Implications of dual practice for universal health coverage.
Making progress towards universal health coverage (UHC) requires that health workers are adequate in numbers, prepared for their jobs and motivated to perform. In establishing the best ways to develop the health workforce, relatively little attention has been paid to the trends and implications of dual practice - concurrent employment in public and private sectors. We review recent research on dual practice for its potential to guide staffing policies in relation to UHC. Many studies describe the characteristics and correlates of dual practice and speculate about impacts, but there is very little evidence that is directly relevant to policy-makers. No studies have evaluated the impact of policies on the characteristics of dual practice or implications for UHC. We address this lack and call for case studies of policy interventions on dual practice in different contexts. Such research requires investment in better data collection and greater determination on the part of researchers, research funding bodies and national research councils to overcome the difficulties of researching sensitive topics of health systems functions.


Sample size for binary logistic prediction models: Beyond events per variable criteria.
Binary logistic regression is one of the most frequently applied statistical approaches for developing clinical prediction models. Developers of such models often rely on an Events Per Variable criterion (EPV), notably EPV ≥10, to determine the minimal sample size required and the maximum number of candidate predictors that can be examined. We present an extensive simulation study in which we studied the influence of EPV, events fraction, number of candidate predictors, the correlations and distributions of candidate predictor variables, area under the ROC curve, and predictor effects on out-of-sample predictive performance of prediction models. The out-of-sample performance (calibration, discrimination and probability prediction error) of developed prediction models was studied before and after regression shrinkage and variable selection. The results indicate that EPV does not have a strong relation with metrics of predictive performance, and is not an appropriate criterion for (binary) prediction model development studies. We show that out-of-sample predictive performance can better be approximated by considering the number of predictors, the total sample size and the events fraction. We propose that the development of new sample size criteria for prediction models should be based on these three parameters, and provide suggestions for improving sample size determination.


Gene probing reveals the widespread distribution, diversity and abundance of isoprene-degrading bacteria in the environment.
Approximately 500 Tg of isoprene are emitted to the atmosphere annually, an amount similar to that of methane, and despite its significant effects on the climate, very little is known about the biological degradation of isoprene in the environment. Isolation and characterisation of isoprene degraders at the molecular level has allowed the development of probes targeting isoA encoding the α-subunit of the isoprene monooxygenase. This enzyme belongs to the soluble diiron centre monooxygenase family and catalyses the first step in the isoprene degradation pathway. The use of probes targeting key metabolic genes is a successful approach in molecular ecology to study specific groups of bacteria in complex environments. Here, we developed and tested a novel isoA PCR primer set to study the distribution, abundance, and diversity of isoprene degraders in a wide range of environments. The new isoA probes specifically amplified isoA genes from taxonomically diverse isoprene-degrading bacteria including members of the genera Rhodococcus, Variovorax, and Sphingopyxis. There was no cross-reactivity with genes encoding related oxygenases from non-isoprene degraders. Sequencing of isoA amplicons from DNA extracted from environmental samples enriched with isoprene revealed that most environments tested harboured a considerable variety of isoA sequences, with poplar leaf enrichments containing more phylogenetically diverse isoA genes. Quantification by qPCR using these isoA probes revealed that isoprene degraders are widespread in the phyllosphere, terrestrial, freshwater and marine environments. Specifically, soils in the vicinity of high isoprene-emitting trees contained the highest number of isoprene-degrading bacteria. This study provides the molecular ecology tools to broaden our knowledge of the distribution, abundance and diversity of isoprene degraders in the environment, which is a fundamental step necessary to assess the impact that microbes have in mitigating the effects of this important climate-active gas.


Parasuicide in psychiatric inpatients: results of a controlled investigation.
A group of 20 psychiatric inpatients who committed parasuicide during their hospital stay were compared with 58 randomly selected control inpatients and with 34 patients who were admitted following a parasuicide. The findings indicate: (1) The frequency of in-hospital parasuicide amounts to 3.2% of all admissions. (2) Apart from the more frequent past suicidal activity of the in-hospital parasuicide group, no really important differences were identified between these and control patients. The broad definition of parasuicide probably accounts for this result. (3) The patients who committed parasuicide before admission represent a diagnostically different, clinically less severely ill population. The differences between both groups of parasuicide patients support the notion of the heterogeneity of the parasuicide population.


Patterns of medical accidents and disputes in the orthodontic field in Korea.
The committee of admitted doctors developed a questionnaire regarding medical dispute and distributed it to 1,600 members of Korean Academy of Orthodontics. The questionnaire consisted of three categories and 56 items covering basic information about the doctors and patients who had experienced medical disputes, the cause and workaround of medical accidents, and methods for taking precautions. The present survey showed a similar proportion of responders who had experienced a medical accident compared to the study in 1997. The primary reason for medical disputes was dissatisfaction with appearance. Many doctors felt that they would likely experience a medical dispute at some point. Most disputes were settled by doctors themselves, usually for an amount of less than 5 million Korean won. For some doctors, medical accidents lead to ongoing psychological problems. Responders felt that continuing education for medical dispute is very necessary. These results reveal a need for the association of orthodontists to lead advancements in education and countermeasures for preventing and managing medical accidents and disputes.


The shape of nuclear falls and their effect on the perception of pitch and prominence: peaks vs. plateaux.
This article investigates the perceptual effect of a high plateau in the intonation contour. Plateaux are flat stretches of contour and have been observed associated with high tones in Standard Southern British (SSB) English. The hypothesis that plateaux may make the accents with which they are associated sound higher in pitch than sharp peaks of the same maximum frequency is tested experimentally. In the first experiment listeners heard pairs of resynthesized utterances where the nuclear accent differed only in shape, not frequency. They indicated which stimulus they thought contained the higher pitched accent. Results showed that plateau-shaped accents sound higher than peaks. In the second experiment the effect of a plateau on prominence relations within an utterance is investigated. Listeners heard resynthesized sentences, and compared two accents. One group indicated which accent sounded higher in pitch and the other indicated which sounded more prominent. Results again indicated that plateau-shaped accents sound higher in pitch and also more prominent; judgments of pitch and prominence were very similar to one another. The results from both experiments indicated that accent shape is a perceptually important variable, although such a fine level of detail is not taken into account by autosegmental-metrical theories of intonation.


The Third National Telemedicine & Telehealth Service Provider Showcase Conference: Advancing Telehealth Partnerships.
As telemedicine and telehealth services are experiencing increasing rates of adoption, industry leaders and healthcare service providers are becoming increasingly focused on human resource issues encountered in the delivery of a broad range of telehealth services. To create a forum for the discussion of many interrelated elements of telehealth service industry, a national conference entitled "Telemedicine & Telehealth Service Provider Showcase" (SPS) Conference was established in 2014, and repeated in 2016 and 2017, in Arizona. These SPS Conferences include thought leaders, telehealth service providers, government administrators, and academicians from leading programs addressing service provider workforce issues. This report summarizes the content of SPS 2017 conference, held in Phoenix, AZ, October 2-3, 2017. The topics covered at SPS 2017 include using telehealth services as a strategic asset; development of appropriate effective partnerships; direct-to-consumer initiatives; important reimbursement, legislative, and regulatory issues (i.e., Centers for Medicare & Medicaid Services [CMS] approaches, financial models, and return on investment [ROI]); marketing; evaluation and applied metrics; remote monitoring and sensors; integration with electronic health records; and overall lessons learned. The content of SPS 2017 is summarized in the body of this report. The SPS 2017 program evaluators included attendees, speakers, and exhibitors. The knowledge attendees gained at SPS 2017 was characterized, by all three groups, as forward-looking and practical. SPS 2017 succeeded in identifying, and focusing on, solutions for issues, challenges, and barriers impacting the rapidly expanding telehealth service segment of the healthcare industry. The growing interest in this annual SPS Conference series apparently reflects, in part, the program committee's successes in identifying practical issues and their potential solutions.


Setting ART initiation targets in response to changing guidelines: The importance of addressing both steady-state and backlog.
Target setting is useful in planning, assessing and improving antiretroviral treatment (ART) programmes. In the past 4 years, the ART initiation environment has been transformed due to the change in eligibility criteria (starting ART at a CD4+ count <350 cells/μl v. <200 cells/μl) and the roll-out of nurse-initiated management of ART. To describe and illustrate the use of a target-setting model for estimating district-based targets in the era of an expanding ART programme and changing CD4+ count thresholds for ART initiation. Using previously described models and data for annual new HIV infections, we estimated both steady-state need for ART initiation and backlog in a North West Province district, accounting for the shift in eligibility. Comparison of actual v. targeted ART initiations was undertaken. The change in CD4+ count threshold adds a once-off group of newly eligible patients to the pool requiring ART - the backlog. The steady-state remains unchanged as it is determined by the annual rate of new HIV infections in previous years. The steady-state need for the district was 639 initiations/month, and the backlog was ~15,388 patients. After the shift in eligibility in September 2011, the steady-state target was exceeded over several months with some backlog addressed. Of the total backlog for this district, 72% remains to be cleared. South Africa has two pools of patients who need ART: the steady-state of HIV-infected patients entering the programme each year, determined by historical infection rates; and the backlog created by the shift in eligibility. The healthcare system needs to build long- term capacity to meet the steady-state need for ART and additional capacity to address the backlog.


Clarifying boundaries of binge eating disorder and psychiatric comorbidity: a latent structure analysis.
Binge eating disorder (BED) presents with substantial psychiatric comorbidity. This latent structure analysis sought to delineate boundaries of BED given its comorbidity with affective and anxiety disorders. A population-based sample of 151 women with BED, 102 women with affective or anxiety disorders, and 259 women without psychiatric disorders was assessed with clinical interviews and self-report-questionnaires. Taxometric analyses were conducted using DSM-IV criteria of BED and of affective and anxiety disorders. The results showed a taxonic structure of BED and of affective and anxiety disorders. Both taxa co-occurred at an above-chance level, but also presented independently with twice-as-large probabilities. Within the BED taxon, diagnostic co-occurrence indicated greater general psychopathology, lower social adaptation, and greater premorbid exposure to parental mood and substance disorder, but not greater eating disorder psychopathology. Eating disorder psychopathology discriminated individuals in the BED taxon from individuals in the affective and anxiety disorders taxon. Diagnostic criteria of BED were more indicative of the BED taxon than were criteria of affective and anxiety disorders. The results show that at the latent level, BED was co-occurring with, yet distinct from, affective and anxiety disorders and was not characterized by an underlying affective or anxiety disorder.


