Management of acute combined fractures of the atlas and axis: A retrospective study of two trauma centers.
Management of combination fractures of the atlas and axis varies from nonoperative immobilization to selective early surgical intervention. In this study, we present our experience in managing these injuries. Electronic databases from two level 1 trauma centers were queried to identify all patients diagnosed with C1-C2 combination fractures from 2009 to present. Patient demographics, fracture characteristics, treatment modality, complications, Frankel scores, and fusion status were collected. Patients were separated into operative and nonoperative cohorts, and comparisons were made between the two groups. Forty-eight patients were included, of which 19 received operative management and 29 were treated nonoperatively. The mean age was 76.1 and 75.3 years, respectively (P = 0.877). Frankel grade distribution was similar on presentation in both groups, with most being neurologically intact. C1 fractures of both the anterior and posterior arch were present in 41.2% patients undergoing fusions compared to 27.6% of patients treated nonoperatively. No significant differences in comorbidities, neurologic deficits, or radiographic measurements were observed across the two groups. This study demonstrates the variety of treatment strategies used for the management of combined C1-C2 fractures. Patients managed operatively tend to have both anterior and posterior C1 arch fractures, while patients managed nonoperatively tend to have either anterior or posterior arch fractures. In general, treatments should be tailored to patients' needs depending on the stability of the fractures, neurological state, and medical comorbidities.


Healthy snacks at the checkout counter: a lab and field study on the impact of shelf arrangement and assortment structure on consumer choices.
The essence of nudging is to adapt the environment in which consumers make decisions to help them make better choices, without forcing certain outcomes upon them. To determine how consumers can effectively be guided to select healthier snacks, we examine the effect of manipulating the assortment structure and shelf layout of an impulse display including both healthy and unhealthy snacks near the checkout counter of a canteen. Both a lab and field study applied a two-factor experimental design manipulating snack offerings both in an on-screen choice environment and a natural environment (hospital staff restaurant). Shelf arrangement (i.e. accessibility) was altered by putting healthy snacks at higher shelves versus lower shelves. Assortment structure (i.e. availability) was altered by offering an assortment that either included 25% or 75% healthy snacks. Participants in the lab study (n = 158) made a choice from a shelf display. A brief survey following snack selection asked participants to evaluate the assortment and their choice. The field experiment took place in a hospital canteen. Daily sales data were collected for a period of four weeks. On completion of the field study, employees (n = 92) filled out a questionnaire about all four displays and rated their attractiveness, healthiness and perceived freedom of choice. The lab study showed a higher probability of healthy snack choice when 75% of the assortment consisted of healthy snacks compared to conditions with 25% healthy snack assortments, even though choices were not rated less satisfying or more restrictive. Regarding shelf display location of healthy snacks, no significant differences were observed. There was also no significant shelf arrangement by assortment structure interactive effect. The field study replicated these findings, in that this assortment structure led to higher sales of healthy snacks. Sales of unhealthy and total snacks were not impacted by manipulations (no main or interaction effects). Employees preferred shelf displays including a larger healthy snack assortment located at top shelves. Employees also felt more freedom in choice when healthy snacks were displayed at top shelves compared to lower shelves. Overall, results suggest that increasing the prominence of healthy snacks by enlarging their availability, while permitting access to unhealthy snacks, is a promising strategy to promote sales. These results point to the importance of nudging strategies to encourage healthier snack patterns.


The CMSSM and NUHM1 after LHC Run 1.
We analyze the impact of data from the full Run 1 of the LHC at 7 and 8 TeV on the CMSSM with [Formula: see text] and [Formula: see text] and the NUHM1 with [Formula: see text], incorporating the constraints imposed by other experiments such as precision electroweak measurements, flavour measurements, the cosmological density of cold dark matter and the direct search for the scattering of dark matter particles in the LUX experiment. We use the following results from the LHC experiments: ATLAS searches for events with [Formula: see text] accompanied by jets with the full 7 and 8 TeV data, the ATLAS and CMS measurements of the mass of the Higgs boson, the CMS searches for heavy neutral Higgs bosons and a combination of the LHCb and CMS measurements of [Formula: see text] and [Formula: see text]. Our results are based on samplings of the parameter spaces of the CMSSM for both [Formula: see text] and [Formula: see text] and of the NUHM1 for [Formula: see text] with 6.8[Formula: see text], 6.2[Formula: see text] and 1.6[Formula: see text] points, respectively, obtained using the MultiNest tool. The impact of the Higgs-mass constraint is assessed using FeynHiggs 2.10.0, which provides an improved prediction for the masses of the MSSM Higgs bosons in the region of heavy squark masses. It yields in general larger values of [Formula: see text] than previous versions of FeynHiggs, reducing the pressure on the CMSSM and NUHM1. We find that the global [Formula: see text] functions for the supersymmetric models vary slowly over most of the parameter spaces allowed by the Higgs-mass and the [Formula: see text] searches, with best-fit values that are comparable to the [Formula: see text] for the best Standard Model fit. We provide 95 % CL lower limits on the masses of various sparticles and assess the prospects for observing them during Run 2 of the LHC.


Electronic aids to daily living: be able to do what you want.
This study explores the experiences of Irish people with high cervical spinal cord injuries living with electronic aids to daily living (EADL) and the meaning attributed to such systems in the context of participation in everyday life. Qualitative methodology using a phenomenological approach was used to explore the phenomenon of living with EADL. Data were collected using four focus groups of users and nonusers of EADL (n = 15). All participants had high cervical spinal cord injuries (C3-5). Groups were video recorded, transcribed verbatim and analysed using descriptive phenomenological analysis. Findings revealed key elements of the meaning of living with EADL. Two key themes, time alone and changed relationships are described. These contribute to the super ordinate theme of autonomy. Findings suggest that participants perceived improvements in both anticipated and actual lived experiences with EADL. Themes are interrelated and together represent a summary of the experience of living with environmental controls. The themes described are similar to those found in other spinal injury studies relating to quality of life. Findings highlight differences in life experiences for those with and without EADL and provides motivation to address this difference. Such insights are valuable for both users and providers of EADL.


Personal and trip characteristics associated with safety equipment use by injured adult bicyclists: a cross-sectional study.
The aim of this study was to estimate use of helmets, lights, and visible clothing among cyclists and to examine trip and personal characteristics associated with their use. Using data from a study of transportation infrastructure and injuries to 690 adult cyclists in Toronto and Vancouver, Canada, we examined the proportion who used bike lights, conspicuous clothing on the torso, and helmets on their injury trip. Multiple logistic regression was used to examine associations between personal and trip characteristics and each type of safety equipment. Bike lights were the least frequently used (20% of all trips) although they were used on 77% of trips at night. Conspicuous clothing (white, yellow, orange, red) was worn on 33% of trips. Helmets were used on 69% of trips, 76% in Vancouver where adult helmet use is required by law and 59% in Toronto where it is not. Factors positively associated with bike light use included night, dawn and dusk trips, poor weather conditions, weekday trips, male sex, and helmet use. Factors positively associated with conspicuous clothing use included good weather conditions, older age, and more frequent cycling. Factors positively associated with helmet use included bike light use, longer trip distances, hybrid bike type, not using alcohol in the 6 hours prior to the trip, female sex, older age, higher income, and higher education. In two of Canada's largest cities, helmets were the most widely used safety equipment. Measures to increase use of visibility aids on both daytime and night-time cycling trips may help prevent crashes.


Evaluating range-expansion models for calculating nonnative species' expansion rate.
Species range shifts associated with environmental change or biological invasions are increasingly important study areas. However, quantifying range expansion rates may be heavily influenced by methodology and/or sampling bias. We compared expansion rate estimates of Roesel's bush-cricket (Metrioptera roeselii, Hagenbach 1822), a nonnative species currently expanding its range in south-central Sweden, from range statistic models based on distance measures (mean, median, 95(th) gamma quantile, marginal mean, maximum, and conditional maximum) and an area-based method (grid occupancy). We used sampling simulations to determine the sensitivity of the different methods to incomplete sampling across the species' range. For periods when we had comprehensive survey data, range expansion estimates clustered into two groups: (1) those calculated from range margin statistics (gamma, marginal mean, maximum, and conditional maximum: ˜3 km/year), and (2) those calculated from the central tendency (mean and median) and the area-based method of grid occupancy (˜1.5 km/year). Range statistic measures differed greatly in their sensitivity to sampling effort; the proportion of sampling required to achieve an estimate within 10% of the true value ranged from 0.17 to 0.9. Grid occupancy and median were most sensitive to sampling effort, and the maximum and gamma quantile the least. If periods with incomplete sampling were included in the range expansion calculations, this generally lowered the estimates (range 16-72%), with exception of the gamma quantile that was slightly higher (6%). Care should be taken when interpreting rate expansion estimates from data sampled from only a fraction of the full distribution. Methods based on the central tendency will give rates approximately half that of methods based on the range margin. The gamma quantile method appears to be the most robust to incomplete sampling bias and should be considered as the method of choice when sampling the entire distribution is not possible.


