling by expression of the RGS domain of the p115-Rho-specific guanine nucleotide exchange factor (p115-RGS) in the PC3 and DU145 cell lines did not reduce cancer cell growth. However, inhibition of G12 signaling with p115-RGS in these cell lines blocked thrombin- and thromboxane A2-stimulated cell invasion. These observations identify the G12 family proteins as important regulators of prostate cancer invasion and suggest that these proteins may be targeted to limit invasion- and metastasis-induced prostate cancer patient mortality.


Relationship of pre-existing antibody to subsequent infection by Mycoplasma pneumoniae in adults.
An extensive study of the epidemiological and serological characteristics of Mycoplasma pneumoniae infection was carried out in a military population. There was an increase in the infection rate at Camp Lejeune during the summer months as indicated by a relative increase in isolations, seroconversions, and hospitalizations for M. pneumoniae pneumonia. Twenty-three percent of the trainees who later became infected had detectable, pre-existing antilipid antibody to M. pneumoniae. When the whole organism was used as antigen, a pre-existing complement fixation (CF) titer of 1:4 or greater correlated with resistance to M. pneumoniae disease as defined by the absence of a fourfold rise in CF antibody, shedding of organisms, and clinical illness. Pre-existing antilipid fraction CF antibody titers of 1:16 or greater correlated with protection against mild and severe M. pneumoniae disease. Antilipid CF antibody titers of 1:4 and 1:8 were related to protection against mild disease but were not associated with protection against pneumonia which required hospitalization. The severity of illness was directly related to the CF antibody response in trainees with acute respiratory disease and pneumonia due to M. pneumoniae. The findings provide a basis for the development of a M. pneumoniae vaccine.


Adjuvant chemotherapy for patients with pathologic node-positive esophageal cancer after induction chemotherapy is associated with improved survival.
The study objectives were to identify variables associated with the use of adjuvant chemotherapy among patients with node-positive esophageal cancer who received induction therapy and to evaluate its relationship with overall survival. Treatment data for patients with esophageal cancer receiving induction chemotherapy ± radiotherapy and esophagectomy were abstracted from the National Cancer Data Base. Pathologic node-positive patients were dichotomized by whether they received 2 or more cycles of adjuvant chemotherapy or none. Kaplan-Meier survival curves were generated, and a Cox proportional hazards model was done to identify factors associated with overall survival. From 2006 to 2012, 3100 patients had pathologic positive nodes after induction therapy and esophagectomy. A total of 2625 patients (84.7%) did not receive adjuvant chemotherapy, and 475 patients (15.3%) did. N3 nodal stage was associated with an increased likelihood of receiving adjuvant chemotherapy (reference: N1, odds ratio, 1.82, 95% confidence interval, 1.15-2.97, P = .01), whereas increasing age (by year, odds ratio, 0.97, confidence interval, 0.96-0.98, P < .001), induction chemoradiation therapy (reference: induction chemotherapy, odds ratio, 0.39, confidence interval, 0.30-0.52, P < .001), and increasing inpatient length of stay after esophagectomy (per day: odds ratio, 0.98, confidence interval, 0.97-0.99, P = .007) were associated with a decreased likelihood. Patients receiving adjuvant chemotherapy had improved overall survival at each pathologic nodal stage: 31.6 months versus 22.7 months for N1 disease (P < .001), 32.4 months versus 19.2 months for N2 disease (P = .035), and 19.5 months versus 10.4 months for N3 disease (P < .001). Adjuvant therapy was independently associated with decreased mortality hazard (hazard ratio, 0.69, 95% confidence interval, 0.57-0.83, P < .001). Patients receiving adjuvant chemotherapy after induction therapy and esophagectomy show a survival benefit at all positive nodal stages. Prospective studies may help further delineate this benefit.


Relevance of chronotype for eating patterns in adolescents.
During adolescence, a shift from morningness to eveningness occurs, yet school continues to start early in the morning. Hence, adolescents are at risk for social jetlag, i.e. a discrepancy between biological and social timing. It remains to be determined whether chronotype associates with daily and daytime-specific eating patterns during this potentially critical period. Therefore, the aim of the present study was to investigate whether chronotype is decisive for daily eating patterns [total energy intake (TEI, kcal), total macronutrient intake (% of TEI), eating occasion frequency (n/day), meal frequency (n/day), snack frequency (n/day), duration of nightly fasting], or daytime-specific eating patterns [morning (before 11 am) energy intake (% of TEI), morning macronutrient intake (% of morning energy intake), regular breakfast skipping (no morning energy intake at least on 2 of 3 days, yes/no), evening (after 6 pm) energy intake (% of TEI), evening macronutrient intake (% of evening energy intake), regular dinner skipping (no evening energy intake at least on 2 of 3 days, yes/no)] in German adolescents. Chronotype was assessed by use of the Munich Chronotype Questionnaire and is defined as the midpoint of sleep corrected for sleep-debt accumulated over the workweek (the later the midpoint of sleep, the later the chronotype). A total of 223 participants (10-18 years) provided 346 questionnaires and concurrent 3-day weighed dietary records. Associations between chronotype and eating patterns were analyzed cross-sectionally using multivariable linear and logistic mixed-effects regression models. Adolescents with earlier and later chronotypes did not differ in their daily eating patterns. With respect to daytime-specific eating patterns, 1 h delay in chronotype was associated with 4.0 (95% CI 2.5-6.6) greater odds of regular breakfast skipping (p < 0.0001). In addition, later chronotype was associated with higher evening energy intake (p = 0.0009). In conclusion, our data show that a later chronotype among adolescents is associated with a shift of food consumption toward later times of the day. Hence, adolescents' eating patterns appear to follow their internal clock rather than socially determined schedules.


Enhanced waterlogging tolerance in barley by manipulation of expression of the N-end rule pathway E3 ligase PROTEOLYSIS6.
Increased tolerance of crops to low oxygen (hypoxia) during flooding is a key target for food security. In Arabidopsis thaliana (L.) Heynh., the N-end rule pathway of targeted proteolysis controls plant responses to hypoxia by regulating the stability of group VII ethylene response factor (ERFVII) transcription factors, controlled by the oxidation status of amino terminal (Nt)-cysteine (Cys). Here, we show that the barley (Hordeum vulgare L.) ERFVII BERF1 is a substrate of the N-end rule pathway in vitro. Furthermore, we show that Nt-Cys acts as a sensor for hypoxia in vivo, as the stability of the oxygen-sensor reporter protein MCGGAIL-GUS increased in waterlogged transgenic plants. Transgenic RNAi barley plants, with reduced expression of the N-end rule pathway N-recognin E3 ligase PROTEOLYSIS6 (HvPRT6), showed increased expression of hypoxia-associated genes and altered seed germination phenotypes. In addition, in response to waterlogging, transgenic plants showed sustained biomass, enhanced yield, retention of chlorophyll, and enhanced induction of hypoxia-related genes. HvPRT6 RNAi plants also showed reduced chlorophyll degradation in response to continued darkness, often associated with waterlogged conditions. Barley Targeting Induced Local Lesions IN Genomes (TILLING) lines, containing mutant alleles of HvPRT6, also showed increased expression of hypoxia-related genes and phenotypes similar to RNAi lines. We conclude that the N-end rule pathway represents an important target for plant breeding to enhance tolerance to waterlogging in barley and other cereals.