Cell-based therapy - navigating troubled waters.
Cells and engineered tissue can be used to treat an increasing number of diseases. This development, together with promising pre-clinical data in regenerative medicine, has raised the expectations of many patients. However, this situation tends to make people vulnerable to the lures of companies that abuse the stem cell promise. The problem is compounded by people's propensity to believe that the healing powers of positive thinking, large sums of money and foreign institutions are greater than those of therapies developed through well-tested, properly constructed, clinical trials.


Modular organization and combinatorial energetics of proline-tyrosine nuclear localization signals.
Proline-tyrosine nuclear localization signals (PY-NLSs) are recognized and transported into the nucleus by human Karyopherin (Kap) beta2/Transportin and yeast Kap104p. Multipartite PY-NLSs are highly diverse in sequence and structure, share a common C-terminal R/H/KX2-5PY motif, and can be subdivided into hydrophobic and basic subclasses based on loose N-terminal sequence motifs. PY-NLS variability is consistent with weak consensus motifs, but such diversity potentially renders comprehensive genome-scale searches intractable. Here, we use yeast Kap104p as a model system to understand the energetic organization of this NLS. First, we show that Kap104p substrates contain PY-NLSs, demonstrating their generality across eukaryotes. Previously reported Kapbeta2-NLS structures explain Kap104p specificity for the basic PY-NLS. More importantly, thermodynamic analyses revealed physical properties that govern PY-NLS binding affinity: (1) PY-NLSs contain three energetically significant linear epitopes, (2) each epitope accommodates substantial sequence diversity, within defined limits, (3) the epitopes are energetically quasi-independent, and (4) a given linear epitope can contribute differently to total binding energy in different PY-NLSs, amplifying signal diversity through combinatorial mixing of energetically weak and strong motifs. The modular organization of the PY-NLS coupled with its combinatorial energetics lays a path to decode this diverse and evolvable signal for future comprehensive genome-scale identification of nuclear import substrates.


Anti-fibrinolytic agents in post partum haemorrhage: a systematic review.
Post partum haemorrhage is a leading cause of maternal death worldwide. It also contributes to maternal morbidity as women may require a hysterectomy to control bleeding, or may require a blood transfusion, which can transmit viral infections. Anti-fibrinolytic agents have been proposed as a treatment for post partum haemorrhage. We conducted a systematic review to assess the effectiveness and safety of anti-fibrinolytic agents in post partum bleeding. All randomised controlled trials of anti-fibrinolytic agents given for bleeding during the postpartum period were included in this review. We searched Medline, PubMed, EMBASE, Cochrane Central Register of Controlled trials, Web of Science, metaRegister of controlled trials, LILACS, Reproductive Health Library, African healthline, POPLINE, MedCarib, CINAHL, Clinicaltrials.gov and the reference lists of eligible trials. Two authors extracted data. Methodological quality was assessed by evaluating allocation concealment. The primary outcome was maternal mortality. Secondary outcomes were blood loss, blood transfusion, hysterectomy, mean haemoglobin concentration, thrombo-embolic events and other adverse effects. We identified three randomised controlled trials involving 461 participants. The trials compared tranexamic acid with no treatment and reported blood loss after delivery. In all three trials, allocation concealment was either inadequate or unclear. The administration of tranexamic acid was associated with a reduction in blood loss of 92 millilitres (95%CI 76 to 109). The most frequently reported adverse effect of tranexamic acid was nausea, although the increase was easily compatible with the play of chance (RR 4.63, 95%CI 0.23 to 95.14). Tranexamic acid may reduce blood loss in post partum haemorrhage. However, the quality of the currently available evidence is poor. Adequately powered, high quality randomised controlled trials are needed.


Effects of treated sewage effluent on immune function in rainbow trout (Oncorhynchus mykiss).
In this study, the immune reactions of rainbow trout (Oncorhynchus mykiss) were examined, after exposure to 10, 30 and 70% of tertiary-treated municipal sewage effluent for 27 days. Exposures were conducted concurrently with and without an immune challenge using intraperitoneal injections of inactivated Aeromonas salmonicida salmonicida. Due to the time required to prepare and analyse samples, fish sampling was conducted over two consecutive days. There was no trout mortality for any of the experimental treatments. The exposure to effluent increased in vitro lymphocyte proliferation, decreased circulating lymphocytes and increased degrading erythrocytes in peripheral blood samples. Circulating lymphocytes were only decreased in the sham-injected, but not in the A. salmonicida-injected group. In addition to effluent effects, circulating lymphocytes and lymphocyte proliferation were decreased on day 2 of sampling as compared to day 1. Concentration-dependent degradation of erythrocytes was only observed on day 2 of sampling. Capture and removal of trout on day 1 of sampling presumably caused low-level stress that affected some results on day 2. Oxidative burst, phagocytosis, lysozyme, leucocyte populations other than lymphocytes and A. salmonicida-specific IgM production were not affected by exposure to effluent, and of these parameters, only oxidative burst and total leucocytes showed sampling day effects. From these results it can be observed, that with the exception of oxidative burst, those variables affected by effluent exposure were also significantly changed by the low-level sampling stress imposed by staggered sampling. Elevated liver mixed-function oxygenase activity as measured by 7-ethoxyresorufin-O-deethylase activity, and increased bile polycyclic aromatic hydrocarbon (PAH) metabolites were observed in response to sewage effluent exposure. As both PAHs and stress are known immune suppressors, it is difficult to conclude whether or not changes in immune parameters due to effluent exposure were caused by the direct action of chemicals, or were due to a general stress response.


High-rate pacing-induced atrial fibrillation effectively reveals properties of spontaneously occurring paroxysmal atrial fibrillation in humans.
Research on paroxysmal atrial fibrillation (AF) assumes that fibrillation induced by rapid pacing adequately reproduces spontaneously occurring paroxysmal AF in humans. We aimed to compare the spectral properties of spontaneous vs. induced AF episodes in paroxysmal AF patients. Eighty-five paroxysmal AF patients arriving in sinus rhythm to the electrophysiology laboratory were evaluated prior to ablation. Atrial fibrillation was induced by rapid pacing from the pulmonary vein-left atrial junctions (PV-LAJ), the coronary sinus (CS), or the high right atrium (HRA). Simultaneous recordings were obtained using multipolar catheters. Off-line power spectral analysis of 5 s bipolar electrograms was used to determine dominant frequency (DF) at recording sites with regularity index >0.2. Sixty-eight episodes were analysed for DF. Comparisons were made between spontaneous (n = 23) and induced (n = 45) AF episodes at each recording site. No significant differences were observed between spontaneous and induced AF episodes in HRA (5.18 ± 0.69 vs. 5.06 ± 0.91 Hz; P = 0.64), CS (5.27 ± 0.69 vs. 5.36 ± 0.76 Hz; P = 0.69), or LA (5.72 ± 0.88 vs. 5.64 ± 0.75 Hz; P = 0.7) regardless of pacing site. Consistent with these results, paired analysis in seven patients with both spontaneous and induced AF episodes, showed no regional DFs differences. Moreover, a left-to-right DF gradient was also present in both spontaneous (PV-LAJ 5.71 ± 0.81 vs. HRA 5.18 ± 0.69 Hz; P = 0.005) and induced (PV-LAJ 5.62 ± 0.72 vs. HRA 5.07 ± 0.91 Hz; P = 0.002) AF episodes, with no differences between them (P = not specific). In patients with paroxysmal AF, high-rate pacing-induced AF adequately mimics spontaneously initiated AF, regardless of induction site.


Exposure to Volatile Organic Compounds and Use of Feminine Hygiene Products Among Reproductive-Aged Women in the United States.
Background: Feminine hygiene products (FHPs) are personal care products widely used by women. A few studies have detected some volatile organic compounds (VOCs) in FHPs, but no previous epidemiological studies have linked use of these products to human exposure to VOCs using biomarkers. Therefore, we evaluated whether the use of FHPs was associated with VOC exposures among reproductive-aged women in the United States. Materials and Methods: Data on 2432 women aged 20-49 years from National Health and Nutrition Examination Survey 2001-2004 were utilized. Self-reported use of feminine products (tampons, sanitary napkins, vaginal douches, sprays, powders, wipes/towelettes, and other products) was obtained from questionnaires. Survey-weighted linear regression models were used to estimate percent changes in VOC whole blood concentrations and 95% confidence intervals (CIs). Results: Black women had significantly more use of vaginal douching and significantly higher whole blood concentrations of 1,4-dichlorobenzene (DCB) (p < 0.0001). After adjusting for confounders, we observed a dose-response relationship between the frequency of vaginal douching in the past 6 months and 1,4-DCB concentrations. Compared with never users, women with occasional use (≤1 time/month) of vaginal douching had 18% (95% CI: -12% to 59%) higher concentrations, and those with frequent use (≥2 time/month) had 81% (95% CI: 2% to 221%) higher concentrations of 1,4-DCB (p for trend = 0.04). Use of feminine powder in the past month was significantly associated with 36% (95% CI: 0.4% to 83%) higher concentrations of ethylbenzene. Conclusions: Our findings suggest that differences in whole blood VOC concentrations might be explained by feminine hygiene practices. The presence of environmental chemicals in FHPs warrants further examination.


Validation of endoscopic activity scores in patients with Crohn's disease based on a post hoc analysis of data from SONIC.
Mucosal healing might alter midterm and long-term outcomes of patients with Crohn's disease (CD) and has become an important end point in clinical trials. However, the minimal degree of mucosal improvement (endoscopic response) required to alter midterm outcomes is not known. We aimed to determine the best definition of endoscopic response by evaluating data on the Simple Endoscopic Score for Crohn's Disease (SES-CD) and the Crohn's Disease Endoscopic Index of Severity (CDEIS) from the Study of Biologic and Immunomodulator Naive Patients in Crohn's Disease (SONIC trial). We analyzed data from 172 patients who participated in the SONIC trial, were found to have endoscopic lesions at baseline, and underwent a second endoscopic examination at week 26 of treatment with infliximab, azathioprine, or both. Mucosal healing was defined as absence of ulcers. A central reader calculated SES-CD and CDEIS results. Different cutoff values were set for endoscopic response based on the SES-CD or CDEIS. The diagnostic ability of these different cutoff values was evaluated using receiver operating characteristic (ROC) curves, positive likelihood ratios (PLR), and negative likelihood ratios (NLR). Corticosteroid-free clinical remission (CFREM) at week 50 was used as a binary classifier. Based on analyses of ROC curves, PLR, and NLR, endoscopic response was defined as a decrease from baseline in SES-CD of at least 50%. At week 26, mucosal healing and endoscopic response were achieved in 48% and 65% of patients, respectively. Mucosal healing at week 26 was associated with CFREM at week 50, with 56% sensitivity, 65% specificity, a PLR of 1.60, and an NLR of 0.67. Endoscopic response at week 26 was associated with CFREM at week 50, with 74% sensitivity, 48% specificity, a PLR of 1.42, and an NLR of 0.54. Endoscopic response, defined as a decrease from baseline in CDEIS of at least 50%, yielded similar results. In patients with CD, mucosal healing and endoscopic response (defined as a decrease from baseline in SES-CD or CDEIS of at least 50%) at week 26 of treatment identified those most likely to be in CFREM at week 50. The ability of the proposed endoscopic response cutoff value to predict midterm CFREM should be validated in an independent, prospective cohort. Its correlation with changes in long-term disease progression still needs to be demonstration. ClinicalTrials.gov, Number: NCT00094458.