Early, Goal-Directed Therapy for Septic Shock - A Patient-Level Meta-Analysis.
After a single-center trial and observational studies suggesting that early, goal-directed therapy (EGDT) reduced mortality from septic shock, three multicenter trials (ProCESS, ARISE, and ProMISe) showed no benefit. This meta-analysis of individual patient data from the three recent trials was designed prospectively to improve statistical power and explore heterogeneity of treatment effect of EGDT. We harmonized entry criteria, intervention protocols, outcomes, resource-use measures, and data collection across the trials and specified all analyses before unblinding. After completion of the trials, we pooled data, excluding the protocol-based standard-therapy group from the ProCESS trial, and resolved residual differences. The primary outcome was 90-day mortality. Secondary outcomes included 1-year survival, organ support, and hospitalization costs. We tested for treatment-by-subgroup interactions for 16 patient characteristics and 6 care-delivery characteristics. We studied 3723 patients at 138 hospitals in seven countries. Mortality at 90 days was similar for EGDT (462 of 1852 patients [24.9%]) and usual care (475 of 1871 patients [25.4%]); the adjusted odds ratio was 0.97 (95% confidence interval, 0.82 to 1.14; P=0.68). EGDT was associated with greater mean (±SD) use of intensive care (5.3±7.1 vs. 4.9±7.0 days, P=0.04) and cardiovascular support (1.9±3.7 vs. 1.6±2.9 days, P=0.01) than was usual care; other outcomes did not differ significantly, although average costs were higher with EGDT. Subgroup analyses showed no benefit from EGDT for patients with worse shock (higher serum lactate level, combined hypotension and hyperlactatemia, or higher predicted risk of death) or for hospitals with a lower propensity to use vasopressors or fluids during usual resuscitation. In this meta-analysis of individual patient data, EGDT did not result in better outcomes than usual care and was associated with higher hospitalization costs across a broad range of patient and hospital characteristics. (Funded by the National Institute of General Medical Sciences and others; PRISM ClinicalTrials.gov number, NCT02030158 .).


Crystal structure of the human 20S proteasome in complex with carfilzomib.
Proteasome inhibition is highly effective as a treatment for multiple myeloma, and recently carfilzomib was granted US FDA approval for the treatment of relapsed and refractory multiple myeloma. Here, we report the X-ray crystal structure of the human constitutive 20S proteasome with and without carfilzomib bound at 2.9 and 2.6 Å, respectively. Our data indicate that the S3 and S4 binding pockets play a pivotal role in carfilzomib's selectivity for chymotrypsin-like sites. Structural comparison with the mouse immunoproteasome crystal structure reveals amino acid substitutions that explain carfilzomib's slight preference for chymotrypsin-like subunits of constitutive proteasomes. In addition, comparison of the human proteasome:carfilzomib complex with the mouse proteasome:PR-957 complex reveals new details that explain why PR-957 is selective for immunoproteasomes. Together, the data presented here support the design of inhibitors for either constitutive or immunoproteasomes, with implications for the treatment of cancers as well as autoimmune and neurodegenerative diseases.


Midwest wrestling study: prediction of minimal weight for high school wrestlers.
This study determined the validity of previously published or newly derived equations to predict fat-free body mass (FFB) in high school wrestlers from the midwestern United States. Five laboratories participated in the data-pooling study (total sample of 860 subjects). Measures included body composition by underwater weighing and anthropometric measurements of body mass, stature, and selected circumferences, diameters, and skinfolds. Cross-validation of selected equations to predict FFB revealed the lowest levels of error from the equations of Lohman, Thorland et al., Katch and McArdle, and Behnke and Wilmore. Modification of the constants in these equations or generation of new equations did not substantially reduce prediction error. Overall, total error for these top equations ranged from 2.44 to 2.59 kg. However, based on observed trends, this error was of lower magnitude with the younger and lighter subjects and of higher magnitude with the older and heavier subjects. We conclude that these equations could be used singularly or collectively to determine FFB, and a minimal weight could then be derived and assigned to a scholastic wrestler.


A critical review of foot orthoses in the rheumatoid arthritic foot.
Foot orthoses are commonly prescribed by health professionals as a form of intervention for the symptomatic foot in rheumatoid arthritis. However, there is a limited evidence base to support the use of foot orthoses in this patient group. This article provides a critical review of the use of foot orthoses in the management of rheumatoid arthritic foot pathologies. A search was conducted in the Cochrane Controlled Trials Register (current issue of the Cochrane Library), Physiotherapy evidence database (PEDro), Medline, The Cumulative Index to Nursing and Allied Health Literature (CINAHL) and Allied and Complementary Medicine (AMED) and from reference lists in journal articles. The language was restricted to English. Searching of the databases was undertaken between December 2004 and March 2005. The results indicated there is no consensus of opinion on the choice of foot orthoses used for the management of pathology in the rheumatoid foot, although there is strong evidence that foot orthoses do reduce pain and improve functional ability. The type of foot orthoses used ranged from simple cushioned insoles to custom-made rigid cast devices. Methodological issues raised included small sample size and poor use of valid and reliable outcome measures. There is limited evidence pertaining to cost-effectiveness. The results indicated a need for further investigation into the most clinically and cost-effective foot orthoses to prescribe in the management of the rheumatoid arthritic foot. This review highlights the need to identify the various types of foot orthoses that are most effective in the management of the established rheumatoid arthritic foot.


Committee Opinion No. 704: Human Papillomavirus Vaccination.
Human papillomavirus (HPV) is associated with anogenital cancer (including cervical, vaginal, vulvar, penile, and anal), oropharyngeal cancer, and genital warts. The HPV vaccination significantly reduces the incidence of anogenital cancer and genital warts. Despite the benefits of HPV vaccines, only 41.9% of girls in the recommended age group, and only 28.1% of males in the recommended age group have received all recom-mended doses. Compared with many other countries, HPV vaccination rates in the United States are unacceptably low. The U.S. Food and Drug Administration has approved three vaccines that are effective at preventing HPV infection. These vaccines cover 2, 4, or 9 HPV serotypes, respectively. Safety data for all three HPV vaccines are reassuring. The HPV vaccines are recommended for girls and boys aged 11-12 years and can be given to females and males up to age 26 years. The Advisory Committee on Immunization Practices and the American College of Obstetricians and Gynecologists recommend routine HPV vaccination for girls and boys at the target age of 11-12 years (but it may be given from the age of 9 years) as part of the adolescent immunization platform in order to help reduce the incidence of anogenital cancer and genital warts associated with HPV infection. Obstetrician-gynecologists and other health care providers should stress to parents and patients the benefits and safety of HPV vaccination and offer HPV vaccines in their offices.


Tailoring the orthopaedic consultation: How perceived patient characteristics influence surgeons' communication.
To investigate whether and how orthopaedic surgeons tailor communication during medical consultations based on perceived patient characteristics. Seven orthopaedic surgeons were repeatedly interviewed following an approach based on ecological momentary assessment. Qualitative content analysis was used to analyse the eighty short interviews. The association between patient characteristics and tailoring approaches was explored in a correspondence analysis of the counted codes. Surgeons estimate patients' competence (illness management and communication abilities), autonomy, and interpersonal behaviour. They report tailoring communication in two-thirds of the consultations. The surgeons' perception was associated with the employment of specific approaches to communication: (1) high patient competence with extensive information provision or no changes in communication, (2) less autonomy and less competence with reassurance and direction, (3) high autonomy with discussions about pace and expectations, and (4) high sociability with communication about personal circumstances and wishes. The surgeon's perception of a patient influences communication during consultations. Future research should address whether these intuitively employed approaches are appropriate, effective, and generalizable to other medical specialists. Tailoring physician-patient communication can improve its quality. The novel approaches identified in this study can be used to formulate and test formal guidelines for tailored communication.


Structure and genetics of the O-antigen of Escherichia coli O169 related to the O-antigen of Shigella boydii type 6.
The O-polysaccharide (O-antigen) of Escherichia coli O169 was studied by sugar analysis along with 1D and 2D (1)H and (13)C NMR spectroscopy. The following structure of the branched hexasaccharide repeating unit was established: [Formula: see text] The O-polysaccharide of E. coli O169 differs from that of Shigella boydii type 6 only in the presence of a side-chain glucose residue. A comparison of the O-antigen biosynthesis gene clusters between the galF to gnd genes in the genomes of the two bacteria revealed their close relationship. The glycosyltransferase gene responsible for the formation of the β-D-Glcp-(1 → 6)-α-D-Galp linkage in the O-antigen was identified in the gene cluster.


Structural and immunological comparison of indigenous human O6-methylguanine-DNA methyltransferase with that encoded by a cloned cDNA.
O6-Methylguanine-DNA methyltransferase, a ubiquitous and unusual DNA repair protein, eliminates mutagenic and cytotoxic O6-alkylguanine from DNA by transferring the alkyl group to one of its cysteine residues in a second-order suicide reaction. This 22-kDa protein was immunoaffinity-purified to homogeneity from cultured human lymphoblasts (CEM-CCRF line) and compared with the O6-methylguanine-DNA methyltransferase purified to homogeneity from Escherichia coli expressing a cloned human cDNA. The cellular and recombinant proteins were identical in size, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of intact molecules and their peptides. Immunoprobing of Western blots with three monoclonal antibodies specific for human cellular O6-methylguanine-DNA methyltransferase further indicated identity of the two proteins. The amino acid sequence of the cellular protein was experimentally determined for 87 out of a total of 207 residues and was found to be identical to that deduced from the cDNA sequence. A unique cysteine residue at position 145 was identified as the methyl acceptor site by autoradiographic analysis of peptides and sequence analysis of 3H-methylated O6-methylguanine-DNA methyltransferase. These observations establish that the cloned O6-methylguanine-DNA methyltransferase cDNA encodes the full-length O6-methylguanine-DNA methyltransferase polypeptide that is normally present in human cells. Moreover, the cellular protein does not appear to be significantly modified by posttranslational processes.


Phase-based, high spatial resolution and distributed, static and dynamic strain sensing using Brillouin dynamic gratings in optical fibers.
A novel technique combining Brillouin phase-shift measurements with Brillouin dynamic gratings (BDGs) reflectometry in polarization-maintaining fibers is presented here for the first time. While a direct measurement of the optical phase in standard BDG setups is impractical due to non-local phase contributions, their detrimental effect is reduced by ~4 orders of magnitude through the coherent addition of Stokes and anti-Stokes reflections from two counter-propagating BDGs in the fiber. The technique advantageously combines the high-spatial-resolution of BDGs reflectometry with the increased tolerance to optical power fluctuations of phasorial measurements, to enhance the performance of fiber-optic strain sensors. We demonstrate a distributed measurement (20cm spatial-resolution) of both static and dynamic (5kHz of vibrations at a sampling rate of 1MHz) strain fields acting on the fiber, in good agreement with theory and (for the static case) with the results of commercial reflectometers.