Hepatic glucose metabolism in late pregnancy: normal versus high-fat and -fructose diet.
Net hepatic glucose uptake (NHGU) is an important contributor to postprandial glycemic control. We hypothesized that NHGU is reduced during normal pregnancy and in a pregnant diet-induced model of impaired glucose intolerance/gestational diabetes mellitus (IGT/GDM). Dogs (n = 7 per group) that were nonpregnant (N), normal pregnant (P), or pregnant with IGT/GDM (pregnant dogs fed a high-fat and -fructose diet [P-HFF]) underwent a hyperinsulinemic-hyperglycemic clamp with intraportal glucose infusion. Clamp period insulin, glucagon, and glucose concentrations and hepatic glucose loads did not differ among groups. The N dogs reached near-maximal NHGU rates within 30 min; mean ± SEM NHGU was 105 ± 9 µmol·100 g liver⁻¹·min⁻¹. The P and P-HFF dogs reached maximal NHGU in 90-120 min; their NHGU was blunted (68 ± 9 and 16 ± 17 µmol·100 g liver⁻¹·min⁻¹, respectively). Hepatic glycogen synthesis was reduced 20% in P versus N and 40% in P-HFF versus P dogs. This was associated with a reduction (>70%) in glycogen synthase activity in P-HFF versus P and increased glycogen phosphorylase (GP) activity in both P (1.7-fold greater than N) and P-HFF (1.8-fold greater than P) dogs. Thus, NHGU under conditions mimicking the postprandial state is delayed and suppressed in normal pregnancy, with concomitant reduction in glycogen storage. NHGU is further blunted in IGT/GDM. This likely contributes to postprandial hyperglycemia during pregnancy, with potential adverse outcomes for the fetus and mother.


Who will need long-term care? Creation and validation of an instrument that identifies older people at risk.
The aim of this study was to create and measure the predictive validity of a screening instrument that identifies older people who are at risk for developing a need for long-term care within a year. This was an observational study, with participants allocated to either a derivation cohort or a validation cohort, in the United States. A nationally representative sample of older community-dwelling Medicare beneficiaries (n = 6,538) participated in the Medicare Current Beneficiary Survey. Questions addressed sociodemographic, functional, health-related, and utilization characteristics in 1991 and 1992, linked to records of Medicare payments for health services during 1991-1992. In the derivation cohort, 14 self-reported characteristics were significant predictors of developing a need for long-term care within 1 year. In the validation cohort, these 14 characteristics identified a high-risk subgroup (18%) that, during the following year, developed a need for long-term care at six times the rate of the low-risk majority. This brief survey instrument identifies a high-risk minority of older people that will, during the following year, develop a need for long-term care at six times the rate of the low-risk majority. This instrument may be useful for targeting at-risk subgroups of older populations to receive interventions designed to preserve functional independence and avert the need for long-term care.


Distinct molecular mechanism for initiating TRAF6 signalling.
Tumour-necrosis factor (TNF) receptor-associated factor 6 (TRAF6) is the only TRAF family member that participates in signal transduction of both the TNF receptor (TNFR) superfamily and the interleukin-1 receptor (IL-1R)/Toll-like receptor (TLR) superfamily; it is important for adaptive immunity, innate immunity and bone homeostasis. Here we report crystal structures of TRAF6, alone and in complex with TRAF6-binding peptides from CD40 and TRANCE-R (also known as RANK), members of the TNFR superfamily, to gain insight into the mechanism by which TRAF6 mediates several signalling cascades. A 40 degrees difference in the directions of the bound peptides in TRAF6 and TRAF2 shows that there are marked structural differences between receptor recognition by TRAF6 and other TRAFs. The structural determinant of the petide TRAF6 interaction reveals a Pro-X-Glu-X-X-(aromatic/acidic residue) TRAF6-binding motif, which is present not only in CD40 and TRANCE-R but also in the three IRAK adapter kinases for IL-1R/TLR signalling. Cell-permeable peptides with the TRAF6-binding motif inhibit TRAF6 signalling, which indicates their potential as therapeutic modulators. Our studies identify a universal mechanism by which TRAF6 regulates several signalling cascades in adaptive immunity, innate immunity and bone homeostasis.


Variations in infant feeding practice are associated with body composition in childhood: a prospective cohort study.
Most studies of infant diet and later body composition focus on milk feeding; few consider the influence of variations in the weaning diet. Our objective was to examine how variations in milk feeding and the weaning diet relate to body composition at 4 yr. A total of 536 children participating in a prospective birth cohort study. Diet was assessed at 6 and 12 months of age. Compliance with weaning guidance was defined by the infant's score for a principal component analysis-defined dietary pattern (infant guidelines) at 12 months. Infants with high infant guidelines scores had diets characterized by high consumption of fruit, vegetables, and home-prepared foods. Body composition was assessed at 4 yr by dual x-ray absorptiometry. Longer duration of breastfeeding was associated with lower fat mass at 4 yr [4.5 kg, 95% confidence interval (CI) of 4.3-4.7 kg, in children breastfed for 12 months or more, compared with 5.0 (95% CI 4.7-5.3) kg in children never breastfed (P = 0.002)] but was not related to body mass index. Children with high infant guidelines scores had a higher lean mass [12.6 (95% CI 12.3-12.9) kg in children in the top quarter of the distribution, compared with 12.0 (95% CI 11.7-12.4) kg in children in the bottom quarter (P = 0.001)]. These associations were independent and were little changed by adjustment for confounding factors. These data suggest that variations in both milk feeding and in the weaning diet are linked to differences in growth and development, and they have independent influences on body composition in early childhood.


Riboflavin offers a targeted strategy for managing hypertension in patients with the MTHFR 677TT genotype: a 4-y follow-up.
We recently reported that the elevated blood pressure (BP) observed in patients with cardiovascular disease who are homozygous for the 677C→T polymorphism (TT genotype) in the gene encoding methylenetetrahydrofolate reductase (MTHFR) was responsive to supplementation with riboflavin-the cofactor for MTHFR. The objective was to investigate the effect of riboflavin on BP targeted at patients with the TT genotype 4 y after initial investigation, during which time major changes in the clinical guidelines for antihypertensive therapy were introduced. A total of 83 patients (representing all 3 genotypes) who participated in a placebo-controlled riboflavin intervention for 16 wk in 2004 agreed to take part. Nested within this follow-up, those with the TT genotype (n = 31) proceeded to intervention with riboflavin (1.6 mg/d for 16 wk) or placebo, conducted in a crossover style whereby the 2004 treatment groups were reversed. At follow-up in 2008, as in 2004, patients with the TT genotype had higher systolic BP (P < 0.01), with a nonsignificant trend noted for higher diastolic BP (P = 0.051). Despite the marked changes in antihypertensive therapy that had occurred, BP remained unchanged in patients with the TT genotype at the time of follow-up. Riboflavin supplementation (administered in 2004 and 2008) produced an overall decrease in systolic (-9.2 ± 12.8 mm Hg; P = 0.001) and diastolic (-6.0 ± 9.9 mm Hg; P = 0.003) BP. Optimizing riboflavin status offers a low-cost targeted strategy for managing elevated BP in this genetically at-risk group. These findings, if confirmed in the general population, could have important implications for the prevention of hypertension.


Participation in the Vascular Quality Initiative is associated with improved perioperative medication use, which is associated with longer patient survival.
Medical management (MM) with antiplatelet (AP) and statin therapy is recommended for most patients undergoing vascular surgery and has been advocated by the Vascular Quality Initiative (VQI). We analyzed the effect of VQI participation on perioperative (preoperative and postoperative) MM use over time and the effect of discharge MM on patient survival. We studied VQI patients treated with MM preoperatively and at discharge from 2005 to 2014, including all elective carotid endarterectomy/carotid stenting (n = 28,092), suprainguinal/infrainguinal bypass (n = 11,362), peripheral vascular interventions (n = 24,476), open/endovascular abdominal aortic aneurysm repair (n = 13,503), and thoracic endovascular aneurysm repair (n = 702). We examined trends of MM use over time, as well as the effect of duration of VQI participation on MM use. Multivariable logistic regression analysis was performed to identify factors associated with MM use. In addition, the Cox proportional hazards model was used to identify factors associated with 5-year survival. MM with AP and statin preoperatively and postoperatively across VQI centers improved from 55% in 2005 to 68% in 2009, with a subsequent overall decline to 62% by 2014, coincident with many new centers with lower MM rates joining VQI in 2010. Longer center participation in VQI was associated with improved perioperative MM overall. This was also noted across all procedure types, with MM increasing from 47% to 82% for aneurysm repairs and 69% to 83% for carotid procedures from 1 to 12 years of participation in VQI. After multivariable adjustment, centers in VQI ≥3 years were 30% more likely to have patients on MM (odds ratio, 1.3, 95% confidence interval [CI], 1.3-1.4). Importantly, discharge on AP and statin therapy was associated with improved 5-year survival, compared with discharge on neither medication (82% [95% CI, 81%-83%] vs 67% [95% CI, 62%-72%]), and an adjusted hazard ratio for death of 0.6 (95% CI, 0.5-0.7; P < .001). Discharge on a single medication was associated with intermediate survival at 5 years (AP only: 77% [95% CI, 75%-79%]; statin only: 73% [95% CI, 68%-77%]). These data demonstrate that MM is associated with improved survival after a number of vascular procedures. Importantly, VQI participation improves the use of MM, demonstrating that involvement in an organized quality effort can affect patient outcomes.


The effect of late gestation foetal hypoglycaemia on cardiovascular and endocrine function in sheep.
An appropriate foetal cardiovascular (CV) response to reduced substrate supply (e.g. oxygen or other nutrients) is vital for growth and development, and may impact on CV control. The prevailing nutritional environment and associated CV changes may influence subsequent CV responses to challenges during late gestation, for example, umbilical cord occlusion (UCO). We investigated the effect of low-circulating glucose on foetal CV control mechanisms and response to UCO. Under general anaesthesia, late gestation foetal sheep (n = 7, 119 days gestational age (dGA), term ∼147 days) were implanted with vascular catheters, a bladder catheter, electrocardiogram electrodes and an umbilical cord occluder. Mean arterial pressure (MAP), heart rate (HR) and kidney function were monitored during maternal saline (MSAL, 125dGA) and insulin (MINS, 126dGA) infusion, and foetal CV responses were assessed during incremental doses of angiotensin II, a 90-s total UCO, and administration of phenylephrine to assess baroreflex function. During MINS infusion, the decrease in maternal and foetal blood glucose was associated with a small but significant decrease in foetal HR and reduced foetal baroreflex sensitivity (P < 0.05). The increase in foetal MAP during a 90-s UCO was greater during hypoglycaemia (P < 0.05). The MAP response to angiotensin II was not affected by hypoglycaemia. Decreased foetal HR and baroreflex sensitivity and increased CV responsiveness to UCO during hypoglycaemia indicates altered CV homoestatic mechanisms. The combination of altered nutrition and a CV challenge, such as UCO, during late gestation may have a cumulative effect on foetal CV function.