Exenatide Improves Bone Quality in a Murine Model of Genetically Inherited Type 2 Diabetes Mellitus.
Type 2 diabetes mellitus (T2DM) is associated with skeletal complications, including an increased risk of fractures. Reduced blood supply and bone strength may contribute to this skeletal fragility. We hypothesized that long-term administration of Exenatide, a glucagon-like peptide-1 receptor agonist, would improve bone architecture and strength of T2DM mice by increasing blood flow to bone, thereby stimulating bone formation. In this study, we used a model of obesity and severe T2DM, the leptin receptor-deficient db/db mouse to assess alterations in bone quality and hindlimb blood flow and to examine the beneficial effects of 4 weeks administration of Exenatide. As expected, diabetic mice showed marked alterations in bone structure, remodeling and strength, and basal vascular tone compared with lean mice. Exenatide treatment improved trabecular bone mass and architecture by increasing bone formation rate, but only in diabetic mice. Although there was no effect on hindlimb perfusion at the end of this treatment, Exenatide administration acutely increased tibial blood flow. While Exenatide treatment did not restore the impaired bone strength, intrinsic properties of the matrix, such as collagen maturity, were improved. The effects of Exenatide on in vitro bone formation were further investigated in primary osteoblasts cultured under high-glucose conditions, showing that Exenatide reversed the impairment in bone formation induced by glucose. In conclusion, Exenatide improves trabecular bone mass by increasing bone formation and could protect against the development of skeletal complications associated with T2DM.


Enhanced refocusing of fat signals using optimized multipulse echo sequences.
Endogenous magnetic resonance contrast based on the localized composition of fat in vivo can provide functional information. We found that the unequal pulse timings of the Uhrig's dynamical decoupling multipulse echo sequences significantly alter the signal intensity compared to conventional, equal-spaced Carr-Purcell-Meiboom-Gill sequences. The signal increases and decreases depending on the tissue and sequence parameters, as well as on the interpulse spacings; particularly strong differences were observed in fatty tissues, which have a highly structured morphology and a wide range of chemical shifts and J-couplings. We found that the predominant mechanism for fat refocusing under multipulse echo sequences is the chemical structure, with stimulated echoes playing a pivotal role. As a result, specialized pulse sequences can be designed to optimize refocusing of the fat chemical shifts and J-couplings, where the degree of refocusing can be tailored to specific types of fats. To determine the optimal time delays, we simulated various Uhrig dynamical decoupling and Carr-Purcell-Meiboom-Gill pulse sequence timings, and these results are compared to experimental results obtained on excised and in vivo fatty tissue. Applications to intermolecular multiple quantum coherence imaging, where the improved echo refocusing translates directly into signal enhancements, are presented as well.


The role of diet in the pathogenesis and management of irritable bowel syndrome (Review).
Most patients with irritable bowel syndrome (IBS) believe that diet plays a significant role in inducing IBS symptoms and desire to know what foods to avoid. It has been found that the intake of calories, carbohydrates, proteins and fat by IBS patients does not differ from that of the background population. IBS patients were found to avoid certain food items that are rich in fermentable oligo-, di- and monosacharides and polyols (FODMAPs), but they did have a high consumption of many other FODMAP-rich food items. The diet of IBS patients was found to consist of a low calcium, magnesium, phosphorus, vitamin B2 and vitamin A content. There is no consistent evidence that IBS patients suffer from food allergy, nor is there documented evidence that food intolerance plays a role in IBS symptoms. Abnormalities in gut hormones have been reported in IBS patients. As gut hormones control and regulate gastrointestinal motility and sensation, this may explain the abnormal gastrointestinal motility and visceral hypersensitivity reported in these patients. Guidance concerning food management which includes individually based restrictions of FODMAP-rich food items and individual evaluation of the effects of protein-, fat- and carbohydrate-rich/poor diets may reduce IBS symptoms.


Control of witchweed Striga hermonthica by intercropping with Desmodium spp., and the mechanism defined as allelopathic.
During investigations into the control of insect damage to maize crops in subsistence farming in Kenya, which involved intercropping with repellent plants, the fodder legumes silverleaf (Desmodium uncinatum) and greenleaf (D. intortum) were also found to reduce dramatically the infestation of maize by parasitic witchweeds such as Striga hermonthica. This effect was confirmed by further field testing and shown to be significantly greater than that observed with other legumes, e.g., cowpea, as were the concomitant yield increases. The mechanism was investigated, and although soil shading and addition of nitrogen fertilizer showed some benefits against S. hermonthica infestation, a putative allelopathic mechanism for D. uncinatum was observed. In screenhouse studies, a highly significant reduction in S. hermonthica infestation was obtained when an aqueous solution, eluting from pots in which D. uncinatum plants were growing, was used to irrigate pots of maize planted in soil seeded with high levels of S. hermonthica. Growth of the parasitic weed was almost completely suppressed. whereas extensive infestation occurred with the control eluate. Laboratory investigations into the allelopathic effect of D. uncinatum, using samples of water-soluble chemical components exuded from cleaned roots, demonstrated that this involved a germination stimulant for S. hermonthica and also an inhibitor for haustorial development.


Gender Differences in Cognition in China and Reasons for Change over Time: Evidence from CHARLS.
In this paper, we model gender differences in cognitive ability in China using a new sample of middle-aged and older Chinese respondents. Modeled after the American Health and Retirement Survey (HRS), CHARLS respondents are 45 years and older and are nationally representative of the Chinese population in this age span. Our measures of cognition in CHARLS rely on two measures that proxy for different dimensions of adult cognition-episodic memory and intact mental status. We relate these cognitive measures to adult health and SES outcomes during the adult years. We find large cognitive differences to the detriment of women that were mitigated by large gender differences in education among these generations of Chinese people. These gender differences in cognition are especially concentrated in the older age groups and poorer communities within the sample. We also investigated historical, geographical, and cultural characteristics of communities to understand how they impact cognition. Economic development and environmental improvement such as having electricity, increases in wage per capita and green coverage ratio generally contribute to higher cognition ability. Women benefit more from the fruits of development -electricity and growth of green coverage ratio are conducive to lessening female disadvantage in cognition.


Host-associated microbiomes are predicted by immune system complexity and climate.
Host-associated microbiomes, the microorganisms occurring inside and on host surfaces, influence evolutionary, immunological, and ecological processes. Interactions between host and microbiome affect metabolism and contribute to host adaptation to changing environments. Meta-analyses of host-associated bacterial communities have the potential to elucidate global-scale patterns of microbial community structure and function. It is possible that host surface-associated (external) microbiomes respond more strongly to variations in environmental factors, whereas internal microbiomes are more tightly linked to host factors. Here, we use the dataset from the Earth Microbiome Project and accumulate data from 50 additional studies totaling 654 host species and over 15,000 samples to examine global-scale patterns of bacterial diversity and function. We analyze microbiomes from non-captive hosts sampled from natural habitats and find patterns with bioclimate and geophysical factors, as well as land use, host phylogeny, and trophic level/diet. Specifically, external microbiomes are best explained by variations in mean daily temperature range and precipitation seasonality. In contrast, internal microbiomes are best explained by host factors such as phylogeny/immune complexity and trophic level/diet, plus climate. Internal microbiomes are predominantly associated with top-down effects, while climatic factors are stronger determinants of microbiomes on host external surfaces. Host immunity may act on microbiome diversity through top-down regulation analogous to predators in non-microbial ecosystems. Noting gaps in geographic and host sampling, this combined dataset represents a global baseline available for interrogation by future microbial ecology studies.


The future of fellowship training in clinical cardiac electrophysiology: Program directors' perspective 2008.
At the Clinical Cardiac Electrophysiology (CCEP) program directors' annual meeting during the 2008 scientific sessions of the Heart Rhythm Society, a consensus emerged for an urgent need to strengthen and rejuvenate fellowship training in clinical cardiac electrophysiology. A writing group of the Heart Rhythm Society Clinical Research and Training Committee was charged with defining these issues. A comprehensive questionnaire designed by the writing group was used to conduct an on-line survey of the 101 CCEP program directors in the United States. Data collected included types of programs, current status of programs, duration of fellowship, teaching responsibilities of faculty, responsibilities of fellows, and volume of electrophysiology procedures. Survey responses were collated and analyzed by the writing group. Given the rapid evolution and increased complexity of current electrophysiology procedures, program directors were of the opinion that 1 year of clinical electrophysiology training may no longer be adequate. A need to strengthen both research and didactic training components of fellowship training was also acknowledged. The number of electrophysiology procedures performed by trainees varied greatly between programs, and standardization of didactic training and procedural volume would be welcomed. Recent trends were recognized that indicate the need for a detailed national work-force analysis in CCEP. Through this national survey, program directors identified specific areas of need for standardization and strengthening of current fellowship training in CCEP. Based on these, specific measures can be taken to ensure the future of CCEP training.


Independent Computerized Cognitive Remediation for Psychosis: An Investigation of Patient Experiences.
Cognitive remediation (CR) training improves cognition and functioning in patients with psychosis. To date, however, few studies have investigated CR from a subjective patient perspective. We recently conducted a randomized control trial demonstrating the effectiveness of a new, low therapist support, computer-based training program. This study aims to assess the service user experience of this program. Twenty CR completers with psychosis were interviewed using both rated and open-ended questions. Thematic analysis identified three broad themes: a) benefits of doing CR, b) costs of doing CR, and c) experience of doing CR. Positive experiences of therapy participation included improved cognition, improved positive self-regard, a development of life skills, and a transfer of benefits to everyday life. Negative experiences included therapy being difficult and tiring, leading to frustration and anxiety. These findings demonstrate the feasibility of low therapist support CR and point to variables that might impact therapy adherence.