Rapid colour shift by reproductive character displacement in Cupido butterflies.
Reproductive character displacement occurs when competition for successful breeding imposes a divergent selection on the interacting species, causing a divergence of reproductive traits. Here, we show that a disputed butterfly taxon is actually a case of male wing colour shift, apparently produced by reproductive character displacement. Using double digest restriction-site associated DNA sequencing and mitochondrial DNA sequencing we studied four butterfly taxa of the subgenus Cupido (Lepidoptera: Lycaenidae): Cupido minimus and the taxon carswelli, both characterized by brown males and females, plus C. lorquinii and C. osiris, both with blue males and brown females. Unexpectedly, taxa carswelli and C. lorquinii were close to indistinguishable based on our genomic and mitochondrial data, despite displaying strikingly different male coloration. In addition, we report and analysed a brown male within the C. lorquinii range, which demonstrates that the brown morph occurs at very low frequency in C. lorquinii. Such evidence strongly suggests that carswelli is conspecific with C. lorquinii and represents populations with a fixed male brown colour morph. Considering that these brown populations occur in sympatry with or very close to the blue C. osiris, and that the blue C. lorquinii populations never do, we propose that the taxon carswelli could have lost the blue colour due to reproductive character displacement with C. osiris. Since male colour is important for conspecific recognition during courtship, we hypothesize that the observed colour shift may eventually trigger incipient speciation between blue and brown populations. Male colour seems to be an evolutionarily labile character in the Polyommatinae, and the mechanism described here might be at work in the wide diversification of this subfamily of butterflies.


Interactions between attention and memory.
Attention and memory cannot operate without each other. In this review, we discuss two lines of recent evidence that support this interdependence. First, memory has a limited capacity, and thus attention determines what will be encoded. Division of attention during encoding prevents the formation of conscious memories, although the role of attention in formation of unconscious memories is more complex. Such memories can be encoded even when there is another concurrent task, but the stimuli that are to be encoded must be selected from among other competing stimuli. Second, memory from past experience guides what should be attended. Brain areas that are important for memory, such as the hippocampus and medial temporal lobe structures, are recruited in attention tasks, and memory directly affects frontal-parietal networks involved in spatial orienting. Thus, exploring the interactions between attention and memory can provide new insights into these fundamental topics of cognitive neuroscience.


Postoperative circadian rhythms and cortisol stress response to two types of cardiac surgery.
Although endocrine stress responses of coronary artery bypass patients have been reported, similar studies have not been conducted in patients undergoing implantation of an automatic cardioverter/defibrillator device. To compare patterns and magnitudes of intra- and postoperative cortisol stress responses in patients undergoing two types of surgery; rate at which postoperative cortisol levels return to presurgical levels; and whether stressors associated with these two procedures affect postoperative circadian rhythms of cortisol, heart rate, and body temperature. A time-series, small-group design was used to investigate 16 coronary artery bypass graft and 9 automatic cardioverter defibrillator patients. For the cortisol stress study, blood samples were obtained pre-, intra-, and postoperatively until day 6. For the circadian rhythm aspect, cortisol circadian rhythm aspect, cortisol levels, heart rate, and oral temperatures were measured every 2 hours for 24 hours beginning on postoperative day 1. Plasma cortisol levels increased significantly toward the latter part of surgery. Generally, no significant group differences were found in cortisol magnitude during and after surgery. Postoperative cortisol levels were significantly elevated from preoperative levels for both groups; however, the automatic cardioverter defibrillator group's cortisol levels declined at a slower rate. The stressors associated with these surgeries were sufficient to disrupt or abolish circadian rhythms for the measured variables. The elevated postoperative cortisol levels and altered circadian rhythms may contribute to increased vulnerability by dampening patients' anti-inflammatory and immunologic responses and adversely affecting their sense of well-being. The clinical significance of these findings is yet to be determined; more research is warranted.


Will consumer-directed health care improve system performance?
Consumer-directed health care plans have attracted attention as a method for managing rising health care spending by giving consumers greater financial control over their health care. However, increased cost-sharing--the principal tool used by these plans to achieve lower spending--may also cause patients to consume less care, even when that care is essential. Research studies have found that lower-income individuals and those with serious health concerns will particularly be at risk, as these consumers bear the burden of higher out-of-pocket costs. Instead of focusing solely on financial incentives, the real goal should be to encourage quality and efficiency among health systems, physicians, and hospitals.


On the use of leaf spectral indices to assess water status and photosynthetic limitations in Olea europaea L. during water-stress and recovery.
Diffusional limitations to photosynthesis, relative water content (RWC), pigment concentrations and their association with reflectance indices were studied in olive (Olea europaea) saplings subjected to water-stress and re-watering. RWC decreased sharply as drought progressed. Following rewatering, RWC gradually increased to pre-stress values. Photosynthesis (A), stomatal conductance (gs), mesophyll conductance (gm), total conductance (gt), photochemical reflectance index (PRI), water index (WI) and relative depth index (RDI) closely followed RWC. In contrast, carotenoid concentration, the carotenoid to chlorophyll ratio, water content reflectance index (WCRI) and structural independent pigment index (SIPI) showed an opposite trend to that of RWC. Photosynthesis scaled linearly with leaf conductance to CO2; however, A measured under non-photorespiratory conditions (A1%O2) was approximately two times greater than A measured at 21% [O2], indicating that photorespiration likely increased in response to drought. A1%O2 also significantly correlated with leaf conductance parameters. These relationships were apparent in saturation type curves, indicating that under non-photorespiratory conditions, CO2 conductance was not the major limitations to A. PRI was significant correlated with RWC. PRI was also very sensitive to pigment concentrations and photosynthesis, and significantly tracked all CO2 conductance parameters. WI, RDI and WCRI were all significantly correlated with RWC, and most notably to leaf transpiration. Overall, PRI correlated more closely with carotenoid concentration than SIPI; whereas WI tracked leaf transpiration more effectively than RDI and WCRI. This study clearly demonstrates that PRI and WI can be used for the fast detection of physiological traits of olive trees subjected to water-stress.


Many-body localization in imperfectly isolated quantum systems.
We use numerical exact diagonalization to analyze which aspects of the many-body localization phenomenon survive in an imperfectly isolated setting, when the system of interest is weakly coupled to a thermalizing environment. We show that widely used diagnostics (such as many-body level statistics and expectation values in exact eigenstates) cease to show signatures of many-body localization above a critical coupling that is exponentially small in the size of the environment. However, we also identify alternative diagnostics for many-body localization, in the spectral functions of local operators. Diagnostics include a discrete spectrum and a hierarchy of energy gaps, including a universal gap at zero frequency. These alternative diagnostics are shown to be robust, and continue to show signatures of many-body localization as long as the coupling to the bath is weaker than the characteristic energy scales in the system. We also examine how these signatures disappear when the coupling to the environment becomes larger than the characteristic energy scales of the system.


Potential solutions to problems posed by the offspring sex ratios of people with parasitic and viral infections.
I have hypothesized that mammalian offspring sex ratios (proportions male at birth) are partially controlled by the hormone levels of both parents around the time of conception. Recently data have been published (privately) which impugn suggestions of mine (based on that hypothesis) relating to the offspring sex ratios of people who are carriers of hepatitis B virus; and (publicly) to hormone profiles of women who are infected with the protozoan parasite Toxoplasma gondii. Here these data are reviewed in the light of data on offspring sex ratios of people infected with hepatitis C, and cytomegalovirus and T. gondii; and of mice with T. gondii and the trichostrongyline nematode Heligmosomoides polygyrus. Three proposals are made viz: 1. Women infected with T. gondii may have high oestrogen levels. This would potentially explain reports of their high offspring sex ratios, long gestations and (perhaps) behavioural traits. 2. Some of these infective organisms may 'prefer' a particular endocrine environment in their host (in the sense that it facilitates replication or transmission or both). 3. Some of these organisms apparently cause changes in that endocrine environment as a consequence of their presence. If both the latter two proposals were correct, it could explain recent puzzling data on the offspring sex ratios of hepatitis B carriers. And if this solution turns out to be correct, it would suggest new endocrine means of combating hepatitis B infection.


Biomphalaria alexandrina in Egypt: past, present and future.
The African species of Biomphalaria appeared as a result of the relatively recent west-to-east trans-Atlantic dispersal of the Biomphalaria glabrata-like taxon. In Egypt, Biomphalaria alexandrina is the intermediate host for Schistosoma mansoni. Biomphalaria alexandrina originated in the area between Alexandria and Rosetta and has historically been confined to the Nile Delta. Schistosoma mansoni reached Egypt via infected slaves and baboons from the Land of Punt through migrations that occurred as early as the Vth Dynasty. The suggestion of the presence of Schistosoma mansoni infection in Lower Egypt during Pharaonic times is discussed despite the fact that that there is no evidence of such infection in Egyptian mummies. It is only recently that Biomphalaria alexandrina colonized the Egyptian Nile from the Delta to Lake Nasser. This change was likely due to the construction of huge water projects, the development of new water resources essential for land reclamation projects and the movement of refugees from the Suez Canal zone to the Delta and vice versa. The situation with respect to Biomphalaria in Egypt has become complicated in recent years by the detection of Biomphalaria glabrata and a hybrid between both species; however, follow-up studies have demonstrated the disappearance of such species within Egypt. The National Schistosoma Control Program has made great strides with respect to the eradication of schistosoma; however, there has unfortunately been a reemergence of Schistosoma mansoni resistant to praziquantel. There are numerous factors that may influence the prevalence of snails in Egypt, including the construction of water projects, the increase in reclaimed areas, global climate change and pollution. Thus, continued field studies in addition to the cooperation of several scientists are needed to obtain an accurate representation of the status of this species. In addition, the determination of the genome sequence for Biomphalaria alexandrina and the use of modern technology will allow for the study of the host-parasite relationship at a molecular level.