How to tap an innocent waveguide.
We study the interaction of a mode propagating in a planar waveguide with a three--dimensional rectangular defect (protrusion or notch) in the structure. The scattering by the defect disturbes the propagation of the mode and light is coupled out of the waveguide. To investigate these phenomena we compute electric field distributions with the Green's tensor technique and show movies with varying defect geometries and different mode polarizations. These calculations should be useful for optimizing specific elements in complex photonic circuits.


Apparent nutrient digestibility and excreta quality in African grey parrots fed two pelleted diets based on coarsely or finely ground ingredients.
A feeding trial was performed to study the influence of particle size in extruded parrot pellets on apparent digestibility and excreta consistency and pH. Two test diets were alternately provided to eight African grey parrots according to a 2 x 2 cross-over design. Both diets were similar in nutrient content and ingredient composition but differed in particle size of the composing particles of individual pellets. Apparent digestibility of macronutrients was studied using the total collection method. Next, the appearance of the excreta was studied by calculation of weight-surface ratio of individual excrements as an objective measurement of consistency. Last, excreta pH was measured directly on fresh excrements and on homogenized 10% excreta solutions. Neither apparent digestibility coefficients nor excreta pH values were significantly different in parrots fed the two diets. However, excreta consistency was significantly (p < 0.05) more solid when fed the coarse diet than when fed with the finely ground diet. The results of this study suggest that excreta consistency can be improved through larger particle size, without adverse effects on nutritive value of the diet.


Simulated Performance of a Xenohybrid Bone Graft (SmartBone®) in the Treatment of Acetabular Prosthetic Reconstruction.
Total hip arthroplasty (THA) is a surgical procedure for the replacement of hip joints with artificial prostheses. Several approaches are currently employed in the treatment of this kind of defect. Overall, the most common method involves using a quite invasive metallic support (a Burch-Schneider ring). Moreover, valid alternatives and less invasive techniques still need to be supported by novel material development. In this work, we evaluated the performance of SmartBone®, a xenohybrid bone graft composed of a bovine bone matrix reinforced with biodegradable polymers and collagen, as an effective support in acetabular prosthesis reconstruction. Specifically, the material's mechanical properties were experimentally determined (E = ~1.25 GPa, Ef = ~0.34 GPa, and Et = ~0.49 GPa) and used for simulation of the hip joint system with a SmartBone® insert. Moreover, a comparison with a similar case treated with a Burch-Schneider ring was also conducted. It was found that it is possible to perform THA revision surgeries without the insertion of an invasive metal support and it can be nicely combined with SmartBone®'s osteointegration characteristics. The material can withstand the loads independently (σmax = ~12 MPa) or be supported by a thinner titanium plate in contact with the bone in the worst cases. This way, improved bone regeneration can be achieved.


Sequential administration of docetaxel followed by cisplatin-vindesine: a pilot study in patients with locally advanced or metastatic non-small cell lung cancer (NSCLC).
In this phase II study, the feasibility and efficacy of sequential chemotherapy were tested with agents shown to be active as monotherapy. Patients with chemotherapy-naive, locally advanced or metastatic non-small cell lung cancer were selected for the study. Treatment involved four cycles of docetaxel (100 mg/m(2) on day 1, every 3 weeks) (sequence A), followed by four cycles of cisplatin-vindesine (cisplatin 120 mg/m(2) on day 1 and vindesine 3 mg/m(2) on days 1, 8, 15, and 22, every 4 weeks) (sequence B). Responding patients received 3 cycles of docetaxel 100 mg/m(2) (day 1, every 3 weeks) as consolidation (sequence C). Thirty-two patients entered the study with thirty being evaluable for efficacy. Four patients showed a partial response and one patient a complete response, resulting in an objective response rate of 16.7 %. The median survival time (intent-to-treat) was 11 months (95 %CI = 8.0-15.4 months) with an estimated 1-year survival rate of 47%. The median time to progression was 17.6 weeks in the evaluable population. Main grade 4 toxicity was neutropenia (21.8 % and 68.2 % of patients in sequence A and B, respectively) while grade 3 peripheral neuropathy was documented in five patients during sequence B. There were no treatment-related deaths. Sequential chemotherapy may show promise for the treatment of advanced non-small cell lung cancer. Given the feasibility of this pilot study, sequential chemotherapy concept should be investigated with newer cisplatin-based regimens using this approach in larger prospective studies.


Role of P-selectin cytoplasmic domain in granular targeting in vivo and in early inflammatory responses.
P-selectin is an adhesion receptor for leukocytes expressed on activated platelets and endothelial cells. The cytoplasmic domain of P-selectin was shown in vitro to contain signals required for both the sorting of this protein into storage granules and its internalization from the plasma membrane. To evaluate in vivo the role of the regulated secretion of P-selectin, we have generated a mouse that expresses P-selectin lacking the cytoplasmic domain (DeltaCT mice). The deletion did not affect the sorting of P-selectin into alpha-granules of platelets but severely compromised the storage of P-selectin in endothelial cells. Unstored P-selectin was proteolytically shed from the plasma membrane, resulting in increased levels of soluble P-selectin in the plasma. The DeltaCT-P-selectin appeared capable of mediating cell adhesion as it supported leukocyte rolling in the mutant mice. However, a secretagogue failed to upregulate leukocyte rolling in the DeltaCT mice, indicating an absence of a releasable storage pool of P-selectin in the endothelium. Furthermore, the neutrophil influx into the inflamed peritoneum was only 30% of the wild-type level 2 h after stimulation. Our results suggest that different sorting mechanisms for P-selectin are used in platelets and endothelial cells and that the storage pool of P-selectin in endothelial cells is functionally important during early stages of inflammation.


Quantitative Evaluation of Intensity Inhomogeneity Correction Methods for Structural MR Brain Images.
The correction of intensity non-uniformity (INU) in magnetic resonance (MR) images is extremely important to ensure both within-subject and across-subject reliability. Here we tackled the problem of objectively comparing INU correction techniques for T1-weighted images, which are the most commonly used in structural brain imaging. We focused our investigations on the methods integrated in widely used software packages for MR data analysis: FreeSurfer, BrainVoyager, SPM and FSL. We used simulated data to assess the INU fields reconstructed by those methods for controlled inhomogeneity magnitudes and noise levels. For each method, we evaluated a wide range of input parameters and defined an enhanced configuration associated with best reconstruction performance. By comparing enhanced and default configurations, we found that the former often provide much more accurate results. Accordingly, we used enhanced configurations for a more objective comparison between methods. For different levels of INU magnitude and noise, SPM and FSL, which integrate INU correction with brain segmentation, generally outperformed FreeSurfer and BrainVoyager, whose methods are exclusively dedicated to INU correction. Nonetheless, accurate INU field reconstructions can be obtained with FreeSurfer on images with low noise and with BrainVoyager for slow and smooth inhomogeneity profiles. Our study may prove helpful for an accurate selection of the INU correction method to be used based on the characteristics of actual MR data.


Long-Term Results of a Phase II Trial of Concomitant Cisplatin-Paclitaxel Chemoradiation in Locally Advanced Cervical Cancer.
The aim of this study was to determine the long-term results of a 7-week schedule of external beam radiation therapy, high dose rate brachytherapy, and weekly cisplatin and paclitaxel in patients with locally advanced carcinoma of the cervix. Thirty-seven patients with International Federation of Gynecology and Obstetrics stages IB2 to IVa cervical cancer were treated with 40 mg/m per week of intravenous cisplatin and 50 mg/m per week of intravenous paclitaxel combined with 45 Gy of pelvic external beam radiation therapy and 28 to 30 Gy of high dose rate brachytherapy. Sixteen patients (43.2%) were able to complete the 6 scheduled cycles of chemotherapy. The median number of weekly chemotherapy cycles administered was 5. Thirty-six (16.2%) of 222 cycles of chemotherapy were not given because of toxicity. The mean dose intensity of cisplatin was 29.6 mg/m per week (95% confidence interval, 27.0-32.1); that of paclitaxel was 40.0 mg/m per week (95% confidence interval, 36.9-43.1). Thirty-four patients (91.8%) completed the planned radiation course in less than 7 weeks. Median radiation treatment length was 43 days. After a median follow-up of 6 years, 7 patients (18.9%) experienced severe (RTOG grade 3 or higher) late toxicity. No fatal events were observed. Ten patients have failed, 1 locally and 9 at distant sites. The 14-year locoregional control rate was 96.7%, and the 14-year freedom from systemic failure rate was 64.6%. Fourteen-year actuarial disease-free survival and overall survival rates were 44.8% and 50%, respectively. This study demonstrates excellent very long-term results and tolerable toxicity although the target weekly dosage of cisplatin and paclitaxel needs to be adjusted in the majority of the patients.


Cardiovascular and stress responses to short-term noise exposures-A panel study in healthy males.
While previous epidemiological studies report adverse effects of long-term noise exposure on cardiovascular health, the mechanisms responsible for these effects are unclear. We sought to elucidate the cardiovascular and stress response to short-term, low (31.5-125Hz) and high (500-2000Hz) frequency noise exposures. Healthy male (n=10) participants were monitored on multiple visits during no noise, low- or high-frequency noise exposure scenarios lasting 40min. Participants were fitted with an ambulatory electrocardiogram (ECG) and blood pressure measures and saliva samples were taken before, during and after noise exposures. ECGs were processed for measures of heart rate variability (HRV): high-frequency power (HF), low-frequency power (LF), the root of the mean squared difference between adjacent normal heart beats (N-N) intervals (RMSSD), and the standard deviation of N-N intervals (SDNN). Systolic blood pressure (SBP), diastolic blood pressure (DPB), and pulse were reported and saliva was analyzed for salivary cortisol and amylase. Multivariate mixed-effects linear regression models adjusted for age were used to identify statistically significant difference in outcomes by no noise, during noise or after noise exposure periods and whether this differed by noise frequency. A total of 658, 205, and 122, HRV, saliva, and blood pressure measurements were performed over 41 person days. Reductions in HRV (LF and RMSSD) were observed during noise exposure (a reduction of 19% (-35,-3.5) and 9.1% (-17,-1.1), respectively). After adjusting for noise frequency, during low frequency noise exposure, HF, LF, and SDNN were reduced (a reduction of 32% (-57,-6.2), 34% (-52,-15), and 16% (-26,-6.1), respectively) and during high frequency noise exposure, a 21% (-39,-2.3) reduction in LF, as compared to during no noise exposure, was found. No significant (p<0.05) changes in blood pressure, salivary cortisol, or amylase were observed. These results suggest that exposure to noise, and in particular, to low-frequency noise, negatively impacts HRV. The frequencies of noise should be considered when evaluating the cardiovascular health impacts of exposure.