Comparison of joint space and end point space robotic training modalities for rehabilitation of interjoint coordination in individuals with moderate to severe impairment from chronic stroke.
We have developed a novel robotic modality called Time Independent Functional Training (TIFT) that provides focused retraining of interjoint coordination after stroke. TIFT was implemented on the ARMin III exoskeleton and provides joint space walls that resist movement patterns that are inconsistent with the targeted interjoint coordination pattern. In a single test session, ten moderate to severely impaired individuals with chronic stroke practiced synchronous shoulder abduction and elbow extension in TIFT and also in a comparison mode commonly used in robotic therapy called end point tunnel training (EPTT). In EPTT, error is limited by forces applied to the hand that are normal to the targeted end point trajectory. The completion percentage of the movements was comparable between modes, but the coordination patterns used by subjects differed between modes. In TIFT, subjects performed the targeted pattern of synchronous shoulder abduction and elbow extension, while in EPTT, movements were completed with compensatory strategies that incorporated the flexor synergy (shoulder abduction with elbow flexion) or the extensor synergy (shoulder adduction with elbow extension). There were immediate effects on free movements, with TIFT resulting in larger improvements in interjoint coordination than EPTT. TIFT's ability to elicit normal coordination patterns merits further investigation into the effects of longer duration training.


Membrane biofilm communities in full-scale membrane bioreactors are not randomly assembled and consist of a core microbiome.
Finding efficient biofouling control strategies requires a better understanding of the microbial ecology of membrane biofilm communities in membrane bioreactors (MBRs). Studies that characterized the membrane biofilm communities in lab-and pilot-scale MBRs are numerous, yet similar studies in full-scale MBRs are limited. Also, most of these studies have characterized the mature biofilm communities with very few studies addressing early biofilm communities. In this study, five full-scale MBRs located in Seattle (Washington, U.S.A.) were selected to address two questions concerning membrane biofilm communities (early and mature): (i) Is the assembly of biofilm communities (early and mature) the result of random immigration of species from the source community (i.e. activated sludge)? and (ii) Is there a core membrane biofilm community in full-scale MBRs? Membrane biofilm (early and mature) and activated sludge (AS) samples were collected from the five MBRs, and 16S rRNA gene sequencing was applied to investigate the bacterial communities of AS and membrane biofilms (early and mature). Alpha and beta diversity measures revealed clear differences in the bacterial community structure between the AS and biofilm (early and mature) samples in the five full-scale MBRs. These differences were mainly due to the presence of large number of unique but rare operational taxonomic units (∼13% of total reads in each MBR) in each sample. In contrast, a high percentage (∼87% of total reads in each MBR) of sequence reads was shared between AS and biofilm samples in each MBR, and these shared sequence reads mainly belong to the dominant taxa in these samples. Despite the large fraction of shared sequence reads between AS and biofilm samples, simulated biofilm communities from random sampling of the respective AS community revealed that biofilm communities differed significantly from the random assemblages (P < 0.001 for each MBR), indicating that the biofilm communities (early and mature) are unlikely to represent a random sample of the AS community. In addition to the presence of unique operational taxonomic units in each biofilm sample (early or mature), comparative analysis of operational taxonomic units and genera revealed the presence of a core biofilm community in the five full-scale MBRs. These findings provided insight into the membrane biofilm communities in full-scale MBRs. More comparative studies are needed in the future to elucidate the factors shaping the core and unique biofilm communities in full-scale MBRs.


Enzymatic Formation of an Artificial Base Pair Using a Modified Purine Nucleoside Triphosphate.
The expansion of the genetic alphabet with additional, unnatural base pairs (UBPs) is an important and long-standing goal in synthetic biology. Nucleotides acting as ligands for the coordination of metal cations have advanced as promising candidates for such an expansion of the genetic alphabet. However, the inclusion of artificial metal base pairs in nucleic acids mainly relies on solid-phase synthesis approaches, and very little is known about polymerase-mediated synthesis. Herein, we report the selective and high yielding enzymatic construction of a silver-mediated base pair (dImC-AgI-dPurP) as well as a two-step protocol for the synthesis of DNA duplexes containing such an artificial metal base pair. Guided by DFT calculations, we also shed light into the mechanism of formation of this artificial base pair as well as into the structural and energetic preferences. The enzymatic synthesis of the dImC-AgI-dPurP artificial metal base pair provides valuable insights for the design of future, more potent systems aiming at expanding the genetic alphabet.


Trehalose: a cryoprotectant that enhances recovery and preserves function of human pancreatic islets after long-term storage.
The scarcity of available tissue for transplantation in diabetes and the need for multiple donors make it mandatory to use an optimal cryopreservation method that allows maximal recovery and preservation of beta-cell function. We have developed a method to cryopreserve islets with excellent survival of endocrine cells. Current methods use DMSO as cryoprotectant. Our method involves introducing both DMSO and the disaccharide trehalose into the cells during cooling. Uptake and release of trehalose occurred during the thermotropic lipid-phase transition measured in pancreatic endocrine cells between 5 degrees and 9 degrees C, using [14C]trehalose. Recovery of adult islets after cryopreservation with 300 mmol/l trehalose was 92 vs. 58% using DMSO alone. In vitro function, in terms of insulin content and release in response to secretagogues, was indistinguishable from fresh islets. Grafts from islets cryopreserved with trehalose contained 14-fold more insulin than grafts from islets cryopreserved without trehalose. Results with human fetal islet-like cell clusters (ICCs) were more pronounced: recovery from cryopreservation was 94%, compared with 42% without trehalose. Complete functionality of fetal cells was also restored; tritiated thymidine incorporation and insulin content and release were similar to fresh tissue. After transplantation in nude mice, there was a 15-fold increase in insulin content of grafts from ICCs cryopreserved with trehalose compared with ICCs cryopreserved without trehalose. Thus, the addition of trehalose to cryopreservation protocols leads to previously unobtainable survival rates of human pancreatic endocrine tissue.


Altered metabolic landscape in IDH-mutant gliomas affects phospholipid, energy, and oxidative stress pathways.
Heterozygous mutations in NADP-dependent isocitrate dehydrogenases (IDH) define the large majority of diffuse gliomas and are associated with hypermethylation of DNA and chromatin. The metabolic dysregulations imposed by these mutations, whether dependent or not on the oncometabolite D-2-hydroxyglutarate (D2HG), are less well understood. Here, we applied mass spectrometry imaging on intracranial patient-derived xenografts of IDH-mutant versus IDH wild-type glioma to profile the distribution of metabolites at high anatomical resolution in situ This approach was complemented by in vivo tracing of labeled nutrients followed by liquid chromatography-mass spectrometry (LC-MS) analysis. Selected metabolites were verified on clinical specimen. Our data identify remarkable differences in the phospholipid composition of gliomas harboring the IDH1 mutation. Moreover, we show that these tumors are characterized by reduced glucose turnover and a lower energy potential, correlating with their reduced aggressivity. Despite these differences, our data also show that D2HG overproduction does not result in a global aberration of the central carbon metabolism, indicating strong adaptive mechanisms at hand. Intriguingly, D2HG shows no quantitatively important glucose-derived label in IDH-mutant tumors, which suggests that the synthesis of this oncometabolite may rely on alternative carbon sources. Despite a reduction in NADPH, glutathione levels are maintained. We found that genes coding for key enzymes in de novo glutathione synthesis are highly expressed in IDH-mutant gliomas and the expression of cystathionine-β-synthase (CBS) correlates with patient survival in the oligodendroglial subtype. This study provides a detailed and clinically relevant insight into the in vivo metabolism of IDH1-mutant gliomas and points to novel metabolic vulnerabilities in these tumors.


Association of Same-Day Discharge With Hospital Readmission After Appendectomy in Pediatric Patients.
Appendectomy is the most common abdominal operation performed in pediatric patients in the United States. Studies in adults have suggested that same-day discharge (SDD) after appendectomy is safe and does not result in higher-than-expected hospital readmissions. To evaluate the influence of SDD on 30-day readmission rates following appendectomy for acute appendicitis in pediatric patients. This retrospective cohort study used the American College of Surgeons National Surgical Quality Improvement Program-Pediatric database to evaluate 30-day readmission rates among pediatric patients who underwent an appendectomy for acute, nonperforated appendicitis. The database provides high-quality surgical outcomes data from more than 80 participating US hospitals, including free-standing pediatric facilities, children's hospitals, specialty centers, children's units within adult hospitals, and general acute care hospitals with a pediatric wing. Patients selected for inclusion (n = 22 771) were between ages 0 and 17 years and underwent appendectomy for uncomplicated appendicitis between January 1, 2012, and December 31, 2015. Patients excluded were those discharged more than 2 days after surgery. Same-day discharge after appendectomy or discharge 1 or 2 days after surgery. The primary outcome was 30-day readmission. Secondary outcomes included surgical-site infections and other wound complications. Of the 20 981 patients, 4662 (22.2%) had SDD and 16 319 (77.8%) were discharged within 1 or 2 days after surgery. The patient cohort included 12 860 boys (61.3%) and 8121 girls (38.7%), with a mean (SD) age of 11.0 (3.56) years. There was no difference in the odds of readmission for patients with SDD compared with those discharged within 2 days (adjusted odds ratio [aOR], 0.82; 95% CI, 0.51-1.04; P = .06; readmission rate, 1.89% vs 2.33%). There was no significant difference in reason for readmission on the basis of discharge timing. Likewise, there was no difference in wound complication rate between patients with SDD and those discharged 1 or 2 days after surgery (aOR 0.75; 95% CI, 0.56-1.01; P = .06). In pediatric patients with acute appendicitis undergoing appendectomy, SDD is not associated with an increase in 30-day hospital readmission rates or wound complications when compared with discharge 1 or 2 days after surgery. Same-day discharge may be an applicable quality metric for the provision of safe and efficient care for pediatric patients with acute, nonperforated appendicitis.