Establishing political priority for global mental health: a qualitative policy analysis.
Mental disorders represent the leading cause of disability worldwide, yet they remain a low global health priority. This paper uses a case study methodology and different data sources (35 interviews and documents) to analyse factors that have shaped the generation of political priority for global mental health over the past three decades (1990-2020) and their changes over time. The Shiffman and Smith framework on determinants of political priority for global health issues is used to organize data into themes: actor power, ideas, political context and issue characteristics. Global mental health has gained political attention, especially over the past decade, yet support remains limited. Findings reveal that actor power is undermined by a fragmented policy community, the absence of one guiding institution or coordination mechanism and little civil society mobilization. Public portrayal of the issue is divided, hampered by the absence of a common understanding by the community and by stigma. Some policy windows have been missed and a strong global governance structure is lacking. Credible indicators and evidence on simple cost-effective solutions, especially in low- and middle-income countries, are scarce. However, opportunities are arising, including an increasing number of leaders and grassroots organizations, multiple arguments for action and integrated solutions resonating with broader audiences, widening political support at the national level, an emerging global governance structure and an expanding evidence base on the scale of the problem and available solutions. The results point to three technical and four political challenges that advocates need to address to increase political support over the next decade.


Stage-specific fluorescence intensity of GFP and mCherry during sporulation In Bacillus Subtilis.
Fluorescent proteins are powerful molecular biology tools that have been used to study the subcellular dynamics of proteins within live cells for well over a decade. Two fluorescent proteins commonly used to enable dual protein labelling are GFP (green) and mCherry (red). Sporulation in the Gram positive bacterium Bacillus subtilis has been studied for many years as a paradigm for understanding the molecular basis for differential gene expression. As sporulation initiates, cells undergo an asymmetric division leading to differential gene expression in the small prespore and large mother cell compartments. Use of two fluorescent protein reporters permits time resolved examination of differential gene expression either in the same compartments or between compartments. Due to the spectral properties of GFP and mCherry, they are considered an ideal combination for co-localisation and co-expression experiments. They can also be used in combination with fluorescent DNA stains such as DAPI to correlate protein localisation patterns with the developmental stage of sporulation which can be linked to well characterised changes in DNA staining patterns. While observing the recruitment of the transcription machinery into the forespore of sporulating Bacillus subtilis, we noticed the occurrence of stage-specific fluorescence intensity differences between GFP and mCherry. During vegetative growth and the initial stages of sporulation, fluorescence from both GFP and mCherry fusions behaved similarly. During stage II-III of sporulation we found that mCherry fluorescence was considerably diminished, whilst GFP signals remained clearly visible. This fluorescence pattern reversed during the final stage of sporulation with strong mCherry and low GFP fluorescence. These trends were observed in reciprocal tagging experiments indicating a direct effect of sporulation on fluorescent protein fluorophores. Great care should be taken when interpreting the results of protein localisation and quantitative gene expression patterns using fluorescent proteins in experiments involving intracellular physiological change. We believe changes in the subcellular environment of the sporulating cell leads to conditions that differently alter the spectral properties of GFP and mCherry making an accurate interpretation of expression profiles technically challenging.


Clinical effectiveness and cost-effectiveness of cholecystectomy compared with observation/conservative management for preventing recurrent symptoms and complications in adults presenting with uncomplicated symptomatic gallstones or cholecystitis: a systematic review and economic evaluation.
Approximately 10-15% of the adult population suffer from gallstone disease, cholelithiasis, with more women than men being affected. Cholecystectomy is the treatment of choice for people who present with biliary pain or acute cholecystitis and evidence of gallstones. However, some people do not experience a recurrence after an initial episode of biliary pain or cholecystitis. As most of the current research focuses on the surgical management of the disease, less attention has been dedicated to the consequences of conservative management. To determine the clinical effectiveness and cost-effectiveness of cholecystectomy compared with observation/conservative management in people presenting with uncomplicated symptomatic gallstones (biliary pain) or cholecystitis. We searched all major electronic databases (e.g. MEDLINE, EMBASE, Science Citation Index, Bioscience Information Service, Cochrane Central Register of Controlled Trials) from 1980 to September 2012 and we contacted experts in the field. Evidence was considered from randomised controlled trials (RCTs) and non-randomised comparative studies that enrolled people with symptomatic gallstone disease (pain attacks only and/or acute cholecystitis). Two reviewers independently extracted data and assessed the risk of bias of included studies. Standard meta-analysis techniques were used to combine results from included studies. A de novo Markov model was developed to assess the cost-effectiveness of the interventions. Two Norwegian RCTs involving 201 participants were included. Eighty-eight per cent of people randomised to surgery and 45% of people randomised to observation underwent cholecystectomy during the 14-year follow-up period. Participants randomised to observation were significantly more likely to experience gallstone-related complications [risk ratio = 6.69; 95% confidence interval (CI) 1.57 to 28.51; p = 0.01], in particular acute cholecystitis (risk ratio = 9.55; 95% CI 1.25 to 73.27; p = 0.03), and less likely to undergo surgery (risk ratio = 0.50; 95% CI 0.34 to 0.73; p = 0.0004), experience surgery-related complications (risk ratio = 0.36; 95% CI 0.16 to 0.81; p = 0.01) or, more specifically, minor surgery-related complications (risk ratio = 0.11; 95% CI 0.02 to 0.56; p = 0.008) than those randomised to surgery. Fifty-five per cent of people randomised to observation did not require an operation during the 14-year follow-up period and 12% of people randomised to cholecystectomy did not undergo the scheduled operation. The results of the economic evaluation suggest that, on average, the surgery strategy costs £1236 more per patient than the conservative management strategy but was, on average, more effective. An increase in the number of people requiring surgery while treated conservatively corresponded to a reduction in the cost-effectiveness of the conservative strategy. There was uncertainty around some of the parameters used in the economic model. The results of this assessment indicate that cholecystectomy is still the treatment of choice for many symptomatic people. However, approximately half of the people in the observation group did not require surgery or suffer complications in the long term indicating that a conservative therapeutic approach may represent a valid alternative to surgery in this group of people. Owing to the dearth of current evidence in the UK setting a large, well-designed, multicentre trial is needed. The study was registered as PROSPERO CRD42012002817. The National Institute for Health Research Health Technology Assessment programme.


Structurally perturbed chiral Bragg reflectors for elliptically polarized light.
The structure of an inorganic chiral medium represented as a stack of identical form-birefringent layers that twist steadily with increasing thickness is perturbed by realigning a fraction of each layer to a fixed direction. Experimental results show that the resulting chiral-birefringent composite medium exhibits Bragg resonance with elliptically polarized light, and simulations indicate that Bragg reflectors can be designed for any polarization including linear.


Generation and maintenance of the respiratory rhythm.
Activities of the phrenic and internal intercostal nerves show that the central nervous rhythm of respiration consists of 3 phases: inspiratory, postinspiratory and expiratory. The discharge patterns of medullary respiratory neurones of the anaesthetized, paralysed cat can be correlated with these phases of the central respiratory cycle, and the postsynaptic activity of individual cells can be analysed to obtain information about the populations of neurones converging upon them. Inferences are drawn about respiratory neurone connectivity and a theory is developed that the respiratory network primarily employs inspiratory-related neurones and that medullary expiratory neurones are less important for the rhythmogenesis of respiration. It is suggested that the inspiratory network consists of a ramp generating excitatory loop network of interneurones whose discharge is brought to an end ('off-switched') by inhibitory late-inspiratory interneurones. The discharge pattern of the latter type of neurone is explained by inhibition arriving from early-inspiratory interneurones. Subsequent to 'off-switching' the ramp generator is assumed to be immediately gated by a very powerful postinspiratory inhibition whereas expiratory activity seems to be disfacilitated at this time. This is the period when 'passive' (stage 1) expiration occurs. Following this interposed postinspiratory phase 'active' (stage 2) expiration may begin, depending on the amount of excitatory inflow to the inspiratory ramp generator. When expiratory neurones are activated the inspiratory system is again synaptically inhibited and the frequency of ventilation is markedly slowed.


Pharmacokinetics of buprenorphine hydrochloride following intramuscular and intravenous administration to American kestrels (Falco sparverius).
To determine the pharmacokinetics of buprenorphine hydrochloride after IM and IV administration to American kestrels (Falco sparverius). 13 healthy 3-year-old captive-bred American kestrels. Buprenorphine hydrochloride (0.6 mg/kg) was administered IM to all birds. Blood samples were collected at 9 times, ranging from 5 minutes to 9 hours after drug administration. Plasma buprenorphine concentrations were measured by use of tandem liquid chromatography-mass spectrometry. Pharmacokinetic parameters were determined by use of least squares linear regression and noncompartmental analysis of naïve pooled data. After a washout period of 2 weeks, the same dose of buprenorphine was administered IV to all birds and blood samples were collected at the same times after drug administration. Maximum plasma buprenorphine concentration was achieved within 5 minutes after IM administration. For IM administration, bioavailability was 94.8% and elimination half-life was 92.1 minutes. For IV administration, steady-state volume of distribution was 4,023.8 mL/kg, plasma clearance was 49.2 mL/min/kg, and elimination half-life was 105.5 minutes. Buprenorphine was rapidly absorbed, and bioavailability was good after IM administration to American kestrels. Plasma buprenorphine concentrations were > 1 ng/mL for 9 hours after both IM and IV administration. These results, in combination with those of a pharmacodynamic study, suggested that the analgesic effects of buprenorphine could last at least 6 to 9 hours in this species. Further investigations of the duration of analgesic effects, multiple-dose protocols, and potential adverse effects of buprenorphine are warranted in American kestrels and other raptors.