Activity level, apoptosis, and development of cachexia in Apc(Min/+) mice.
Criteria for diagnosing cachexia in adults include unintentional loss in body weight, decreased strength, fatigue, anorexia, and low muscle mass. Cachexia is also associated with systemic inflammation, altered metabolism, and anemia. The Apc(Min/+) mouse is a model of cachexia directly related to intestinal tumor burden and subsequent chronic inflammation. These mice also demonstrate muscle weakness, fatigue, decreased volitional activity, and elevated circulating IL-6 levels. The purpose of this study was to determine the time course of changes in physical activity and their relationship to anemia, muscle apoptosis, and muscle mass and body mass loss during cachexia. A subset of male Apc(Min/+) mice were given access to voluntary activity wheels from 5 to 26 wk of age, while sedentary male Apc(Min/+) mice were housed in cages lacking wheels. At the study's end mice were stratified by cachectic symptoms. Severely cachectic mice had decreased wheel running performance at 15 wk of age, while anemia and body weight loss were not present until 18 wk of age. Severely cachectic mice had lower hemoglobin levels compared with mildly cachectic mice at 13, 18, and 22 wk of age. Severely cachectic mice also demonstrated threefold more BCL2-associated X protein (BAX) protein in the gastrocnemius muscle at 26 wk of age compared with mildly cachectic mice. In sedentary Apc(Min/+) mice at 26 wk of age anemia was present, and markers of apoptosis were induced in severely cachectic muscle. Proapoptotic protein expression was induced in both red and white portions of gastrocnemius muscle as well as in soleus muscle of severely cachectic mice compared with mildly cachectic mice. These data demonstrate that decrements in wheel running performance precede loss of body mass and that inherent muscle oxidative capacity is not protective against muscle apoptosis.


Effects of the P20 protein from Bacillus thuringiensis israelensis on insecticidal crystal protein Cry4Ba.
The accessory protein P20 from Bacillus thuringiensis israelensis has been defined as an important molecular chaperone for forming crystal Cyt1Aa, and enhancing Cry11Aa and Cry4Aa expression. To investigate its putative role in Cry4Ba delta-endotoxin production and toxicity, a p20 gene was cloned and introduced into B. thuringiensis recombinant strain expressing cry4Ba type gene (cry4BLB). The delta-endotoxin synthesis was enhanced by 262%. The generated inclusions were assayed against third instar larvae of Aedes aegypti. The combination of P20 protein with Cry4BLB delta-endotoxin led to a stable mortality rate of 25% with doses ranging from 0.2 mg l(-1) to 0.6 mg l(-1). Cry4BLB crystals produced in the presence of P20 were much less soluble than those produced by the control strain lacking P20 at pH lower than or equal to 10.5. The observed toxicity perturbation correlates with a decrease of Cry4BLB inclusions solubility. The presence of P20 protein has affected Cry4BLB crystallization and altered greatly its solubility properties. Cry4Ba effectiveness against A. aegypti larvae is related to the solubilization step in larval guts environment.


Whole-Genome Sequence Data Suggest Environmental Adaptation of Ethiopian Sheep Populations.
Great progress has been made over recent years in the identification of selection signatures in the genomes of livestock species. This work has primarily been carried out in commercial breeds for which the dominant selection pressures are associated with artificial selection. As agriculture and food security are likely to be strongly affected by climate change, a better understanding of environment-imposed selection on agricultural species is warranted. Ethiopia is an ideal setting to investigate environmental adaptation in livestock due to its wide variation in geo-climatic characteristics and the extensive genetic and phenotypic variation of its livestock. Here, we identified over three million single nucleotide variants across 12 Ethiopian sheep populations and applied landscape genomics approaches to investigate the association between these variants and environmental variables. Our results suggest that environmental adaptation for precipitation-related variables is stronger than that related to altitude or temperature, consistent with large-scale meta-analyses of selection pressure across species. The set of genes showing association with environmental variables was enriched for genes highly expressed in human blood and nerve tissues. There was also evidence of enrichment for genes associated with high-altitude adaptation although no strong association was identified with hypoxia-inducible-factor (HIF) genes. One of the strongest altitude-related signals was for a collagen gene, consistent with previous studies of high-altitude adaptation. Several altitude-associated genes also showed evidence of adaptation with temperature, suggesting a relationship between responses to these environmental factors. These results provide a foundation to investigate further the effects of climatic variables on small ruminant populations.


Spin-orbit coupling in the Mott insulator Ca(2)RuO(4).
O 1s x-ray absorption study of the Mott insulator Ca(2)RuO(4) shows that the orbital population of the 4d t(2g) band dramatically changes with temperature. In addition, spin-resolved circularly polarized photoemission study of Ca(2)RuO(4) shows that a substantial orbital angular momentum is induced in the Ru 4d t(2g) band. Based on the experimental results and model Hartree-Fock calculations, we argue that the cooperation between the strong spin-orbit coupling in the Ru 4d t(2g) band and the small distortion of the RuO(6) octahedra causes the interesting changeover of the spin and orbital anisotropy as a function of temperature.


Enlargement of the Sylvian aqueduct: a sequel of head injuries.
A review of pneumoencephalography and clinical data in a large group of patients has shown that severe head injury may be followed by a consistent clinical-radiological syndrome. A prolonged period of unconsciousness is a characteristic initial feature of this syndrome. Clinically there is ataxia and dysarthria and, often, abnormal movements and oculomotor dysfunction. The characteristic radiological feature is an enlargement of the aqueduct of Sylvius. The clinical picture, together with the enlargement of the aqueduct, suggests that loss of neuronal and axonal substance in the midbrain is the probable pathology.


Interactive effects of land use history and natural disturbance on seedling dynamics in a subtropical forest.
Human-impacted forests are increasing in extent due to widespread regrowth of secondary forests on abandoned lands. The degree and speed of recovery from human disturbance in these forests will determine their value in terms of biodiversity conservation and ecosystem function. In areas subject to periodic, severe natural disturbances, such as hurricanes, it has been hypothesized that human and natural disturbance may interact to either erase or preserve land use legacies. To increase understanding of how interactions between human and natural disturbance influence forest regeneration and recovery, we monitored seedlings in a human- and hurricane-impacted forest in northeastern Puerto Rico over a approximately 10-yr period and compared seedling composition and dynamics in areas that had experienced high- and low-intensity human disturbance during the first half of the 20th century. We found that land use history significantly affected the composition and diversity of the seedling layer and altered patterns of canopy openness and seedling dynamics following hurricane disturbance. The area that had been subject to high-intensity land use supported a higher density, but lower diversity, of species. In both land use history categories, the seedling layer was dominated by the same two species, Prestoea acuminata var. montana and Guarea guidonia. However, seedlings of secondary-successional species tended to be more abundant in the high-intensity land use area, while late-successional species were more abundant in the low-intensity area, consistent with patterns of adult tree distributions. Seedlings of secondary-forest species showed greater increases in growth and survival following hurricane disturbance compared to late-successional species, providing support for the hypothesis that hurricanes help preserve the signature of land use history. However, the increased performance of secondary-forest species occurred predominantly in the low-intensity land use area, suggesting that hurricanes act to homogenize differences in species composition between areas with differing land use histories by increasing secondary-forest species regeneration in areas that experienced little direct human disturbance. Our results suggest that, through effects on seedling dynamics, hurricanes may extend the signature of land use history beyond the average recovery time of forests not subject to intense natural disturbance events.


Diagnosis of Encephalitozoon cuniculi infection by western blot and the use of cross-reactive antigens for the possible detection of microsporidiosis in humans.
Microsporidia are very primitive, eukaryotic, obligate, intracellular, protozoan parasites. Encephalitozoon cuniculi, a microsporidian originally described from a rabbit infection, has been described in humans as well as in many species of laboratory animals. We report the detection of E. cuniculi by Western blotting in a rabbit with torticollis that was obtained from an Encephalitozoon-free colony. Cross-reactivity of this serum was observed with antigens prepared from several genera of microsporidia. Identical Western blotting patterns were obtained with sera obtained from a rabbit immunized with E. cuniculi that was purified from tissue culture cells. In addition, we were able to demonstrate cross-reactivity between E. cuniculi rabbit antisera and Enterocytozoon bieneusi antigens by indirect immunofluorescent assay techniques in human intestinal biopsy samples. These cross-reactions between microsporidia may be useful in developing diagnostic tests for non-cultivatable microsporidia such as Enterocytozoon bieneusi.


The accuracy of magnetic resonance imaging (MRI) in detecting meniscal pathology.
The purpose of this study was to determine the accuracy of Magnetic Resonance Imaging (MRI) scanning in the detection of meniscal pathology in a district general hospital. We retrospectively analysed a single-surgeon series of 240 knee arthroscopic investigations for all indications. The arthroscopic reports included an outline diagram of the meniscus upon which the surgeon could record his operative findings. 112 of these patients had also had a recent MRI scan. We compared the MRI findings with the arthroscopy findings. 66 patients had a positive MRI scan. 64 of these were found to have a meniscal tear at surgery. 37 MRI scans were reported as "no tear", of which four were found to have a meniscal tear at surgery. Nine MRI scans were descriptive, e.g. "signal change, possible tear", or "tear cannot be ruled out." These tended to correspond with equivocal arthroscopic findings of "degeneration" or "fibrillation". In our series of 112 patients with meniscal pathology, MRI scanning was 90.5% sensitive, 89.5% specific and 90.1% accurate. False positive MRI scans may lead to unnecessary surgery. Patients with negative MRI scans had a mean delay to surgery of 33 weeks compared to 18 weeks for patients with positive MRI scans. Patients with false negative MRI results may wait longer for their surgery. Two of the false negative MRI scan reports clearly showed meniscus tears, which were not identified by the reporting radiologist. In our series, the MRI scan itself was more accurate than the reporting. It is important to have an experienced musculoskeletal radiologist to minimise the number of missed meniscal tears. It is also important for the surgeon to review the MRI scan itself, as well as the report.