Mosquito vectors and the spread of cancer: an overlooked connection?
Mosquitoes (Diptera: Culicidae) represent a key threat for millions of humans and animals worldwide, vectoring important pathogens and parasites, including malaria, dengue, filariasis, and Zika virus. Besides mosquito-borne diseases, cancers figure among the leading causes of mortality worldwide. It is expected that annual cancer cases will rise from 14 million in 2012 to 22 million within the next two decades. Notably, there are few contrasting evidences of the relationship between cancer and mosquito-borne diseases, with special reference to malaria. However, analogies at the cellular level for the two diseases were reported. Recently, a significant association of malaria incidence with all cancer mortality in 50 USA states was highlighted and may be explained by the ability of Plasmodium to induce suppression of the immune system. However, it was hypothesized that Anopheles vectors may transmit obscure viruses linked with cancer development. The possible activation of cancer pathways by mosquito feeding events is not rare. For instance, the hamster reticulum cell sarcoma can be transmitted through the bites of Aedes aegypti by a transfer of tumor cells. Furthermore, mosquito bites may influence human metabolic pathways following different mechanisms, leading to other viral infections and/or oncogenesis. Hypersensitivity to mosquito bites is routed by a unique pathogenic mechanism linking Epstein-Barr virus infection, allergy, and oncogenesis. During dengue virus infection, high viral titers, macrophage infiltration, and tumor necrosis factor alpha production in the local tissues are the three key important events that lead to hemorrhage. Overall, basic epidemiological knowledge on the relationships occurring between mosquito vector activity and the spread of cancer is urgently needed, as well as detailed information about the ability of Culicidae to transfer viruses or tumor cells among hosts over time. Current evidences on nanodrugs with multipotency against mosquito-borne diseases and cancers are reviewed, with peculiar attention to their mechanisms of action.


Ellagic acid and rosmarinic acid attenuate doxorubicin-induced testicular injury in rats.
The anticancer drug doxorubicin causes testicular toxicity as an undesirable effect. The present study was undertaken to investigate the possible protection of ellagic acid and rosmarinic acid during doxorubicin administration. For this purpose eight groups of male Sprague-Dawley rats were used (n = 10), one group received vehicle served as control, and other groups received 5 mg/kg doxorubicin twice a week for 2 weeks for a cumulative dose of 20 mg/kg, ellagic acid (10 mg/kg/day, 14 consecutive days p.o.), rosmarinic acid (75 mg/kg/day, 14 consecutive days p.o.), ellagic acid and rosmarinic acid. The latter three regimens were given to control and doxorubicin-received rats. Doxorubicin decreased testicular relative weight, sperm count, motility, serum testosterone, testicular glycogen, and sialic acid with increased incidence of histopathological changes, oxidative stress, tumor necrosis factor-alpha, as well as cholinesterase activity. Conversely, ellagic and rosmarinic acid treatment ameliorated such damage, thus showing the possibility to use as an adjuvant during doxorubicin treatment.


Characterization of economic requirements for a "carbon-debt-free country".
In recent years, greenhouse gas emission controls that incorporate the supply chains of products and services, thereby emphasizing the role of consumers rather than producers, have been drawing increasing attention. A country's consumption-based emissions, including those due to global supply chains, reflect the total emissions on which the national economy relies. To design effective emissions control strategies there is therefore an urgent need for countries to elucidate the structural relationship between their domestic economy and emissions occurring through global supply chains. Here we consider the structural characteristics of consumption-based emissions in Japan, which in 2005 totaled 1675 Mt CO(2)eq. Outside the country the Japanese economy generated global emissions of 541 Mt CO(2)eq, 35.7% of which were UNFCCC Annex I emissions and 64.3% were non-Annex I and other emissions. This figure of 64.3% reveals that Japan is actually relying to a considerable degree on emissions that are subject to no international obligations. We identify key economic contributors to consumption-based emissions at the commodity level and specify items of household expenditure that are effective options for both financial savings and emissions reduction. We then discuss the importance of emissions control for evolving toward a "carbon-debt-free country".


Quantifying hybridization in realistic time.
Recently, numerous practical and theoretical studies in evolutionary biology aim at calculating the extent to which reticulation-for example, horizontal gene transfer, hybridization, or recombination-has influenced the evolution for a set of present-day species. It has been shown that inferring the minimum number of hybridization events that is needed to simultaneously explain the evolutionary history for a set of trees is an NP-hard and also fixed-parameter tractable problem. In this article, we give a new fixed-parameter algorithm for computing the minimum number of hybridization events for when two rooted binary phylogenetic trees are given. This newly developed algorithm is based on interleaving-a technique using repeated kernelization steps that are applied throughout the exhaustive search part of a fixed-parameter algorithm. To show that our algorithm runs efficiently to be applicable to a wide range of practical problem instances, we apply it to a grass data set and highlight the significant improvements in terms of running times in comparison to an algorithm that has previously been implemented.


Universal endogenous gene controls for bisulphite conversion in analysis of plant DNA methylation.
Accurate analysis of DNA methylation by bisulphite sequencing depends on the complete conversion of all cytosines into uracil. Until now there has been no standard or universal gene identified as an endogenous control to monitor the conversion frequency in plants. Here, we report the development of PCR based assays for one nuclear gene IND (INDEHISCENT) and two mitochondrial genes, NAD (NICOTINAMIDE ADENINE DINUCLEOTIDE) and ATP1 (ATPase SUBUNIT 1). We demonstrated their efficacy as bisulphite conversion controls in Brassica and other plant taxa. The target regions amplified by four primer pairs were found to be consistently free from DNA methylation. Primer pairs for IND.a and NAD were effective within Brassica species, whereas two primer pairs for ATP1 provided reliable controls across a representative range of dicot and monocot angiosperm species. These primer sets may therefore be adopted as controls in plant methylation analysis for a wide range of studies.


Spatial positive feedback at the onset of mitosis.
Mitosis is triggered by the activation of Cdk1-cyclin B1 and its translocation from the cytoplasm to the nucleus. Positive feedback loops regulate the activation of Cdk1-cyclin B1 and help make the process irreversible and all-or-none in character. Here we examine whether an analogous process, spatial positive feedback, regulates Cdk1-cyclin B1 redistribution. We used chemical biology approaches and live-cell microscopy to show that nuclear Cdk1-cyclin B1 promotes the translocation of Cdk1-cyclin B1 to the nucleus. Mechanistic studies suggest that cyclin B1 phosphorylation promotes nuclear translocation and, conversely, nuclear translocation promotes cyclin B1 phosphorylation, accounting for the feedback. Interfering with the abruptness of Cdk1-cyclin B1 translocation affects the timing and synchronicity of subsequent mitotic events, underscoring the functional importance of this feedback. We propose that spatial positive feedback ensures a rapid, complete, robust, and irreversible transition from interphase to mitosis and suggest that bistable spatiotemporal switches may be widespread in biological regulation.


Antibodies against domain I of β2-glycoprotein I: the one and only?
The antiphospholipid syndrome (APS) is diagnosed by the occurrence of thrombosis and/or specific pregnancy morbidity. However, the diagnosis of APS is not easy and is hampered by several problems including high prevalence of clinical symptoms and high variability between different assays resulting in a high false-positive rate. Currently APS can be diagnosed for example by detecting anti-β2-glycoprotein I antibodies by ELISA. It has been reported that β2-glycoprotein I (β2GPI) changes its conformation from a native to an active form and thereby it opens up enabling antibodies to bind a specific epitope. We amongst others have shown that epitope glycine40-arginine43 of domain I of β2GPI is predominantly responsible for binding thrombosis related antibodies. Antibodies with affinity towards other epitopes have not been associated with thrombosis. Despite these results the question remains whether these domain I antibodies are the only antibodies of importance for the detection of APS.


Obesity is associated with the future risk of heavy truck crashes among newly recruited commercial drivers.
This study estimates the dose-response relationship between body mass index (BMI) and crash risk in operators of heavy commercial motor vehicles. Intake data were collected during the first two weeks of instruction from 744 new truck drivers training for their commercial driver's licenses at a school operated by the cooperating trucking firm. Drivers were then followed prospectively on the job using the firm's operational data for two years, or until employment separation, whichever came first. Multivariate Poisson regression and Cox proportional hazards models were used to estimate the relationship between crash risk and BMI, controlling for demographic characteristics and for variations in the exposure to risks on the road. Results from the Poisson regression, which used cumulative miles driven as an exposure control, indicated that compared to normal BMI (18.5<BMI<25) the risk ratio (RR) for all crashes was significantly higher for drivers in the combined obesity Classes II and III: RR=1.55 (95% CI 1.24-1.94). Similarly, the multivariate Cox proportional hazard model (controlling for miles and job type on a week-by-week basis) showed that crash risk was significantly higher compared to normal BMI for the same combined obesity Classes II and III: RR=1.54 (95% CI 1.13-2.10). The results of this prospective study establish an association between obesity and crash risk and have important implications for driver health and public safety.


Dietary whey protein increases liver and skeletal muscle glycogen levels in exercise-trained rats.
We investigated the effect of different types of dietary protein on glycogen content in liver and skeletal muscle of exercise-trained rats. Twenty-four male Sprague-Dawley rats (approximately 100 g; n 6 per group) were divided into sedentary or exercise-trained groups with each group being fed either casein or whey protein as the source of dietary protein. Rats in the exercised groups were trained during 2 weeks using swimming exercise for 120 min/d, 6 d/week. Exercise training resulted in an increase in the skeletal muscle glycogen content. Furthermore, the whey protein group significantly increased the skeletal muscle glycogen content compared with the casein group. The increase in glycogen content in liver was significantly greater in rats fed the whey protein diet compared with those fed the casein diet. We also found that the whey protein diet increased the activity of liver glucokinase, whereas it decreased the activities of 6-phosphofructokinase and pyruvate kinase compared with the casein diet. However, hepatic total glycogen synthase activity and mRNA expression were similar with the two diets. In the skeletal muscle, whey protein decreased only 6-phosphofructokinase activity compared with casein. Total glycogen synthase activity in the skeletal muscle in the whey protein group was significantly higher than that in the casein group. The present study is the first to demonstrate that a diet based on whey protein may increase glycogen content in liver and skeletal muscle of exercise-trained rats. We also observed that whey protein regulated glycogen metabolism in these two tissues by different mechanisms.