Molecular epidemiology of health care-associated transmission of hepatitis B and C viruses.
The term "molecular epidemiology" has been ascribed to a host of different activities that involve gene-sequence analysis. Some examples of molecular epidemiology include modeling exercises of phylogenetic trees to reconstruct epidemics; studies of the evolution of hepatitis C virus (HCV); rates of nucleotide substitution in the hepatitis B virus (HBV) surface (S) gene; variations in the core promoter/pre-core/core region of HBV genotype C from different sources; analysis of HBV surface antigen mutations; molecular clock analyses of the short-term evolution of HCV; and analyses of clades and surface antigen polymorphisms of HBV. However, for most epidemiologists molecular epidemiology of viral hepatitis usually refers to studies of gene-sequence homology in HBV or HCV recovered from people in the community or an institution that allows better characterization and assignment of related clusters of infection.


Management Options for Irritable Bowel Syndrome.
Irritable bowel syndrome (IBS) is associated with diverse pathophysiologic mechanisms. These mechanisms include increased abnormal colonic motility or transit, intestinal or colorectal sensation, increased colonic bile acid concentration, and superficial colonic mucosal inflammation, as well as epithelial barrier dysfunction, neurohormonal up-regulation, and activation of secretory processes in the epithelial layer. Novel approaches to treatment include lifestyle modification, changes in diet, probiotics, and pharmacotherapy directed to the motility, sensation, and intraluminal milieu of patients with IBS. Despite recent advances, there is a need for development of new treatments to relieve pain in IBS without deleterious central or other adverse effects.


Mortality from external causes in Africa and Asia: evidence from INDEPTH Health and Demographic Surveillance System Sites.
Mortality from external causes, of all kinds, is an important component of overall mortality on a global basis. However, these deaths, like others in Africa and Asia, are often not counted or documented on an individual basis. Overviews of the state of external cause mortality in Africa and Asia are therefore based on uncertain information. The INDEPTH Network maintains longitudinal surveillance, including cause of death, at population sites across Africa and Asia, which offers important opportunities to document external cause mortality at the population level across a range of settings. To describe patterns of mortality from external causes at INDEPTH Network sites across Africa and Asia, according to the WHO 2012 verbal autopsy (VA) cause categories. All deaths at INDEPTH sites are routinely registered and followed up with VA interviews. For this study, VA archives were transformed into the WHO 2012 VA standard format and processed using the InterVA-4 model to assign cause of death. Routine surveillance data also provide person-time denominators for mortality rates. A total of 5,884 deaths due to external causes were documented over 11,828,253 person-years. Approximately one-quarter of those deaths were to children younger than 15 years. Causes of death were dominated by childhood drowning in Bangladesh, and by transport-related deaths and intentional injuries elsewhere. Detailed mortality rates are presented by cause of death, age group, and sex. The patterns of external cause mortality found here generally corresponded with expectations and other sources of information, but they fill some important gaps in population-based mortality data. They provide an important source of information to inform potentially preventive intervention designs.


Migration experiences, employment status and psychological distress among Somali immigrants: a mixed-method international study.
The discourse about mental health problems among migrants and refugees tends to focus on adverse pre-migration experiences; there is less investigation of the environmental conditions in which refugee migrants live, and the contrasts between these situations in different countries. This cross-national study of two samples of Somali refugees living in London (UK) and Minneapolis, Minnesota, (USA) helps to fill a gap in the literature, and is unusual in being able to compare information collected in the same way in two cities in different countries. There were two parts to the study, focus groups to gather in-depth qualitative data and a survey of health status and quantifiable demographic and material factors. Three of the focus groups involved nineteen Somali professionals and five groups included twenty-eight lay Somalis who were living in London and Minneapolis. The quantitative survey was done with 189 Somali respondents, also living in London and Minneapolis. We used the MINI International Neuropsychiatric Interview (MINI) to assess ICD-10 and DSM-IV mental disorders. The overall qualitative and quantitative results suggested that challenges to masculinity, thwarted aspirations, devalued refugee identity, unemployment, legal uncertainties and longer duration of stay in the host country account for poor psychological well-being and psychiatric disorders among this group. The use of a mixed-methods approach in this international study was essential since the quantitative and qualitative data provide different layers and depth of meaning and complement each other to provide a fuller picture of complex and multi-faceted life situations of refugees and asylum seekers. The comparison between the UK and US suggests that greater flexibility of access to labour markets for this refugee group might help to promote opportunities for better integration and mental well-being.


Integrated community case management of fever in children under five using rapid diagnostic tests and respiratory rate counting: a multi-country cluster randomized trial.
Evidence on the impact of using diagnostic tests in community case management of febrile children is limited. This effectiveness trial conducted in Burkina Faso, Ghana, and Uganda, compared a diagnostic and treatment package for malaria and pneumonia with presumptive treatment with anti-malarial drugs; artemisinin combination therapy (ACT). We enrolled 4,216 febrile children between 4 and 59 months of age in 2009-2010. Compliance with the malaria rapid diagnostic test (RDT) results was high in the intervention arm across the three countries, with only 4.9% (17 of 344) of RDT-negative children prescribed an ACT. Antibiotic overuse was more common: 0.9% (4 of 446) in Uganda, 38.5% (114 of 296) in Burkina Faso, and 44.6% (197 of 442) in Ghana. Fever clearance was high in both intervention and control arms at both Day 3 (97.8% versus 96.9%, P = 0.17) and Day 7 (99.2% versus 98.8%, P = 0.17). The use of diagnostic tests limits overuse of ACTs. Its impact on antibiotic overuse and on fever clearance is uncertain.


The Haemophilus influenzae Sap transporter mediates bacterium-epithelial cell homeostasis.
Nontypeable Haemophilus influenzae (NTHI) is a commensal inhabitant of the human nasopharynx and a causative agent of otitis media and other diseases of the upper and lower human airway. During colonization within the host, NTHI must acquire essential nutrients and evade immune attack. We previously demonstrated that the NTHI Sap transporter, an inner membrane protein complex, mediates resistance to antimicrobial peptides and is required for heme homeostasis. We hypothesized that Sap transporter functions are critical for NTHI interaction with the host epithelium and establishment of colonization. Thus, we cocultured the parent or the sapA mutant on polarized epithelial cells grown at an air-liquid interface, as a physiological model of NTHI colonization, to determine the contribution of the Sap transporter to bacterium-host cell interactions. Although SapA-deficient NTHI was less adherent to epithelial cells, we observed a significant increase in invasive bacteria compared to the parent strain. Upon internalization, the sapA mutant appeared free in the cytoplasm, whereas the parent strain was primarily found in endosomes, indicating differential subcellular trafficking. Additionally, we observed reduced inflammatory cytokine production by the epithelium in response to the sapA mutant strain compared to the parental strain. Furthermore, chinchilla middle ears challenged with the sapA mutant demonstrated a decrease in disease severity compared to ears challenged with the parental strain. Collectively, our data suggest that NTHI senses host environmental cues via Sap transporter function to mediate interaction with host epithelial cells. Epithelial cell invasion and modulation of host inflammatory cytokine responses may promote NTHI colonization and access to essential nutrients.


Transfusion practices and complications in thalassemia.
The severe forms of thalassemia are the most common inherited anemias managed with regular blood transfusion therapy. Transfusion policies and complications are critical to quality of life and survival, but there is a lack of standardized care. A survey of 58 items was completed in 2016 by 11 centers in California, Washington, Oregon, Nevada, and Arizona providing long-term care for thalassemia. The questionnaire addressed demographic information, transfusion practices and complications, and educational needs. The centers followed 717 patients with β-thalassemia (314, 43.8%) or α-thalassemia (394, 55%). One-third (34.7%) of patients were transfusion-dependent. Indications and goals of transfusion therapy differed between centers. Prestorage leukoreduction was universal, while routine irradiation of units was limited to one site. Red blood cell antigen phenotype was determined before the first transfusion and patients received Rh/Kell-matched units. However, more than half of the transfused patients had received blood at multiple hospitals within or outside the United States. Alloantibodies were seen in 16.9% of transfused group, but management of such patients was variable. Unusual or emerging transfusion-transmitted pathogens were not observed. Multiple educational needs were recognized, with iron overload as the biggest challenge; the approach to iron chelation varied within the group. This study identified many patients not included in earlier surveys limited to major national centers, suggesting that the thalassemia population in the United States is vastly underestimated. Lack of evidence-based guidelines is a barrier to optimal care, which should be addressed through regional consortia of thalassemia centers.


Resistant starch and exercise independently attenuate weight regain on a high fat diet in a rat model of obesity.
Long-term weight reduction remains elusive for many obese individuals. Resistant starch (RS) and exercise may be useful for weight maintenance. The effects of RS, with or without exercise, on weight regain was examined during relapse to obesity on a high carbohydrate, high fat (HC/HF) diet. Obesity-prone rats were fed ad libitum for 16 weeks then weight reduced on a low fat diet to induce a 17% body weight loss (weight reduced rats). Weight reduced rats were maintained on an energy-restricted low fat diet for 18 weeks, with or without a daily bout of treadmill exercise. Rats were then allowed free access to HC/HF diet containing low (0.3%) or high (5.9%) levels of RS. Weight regain, energy balance, body composition, adipocyte cellularity, and fuel utilization were monitored as rats relapsed to obesity and surpassed their original, obese weight. Both RS and exercise independently attenuated weight regain by reducing the energy gap between the drive to eat and suppressed energy requirements. Exercise attenuated the deposition of lean mass during relapse, whereas its combination with RS sustained lean mass accrual as body weight returned. Early in relapse, RS lowered insulin levels and reduced the deposition of fat in subcutaneous adipose tissue. Exercise cessation at five weeks of relapse led to increased weight gain, body fat, subcutaneous adipocytes, and decreased lean mass; all detrimental consequences to overall metabolic health. These data are the first to show the complimentary effects of dietary RS and regular exercise in countering the metabolic drive to regain weight following weight loss and suggest that exercise cessation, in the context of relapse on a HC/HF diet, may have dire metabolic consequences.