Luteolin Targets the Toll-Like Receptor Signaling Pathway in Prevention of Hepatic and Adipocyte Fibrosis and Insulin Resistance in Diet-Induced Obese Mice.
This study was to investigate the protective role of luteolin on inflammation-mediated metabolic diseases, focusing on the role of luteolin in the modulation of the Toll-like receptor (TLR) signaling pathway. C57BL/6J mice were fed a normal, high-fat, or high-fat + 0.005% (w/w) luteolin diet for 16 weeks. Luteolin improved chronic low-grade inflammation by modulating the TLR signaling pathway, resulting in reduced pro-inflammatory cytokines and macrophage accumulation. A positive relationship was detected between gene expressions of Tlr5, Map2k7, Mapk12, Mapk13, and Mapk9 and lipogenesis in epididymal white adipose tissue (eWAT) of luteolin-treated mice, which was linked to attenuation of hepatic lipotoxicity by increasing free fatty acid (FFA) flux to the WAT. Luteolin prevented fibrosis by decreasing extracellular matrix accumulation and cathepsin gene expressions, while enhancing the hepatic antioxidant system. Emr1 and Ccl7, important markers inducing low-grade inflammation, were affected by advanced age and greater body weight, which were normalized by luteolin treatment. Luteolin improved insulin resistance by normalizing pancreatic islet dysfunction and differentially modulating the plasma glucagon-like peptide-1 and gastric inhibitory polypeptide levels. Our results suggest that luteolin ameliorates diet-induced obesity and its comorbidities. Overall, this study provides novel insights into the effect of luteolin on the links among adiposopathy, insulin resistance, hepatic steatosis, and fibrosis.


A guide to the effects of a large portion of the residues of triosephosphate isomerase on catalysis, stability, druggability, and human disease.
Triosephosphate isomerase (TIM) is a ubiquitous enzyme, which appeared early in evolution. TIM is responsible for obtaining net ATP from glycolysis and producing an extra pyruvate molecule for each glucose molecule, under aerobic and anaerobic conditions. It is placed in a metabolic crossroad that allows a quick balance of the triose phosphate aldolase produced by glycolysis, and is also linked to lipid metabolism through the alternation of glycerol-3-phosphate and the pentose cycle. TIM is one of the most studied enzymes with more than 199 structures deposited in the PDB. The interest for this enzyme stems from the fact that it is involved in glycolysis, but also in aging, human diseases and metabolism. TIM has been a target in the search for chemical compounds against infectious diseases and is a model to study catalytic features. Until February 2017, 62% of all residues of the protein have been studied by mutagenesis and/or using other approaches. Here, we present a detailed and comprehensive recompilation of the reported effects on TIM catalysis, stability, druggability and human disease produced by each of the amino acids studied, contributing to a better understanding of the properties of this fundamental protein. The information reviewed here shows that the role of the noncatalytic residues depend on their molecular context, the delicate balance between the short and long-range interactions in concerted action determining the properties of the protein. Each protein should be regarded as a unique entity that has evolved to be functional in the organism to which it belongs. Proteins 2017; 85:1190-1211. © 2017 Wiley Periodicals, Inc.


International policies to reduce plastic marine pollution from single-use plastics (plastic bags and microbeads): A review.
Marine plastic pollution has been a growing concern for decades. Single-use plastics (plastic bags and microbeads) are a significant source of this pollution. Although research outlining environmental, social, and economic impacts of marine plastic pollution is growing, few studies have examined policy and legislative tools to reduce plastic pollution, particularly single-use plastics (plastic bags and microbeads). This paper reviews current international market-based strategies and policies to reduce plastic bags and microbeads. While policies to reduce microbeads began in 2014, interventions for plastic bags began much earlier in 1991. However, few studies have documented or measured the effectiveness of these reduction strategies. Recommendations to further reduce single-use plastic marine pollution include: (i) research to evaluate effectiveness of bans and levies to ensure policies are having positive impacts on marine environments; and (ii) education and outreach to reduce consumption of plastic bags and microbeads at source.


The mutacins of Streptococcus mutans: regulation and ecology.
Streptococcus mutans is generally recognized as a causative agent of human dental caries. The production of mutacins (bacteriocins) by S. mutans is considered to be an important factor in the colonization and establishment of S. mutans in the dental biofilm. Two types of mutacins have been characterized: the lantibiotics and the non-lantibiotics. The lantibiotics generally have a wider spectrum of activity than the non-lantibiotics, which make them attractive targets for development into new antimicrobial modalities. The non-lantibiotics are much more prevalent among strains of S. mutans and play a significant role in both community-level and population-level interactions in the dental biofilm. These interactions are directly mediated through the ComCDE two-component system and the newly characterized LytTR Regulation Systems HdrRM and BrsRM. These systems coordinate natural competence development and mutacin production as a means to acquire transforming DNA either by killing closely related streptococcal species in the vicinity of S. mutans, or through an altruistic suicide mechanism among a subpopulation of competent cells within the S. mutans community. As more S. mutans strains are sequenced, it is anticipated that additional mutacins with novel functions will be discovered, which may yield further insights into the ecological role of mutacins within the oral biofilm.


Palmitoylated calnexin is a key component of the ribosome-translocon complex.
A third of the human genome encodes N-glycosylated proteins. These are co-translationally translocated into the lumen/membrane of the endoplasmic reticulum (ER) where they fold and assemble before they are transported to their final destination. Here, we show that calnexin, a major ER chaperone involved in glycoprotein folding is palmitoylated and that this modification is mediated by the ER palmitoyltransferase DHHC6. This modification leads to the preferential localization of calnexin to the perinuclear rough ER, at the expense of ER tubules. Moreover, palmitoylation mediates the association of calnexin with the ribosome-translocon complex (RTC) leading to the formation of a supercomplex that recruits the actin cytoskeleton, leading to further stabilization of the assembly. When formation of the calnexin-RTC supercomplex was affected by DHHC6 silencing, mutation of calnexin palmitoylation sites or actin depolymerization, folding of glycoproteins was impaired. Our findings thus show that calnexin is a stable component of the RTC in a manner that is exquisitely dependent on its palmitoylation status. This association is essential for the chaperone to capture its client proteins as they emerge from the translocon, acquire their N-linked glycans and initiate folding.


Ancient fig wasps indicate at least 34 Myr of stasis in their mutualism with fig trees.
Fig wasps and fig trees are mutually dependent, with each of the 800 or so species of fig trees (Ficus, Moraceae) typically pollinated by a single species of fig wasp (Hymenoptera: Agaonidae). Molecular evidence suggests that the relationship existed over 65 Ma, during the Cretaceous. Here, we record the discovery of the oldest known fossil fig wasps, from England, dated at 34 Ma. They possess pollen pockets that contain fossil Ficus pollen. The length of their ovipositors indicates that their host trees had a dioecious breeding system. Confocal microscopy and scanning electron microscopy reveal that the fossil female fig wasps, and more recent species from Miocene Dominican amber, display the same suite of anatomical characters associated with fig entry and pollen-carrying as modern species. The pollen is also typical of modern Ficus. No innovations in the relationship are discernible for the last tens of millions of years.


Readthrough acetylcholinesterase (AChE-R) and regulated necrosis: pharmacological targets for the regulation of ovarian functions?
Proliferation, differentiation and death of ovarian cells ensure orderly functioning of the female gonad during the reproductive phase, which ultimately ends with menopause in women. These processes are regulated by several mechanisms, including local signaling via neurotransmitters. Previous studies showed that ovarian non-neuronal endocrine cells produce acetylcholine (ACh), which likely acts as a trophic factor within the ovarian follicle and the corpus luteum via muscarinic ACh receptors. How its actions are restricted was unknown. We identified enzymatically active acetylcholinesterase (AChE) in human ovarian follicular fluid as a product of human granulosa cells. AChE breaks down ACh and thereby attenuates its trophic functions. Blockage of AChE by huperzine A increased the trophic actions as seen in granulosa cells studies. Among ovarian AChE variants, the readthrough isoform AChE-R was identified, which has further, non-enzymatic roles. AChE-R was found in follicular fluid, granulosa and theca cells, as well as luteal cells, implying that such functions occur in vivo. A synthetic AChE-R peptide (ARP) was used to explore such actions and induced in primary, cultured human granulosa cells a caspase-independent form of cell death with a distinct balloon-like morphology and the release of lactate dehydrogenase. The RIPK1 inhibitor necrostatin-1 and the MLKL-blocker necrosulfonamide significantly reduced this form of cell death. Thus a novel non-enzymatic function of AChE-R is to stimulate RIPK1/MLKL-dependent regulated necrosis (necroptosis). The latter complements a cholinergic system in the ovary, which determines life and death of ovarian cells. Necroptosis likely occurs in the primate ovary, as granulosa and luteal cells were immunopositive for phospho-MLKL, and hence necroptosis may contribute to follicular atresia and luteolysis. The results suggest that interference with the enzymatic activities of AChE and/or interference with necroptosis may be novel approaches to influence ovarian functions.


Landscape connectivity influences gene flow in a roe deer population inhabiting a fragmented landscape: an individual-based approach.
Changes in agricultural practices and forest fragmentation can have a dramatic effect on landscape connectivity and the dispersal of animals, potentially reducing gene flow within populations. In this study, we assessed the influence of woodland connectivity on gene flow in a traditionally forest-dwelling species--the European roe deer--in a fragmented landscape. From a sample of 648 roe deer spatially referenced within a study area of 55 x 40 km, interindividual genetic distances were calculated from genotypes at 12 polymorphic microsatellite loci. We calculated two geographical distances between each pair of individuals: the Euclidean distance (straight line) and the 'least cost distance' (the trajectory that maximizes the use of wooded corridors). We tested the correlation between genetic pairwise distances and the two types of geographical pairwise distance using Mantel tests. The correlation was better using the least cost distance, which takes into account the distribution of wooded patches, especially for females (the correlation was stronger but not significant for males). These results suggest that in a fragmented woodland area roe deer dispersal is strongly linked to wooded structures and hence that gene flow within the roe deer population is influenced by the connectivity of the landscape.