The associations of blood pressure parameters with all-cause and cardiovascular mortality in peritoneal dialysis patients: a cohort study in China.
It remains controversial to claim blood pressure (BP) as a leading risk factor for high risk of death in peritoneal dialysis patients, and less is known about the relationship between BP and mortality in Chinese peritoneal dialysis patients. From Zhejiang Renal Data System in China, we collected data on patients treated and followed up at 98 peritoneal dialysis centres from 2008 to 2016. The associations of BP parameters [SBP, DBP, mean arterial pressure (MAP) and pulse pressure (PP)] with all-cause and cardiovascular mortality were examined. We fitted Cox models for mortality with penalized splines using nonparametric smoothers. Several sensitivity analyses were performed to confirm the robustness of our primary findings. A total of 7335 Chinese peritoneal dialysis patients were included. During a median follow-up of 35.8 months, 1281 (17.5%) patients died. SBP, DBP, MAP follow a U-shaped pattern of both all-cause and cardiovascular mortality. PP presents a reverse L-shaped association with all-cause mortality. Either a higher (SBP >141, DBP >85 or MAP >102 mmHg) or lower (SBP <119, DBP <67 or MAP <88 mmHg) BP tends to have a significantly higher all-cause and cardiovascular mortality risk. Higher PP (>60 mmHg) is related to a higher risk of all-cause mortality, but not cardiovascular mortality. These associations remain the same in our competing risk analysis and subgroup analyses. These data indicate U-shaped associations of SBP, DBP and MAP with all-cause mortality and cardiovascular mortality, respectively, and a reverse L-shaped association of PP with all-cause mortality. Further studies are needed to reliably establish the optimal BP targets for better hypertension control in peritoneal dialysis patients.


Sodium-hydrogen exchange and its role in controlling contractility during acidosis in cardiac muscle.
Intracellular pH (pHi) and Na (aina) were recorded in isolated sheep cardiac Purkinje fibres using ion-selective microelectrodes while simultaneously recording twitch tension. A fall of pHi stimulated acid-extrusion via sarcolemmal Na-H exchange but the extrusion was inhibited by reducing extracellular pH (pHo), indicating an inhibitory effect of external H ions upon the exchanger. Intracellular acidosis can reduce contraction by directly reducing myofibrillar Ca2+ sensitivity. The activation of Na-H exchange at low pHi can offset this direct inhibitory effect of H+ ions since exchange-activation elevates aina which then indirectly elevates Ca2+i (via Na-Ca exchange) thus tending to restore tension. This protection of contraction during intracellular acidosis can be removed if extracellular pH is also allowed to fall since, under these conditions, Na-H exchange is inhibited.


Pharmacokinetics and pharmacodynamics of LGD-3303 [9-chloro-2-ethyl-1-methyl-3-(2,2,2-trifluoroethyl)-3H-pyrrolo-[3,2-f]quinolin-7(6H)-one], an orally available nonsteroidal-selective androgen receptor modulator.
Selective androgen receptor modulators (SARMs) are a new class of molecules in development to treat a variety of diseases. SARMs maintain the beneficial effects of androgens, including increased muscle mass and bone density, while having reduced activity on unwanted side effects. The mechanisms responsible for the tissue-selective activity of SARMs are not fully understood, and the pharmacokinetic (PK)/pharmacodynamic (PD) relationships are poorly described. Tissue-specific compound distribution potentially could be a mechanism responsible for apparent tissue selectivity. We examined the PK/PD relationship of a novel SARM, LGD-3303 [9-chloro-2-ethyl-1-methyl-3-(2,2,2-trifluoroethyl)-3H-pyrrolo[3,2-f]quinolin-7(6H)-one], in a castrated rat model of androgen deficiency. LGD-3303 has potent activity on levator ani muscle but is a partial agonist on the preputial gland and ventral prostate. LGD-3303 never stimulated ventral prostate above intact levels despite increasing plasma concentrations of compound. Tissue-selective activity was maintained when LGD-3303 was dosed orally or by continuous infusion, two routes of administration with markedly different time versus exposure profiles. Despite the greater muscle activity relative to prostate activity, local tissue concentrations of LGD-3303 were higher in the prostate than in the levator ani muscle. LGD-3303 has SARM properties that are independent of its pharmacokinetic profile, suggesting that the principle mechanism for tissue-selective activity is the result of altered molecular interactions at the level of the androgen receptor.


The influence of antibiotics and other factors on reconsultation for acute lower respiratory tract illness in primary care.
Antibiotics are prescribed to the majority of patients consulting their general practitioner (GP) for lower respiratory tract illness (LRTi). A common reason for prescription is the belief that antibiotics reduce re-attendance; a motive supported by the high reconsultation rates for this largely self-limiting illness. Information about reconsultation following treatment of LRTi, and the factors that influence it, is scarce. To explore factors associated with reconsultation after initial management of LRTi. Analysis of data collected prospectively during presentation of acute LRTi in primary care. Seventy-six per cent of 518 patients were prescribed antibiotics, and 30% reconsulted for similar symptoms within the next 28 days (29% of those who were given antibiotics and 33% of those who were not). Forty-one per cent of patients who had seen their GP 15 or more times in the previous two years reconsulted, compared with 13% of those who had made fewer than five visits. Reconsultation was more common in patients with a history of underlying disease (38.6% versus 24.3%) and in patients who reported dyspnoea (41.5% versus 24.3%). Reconsultation is common in acute LRTi and is associated with a heightened consulting habit prior to the index consultation, the presence of previous ill health, and dyspnoea. It appears not to be influenced by prescribing antibiotics.


Adaptive response to ionizing radiation induced by low dose of gamma ray in human hepatoma cell lines.
When cells are exposed to a low dose of a mutagenic or clastogenic agent, they often become less sensitive to the effects of a higher dose administered subsequently. Such adaptive responses were first described in Escherichia coli. Studies on mammalian cells have been limited to human lymphocytes exposed to low doses of an alkylating agent. In this study, the adaptive response to 1 cGy of gamma rays was investigated in human tumor cells using two human hepatoma cell lines, Hep G2 and Hep 3B. Experiments were carried out by delivering 1 cGy followed by 50 cGy of gamma radiation and chromatid breaks were scored as an endpoint. The results of this study indicate that prior exposure to 1 cGy of gamma rays reduces the number of chromatid breaks induced by subsequent higher doses (50 cGy). The time necessary for the expression of the adaptive response was determined by varying the time interval between the two doses from 1 hour to 72 hours. In G2 chromatids, the adaptive response was observed both at short time intervals, as early as 1 hour, and at long time intervals. In S chromatids, however, the adaptive response was shown only at long time intervals. When 3-aminobenzamide, an inhibitor of poly (ADP-ribose) polymerase, was added after 50 cGy, adaptive responses were abolished in all the experimental groups. Therefore, it is suggested that the adaptive response can be observed in human hepatoma cell lines, which is first documented through this study.(ABSTRACT TRUNCATED AT 250 WORDS)


Sustained Adrenergic Signaling Promotes Intratumoral Innervation through BDNF Induction.
Mounting clinical and preclinical evidence supports a key role for sustained adrenergic signaling in the tumor microenvironment as a driver of tumor growth and progression. However, the mechanisms by which adrenergic neurotransmitters are delivered to the tumor microenvironment are not well understood. Here we present evidence for a feed-forward loop whereby adrenergic signaling leads to increased tumoral innervation. In response to catecholamines, tumor cells produced brain-derived neurotrophic factor (BDNF) in an ADRB3/cAMP/Epac/JNK-dependent manner. Elevated BDNF levels in the tumor microenvironment increased innervation by signaling through host neurotrophic receptor tyrosine kinase 2 receptors. In patients with cancer, high tumor nerve counts were significantly associated with increased BDNF and norepinephrine levels and decreased overall survival. Collectively, these data describe a novel pathway for tumor innervation, with resultant biological and clinical implications.Significance: Sustained adrenergic signaling promotes tumor growth and metastasis through BDNF-mediated tumoral innervation. Cancer Res; 78(12); 3233-42. ©2018 AACR.


Relationships between physical activity and physical capacity in adolescent females and bone mass in adulthood.
This study investigates whether physical activity and physical performance in adolescence are positively related to adult bone mineral density (BMD). In 1974, physical activity, endurance, and muscular strength were measured in 204 randomly selected female students, age 16.1 +/- 0.3 year (range 15-17 years). Twenty years later, 36 of the women volunteered to undergo a measurement of their BMD. Women who were members in a sports club in adolescence had significantly higher adult BMD (mean differences of 5% to 17% depending on site) compared with subjects who were not engaged in a sports club. Furthermore, women with persistent weight-bearing activity in adulthood had significantly higher BMD compared with women who had stopped being active or had never been active. The differences ranged between 5% and 19% with the highest difference found in trochanter BMD. Stepwise regression analyses showed that membership in a sports club at baseline was a significant independent predictor of BMD in the total body, lumbar spine, legs, trochanter, and femoral neck, explaining 17-26% of the variation in BMD. Change in body weight was a strong independent predictor of BMD of the total body and arms, explaining 8% of the variation in both sites. In addition, running performance at baseline was an independent predictor of total body BMD, whereas the two-hand lift performance significantly predicted BMD of the total body, legs and trochanter. The hanging leg-lift and handgrip were both significant predictors of arm BMD. In conclusion, membership in a sports club and site-specific physical performance in adolescence together with the change in body weight were significantly associated with adult BMD in premenopausal women.


Ultrastructural morphometry of hypertensive medial damage in lenticulostriate and other arteries.
Characteristic medial damage occurs in the distal lenticulostriate arteries in patients with hypertensive cerebral hemorrhage. Medical changes were studied in the proximal and distal lenticulostriate arteries and cortical circumflex arterioles. Additionally, intramyocardial coronary, gastric submucosal and renal interlobular arterioles were examined in hypertensive patients. Hypertensive medial damage consisted of irregular atrophy and paucity of smooth muscle with accumulation of nonfatty debris and basement membranes. These changes were diffuse, extensive and not uniform, and were unrelated to atherosclerosis or bifurcation. By electron microscopic morphometry smooth muscle occupied 28% of the media in the distal lenticulostriate arteries of hypertensive patients as compared with 80% in controls. The proximal lenticulostriate and cortical branches in hypertensive patients showed values higher than 50%, as did the coronary, gastric and renal arterioles. Massive primary hemorrhages are prevalent in the area supplied by the distal lenticulostriate artery, 150-660 microns in diameter. Fibrinoid necrosis and microaneurysm occur in more peripheral arterioles, 70-200 microns. Thus, hypertensive medial damage with significant reduction of the smooth muscle area to less than 50% of the total media is viewed as an additional important factor which predisposes to sudden arterial rupture and massive cerebral hemorrhage.