Tobacco control policy development in the European Union: do political factors matter?
There has been much variation between European countries in the development of tobacco control policy. Not much is known about the factors that shape this variation. This study aimed to assess the role of political factors in tobacco control policy development. We used data from 11 European countries from 1996 to 2010. Multilevel regression modelling was used to investigate associations between the Tobacco Control Scale (TCS) and indicators of left-wing government and government effectiveness (policy formulation, implementation and enforcement), with control for confounders. An association was found between left-wing government and TCS over the period 1996-2003, but not over the whole period. The association between government effectiveness and TCS was significant and negative over the whole period, but positive between 2001 and 2005. Residual analysis showed that TCS scores from 2002 onwards were higher than expected based on the political factors. Associations varied among the five subscales of the TCS. The results suggest that, on the whole, national political factors have had only a minor influence on the large variations in tobacco control policy development in Europe over the last 15 years. However, there are indications that left-wing governments were important for early adoption of tobacco control policy, and high government effectiveness was important in the phase of adopting innovative policies. However, since 2002, with the advent of international treaties, the influence of national politics has diminished.


Does human skin truly behave as an array of helical antennae in the millimeter and terahertz wave ranges?
The sweat ducts of the human perspiration system are helically shaped tubes, filled with a conductive aqueous solution. Recent studies have claimed that these ducts act as an array of low-Q helical antennae and are dominant in shaping the spectral response in the subterahertz region. Using local homogenization theory for the skin embedded with sweat ducts, we found that multiple interference effects from the skin layers play the major role in determining the skin electromagnetic characteristics in the millimeter and terahertz regions without the need for the assumption of the sweat ducts acting as low-Q helical antennae.


LY75 Ablation Mediates Mesenchymal-Epithelial Transition (MET) in Epithelial Ovarian Cancer (EOC) Cells Associated with DNA Methylation Alterations and Suppression of the Wnt/β-Catenin Pathway.
Growing evidence demonstrates that epithelial-mesenchymal transition (EMT) plays an important role in epithelial ovarian cancer (EOC) progression and spreading; however, its molecular mechanisms remain poorly defined. We have previously shown that the antigen receptor LY75 can modulate EOC cell phenotype and metastatic potential, as LY75 depletion directed mesenchymal-epithelial transition (MET) in EOC cell lines with mesenchymal phenotype. We used the LY75-mediated modulation of EMT as a model to investigate for DNA methylation changes during EMT in EOC cells, by applying the reduced representation bisulfite sequencing (RRBS) methodology. Numerous genes have displayed EMT-related DNA methylation patterns alterations in their promoter/exon regions. Ten selected genes, whose DNA methylation alterations were further confirmed by alternative methods, were further identified, some of which could represent new EOC biomarkers/therapeutic targets. Moreover, our methylation data were strongly indicative for the predominant implication of the Wnt/β-catenin pathway in the EMT-induced DNA methylation variations in EOC cells. Consecutive experiments, including alterations in the Wnt/β-catenin pathway activity in EOC cells with a specific inhibitor and the identification of LY75-interacting partners by a proteomic approach, were strongly indicative for the direct implication of the LY75 receptor in modulating the Wnt/β-catenin signaling in EOC cells.


Vitamins and mast cells.
The immune system is a highly complex, intricately regulated group of cells whose integrated function is essential to health. The mast cell inflammatory response is characterized by an early phase with massive discharge of mediators stored in cytoplasmic secretory granules. Through multigranular/compound exocytosis and a late phase that involves generation of arachidonic acid metabolites and de novo synthesis of cytokines/chemokines and growth factors. Vitamins have been shown to have a protective effect on the body's immune cells. Vitamin C and E are necessary in allergic disease treatment where mast cells are involved. In addition, ascorbic acid and pyridoxine are useful compounds for the treatment of inflammatory disorder of the respiratory airways. Here we revisited the inter-relationship between vitamins and mast cells.


Spectroscopic properties of rubber oxygenase RoxA from Xanthomonas sp., a new type of dihaem dioxygenase.
Natural rubber [poly-(cis-1,4-isoprene)] is cleaved to 12-oxo-4,8-dimethyltrideca-4,8-diene-1-al (ODTD) by rubber oxygenase A (RoxA) isolated from Xanthomonas sp. RoxA has two c-type haem centres that show two distinct alpha-bands at 549 and 553 nm in the dithionite-reduced state. A well-resolved midpoint potential (E(0)') of -65 mV was determined for one haem by spectrophotometric titrations in the absence of dioxygen with dithionite and ferricyanide as reductant and oxidant, respectively. The midpoint potential of the second haem was not resolvable (E(0)' about -130 to -160 mV). One of the two haems was reduced by NADH (549 nm alpha-band), similar to bacterial dihaem peroxidases. Evidence for an electron transfer between the two haems was provided by slow reduction of the second haem (553 nm alpha-band) upon incubation of the partially reduced enzyme at room temperature. Addition of imidazole or related compounds to RoxA led to UV/vis spectral features similar to those observed for partially reduced RoxA. Notably, reduction of RoxA with dithionite or NADH, or binding of compounds such as imidazole, resulted in a reversible inactivation of the enzyme, unlike dihaem peroxidases. In line with this result, RoxA did not show any peroxidase activity. EPR spectra of RoxA as isolated showed two low-spin Fe(III) haem centres, with apparent g-values of 3.39, 3.09, 2.23, 1.92 and 1.50. A weak signal in the g=6 region resulting from a high-spin Fe(III) haem was also observed with a preparation-dependent intensity that disappeared in the presence of imidazole. Attempts to provide spectroscopic evidence for binding of the natural substrate (polyisoprene latex) to RoxA failed. However, experimental data are presented that RoxA is able to subtract redox equivalents from its substrate or from model compounds. In conclusion, RoxA is a novel type of dihaem dioxygenase with features clearly different from classical cytochrome c peroxidases.


Whole-body MRI of patients with polymyalgia rheumatica identifies a distinct subset with complete patient-reported response to glucocorticoids.
To determine whether whole-body MRI defines clinically relevant subgroups within polymyalgia rheumatica (PMR) including glucocorticoid responsiveness. 22 patients with PMR and 16 with rheumatoid arthritis (RA), untreated and diagnosed by consultant rheumatologists, underwent whole-body, multiple-joint MRI, scored by two experts. Patients with PMR reported whether they felt 'back to normal' on glucocorticoid therapy and were followed for a median of 2 years. All patients with PMR were deemed to respond to glucocorticoids clinically. A characteristic pattern of symmetrical, extracapsular inflammation, adjacent to greater trochanter, acetabulum, ischial tuberosity and/or symphysis pubis, was observed in 14/22 of the PMR cases. In PMR, this pattern was associated with complete glucocorticoid response (p=0.01), higher pretreatment C-reactive protein (CRP) and serum interleukin-6 (IL-6), and better post-treatment fatigue and function. Only 1/14 in the extracapsular group could stop glucocorticoids within 1 year, compared with 4/7 of the others. A score derived from the five sites discriminating best between PMR and RA correlated with IL-6 (p<0.002). IL-6 levels ≥16.8 pg/mL had 86% sensitivity and 86% specificity for the extracapsular MRI pattern. A subset of patients with rheumatologist-diagnosed PMR had a characteristic, extracapsular pattern of MRI inflammation, associated with elevated IL-6/CRP and with complete patient-reported glucocorticoid responsiveness.


New functions for DNA binding domains.
How the same transcription factor can be both an activator and a repressor of transcription is a biological mystery. Rao describes work that shows that the structure of the transcription factor may actually change depending on the DNA sequence bound by the factor. This change in structure may influence the binding of activators or repressors of transcription, thus allowing a single transcription factor to have two different activities.


Can safety assurance procedures in the food industry be used to evaluate a medical screening programme? The application of the Hazard Analysis and Critical Control Point system to an antenatal serum screening programme for Down's syndrome. Stage 2: overcoming the hazards in programme delivery.
An adaptation of the first stage of the systematic qualitative tool, the Hazard Analysis and Critical Control Point (HACCP) system from the food industry, was used to identify problems in the delivery of a district-wide antenatal screening programme for Down's syndrome. The aim of the second stage, described here, was to develop specific solutions (termed controls) for these problems. The views of staff delivering the programme and of members of the Down's screening steering group were used to identify control measures. The steering group members were also asked to identify which controls were critical to a risk management strategy, i.e. whether a control was essential to eliminate the hazard or if other control measures operating further down-line would be sufficient. The control measures were piloted both to determine if they were operationally feasible and whether they were practical in the context of staff workload and attitude. Two main control measures were the development of a rolling education programme for staff and of a written protocol with a checklist to verify stepwise delivery of the Down's screening programme. The checklist was designed to be included in the notes of every pregnant woman to enable audit of the offer of screening and delivery of each step of the screening programme. The control measures were all tailored specifically to overcome the problems reported by staff during the first stage of the work, the hazard analysis. The HACCP system proved useful in confirming the existence of problems in delivery of a Down's serum screening programme and in determining solutions tailored to overcome those problems. Future work is required to monitor and evaluate the usefulness of the control measures implemented.


Reversal of neuropathy phenotypes in conditional mouse model of Charcot-Marie-Tooth disease type 2E.
Mutations in the gene encoding for the neurofilament light subunit (NF-L) are responsible for Charcot-Marie-Tooth (CMT) neuropathy type 2E. To address whether CMT2E disease is potentially reversible, we generated a mouse model with conditional doxycycline-responsive gene system that allows repression of mutant hNF-LP22S transgene expression in adult neurons. The hNF-LP22S;tTa transgenic (tg) mice recapitulated key features of CMT2E disease, including aberrant hindlimb posture, motor deficits, hypertrophy of muscle fibres and loss of muscle innervation without neuronal loss. Remarkably, a 3-month treatment of hNF-LP22S;tTa mice with doxycycline after onset of disease efficiently down-regulated expression of hNF-LP22S and it caused reversal of CMT neurological phenotypes with restoration of muscle innervation and of neurofilament protein distribution along the sciatic nerve. These data suggest that therapeutic approaches aimed at abolishing expression or neutralizing hNF-L mutants might not only halt the progress of CMT2E disease, but also revert the disabilities.