Extended weekly dose-dense paclitaxel/carboplatin is feasible and active in heavily pre-treated platinum-resistant recurrent ovarian cancer.
There is increasing evidence of the efficacy of dose-dense therapy in the management of platinum-resistant/refractory ovarian cancer. We report our experience of extended weekly carboplatin and paclitaxel in this population group. Twenty patients with platinum-resistant/refractory ovarian cancer received carboplatin AUC 3 and paclitaxel 70 mg m(-2) on day 1, 8, 15 q 4 weekly for six planned cycles. Toxicity was assessed using Common Toxicity Criteria. Response was evaluated using radiological and CA125 criteria. Median age was 61 years (range 40-74 years). Median number of prior therapies is three (range 1-8). Response rate was 60% by radiological criteria (RECIST) and 76% by CA125 assessment. Grade 3 toxicities consisted of neutropenia (29% of patients) and anaemia (5%). One patient experienced grade 4 neutropenia. No grade 3/4 thombocytopaenia was reported. Fatigue, nausea and peripheral neuropathy were the most frequent non-hematological side effects. Median progression-free survival was 7.9 months and overall survival was 13.3 months. The dynamics of response to dose-dense therapy were as rapid as with front-line therapy within the same patient. This dose-dense regimen can be extended to at least 18 weekly cycles over 6 months and is well tolerated with high response rates in heavily pre-treated, platinum-resistant ovarian cancer. It forms a highly active and tolerable cytotoxic scaffold to which molecular-targeted therapies can be added in platinum-resistant ovarian cancer.


Acinetobacter baumannii outer membrane protein A targets the nucleus and induces cytotoxicity.
Acinetobacter baumannii is an emerging opportunistic pathogen responsible for healthcare-associated infections. The outer membrane protein A of A. baumannii (AbOmpA) is the most abundant surface protein that has been associated with the apoptosis of epithelial cells through mitochondrial targeting. The nuclear translocation of AbOmpA and the subsequent pathology on host cells were further investigated. AbOmpA directly binds to eukaryotic cells. AbOmpA translocates to the nucleus by a novel monopartite nuclear localization signal (NLS). The introduction of rAbOmpA into the cells or a transient expression of AbOmpA-EGFP causes the nuclear localization of these proteins, while the fusion proteins of AbOmpADeltaNLS-EGFP and AbOmpA with substitutions in residues lysine to alanine in the NLS sequences represent an exclusively cytoplasmic distribution. The nuclear translocation of AbOmpA induces cell death in vitro. Furthermore, the microinjection of rAbOmpA into the nucleus of Xenopus laevis embryos fails to develop normal embryogenesis, thus leading to embryonic death. We propose a novel pathogenic mechanism of A. baumannii regarding the nuclear targeting of the bacterial structural protein AbOmpA.


[Splenectomy in idiopathic thrombocytopenic purpura].
Between January 1976 and December 1987, 44 patients with idiopathic thrombocytopenic purpura were submitted to splenectomy. This study analyzes the results of treatment and the usefulness of several prognostic markers. There were 38 female patients and six males with a median age of 28.5 years (range 11 to 66 years). Forty two out of 44 patients received preoperative steroids. Seventy five percent achieved a satisfactory response to this treatment but 31 relapsed (in eight recurrence was observed when the steroid dose was tapered). Eleven patients persisted symptomatic notwithstanding medical treatment. In two cases splenectomy was performed as an emergency procedure, and in one patient who was already in remission with previous steroid treatment, splenectomy was performed for other medical reasons. Operative mortality rate was 4 percent, and five patients developed postoperative complications (three had pneumonia, one pulmonary thromboembolism and one would infection). Eighty four percent of the patients were free of disease at one year followup survival at 10 year period was 80 percent. No correlation was found between age, duration of symptoms, previous response to steroids or time-period of medical treatment with the results obtained with splenectomy.


Pathophysiology and pharmacological treatment of gastroesophageal reflux disease.
Gastroesophageal reflux disease (GERD) is one of the most common diagnoses in a gastroenterologist's practice. Gastroesophageal reflux describes the retrograde movement of gastric contents through the lower esophageal sphincter (LES) to the esophagus. It is a common, normal phenomenon which may occur with or without accompanying symptoms. Symptoms associated with GERD include heartburn, acid regurgitation, noncardiac chest pain, dysphagia, globus pharyngitis, chronic cough, asthma, hoarseness, laryngitis, chronic sinusitis and dental erosions. The introduction of fiberoptic instruments and ambulatory devices for continuous monitoring of esophageal pH (24-hour pH monitoring) has led to great improvement in the ability to diagnose reflux disease and reflux-associated complications. The development of pathological reflux and GERD can be attributed to many factors. Pathophysiology of GERD includes incompetent LES because of a decreased LES pressure, transient lower esophageal sphincter relaxations (TLESRs) and deficient or delayed esophageal acid clearance. Uncomplicated GER may be treated by modification of life style and eating habits in an early stage of GERD. The various agents currently used for treatment of GERD include mucoprotective substances, antacids, H(2) blockers, prokinetics and proton pump inhibitors. Although these drugs are effective, they do not necessarily influence the underlying causes of the disease by improving the esophageal clearance, increasing the LESP or reducing the frequency of TLESRs. The following article gives an overview regarding current concepts of the pathophysiology and pharmacological treatment of GERD.


Analysis of neurodegenerative disease-causing genes in dementia with Lewy bodies.
Dementia with Lewy bodies (DLB) is a clinically heterogeneous disorder with a substantial burden on healthcare. Despite this, the genetic basis of the disorder is not well defined and its boundaries with other neurodegenerative diseases are unclear. Here, we performed whole exome sequencing of a cohort of 1118 Caucasian DLB patients, and focused on genes causative of monogenic neurodegenerative diseases. We analyzed variants in 60 genes implicated in DLB, Alzheimer's disease, Parkinson's disease, frontotemporal dementia, and atypical parkinsonian or dementia disorders, in order to determine their frequency in DLB. We focused on variants that have previously been reported as pathogenic, and also describe variants reported as pathogenic which remain of unknown clinical significance, as well as variants associated with strong risk. Rare missense variants of unknown significance were found in APP, CHCHD2, DCTN1, GRN, MAPT, NOTCH3, SQSTM1, TBK1 and TIA1. Additionally, we identified a pathogenic GRN p.Arg493* mutation, potentially adding to the diversity of phenotypes associated with this mutation. The rarity of previously reported pathogenic mutations in this cohort suggests that the genetic overlap of other neurodegenerative diseases with DLB is not substantial. Since it is now clear that genetics plays a role in DLB, these data suggest that other genetic loci play a role in this disease.


Regulation of LIM-domain-binding 1 protein expression by ubiquitination of Lys134.
LDB1 (LIM-domain-binding 1) is a cofactor that participates in formation of transcriptional regulatory complexes involving transcription factors containing LIM domains as well as other factors. The amount of LDB1 protein in cells has previously been shown to be modulated by RNF12 (RING finger protein 12). RNF12 is an E3 ubiquitin ligase that can target LDB1 for poly-ubiquitination and degradation via the proteasome. We find that in HEK (human embryonic kidney)-293 cells expression of RNF12 leads to mono-ubiquitination of LDB1 and increased levels of LDB1 protein. Mutagenesis studies identified Lys134 of LDB1 as the residue that is mono-ubiquitinated by RNF12. Mutation of Lys134 of LDB1 to arginine blocks the formation of mono-ubiquitinated LDB1 and surprisingly also increases LDB1 protein expression in HEK-293 cells. This leads to a model in which Lys134 of LDB1 can be either mono-ubiquitinated, leading to stabilization, or poly-ubiquitinated, leading to degradation by the proteasome pathway. We also find that ubiquitin-LDB1 fusion proteins are stabilized in HEK-293 cells, offering further evidence that mono-ubiquitination stabilizes LDB1 in these cells. Expression in Xenopus laevis embryos of an LDB1 protein in which Lys134 is replaced with arginine leads to enhanced expression of the mutant protein as compared with the wild-type protein. These findings provide evidence that modification of Lys134 can play a major role in regulating LDB1 expression.


Symptomatic relevance of intravertebral cleft in patients with osteoporotic vertebral fracture.
The present study was designed to determine clinical and radiographic characteristics of unhealed osteoporotic vertebral fractures (OVFs) and the role of fracture mobility and an intravertebral cleft in the regulation of pain symptoms in patients with an OVF. Patients who had persistent low-back pain for 3 months or longer and a collapsed thoracic or lumbar vertebra that had an intervertebral cleft and abnormal mobility were referred to as having unhealed OVFs. Twenty-four patients with an unhealed OVF and 30 patients with an acute OVF were compared with regard to several clinical and radiographic features including the presence of an intravertebral fluid sign. Subsequently, the extent of dynamic mobility of the fractured vertebra was analyzed for correlation with the patients' age, duration of symptoms, back pain visual analog scale (VAS) score, and performance status. Finally, in cases of unhealed OVFs, the subgroup of patients with positive fluid signs was compared with the subgroup of patients with negative fluid signs. Patients with an unhealed OVF were more likely to have a crush-type fracture, shorter vertebral height of the fractured vertebra, and a fracture with a positive fluid sign than those with an acute OVF. The extent of dynamic mobility of the vertebra correlated significantly with the VAS score in patients with an unhealed OVF. In addition, a significant correlation with the extent of dynamic vertebral mobility with performance status was seen in patients with an unhealed OVF and those with an acute OVF. Of the 24 patients with an unhealed OVF, 14 had a positive fluid sign in the affected vertebra. Patients with a positive fluid sign exhibited a statistically significantly greater extent of dynamic vertebral mobility, a higher VAS score, a higher performance status grade, and a greater likelihood of having a crush-type fracture than those with a negative fluid sign. All but 1 patient with an unhealed OVF and a positive fluid sign had an Eastern Cooperative Oncology Group Performance Status Grade 3 or 4 (bedridden most or all of the time). In sharp contrast, all 10 patients with an unhealed OVF and a negative fluid sign were Grade 1 or 2. Unhealed OVFs form a group of fractures that are distinct from acute OVFs regarding radiographic morphometry and contents of the intravertebral cleft. Dynamic vertebral mobility serves as a primal pain determinant in patients with an unhealed OVF and potentially in those with an acute OVF. Fluid accumulation in the intravertebral cleft of unhealed OVFs likely reflects long-term bedridden positioning of the patients in daily activity.