Patient-oriented and performance-based outcomes after knee autologous chondrocyte implantation: a timeline for the first year of recovery.
It is well established that autologous chondrocyte implantation (ACI) can require extended recovery postoperatively; however, little information exists to provide clinicians and patients with a timeline for anticipated function during the first year after ACI. To document the recovery of functional performance of activities of daily living after ACI. ACI patients (n = 48, 29 male; 35.1 ± 8.0 y). All patients completed functional tests (weight-bearing squat, walk-across, sit-to-stand, step-up/over, and forward lunge) using the NeuroCom long force plate (Clackamas, OR) and completed patient-reported outcome measures (International Knee Documentation Committee Subjective Knee Evaluation Form, Lysholm, Western Ontario and McMaster Osteoarthritis Index [WOMAC], and 36-Item Short-Form Health Survey) preoperatively and 3, 6, and 12 mo postoperatively. A covariance pattern model was used to compare performance and self-reported outcome across time and provide a timeline for functional recovery after ACI. Participants demonstrated significant improvement in walk-across stride length from baseline (42.0% ± 8.9% height) at 6 (46.8% ± 8.1%) and 12 mo (46.6% ± 7.6%). Weight bearing on the involved limb during squatting at 30°, 60°, and 90° was significantly less at 3 mo than presurgery. Step-up/over time was significantly slower at 3 mo (1.67 ± 0.69 s) than at baseline (1.49 ± 0.33 s), 6 mo (1.51 ± 0.36 s), and 12 mo (1.40 ± 0.26 s). Step-up/over lift-up index was increased from baseline (41.0% ± 11.3% body weight [BW]) at 3 (45.0% ± 11.7% BW), 6 (47.0% ± 11.3% BW), and 12 mo (47.3% ± 11.6% BW). Forward-lunge time was decreased at 3 mo (1.51 ± 0.44 s) compared with baseline (1.39 ± 0.43 s), 6 mo (1.32 ± 0.05 s), and 12 mo (1.27 ± 0.06). Similarly, forward-lunge impact force was decreased at 3 mo (22.2% ± 1.4% BW) compared with baseline (25.4% ± 1.5% BW). The WOMAC demonstrated significant improvements at 3 mo. All patient-reported outcomes were improved from baseline at 6 and 12 mo postsurgery. Patients' perceptions of improvements may outpace physical changes in function. Decreased function for at least the first 3 mo after ACI should be anticipated, and improvement in performance of tasks requiring weight-bearing knee flexion, such as squatting, going down stairs, or lunging, may not occur for a year or more after surgery.


Integration or fragmentation? Racial diversity and the American future.
Over the next generation or two, America's older, largely white population will increasingly be replaced by today's disproportionately poor minority children. All future growth will come from populations other than non-Hispanic whites as America moves toward a majority-minority society by 2043. This so-called Third Demographic Transition raises important implications about changing racial boundaries in the United States, that is, about the physical, economic, and sociocultural barriers that separate different racial and ethnic groups. America's racial transformation may place upward demographic pressure on future poverty and inequality as today's disproportionately poor and minority children grow into adult roles. Racial boundaries will be reshaped by the changing meaning of race and ethnicity, shifting patterns of racial segregation in neighborhoods and the workplace, newly integrating (or not) friendship networks, and changing rates of interracial marriage and childbearing. The empirical literature provides complicated lessons and offers few guarantees that growing racial diversity will lead to a corresponding breakdown in racial boundaries-that whites and minorities will increasingly share the same physical and social spaces or interact as coequals. How America's older population of elected officials and taxpayers responds today to America's increasingly diverse population will provide a window to the future, when today's children successfully transition (or not) into productive adult roles. Racial and ethnic inclusion will be reshaped by changing ethnoracial inequality, which highlights the need to invest in children-now.


Virological success after 12 and 24 months of antiretroviral therapy in sub-Saharan Africa: Comparing results of trials, cohorts and cross-sectional studies using a systematic review and meta-analysis.
UNAIDS recently defined the 90-90-90 target as a way to end the HIV epidemic. However, the proportion of virological success following antiretroviral therapy (ART) may not be as high as the anticipated 90%, and may in fact be highly heterogeneous. We aimed to describe the proportion of virological success in sub-Saharan Africa and to identify factors associated with the proportion of virological success. We performed a systematic review and meta-analysis focusing on the proportion of patients in sub-Saharan Africa who demonstrate virological success at 12 and 24 months since ART initiation, as well as at 6 and 36 months, where possible. Programme factors associated with the proportion of virological success were identified using meta-regression. Analyses were conducted using both on-treatment (OT) and intention-to-treat (ITT) approaches. Eighty-five articles were included in the meta-analysis, corresponding to 125 independent study populations. Using an on-treatment approach, the proportions (95% confidence interval (CI)) of virological success at 12 (n = 64) and at 24 (n = 32) months since ART initiation were 87.7% (81.3-91.0) and 83.7% (79.8-87.6), respectively. Univariate analysis indicated that the proportion of virological success was not different by study design. Multivariate analysis at 24 months showed that the proportion of virological success was significantly larger in studies conducted in public sector sites than in other sites (p = 0.045). Using an ITT approach, the proportions (95% CI) of virological success at 12 (n = 50) and at 24 (n = 20) months were 65.4% (61.8-69.1) and 56.8% (51.3-62.4), respectively. At 12 months, multivariate analysis showed that the proportion of success was significantly lower in cohort studies than in trials (63.0% vs. 71.1%; p = 0.017). At 24 months, univariate analysis demonstrated that the proportion of success was also lower in cohorts. Regardless of the time following ART initiation, and of the threshold, proportions of virological success were highly variable. Evidence from this review suggests that the new international target of 90% of patients controlled is not yet being achieved, and that in order to improve the virological outcome, efforts should be made to improve retention in care.


Spatio-temporal feature-extraction techniques for isolated gesture recognition in Arabic sign language.
This paper presents various spatio-temporal feature-extraction techniques with applications to online and offline recognitions of isolated Arabic Sign Language gestures. The temporal features of a video-based gesture are extracted through forward, backward, and bidirectional predictions. The prediction errors are thresholded and accumulated into one image that represents the motion of the sequence. The motion representation is then followed by spatial-domain feature extractions. As such, the temporal dependencies are eliminated and the whole video sequence is represented by a few coefficients. The linear separability of the extracted features is assessed, and its suitability for both parametric and nonparametric classification techniques is elaborated upon. The proposed feature-extraction scheme was complemented by simple classification techniques, namely, K nearest neighbor (KNN) and Bayesian, i.e., likelihood ratio, classifiers. Experimental results showed classification performance ranging from 97% to 100% recognition rates. To validate our proposed technique, we have conducted a series of experiments using the classical way of classifying data with temporal dependencies, namely, hidden Markov models (HMMs). Experimental results revealed that the proposed feature-extraction scheme combined with simple KNN or Bayesian classification yields comparable results to the classical HMM-based scheme. Moreover, since the proposed scheme compresses the motion information of an image sequence into a single image, it allows for using simple classification techniques where the temporal dimension is eliminated. This is actually advantageous for both computational and storage requirements of the classifier.


Prestroke weight loss is associated with poststroke mortality among men in the Honolulu-Asia Aging Study.
To examine baseline prestroke weight loss and poststroke mortality among men. Longitudinal study of late-life prestroke body mass index (BMI), weight loss, and BMI change (midlife to late life) with up to 8-year incident stroke and mortality follow-up. Community-based aging study data. Japanese-American men (N=3581; age range, 71-93y) who were stroke free at baseline. Not applicable. Poststroke mortality: 30 days poststroke, analyzed with stepwise multivariable logistic regression; and long-term poststroke (up to 8y), analyzed with stepwise multivariable Cox regression. Weight loss (4.5kg decrements) was associated with increased 30-day poststroke mortality (adjusted odds ratio=1.48; 95% confidence interval [CI], 1.14-1.92), long-term mortality after incident stroke (all types, n=225; adjusted hazards ratio (aHR)=1.25; 95% CI, 1.09-1.44), and long-term mortality after incident thromboembolic stroke (n=153; aHR=1.19; 95% CI, 1.01 to 1.40). Men with overweight/obese late-life BMI (≥25kg/m(2), compared with healthy/underweight BMI) had increased long-term mortality after incident hemorrhagic stroke (n=54; aHR=2.27; 95% CI, 1.07-4.82). Neither desirable nor excessive BMI reductions (vs no change/increased BMI) were associated with poststroke mortality. In the overall sample (N=3581), nutrition factors associated with increased long-term mortality included the following: (1) weight loss (10lb decrements; aHR=1.15; 95% CI, 1.09-1.21), (2) underweight BMI (vs healthy BMI; aHR=1.76; 95% CI, 1.40-2.20), and (3) both desirable and excessive BMI reductions (vs no change or gain, separate model from weight loss and BMI; aHR range, 1.36-1.97; P<.001). Although obesity is a risk factor for stroke incidence, prestroke weight loss was associated with increased poststroke (all types and thromboembolic) mortality. Overweight/obese late-life BMI was associated with increased posthemorrhagic stroke mortality. Desirable and excessive BMI reductions were not associated with poststroke mortality. Weight loss, underweight late-life BMI, and any BMI reduction were all associated with increased long-term mortality in the overall sample.


Peptides for tumor-specific drug targeting: state of the art and beyond.
Important elements for an efficient tumor targeted delivery are cancer-specific de novo- or over-expression of target receptors and their availability on the tumor cell surface. Peptides can be designed to selectively bind to such receptors and act as tumor-targeting ligands, as has been revealed in several preclinical studies. Notably, the amino acid sequences of these peptides can be chemically modified to prevent enzymatic degradation and improve the stability of the peptide. Furthermore, tumor-targeting peptides can be conjugated to the surface of nanosized drug carriers for the selective delivery of anticancer drugs to tumors. In this review, tumor receptors for which targeting peptides have been identified are described, and their targeting potential is considered. Various chemical modifications of peptides are thoroughly described, and targeting peptides as well as peptide-functionalized nanocarriers are critically discussed. The limitations of active targeting nanocarriers are evaluated in detail, and an outlook on the potential of tumor-targeting peptides and their clinical applications is provided.