Associations of foot and ankle characteristics with knee symptoms and function in individuals with patellofemoral osteoarthritis.
Foot and ankle characteristics are associated with patellofemoral pain (PFP) and may also relate to patellofemoral osteoarthritis (PFOA). A greater understanding of these characteristics and PFOA, could help to identify effective targeted treatments. To determine whether foot and ankle characteristics are associated with knee symptoms and function in individuals with PFOA. For this cross-sectional study we measured weightbearing ankle dorsiflexion range of motion, foot posture (via the Foot Posture Index [FPI]), and midfoot mobility (via the Foot Measurement Platform), and obtained patient-reported outcomes for knee symptoms and function (100 mm visual analogue scales, Anterior Knee Pain Scale [AKPS], Knee injury and Osteoarthritis Outcome Score, repeated single step-ups and double-leg sit-to-stand to knee pain onset). Pearson's r with significance set at p < 0.05 was used to determine the association between foot and ankle charateristics, with knee symptoms and function, adjusting for age. 188 participants (126 [67%] women, mean [SD] age of 59.9 [7.1] years, BMI 29.3 [5.6] kg/m2) with symptomatic PFOA were included in this study. Lower weightbearing ankle dorsiflexion range of motion had a small significant association with higher average knee pain (partial r = - 0.272, p < 0.001) and maximum knee pain during stair ambulation (partial r = - 0.164, p = 0.028), and lower scores on the AKPS (indicative of greater disability; partial r = 0.151, p = 0.042). Higher FPI scores (indicating a more pronated foot posture) and greater midfoot mobility (foot mobility magnitude) were significantly associated with fewer repeated single step-ups (partial r = - 0.181, p = 0.023 and partial r = - 0.197, p = 0.009, respectively) and double-leg sit-to-stands (partial r = - 0.202, p = 0.022 and partial r = - 0.169, p = 0.045, respectively) to knee pain onset, although the magnitude of these relationships was small. The amount of variance in knee pain and disability explained by the foot and ankle characteristics was small (R2-squared 2 to 8%). Lower weightbearing ankle dorsiflexion range of motion, a more pronated foot posture, and greater midfoot mobility demonstrated small associations with worse knee pain and greater disability in individuals with PFOA. Given the small magnitude of these relationships, it is unlikely that interventions aimed solely at addressing foot and ankle mobility will have substantial effects on knee symptoms and function in this population. The RCT was prospectively registered on 15 March 2017 with the Australia and New Zealand Clinical Trials Registry ( ANZCTRN12617000385347 ).


Does availability of informal care within the household impact hospitalisation?
This study assesses the effect of having informal support available at home on inpatient care use in Switzerland. The main contributions are to consider the availability of care regardless of its source, measured by multiple-adult living arrangements, and to examine this effect by type of inpatient care and source of potential support. A two-part model with region and time fixed effects is estimated to determine the impact of informal care availability on the likelihood of hospitalisation and length of stay, conditional on hospitalisation. The analysis is conducted on a sample of individuals aged 18+ from four waves of the Swiss Household Panel survey (2004-2007). Overall, availability of informal care has no impact on the likelihood of hospitalisation but does significantly reduce length of stay by 1.9 days. Available support has no effect on the shortest stays (up to 10 days), but has a significant impact on acute care stays up to 30 days and longer stays. Additionally, the effect does not significantly vary whether the source of informal support is a spouse only, a spouse and other adults, or other adults only. These results indicate that social changes leading to an expansion in the proportion of one-person households may increase future inpatient care use.


Drug-gene interactions between genetic polymorphisms and antihypertensive therapy.
Genetic factors may influence the response to antihypertensive medication. A number of studies have investigated genetic polymorphisms as determinants of cardiovascular response to antihypertensive drug therapy. In most candidate gene studies, no such drug-gene interactions were found. However, there is observational evidence that hypertensive patients with the 460 W allele of the alpha-adducin gene have a lower risk of myocardial infarction and stroke when treated with diuretics compared with other antihypertensive therapies. With regard to blood pressure response, interactions were found between genetic polymorphisms for endothelial nitric oxide synthase and diuretics, the alpha-adducin gene and diuretics, the alpha-subunit of G protein and beta-adrenoceptor antagonists, and the ACE gene and angiotensin II type 1 (AT(1)) receptor antagonists. Other studies found an interaction between ACE inhibitors and the ACE insertion/deletion (I/D) polymorphism, which resulted in differences in AT(1) receptor mRNA expression, left ventricular hypertrophy and arterial stiffness between different genetic variants. Also, drug-gene interactions between calcium channel antagonists and ACE I/D polymorphism regarding arterial stiffness have been reported. Unfortunately, the quality of these studies is quite variable. Given the methodological problems, the results from the candidate gene studies are still inconclusive and further research is necessary.


Autoantibodies against the fibrinolytic receptor, annexin 2, in antiphospholipid syndrome.
The association of thrombosis and gestational morbidity with antiphospholipid antibodies is termed antiphospholipid syndrome (APS). Annexin 2 (A2) is a profibrinolytic endothelial cell surface receptor that binds plasminogen, its tissue activator (tPA), and beta(2)-glycoprotein I (beta2GPI), the main antigen for antiphospholipid antibodies. Here, we evaluate A2 as a target antigen in APS. Serum samples from 434 individuals (206 patients with systemic lupus erythematosus without thrombosis, 62 with APS, 21 with nonautoimmune thrombosis, and 145 healthy individuals) were analyzed by enzyme-linked immunosorbent assay (ELISA) and immunoblot for antiphospholipid and A2 antibodies. Anti-A2 antibodies (titer > 3 SDs) were significantly more prevalent in patients with APS (22.6%; venous, 17.5%; arterial, 34.3%; and mixed thrombosis, 40.4%) than in healthy individuals (2.1%, P < .001), patients with nonautoimmune thrombosis (0%, P = .017), or patients with lupus without thrombosis (6.3%, P < .001). Anti-A2 IgG enhanced the expression of tissue factor on endothelial cells (6.4-fold +/- 0.13-fold SE), blocked A2-supported plasmin generation in a tPA-dependent generation assay (19%-71%) independently of beta2GPI, and inhibited cell surface plasmin generation on human umbilical vein endothelial cells (HUVECs) by 34% to 83%. We propose that anti-A2 antibodies contribute to the prothrombotic diathesis in antiphospholipid syndrome.


Current status and future perspective of laparoscopic surgery in hepatobiliary disease.
Recent advances in minimally invasive surgery include laparoscopic and robotic surgery. These surgical techniques have changed the paradigm of surgical treatment for hepatobiliary diseases. Minimally invasive surgery has the advantages of minimal wound extension for cosmetic effect, early postoperative recovery, and few postoperative complications in patients. For laparoscopic liver resection, the indications have been expanded and oncological outcome was proven to be similar with open surgery in the malignant disease. Laparoscopic cholecystectomy is a classical operation for benign gallbladder diseases and the effort to decrease the surgical wound resulted to perform single incision laparoscopic cholecystectomy. For choledochal cyst, laparoscopic surgery is applied gradually despite of the difficulties associated with anastomosis, and robotic surgery for hepatobiliary disease is also performed for more minimally invasive surgery; however, while admitting the advantage of robotic surgery, robotic technology should be improved for development of more convenient and cheaper instrument and continuous efforts to enhance surgical technique to overcome long operation is necessary. In this review, the status and future perspectives of minimally invasive surgery for hepatobiliary diseases are summarized and discussed.


In vivo cardiac reprogramming contributes to zebrafish heart regeneration.
Despite current treatment regimens, heart failure remains the leading cause of morbidity and mortality in the developed world due to the limited capacity of adult mammalian ventricular cardiomyocytes to divide and replace ventricular myocardium lost from ischaemia-induced infarct. Hence there is great interest to identify potential cellular sources and strategies to generate new ventricular myocardium. Past studies have shown that fish and amphibians and early postnatal mammalian ventricular cardiomyocytes can proliferate to help regenerate injured ventricles; however, recent studies have suggested that additional endogenous cellular sources may contribute to this overall ventricular regeneration. Here we have developed, in the zebrafish (Danio rerio), a combination of fluorescent reporter transgenes, genetic fate-mapping strategies and a ventricle-specific genetic ablation system to discover that differentiated atrial cardiomyocytes can transdifferentiate into ventricular cardiomyocytes to contribute to zebrafish cardiac ventricular regeneration. Using in vivo time-lapse and confocal imaging, we monitored the dynamic cellular events during atrial-to-ventricular cardiomyocyte transdifferentiation to define intermediate cardiac reprogramming stages. We observed that Notch signalling becomes activated in the atrial endocardium following ventricular ablation, and discovered that inhibiting Notch signalling blocked the atrial-to-ventricular transdifferentiation and cardiac regeneration. Overall, these studies not only provide evidence for the plasticity of cardiac lineages during myocardial injury, but more importantly reveal an abundant new potential cardiac resident cellular source for cardiac ventricular regeneration.


White matter alterations to cingulum and fornix following very preterm birth and their relationship with cognitive functions.
Very preterm birth (VPT; <32 weeks of gestation) has been associated with impairments in memory abilities and functional neuroanatomical brain alterations in medial temporal and fronto-parietal areas. Here we investigated the relationship between structural connectivity in memory-related tracts and various aspects of memory in VPT adults (mean age 19) who sustained differing degrees of perinatal brain injury (PBI), as assessed by neonatal cerebral ultrasound. We showed that the neurodevelopmental consequences of VPT birth persist into young adulthood and are associated with neonatal cranial ultrasound classification. At a cognitive level, VPT young adults showed impairments specific to effective organization of verbal information and visuospatial memory, whereas at an anatomical level they displayed reduced volume of memory-related tracts, the cingulum and the fornix, with greater alterations in those individuals who experienced high-grade PBI. When investigating the association between these tracts and memory scores, perseveration errors were associated with the volume of the fornix and dorsal cingulum (connecting medial frontal and parietal lobes). Visuospatial memory scores were associated with the volume of the ventral cingulum (connecting medial parietal and temporal lobes). These results suggest that structural connectivity alterations could underlie memory difficulties in preterm born individuals.