Acquisition of virulence factors in livestock-associated MRSA: Lysogenic conversion of CC398 strains by virulence gene-containing phages.
Staphylococcus aureus MRSA strains belonging to the clonal complex 398 (CC398) are highly prevalent in livestock and companion animals but may also cause serious infections in humans. CC398 strains in livestock usually do not possess well-known virulence factors that can be frequently found in other MRSA sequence types (ST). Since many staphylococcal virulence genes are residing on the genomes of temperate phages, the question arises why livestock-associated (LA-) CC398 strains are only rarely infected by those phages. We isolated and characterized four temperate phages (P240, P282, P630 and P1105) containing genes of the immune evasion cluster (IEC) and/or for the Panton-Valentine leucocidin (PVL). Sequence analysis of the phage genomes showed that they are closely related to known phages and that the DNA region encoding lysis proteins, virulence factors and the integrase exhibits numerous DNA repeats which may facilitate genomic rearrangements. All phages lysed and lysogenized LA-CC398 strains. Integration of IEC phage P282 was detected at ten sites of the hosts' chromosome. The prophages were stably inherited in LA-CC398 and enterotoxin A, staphylokinase and PVL toxin were produced. The data demonstrate that lysogenic conversion of LA-CC398 strains by virulence-associated phages may occur and that new pathotypes may emerge by this mechanism.


Effect of soy- and whey protein-isolate supplemented diet on the redox parameters of trained mice.
A number of clinical trials have successfully been performed using whey and/or soy proteins in the treatment of many diseases. They both have antioxidant properties, which appears to be a factor in aerobic physical performance as well. In addition, these are the most often used supplements that sportsmen take to increase their performance. To investigate the effect of whey and soy protein supplementation on redox parameters in the muscle, on body weight, and body composition in swimming-trained and non-trained animals. The effect of whey and soy protein-isolate supplementation on muscle redox parameters, body weight, and body composition in trained and non-trained mice was investigated after a single exhaustive bout of exercise. Steady state free radical concentration measured using electron spin resonance (ESR) spectroscopy, reduced and oxidized glutathione ratio, thiobarbituric acid-reactive substances (TBARS), and protein carbonyl levels of the red leg muscle were measured. Free radical concentrations and glutathione composition of the tissue indicated that whey protein supplementation of the regular diet was able to prevent oxidative stress regardless of training. Soy protein supplementation decreased TBARS only in the muscle of untrained animals, while training per se lowered protein damage in all investigated groups. A mixture of soy and whey protein supplementation resulted in leaner animals after training, but had no synergistic effect on either of the measured redox parameters. Athletes consuming these supplements could train with higher exercise intensity. The antioxidant effect of the two proteins is based on different mechanisms of action.


An international survey of patients with cervical dystonia.
This was an international survey undertaken to assess cervical dystonia (CD) patients own perceptions of their illness and its management. A total of 1,071 self-identified respondents with CD in 38 countries completed the online survey between March and December 2012. The mean time since diagnosis was 9.6 years and over half (54%) of patients surveyed were not diagnosed in the first year. When asked how the symptoms of CD affected them, two-thirds (66%) of patients reported they experienced a lot of pain, and 61% said that they suffered depression and mood alterations; only 7% reported no impact on their lives. Despite problems with the diagnosis, almost 70% of respondents reported being satisfied with the overall relationship with their doctor. Patient treatment expectations were high, with 63% expecting freedom from spasms and 62% expecting freedom from pain. Over half (53%) expected to be able to return to a normal routine (53%). The most common treatment reported was botulinum toxin (BoNT) (86%), followed by oral medication (58%) and physiotherapy/physical therapy (37%). Among patients treated on BoNT, 56% were fairly/very satisfied, 25% were fairly/very dissatisfied and 20% were neither satisfied nor dissatisfied with the outcome. In conclusion, this international survey highlights the broad impact of CD on several aspects of patient life. Taken overall, the survey suggests that that patients need to be better informed about their condition, treatments available and the limitations of those treatments. It may be that realistically managing patient expectations of treatment would reduce the dissatisfaction of some patients.


Type 2 diabetes risk alleles demonstrate extreme directional differentiation among human populations, compared to other diseases.
Many disease-susceptible SNPs exhibit significant disparity in ancestral and derived allele frequencies across worldwide populations. While previous studies have examined population differentiation of alleles at specific SNPs, global ethnic patterns of ensembles of disease risk alleles across human diseases are unexamined. To examine these patterns, we manually curated ethnic disease association data from 5,065 papers on human genetic studies representing 1,495 diseases, recording the precise risk alleles and their measured population frequencies and estimated effect sizes. We systematically compared the population frequencies of cross-ethnic risk alleles for each disease across 1,397 individuals from 11 HapMap populations, 1,064 individuals from 53 HGDP populations, and 49 individuals with whole-genome sequences from 10 populations. Type 2 diabetes (T2D) demonstrated extreme directional differentiation of risk allele frequencies across human populations, compared with null distributions of European-frequency matched control genomic alleles and risk alleles for other diseases. Most T2D risk alleles share a consistent pattern of decreasing frequencies along human migration into East Asia. Furthermore, we show that these patterns contribute to disparities in predicted genetic risk across 1,397 HapMap individuals, T2D genetic risk being consistently higher for individuals in the African populations and lower in the Asian populations, irrespective of the ethnicity considered in the initial discovery of risk alleles. We observed a similar pattern in the distribution of T2D Genetic Risk Scores, which are associated with an increased risk of developing diabetes in the Diabetes Prevention Program cohort, for the same individuals. This disparity may be attributable to the promotion of energy storage and usage appropriate to environments and inconsistent energy intake. Our results indicate that the differential frequencies of T2D risk alleles may contribute to the observed disparity in T2D incidence rates across ethnic populations.


Short-term effects of electroconvulsive treatment on the uptake of 99mTc-exametazime into brain in major depression shown with single photon emission tomography.
Fifteen patients with major depression who were being treated with bilateral electroconvulsive treatment (ECT) were investigated before and 45 min after a single ECT using split-dose Single Photon Emission Tomography (SPET or SPECT) with 99mTc-Exametazime. All patients suffered from unipolar depressive illness and were rated on the Newcastle scale and with the 17-item Hamilton scale. They completed tests of orientation and verbal memory on the day of ECT. For comparison, verbal memory was also tested on the preceding day. The uptake of 99mTc-Exametazime was expressed relative to calcarine/occipital cortex. Significant decreases in tracer uptake were confined to the inferior anterior cingulate cortex. The changes were correlated with the severity of depressive symptoms and more weakly with decrements of memory function produced by ECT; there was no significant correlation with stimulus intensity or electroencephalographic measures of seizure duration.


CHRFAM7A, a human-specific and partially duplicated α7-nicotinic acetylcholine receptor gene with the potential to specify a human-specific inflammatory response to injury.
Conventional wisdom presumes that the α7nAChR product of CHRNA7 expression mediates the ability of the vagus nerve to regulate the inflammatory response to injury and infection. Yet, 15 years ago, a 2nd structurally distinct and human-specific α7nAChR gene was discovered that has largely escaped attention of the inflammation research community. The gene, originally called dupα7nAChR but now known as CHRFAM7A, has been studied exhaustively in psychiatric research because of its association with mental illness. However, dupα7nAChR/CHRFAM7A expression is relatively low in human brain but elevated in human leukocytes. Furthermore, α7nAChR research in human tissues has been confounded by cross-reacting antibodies and nonspecific oligonucleotide primers that crossreact in immunoblotting, immunohistochemistry, and RT-PCR. Yet, 3 independent reports show the human-specific CHRFAM7A changes cell responsiveness to the canonical α7nAChR/CHRNA7 ion-gated channel. Because of its potential for the injury research community, its possible significance to human leukocyte biology, and its relevance to human inflammation, we review the discovery and structure of the dupα7nAChR/CHRFAM7A gene, the distribution of its mRNA, and its biologic activities and then discuss its possible role(s) in specifying human inflammation and injury. In light of emerging concepts that point to a role for human-specific genes in complex human disease, the existence of a human-specific α7nAChR regulating inflammatory responses in injury underscores the need for caution in extrapolating findings in the α7nAChR literature to man. To this end, we discuss the translational implications of a uniquely human α7nAChR-like gene on new drug target discovery and therapeutics development for injury, infection, and inflammation.


Nrf2 activation by sulforaphane restores the age-related decrease of T(H)1 immunity: role of dendritic cells.
The decrease in cellular immunity with aging is of considerable public health importance. Recent studies suggest that the redox equilibrium of dendritic cells (DCs) is a key factor in maintaining protective cellular immunity and that a disturbance of this homeostatic mechanism could contribute to immune senescence. We sought (1) to elucidate the role of DC redox equilibrium in the decrease of contact hypersensitivity (CHS) and T(H)1 immunity during aging and (2) to determine how restoration of glutathione (GSH) levels by the Nrf2-mediated antioxidant defense pathway affects this decrease. We assessed the effect of Nrf2 deficiency and boosting of GSH levels by the Nrf2 agonist sulforaphane or the thiol precursor N-acetyl cysteine (NAC) on the CHS response to contact antigens in old mice. We studied the effect of SFN and NAC on restoring T(H)1 immunity by treating DCs ex vivo before adoptive transfer and in vivo challenge. Aging was associated with a decreased CHS response that was accentuated by Nrf2 deficiency. Systemic SFN treatment reversed this decrease through Nrf2-mediated antioxidant enzyme expression and GSH synthesis. Adoptive transfer of DCs from old animals induced a weakened CHS response in recipient animals. Treatment of DCs from old animals with SFN or NAC ex vivo restored the in vivo challenge response. SFN and NAC upregulate T(H)1 immunity in aging through a restoration of redox equilibrium.


[Effects of fenvalerate and phoxim insecticides on glutamate and gamma-aminobutyric acid immunoreactive cells in the central nervous system of rats].
In order to study the neurotoxic effect and the mechanism of fenvalerate and phoxim insecticides, rats were orally treated with fenvalerate (20 mg/kg BW), phoxim (160 mg/kgBW) and fenvalerate (20 mg/kgBW) plus phoxim (160 mg/kgBW). The glutamate (Glu) and gamma-aminobutyric acid(GABA) immunoreactive cells of the central nervous system of rats were observed by immunohistochemical method. The results showed that the total number, percentage of positive area and integral optical density of Glu immunoreactive cells in cerebral cortex, hippocampus and striate exhibited a characteristic decrease in the groups fed with fenvalerate and fenvalerate plus phoxim, while those of GABA were enhanced to certain extents. No obvious changes in Glu or GABA were observed in the phoxim group as compared with the controls, and no significant difference was observed between the fenvalerate group and the fenvalerate plus phoxim group. The results suggest that the disturbance of Glu and GABA metabolism may play an important role in the development of pyrethroid neurotoxicity, and there is no interaction between phoxim and fenvalerate pesticides on Glu and GABA activities.