Perceptions of What Is Important for Appetite-An Interview Study With Older People Having Food Distribution.
The proportion of older people in the population increases and more and more continue living in their own homes. Appetite among the elderly people is important to their nutrition and health. The increased risk of unintended weight loss and malnutrition is linked to food distribution among home-living elderly people. The aim was to describe experiences and perceptions of what matters to appetite among home-living elderly people having food distribution. The design was qualitative where interview data were collected among 13 interviewees in 2017 to 2018. Data were analyzed using qualitative content analysis. The results are presented in three domains: the food, the meal situation, and the adaptation to meal service with categories and themes responding to each domain. The six themes related to appetite among the elderly people concerned the following: eating tasty, savory, and culturally adapted food; eating healthy and sustainable food; eating alone or together with others; eating in a pleasant meal environment; having choices to make about the meal; and last, accepting disabilities and increased dependency. One conclusion is that many aspects should be taken into consideration when promoting appetites of people who also get food distribution. It is highly individual and an understanding of which aspects are relevant must be considered; consequently, person-centered care is suggested to promote appetite.


Evaluation of the microclimate in poultry transport module drawers during the marketing process of end-of-lay hens from farm to slaughter.
Changes in module drawer temperature and relative humidity were monitored for 24 commercial loads of hens. Mathematical models revealed significant differences in predicted drawer temperature depending on their location and the outside environmental temperature. Higher predicted temperatures were found in uppermost drawers of the top modules at the front of the lorry, and lower temperatures in drawers on the outer sides of modules and in those drawers in modules next to the back of the lorry in both the upper and lower modules during transport. In the lairage, drawer temperature generally decreased, except in drawers at the top of modules where temperatures increased. Temperature increases were most often recorded in modules which had been located at the rear of the lorry, which were generally cooler during transport. End-of-lay hens would appear to be exposed to a greater risk of cold stress rather than heat stress in the UK. Inspection of birds during transport, or upon arrival, should be directed to the bottom and side drawers of a load when looking for cold stress, and the top row of drawers (centre) of the top modules when looking for heat stress. The frequency of inspections should increase at times of high ambient temperature while the birds are being held in lairages. Adjusting the numbers of birds loaded per drawer according to bird condition and weather appears to be an effective mitigation strategy which is already in use commercially.


Orthotopic xenografts of human melanoma and colonic and ovarian carcinoma in sheep to evaluate radioimmunotherapy.
Extrapolation to humans from experimental radioimmunotherapy in nude mouse xenograft models is confounded by large relative tumour size and small volume of distribution in mice allowing tumour uptake of radiolabelled antibodies unattainable in patients. Our large animal model of human tumours in cyclosporin-immunosuppressed sheep demonstrated tumour uptake of targeted radiolabelled monoclonal antibodies comparable with uptakes reported in clinical trials. Sheep immunosuppression with daily intravenous cyclosporin augmented by oral ketoconazole maintained trough blood levels of cyclosporin within the range 1000-1500 ng ml(-1). Human tumour cells were transplanted orthotopically by inoculation of 10(7) cells: SKMEL melanoma subcutaneously; LS174T and HT29 colon carcinoma into bowel, peritoneum and liver; and JAM ovarian carcinoma into ovary and peritoneum. Tumour xenografts grew at all sites within 3 weeks of inoculation, preserving characteristic morphology without evidence of necrosis or host rejection. Lymphatic metastasis was demonstrated in regional nodes draining xenografts of melanoma and ovarian carcinoma. Colonic LS1 74T xenografts produced mucin and carcinoembryonic antigen (CEA). The anti-CEA IgG1 monoclonal antibody A5B7 was radiolabelled with iodine-131 and administered intravenously to sheep. Peak uptake at 5 days in orthotopic human tumour transplants in gut was 0.027% DI g(-1) (percentage of injected dose per gram) and 0.034% DI g(-1) in hepatic metastases with tumour to blood ratios of 2-2.5. Non-specific tumour uptake in melanoma was 0.003% DI g(-1). Uptake of radiolabelled monoclonal antibody in human tumours in our large animal model is comparable with that observed in patients and may be more realistic than nude mice xenografts for prediction of clinical efficacy of radioimmunotherapy.


Perspectives on faculty clinical practice: views of occupational therapy curriculum chairpersons and faculty members.
This study investigated whether and how faculty clinical practice (FCP) was being implemented in occupational therapy professional education programs. Chairpersons and faculty members from all accredited entry-level occupational therapy curricula in the United States were asked to complete questionnaires about their involvement in FCP. The chairperson questionnaire consisted of primarily closed-ended questions that addressed the organization of faculty practice within their programs, whereas the faculty questionnaire was primarily open-ended and addressed faculty members' individual involvement in FCP as well as the perceived benefits and drawbacks. Responses were analyzed for 39 program chairpersons and 136 faculty members were analyzed. Twenty-five chairperson respondents reported that their educational program had FCP, and 44 faculty respondents indicated involvement in FCP during the past year. FCP was more prevalent in public-funded institutions and academic health centers. Faculty members primarily engaged in FCP in order to stay current, enhance teaching, and develop networks. They reported increased credibility with students as an important benefit of clinical practice. The primary reasons faculty members did not engage in clinical practice were insufficient time and institutional policies. Although many faculty members in occupational therapy education programs value clinical practice as a part of their educator role, it is necessary to negotiate responsibilities and rewards to prevent role overload and comply with institutional policies.


[The obligation to inform the patient: issues on the right to be informed].
This article aims to analyze, from a legal perspective, the boundaries of the obligation imposed on health care providers to inform the patient. This requirement originated and was developed as an ethical issue. However, with the newly approved law regulating the rights and duties of patients, the obligation to inform can be viewed from prisms and principles that differ from those governing medical ethics. With this purpose, we will focus on the comparative experience, which will allow us to evaluate the responsibility of health care providers when this duty is breached. We will try to answer the following questions: Which medical information must be informed to the patient? When should the doctor inform the patient? In which form should this information be provided?.


Impact of primary local treatment on the development of distant metastases or death through locoregional recurrence in young breast cancer patients.
In this study, we tested the hypothesis whether breast conserving therapy (BCT) compared with mastectomy is associated with a negative outcome in terms of distant metastases or death (DMD) and investigated the relation between locoregional recurrence (LRR) and DMD in young breast cancer (BC) patients. This study included a consecutive series of 536 patients ≤40 years of age at diagnosis with pathological T1N0-3M0 BC, treated between 1989 and 2005. A multistate survival model was used to evaluate the influences of local treatment and LRR on DMD, adjusted for potential prognostic factors. Patients were treated with mastectomy (N = 213) or BCT (N = 323). Median age at diagnosis was 36.3 years, with a median follow-up of 9.0 years. The 10-year actuarial cumulative incidence of DMD was 30.6 % after mastectomy and 26.3 % after BCT (P = 0.04). In total, 81 (15 %) LRRs were observed. After BCT, patients had a threefold higher risk of LRR than after mastectomy (HR 2.9; 95 % CI 1.6-5.3). Patients with LRR had a higher risk of DMD compared with patients without LRR (HR 5.5; 95 % CI 2.1-14.5). However, BCT was not negatively associated with DMD-after-LRR (HR 0.47; 95 % CI 0.2-1.1, BCT vs mastectomy). In conclusion, although LRR significantly affected DMD, the increased risk of LRR after BCT compared with mastectomy did not lead to a worse DMD outcome in BC patients ≤40 years of age.


Diagnostic value of pediatric blood culture bottles for acute postoperative endophthalmitis.
To report our experience using conventional culture methods (CM) and pediatric blood culture bottles (PBCBs) for vitreous sample culture of acute postoperative endophthalmitis. A retrospective study was conducted at the Department of Ophthalmology, Hospital das Clinicas, HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Sao Paulo, BR, from January 2010 to December 2015, and it included 54 patients with clinically suspected acute postoperative endophthalmitis. Vitreous samples were obtained by vitreous tap or vitrectomy. Samples from January 2010 to December 2011 were cultivated in CM, whereas samples from January 2012 to December 2015 were inoculated in PBCBs. The measured outcome was the yield of positive cultures. Twenty cases were included in the CM group, and 34 cases were included in the PBCB group. The yield of positive cultures in PBCBs (64.7%) was significantly higher than that in conventional CM (35%, p=0.034). Staphylococcus epidermidis and Streptococcus viridans were the two most commonly found agents. PBCBs can be used successfully in clinically suspected endophthalmitis. The method showed a higher yield of positive cultures than the conventional method. This technique appears to have several advantages over the traditional method: it saves time, as only one medium needs to be inoculated; transportation to a laboratory is easier than in the traditional method, and there is no need to maintain a supply of fresh agar media. The use of PBCBs may be recommended as the primary method for microbiological diagnosis and is especially suitable for office settings and remote clinics.


Tunable in-plane torsional strength of surface functionalized two dimensional nanomaterials.
In this paper, the in-plane torsional properties of two dimensional nanomaterials are revealed to be tunable by surface functionalization using molecular dynamics simulations. The torsional strengths of both graphene and graphyne under circular shearing are found to first decrease and then increase anomalously with the increase of the hydrogenation ratio. The minimum strength of such U-shaped strength evolution appears at different hydrogenation ratios H for graphene (H = 50%) and graphyne (H = 70%), demonstrating a correlation between hydrogenation and the unit lattices of 2D nanomaterials. By studying the torsional characteristics of a graphene annulus with varying outer-to-inner boundary radius ratios, the anomalous increase of torsional strength at a high hydrogenation ratio is revealed to be the result of wrinkling deformation as well as chemical bond hybridization. Further analyses of the local stress distribution are carried out for understanding the mechanism of surface hydrogenation tuned in-plane torsional strength of graphene and graphyne. Torsional strength of the graphene annulus with an elliptical outer boundary also shows a U-shaped evolution with hydrogenation ratio, while the shape effect of the graphene annulus can smooth the change of the torsional characteristics with hydrogenation. Our results provide novel insights into the development of 2D nanomaterial-based annular devices.


Asking sensitive questions in conservation using Randomised Response Techniques.
Conservation increasingly seeks knowledge of human behaviour. However, securing reliable data can be challenging, particularly if the behaviour is illegal or otherwise sensitive. Specialised questioning methods such as Randomised Response Techniques (RRTs) are increasingly used in conservation to provide greater anonymity, increase response rates, and reduce bias. A rich RRT literature exists, but successfully navigating it can be challenging. To help conservationists access this literature, we summarise the various RRT designs available and conduct a systematic review of empirical applications of RRTs within (n = 32), and beyond conservation (n = 66). Our results show increased application of RRTs in conservation since 2000. We compare the performance of RRTs against known prevalence of the sensitive behaviour and relative to other questioning techniques to assess how successful RRTs are at reducing bias (indicated by securing higher estimates). Findings suggest that RRT applications in conservation were less likely than those in other disciplines to provide prevalence estimates equal to, or higher than those derived from direct questions. Across all disciplines, we found reports of non-compliance with RRT instructions were common, but rarely accounted for in study design or analysis. For the first time, we provide conservationists considering RRTs with evidence on what works, and provide guidance on how to develop robust designs suitable for conservation research contexts. We highlight when alternate methods should be used, how to increase design efficiency and improve compliance with RRT instructions. We conclude RRTs are a useful tool, but their performance depends on careful design and implementation.


FSH directly regulates chondrocyte dedifferentiation and cartilage development.
Previous studies suggest that postmenopausal osteoarthritis is linked to a decrease in estrogen levels. However, whether follicle-stimulating hormone (FSH), the upstream hormone of estrogen, affects cartilage destruction and thus contributes to the onset of osteoarthritis has never been explored. To evaluate the potential involvement of FSH in joint degeneration and to identify the molecular mechanisms through which FSH influences chondrocytes, mouse cartilage chondrocytes and the ATDC5 chondrocyte cell line were treated with FSH and inhibitors of intracellular signaling pathways. We observed that FSH induces chondrocyte dedifferentiation by decreasing type II collagen (Coll-II) synthesis. Chondrocyte cytoskeleton reorganization was also observed after FSH treatment. The FSH-induced decrease in Coll-II was rescued by ERK-1/2 inhibition but aggravated by p38 inhibition. In addition, knocking down the FSH receptor (Fshr) by using Fshr siRNA abolished chondrocyte dedifferentiation, as indicated by the increased expression of Coll-II. Inhibition of the protein Gαi by pertussis toxin (PTX) also restored FSH-inhibited Coll-II, suggesting that Gαi is downstream of FSHR in chondrocyte dedifferentiation. FSHβ antibody blockade prevented cartilage destruction and cell loss in mice. Moreover, decreased Coll-II staining due to the progression of aging could be rescued by blocking FSH. Thus, we suggest that high circulating FSH, independent of estrogen, is an important regulator in chondrocyte dedifferentiation and cartilage destruction.


The politics of comparative effectiveness research: lessons from recent history.
Efforts to support and use comparative effectiveness research (CER), some more successful than others, have been promulgated at various times over the last forty years. Following a resurgence of interest in CER, recent health care reforms provided substantial support to strengthen its role in US health care. While CER has generally captured bipartisan support, detractors have raised concerns that it will be used to ration services and heighten government control over health care. Such concerns almost derailed the initiative during passage of the health care reform legislation and are still present today. Given recent investments in CER and the debates surrounding its development, the time is ripe to reflect on past efforts to introduce CER in the United States. This article examines previous initiatives, highlighting their prescribed role in US health care, the reasons for their success or failure, and the political lessons learned. Current CER initiatives have corrected for many of the pitfalls experienced by previous efforts. However, past experiences point to a number of issues that must still be addressed to ensure the long-term success and sustainability of CER, including adopting realistic aims about its impact, demonstrating the impact of Patient-Centered Outcomes Research Institute (PCORI) and communicating the benefits of CER, and maintaining strong political and stakeholder support.


Neuroimaging in epilepsy.
Epilepsy neuroimaging is important for detecting the seizure onset zone, predicting and preventing deficits from surgery and illuminating mechanisms of epileptogenesis. An aspiration is to integrate imaging and genetic biomarkers to enable personalized epilepsy treatments. The ability to detect lesions, particularly focal cortical dysplasia and hippocampal sclerosis, is increased using ultra high-field imaging and postprocessing techniques such as automated volumetry, T2 relaxometry, voxel-based morphometry and surface-based techniques. Statistical analysis of PET and single photon emission computer tomography (STATISCOM) are superior to qualitative analysis alone in identifying focal abnormalities in MRI-negative patients. These methods have also been used to study mechanisms of epileptogenesis and pharmacoresistance.Recent language fMRI studies aim to localize, and also lateralize language functions. Memory fMRI has been recommended to lateralize mnemonic function and predict outcome after surgery in temporal lobe epilepsy. Combinations of structural, functional and post-processing methods have been used in multimodal and machine learning models to improve the identification of the seizure onset zone and increase understanding of mechanisms underlying structural and functional aberrations in epilepsy.


Impaired arachidonic (20:4n-6) and docosahexaenoic (22:6n-3) acid synthesis by phenylalanine metabolites as etiological factors in the neuropathology of phenylketonuria.
The recent literature on polyunsaturated fatty acid metabolism in phenylketonuria (PKU) is critically analyzed. The data suggest that developmental impairment of the accretion of brain arachidonic (20:4n-6) and docosahexaenoic (22:6n-3, DHA) acids is a major etiological factor in the microcephaly and mental retardation of uncontrolled PKU and maternal PKU. These fatty acids appear to be synthesized by the recently elucidated carnitine-dependent, channeled, mitochondrial fatty acid desaturases for which alpha-tocopherolquinone (alpha-TQ) is an essential enzyme cofactor. alpha-TQ can be synthesized either de novo or from alpha-tocopherol. The fetus and newborn would primarily rely on de novo alpha-TQ synthesis for these mitochondrial desaturases because of low maternal transfer of alpha-tocopherol. Homogentisate, a pivotal intermediate in the de novo pathway of alpha-TQ synthesis, is synthesized by 4-hydroxyphenylpyruvate dioxygenase. The major catabolic products of excess phenylalanine, viz. phenylpyruvate and phenyllactate, are proposed to inhibit alpha-TQ synthesis at the level of the dioxygenase reaction by competing with its 4-hydroxyphenylpyruvate substrate, thus leading to a developmental impairment of 20:4n-6 and 22:6n-3 synthesis in uncontrolled PKU and fetuses of PKU mothers. The data suggest that dietary supplementation with carnitine, 20:4n-6, and 22:6n-3 may have therapeutic value for PKU mothers and for PKU patients who have been shown to have a low plasma status of these essential metabolites.


Congenital lipoid adrenal hyperplasia caused by a novel splicing mutation in the gene for the steroidogenic acute regulatory protein.
Steroidogenic acute regulatory protein (StAR) plays a crucial role in the transport of cholesterol from the cytoplasm to the inner mitochondrial membrane, facilitating its conversion to pregnenolone by cytochrome P450scc. Its essential role in steroidogenesis was demonstrated after observing that StAR gene mutations gave rise to a potentially lethal disease named congenital lipoid adrenal hyperplasia, in which virtually no steroids are produced. We report here a 2-month-old female patient, karyotype 46XY, who presented with growth failure, convulsions, dehydration, hypoglycemia, hyponatremia, hypotension, and severe hyperpigmentation suggestive of adrenal insufficiency. Serum cortisol, 17OH-progesterone, dehydroepiandrosterone sulfate, testosterone, 17OH-pregnenolone, and aldosterone levels were undetectable in the presence of high ACTH and plasma renin activity levels. Immunohistochemical analysis of testis tissues revealed the absence of StAR protein. Molecular analysis of StAR gene demonstrated a homozygous G to T mutation within the splice donor site of exon 1 (IVS1 + 1G>T). Her parents and one brother were heterozygous for this mutation. In vitro analysis of the mutation was performed in COS cells transfected with minigenes coding regions spanning exon-intron 1 to 3 carrying the mutant and the wild-type sequences. RT-PCR analyses of the mutant gene showed an abnormal mRNA transcript of 2430 bp (normal size 433 bp). Sequence analysis of the mutant mRNA demonstrated the retention of intron 1. Immunolocalization of the StAR minigene product detected the peptide in the mitochondria of COS cells transfected with the wild-type minigene but not in those transfected with the mutant minigene. We conclude that this mutation gives rise to a truncated StAR protein, which lacks an important N-terminal region and the entire lipid transfer domain.


Structure and function in rhodopsin: replacement by alanine of cysteine residues 110 and 187, components of a conserved disulfide bond in rhodopsin, affects the light-activated metarhodopsin II state.
A disulfide bond that is evidently conserved in the guanine nucleotide-binding protein-coupled receptors is present in rhodopsin between Cys-110 and Cys-187. We have replaced these two cysteine residues by alanine residues and now report on the properties of the resulting rhodopsin mutants. The mutant protein C110A/C187A expressed in COS cells resembles wild-type rhodopsin in the ground state. It folds correctly to bind 11-cis-retinal and form the characteristic rhodopsin chromophore. It is inert to hydroxylamine in the dark, and its stability to dark thermal decay is reduced, relative to that of the wild type, by a delta delta G not equal to of only -2.9 kcal/mol. Further, the affinities of the mutant and wild-type rhodopsins to the antirhodopsin antibody rho4D2 are similar, both in the dark and in light. However, the metarhodopsin II (MII) and MIII photointermediates of the mutant are less stable than those formed by the wild-type rhodopsin. Although the initial rates of transducin activation are the same for both mutant and wild-type MII intermediates at 4 degrees C, at 15 degrees C the MII photointermediate in the mutant decays more than 20 times faster than in wild type. We conclude that the disulfide bond between Cys-110 and Cys-187 is a key component in determining the stability of the MII structure and its coupling to transducin activation.


Palladium/IzQO-Catalyzed Coordination-Insertion Copolymerization of Ethylene and 1,1-Disubstituted Ethylenes Bearing a Polar Functional Group.
Coordination-insertion copolymerization of ethylene with 1,1-disubstituted ethylenes bearing a polar functional group, such as methyl methacrylate (MMA), is a long-standing challenge in catalytic polymerization. The major obstacle for this process is the huge difference in reactivity of ethylene versus 1,1-disubstituted ethylenes toward both coordination and insertion. Herein we report the copolymerization of ethylene and 1,1-disubstituted ethylenes by using an imidazo[1,5-a]quinolin-9-olate-1-ylidene-supported palladium catalyst. Various types of 1,1-disubstituted ethylenes were successfully incorporated into the polyethylene chain. In-depth characterization of the obtained copolymers and mechanistic inferences drawn from stoichiometric reactions of alkylpalladium complexes with methyl methacrylate and ethylene indicate that the copolymerization proceeds by the same coordination-insertion mechanism that has been postulated for ethylene.