The balanced mind: the variability of task-unrelated thoughts predicts error monitoring.
Self-generated thoughts unrelated to ongoing activities, also known as "mind-wandering," make up a substantial portion of our daily lives. Reports of such task-unrelated thoughts (TUTs) predict both poor performance on demanding cognitive tasks and blood-oxygen-level-dependent (BOLD) activity in the default mode network (DMN). However, recent findings suggest that TUTs and the DMN can also facilitate metacognitive abilities and related behaviors. To further understand these relationships, we examined the influence of subjective intensity, ruminative quality, and variability of mind-wandering on response inhibition and monitoring, using the Error Awareness Task (EAT). We expected to replicate links between TUT and reduced inhibition, and explored whether variance in TUT would predict improved error monitoring, reflecting a capacity to balance between internal and external cognition. By analyzing BOLD responses to subjective probes and the EAT, we dissociated contributions of the DMN, executive, and salience networks to task performance. While both response inhibition and online TUT ratings modulated BOLD activity in the medial prefrontal cortex (mPFC) of the DMN, the former recruited a more dorsal area implying functional segregation. We further found that individual differences in mean TUTs strongly predicted EAT stop accuracy, while TUT variability specifically predicted levels of error awareness. Interestingly, we also observed co-activation of salience and default mode regions during error awareness, supporting a link between monitoring and TUTs. Altogether our results suggest that although TUT is detrimental to task performance, fluctuations in attention between self-generated and external task-related thought is a characteristic of individuals with greater metacognitive monitoring capacity. Achieving a balance between internally and externally oriented thought may thus aid individuals in optimizing their task performance.


Reward-related FMRI activation of dopaminergic midbrain is associated with enhanced hippocampus-dependent long-term memory formation.
Long-term potentiation in the hippocampus can be enhanced and prolonged by dopaminergic inputs from midbrain structures such as the substantia nigra. This improved synaptic plasticity is hypothesized to be associated with better memory consolidation in the hippocampus. We used a condition that reliably elicits a dopaminergic response, reward anticipation, to study the relationship between activity of dopaminergic midbrain areas and hippocampal long-term memory in healthy adults. Pictures of object drawings that predicted monetary reward were associated with stronger fMRI activity in reward-related brain areas, including the substantia nigra, compared with non-reward-predicting pictures. Three weeks later, recollection and source memory were better for reward-predicting than for non-reward-predicting pictures. FMRI activity in the hippocampus and the midbrain was higher for reward-predicting pictures that were later recognized compared with later forgotten pictures. These data are consistent with the hypothesis that activation of dopaminergic midbrain regions enhances hippocampus-dependent memory formation, possibly by enhancing consolidation.


CCL5/RANTES contributes to hypothalamic insulin signaling for systemic insulin responsiveness through CCR5.
Many neurodegenerative diseases are accompanied by metabolic disorders. CCL5/RANTES, and its receptor CCR5 are known to contribute to neuronal function as well as to metabolic disorders such as type 2 diabetes mellitus, obesity, atherosclerosis and metabolic changes after HIV infection. Herein, we found that the lack of CCR5 or CCL5 in mice impaired regulation of energy metabolism in hypothalamus. Immunostaining and co-immunoprecipitation revealed the specific expression of CCR5, associated with insulin receptors, in the hypothalamic arcuate nucleus (ARC). Both ex vivo stimulation and in vitro tissue culture studies demonstrated that the activation of insulin, and PI3K-Akt pathways were impaired in CCR5 and CCL5 deficient hypothalamus. The inhibitory phosphorylation of insulin response substrate-1 at Ser302 (IRS-1S302) but not IRS-2, by insulin was markedly increased in CCR5 and CCL5 deficient animals. Elevating CCR5/CCL5 activity induced GLUT4 membrane translocation and reduced phospho-IRS-1S302 through AMPKα-S6 Kinase. Blocking CCR5 using the antagonist, MetCCL5, abolished the de-phosphorylation of IRS-1S302 and insulin signal activation. In addition, intracerebroventricular delivery of MetCCL5 interrupted hypothalamic insulin signaling and elicited peripheral insulin responsiveness and glucose intolerance. Taken together, our data suggest that CCR5 regulates insulin signaling in hypothalamus which contributes to systemic insulin sensitivity and glucose metabolism.


Towards standard setting for patient-reported outcomes in the NHS homeopathic hospitals.
We report findings from a pilot data collection study within a programme of quality assurance, improvement and development across all five homeopathic hospitals in the UK National Health Service (NHS). (1) To pilot the collection of clinical data in the homeopathic hospital outpatient setting, recording patient-reported outcome since first appointment; (2) to sample the range of medical complaints that secondary-care doctors treat using homeopathy, and thus identify the nature and complexity of complaints most frequently treated nationally; (3) to present a cross section of outcome scores by appointment number, including that for the most frequently treated medical complaints; (4) to explore approaches to standard setting for homeopathic practice outcome in patients treated at the homeopathic hospitals. A total of 51 medical practitioners took part in data collection over a 4-week period. Consecutive patient appointments were recorded under the headings: (1) date of first appointment in the current series; (2) appointment number; (3) age of patient; (4) sex of patient; (5) main medical complaint being treated; (6) whether other main medical complaint(s); (7) patient-reported change in health, using Outcome Related to Impact on Daily Living (ORIDL) and its derivative, the ORIDL Profile Score (ORIDL-PS; range, -4 to +4, where a score <or=-2 or >or=+2 indicates an effect on the quality of a patient's daily life); (8) receipt of other complementary medicine for their main medical complaint. The distribution of patient age was bimodal: main peak, 49 years; secondary peak, 6 years. Male:female ratio was 1:3.5. Data were recorded on a total of 1797 individual patients: 195 first appointments, 1602 follow-ups (FUs). Size of clinical service and proportion of patients who attended more than six visits varied between hospitals. A total of 235 different medical complaints were reported. The 30 most commonly treated complaints were (in decreasing order of frequency): eczema; chronic fatigue syndrome (CFS); menopausal disorder; osteoarthritis; depression; breast cancer; rheumatoid arthritis; asthma; anxiety; irritable bowel syndrome; multiple sclerosis; psoriasis; allergy (unspecified); fibromyalgia; migraine; premenstrual syndrome; chronic rhinitis; headache; vitiligo; seasonal allergic rhinitis; chronic intractable pain; insomnia; ulcerative colitis; acne; psoriatic arthropathy; urticaria; ovarian cancer; attention-deficit hyperactivity disorder (ADHD); epilepsy; sinusitis. The proportion of patients with important co-morbidity was higher in those seen after visit 6 (56.9%) compared with those seen up to and including that point (40.7%; P<0.001). The proportion of FU patients reporting ORIDL-PS>or=+2 (improvement affecting daily living) increased overall with appointment number: 34.5% of patients at visit 2 and 59.3% of patients at visit 6, for example. Amongst the four most frequently treated complaints, the proportion of patients that reported ORIDL-PS>or=+2 at visit numbers greater than 6 varied between 59.3% (CFS) and 73.3% (menopausal disorder). We have successfully piloted a process of national clinical data collection using patient-reported outcome in homeopathic hospital outpatients, identifying a wide range and complexity of medical complaints treated in that setting. After a series of homeopathy appointments, a high proportion of patients, often representing "effectiveness gaps" for conventional medical treatment, reported improvement in health affecting their daily living. These pilot findings are informing our developing programme of standard setting for homeopathic care in the hospital outpatient context.


Axes establishment during eye morphogenesis in Xenopus by coordinate and antagonistic actions of BMP4, Shh, and RA.
We have examined the roles of BMP4, Shh, and retinoic acid in establishing the proximal-distal and dorsal-ventral axes in the developing Xenopus eye. Misexpression of BMP4 caused the absence of an optic stalk and the expansion of dorsal and distal markers, tbx2/3/5, and pax6, at the expense of ventral and proximal markers vax2 and pax2. When Shh or Noggin, an antagonist of BMPs, was misexpressed, the reverse expression patterns of these marker genes were observed. These results suggest that BMP4 is involved in the specification of not only dorsal in the optic cup but also distal in the optic vesicle. Because Shh did not suppress bmp4 expression, unlike Noggin, Shh and BMP4 may antagonistically regulate common downstream genes in developing eye. We also found the difference between the effects of Shh and retinoic acid, another possible ventralizing factor, suggesting that Shh could promote ventralization independently of retinoic acid. These findings provide important clues to the coordinate and antagonistic actions of BMP4, Shh, and retinoic acid in axes specifications of Xenopus eyes.


The effects of load carriage and muscle fatigue on lower-extremity joint mechanics.
Military personnel are commonly afflicted by lower-extremity overuse injuries. Load carriage and muscular fatigue are major stressors during military basic training. To examine effects of load carriage and muscular fatigue on lower-extremity joint mechanics during walking. Eighteen men performed the following tasks: unloaded walking, walking with a 32-kg load, fatigued walking with a 32-kg load, and fatigued walking. After the second walking task, muscle fatigue was elicited through a fatiguing protocol consisting of metered step-ups and heel raises with a 16-kg load. Each walking task was performed at 1.67 m x s(-1) for 5 min. Walking movement was tracked by a VICON motion capture system at 120 Hz. Ground reaction forces were collected by a tandem force instrumented treadmill (AMTI) at 2,400 Hz. Lower-extremity joint mechanics were calculated in Visual 3D. There was no interaction between load carriage and fatigue on lower-extremity joint mechanics (p > .05). Both load carriage and fatigue led to pronounced alterations of lower-extremity joint mechanics (p < .05). Load carriage resulted in increases of pelvis anterior tilt, hip and knee flexion at heel contact, and increases of hip, knee, and ankle joint moments and powers during weight acceptance. Muscle fatigue led to decreases of ankle dorsiflexion at heel contact, dorsiflexor moment, and joint power at weight acceptance. In addition, muscle fatigue increased demand for hip extensor moment and power at weight acceptance. Statistically significant changes in lower-extremity joint mechanics during loaded and fatigued walking may expose military personnel to increased risk for overuse injuries.