Dissociative effects of methylphenidate in nonhuman primates: trade-offs between cognitive and behavioral performance.
Low doses of methylphenidate reduce hyperactivity and improve attention in individuals with attention deficit hyperactivity disorder (ADHD) as well as in healthy humans and animals. Despite its extensive use, relatively little is known about its mechanisms of action. This study investigated the effects of methylphenidate on working memory performance, impulsivity, response accuracy and precision, and the ability to stay on task in rhesus monkeys using an oculomotor delayed response task. Methylphenidate affected task performance in an inverted-U manner in all three subjects tested. The improvements resulted from a reduction in premature responses and, importantly, not from improvement in the memory of target location. The length of time subjects participated in each session was also affected dose dependently. However, the dose at which the length of participation was maximally increased significantly impaired performance on the working memory task. This dissociation of effects has implications for the treatment of ADHD, for the nonprescription use of methylphenidate for cognitive enhancement, and for furthering the basic understanding of the neural substrate underlying these processes.


Time-location analysis for exposure assessment studies of children using a novel global positioning system instrument.
Global positioning system (GPS) technology is used widely for business and leisure activities and offers promise for human time-location studies to evaluate potential exposure to environmental contaminants. In this article we describe the development of a novel GPS instrument suitable for tracking the movements of young children. Eleven children in the Seattle area (2-8 years old) wore custom-designed data-logging GPS units integrated into clothing. Location data were transferred into geographic information systems software for map overlay, visualization, and tabular analysis. Data were grouped into five location categories (in vehicle, inside house, inside school, inside business, and outside) to determine time spent and percentage reception in each location. Additional experiments focused on spatial resolution, reception efficiency in typical environments, and sources of signal interference. Significant signal interference occurred only inside concrete/steel-frame buildings and inside a power substation. The GPS instruments provided adequate spatial resolution (typically about 2-3 m outdoors and 4-5 m indoors) to locate subjects within distinct microenvironments and distinguish a variety of human activities. Reception experiments showed that location could be tracked outside, proximal to buildings, and inside some buildings. Specific location information could identify movement in a single room inside a home, on a playground, or along a fence line. The instrument, worn in a vest or in bib overalls, was accepted by children and parents. Durability of the wiring was improved early in the study to correct breakage problems. The use of GPS technology offers a new level of accuracy for direct quantification of time-location activity patterns in exposure assessment studies.


Serum selenium status in Graves' disease with and without orbitopathy: a case-control study.
Selenium is effective in improving quality of life and reducing the progression of active Graves' orbitopathy. The effect of correcting relative selenium deficiency on improving Graves' orbitopathy is unknown, as baseline selenium levels have not previously been measured. The study aims to determine whether serum selenium levels are reduced in patients with Graves' disease with orbitopathy (GO) compared with without orbitopathy (GD). A prospective, case-control study performed between 2009 and 2012 at endocrine and ophthalmology clinics in Australia. A total of 198 patients with Graves' disease participated in the study: 101 with Graves' orbitopathy and 97 without Graves' orbitopathy. Serum selenium levels in both groups. Mean serum selenium levels were significantly lower in GO (1·10 ± 0·18 μm) than in GD (1·19 ± 0·20 μm) (P = 0·001). Mean selenium levels appeared to decrease in parallel with increasing severity of GO; selenium level was 1·19 ± 0·20 μm in GD, 1·10 ± 0·19 μm in moderate-to-severe GO and 1·09 ± 0·17 μm in sight-threatening GO (P = 0·003). Serum selenium levels remained significantly lower in GO after adjusting for age, smoking status, thyroidectomy, radioactive iodine treatment and residential location. Serum selenium levels are lower in patients with GO compared with GD in an Australian study population with marginal selenium status. Relative selenium deficiency may be an independent risk factor for orbitopathy in patients with Graves' disease.


Duplications and losses in gene families of rust pathogens highlight putative effectors.
Rust fungi are a group of fungal pathogens that cause some of the world's most destructive diseases of trees and crops. A shared characteristic among rust fungi is obligate biotrophy, the inability to complete a lifecycle without a host. This dependence on a host species likely affects patterns of gene expansion, contraction, and innovation within rust pathogen genomes. The establishment of disease by biotrophic pathogens is reliant upon effector proteins that are encoded in the fungal genome and secreted from the pathogen into the host's cell apoplast or within the cells. This study uses a comparative genomic approach to elucidate putative effectors and determine their evolutionary histories. We used OrthoMCL to identify nearly 20,000 gene families in proteomes of 16 diverse fungal species, which include 15 basidiomycetes and one ascomycete. We inferred patterns of duplication and loss for each gene family and identified families with distinctive patterns of expansion/contraction associated with the evolution of rust fungal genomes. To recognize potential contributors for the unique features of rust pathogens, we identified families harboring secreted proteins that: (i) arose or expanded in rust pathogens relative to other fungi, or (ii) contracted or were lost in rust fungal genomes. While the origin of rust fungi appears to be associated with considerable gene loss, there are many gene duplications associated with each sampled rust fungal genome. We also highlight two putative effector gene families that have expanded in Cqf that we hypothesize have roles in pathogenicity.


Left ventricular remodelling and improvement in Freestyle stentless valve haemodynamics.
To assess how left ventricular (LV) hypertrophy, geometry and function change after stentless aortic valve replacement for aortic stenosis, and to elucidate the physiological mechanism of the improvement in stentless valve haemodynamics. 81 patients with aortic stenosis (age 75 +/- 6 years, 47 male) underwent aortic valve replacement (plus CABG in 33 patients) with a Freestyle stentless porcine valve (mean size 23 +/- 2 mm). They were prospectively investigated by Doppler echocardiography at 2 weeks, 3-6, 12, and 24 months after operation. Two hundred and forty-six echocardiograms were obtained and analysed. Aortic valve performance was assessed from its effective orifice area (EOA), the transvalvular increase in mean flow velocity (delta mV), the deceleration time of aortic flow velocity, and mean pressure drop (mPG). LV hypertrophy was assessed from LV mass index; LV geometry, from the ratio of wall thickness to the radius (T/R ratio) and LV function, from stroke volume index (LVSVI) and myocardial stroke work (SW). By 2 years after operation, LV mass index had fallen from 162 +/- 64 to 109 +/- 36, g/m2, and T/R ratio from 0.61 +/- 0.25 to 0.43 +/- 0.10. LVSVI increased from 29.4 +/- 10 to 42 +/- 17, ml/m2, and myocardial SW from 3.1 +/- 1.6 to 5.2 +/- 2.2, mJ/cm3 (all P < 0.001 by ANOVA), while LV outflow tract diameter remained unchanged. At the same time, stentless valve EOA increased from 1.59 +/- 0.75 to 2.2 +/- 0.72, cm2, and delta mV (from 82 +/- 31 to 49 +/- 24, cm/s) and mPG (from 9.7 +/- 5.0 to 5.2 +/- 3.7 mmHg) both fell significantly (all P < 0.001 by ANOVA): as the deceleration time of aortic flow velocity increased from 153.6 +/- 64.1 to 202.7 +/- 37.6 ms (P < 0.001 by ANOVA). After stentless aortic valve replacement, LV mass index and wall thickness both fall towards normal, and myocardial stroke work increases. These ventricular remodelling processes are accompanied by a more physiological flow jet at valve cusp level, which permits a greater stroke volume to be ejected with a smaller transvavular velocity increase, so that effective orifice area increases.


[Hippocrates commentaries in modern age].
The Hippocratic commentaries represent a genre utilised not only by university teachers and so-called "medical philologists", but also by working doctors, personal physicians, and other medical practitioners. As such, the genre of the commentary is located at a crucial point of intersection between medical history, theory, and practice, as well as between orthodox and "alternative" conceptions of medicine; the commentaries are thus ideally placed to reflect the diversity of early modern medicine as well as the larger cultural context in which it was practised and debated. In this paper, some general remarks on researching Hippocrates commentaries are presented and followed by two test cases: the commentaries on the Hippocratic Letters and on the Hippocratic Oath.


N-Labelled proteins by cell-free protein synthesis. Strategies for high-throughput NMR studies of proteins and protein-ligand complexes.
[(15)N]-heteronuclear single quantum coherence (HSQC) spectra provide a readily accessible fingerprint of [(15)N]-labelled proteins, where the backbone amide group of each nonproline amino acid residue contributes a single cross-peak. Cell-free protein synthesis offers a fast and economical route to enhance the information content of [(15)N]-HSQC spectra by amino acid type selective [(15)N]-labelling. The samples can be measured without chromatographic protein purification, dilution of isotopes by transaminase activities are suppressed, and a combinatorial isotope labelling scheme can be adopted that combines reduced spectral overlap with a minimum number of samples for the identification of all [(15)N]-HSQC cross-peaks by amino acid residue type. These techniques are particularly powerful for tracking [(15)N]-HSQC cross-peaks after titration with unlabelled ligand molecules or macromolecular binding partners. In particular, combinatorial isotope labelling can provide complete cross-peak identification by amino acid type in 24 h, including protein production and NMR measurement.


HIV drug resistance testing by high-multiplex "wide" sequencing on the MiSeq instrument.
Limited access to HIV drug resistance testing in low- and middle-income countries impedes clinical decision-making at the individual patient level. An efficient protocol to address this issue must be established to minimize negative therapeutic outcomes for HIV-1-infected individuals in such settings. This is an observational study to ascertain the potential of newer genomic sequencing platforms, such as the Illumina MiSeq instrument, to provide accurate HIV drug resistance genotypes for hundreds of samples simultaneously. Plasma samples were collected from Canadian patients during routine drug resistance testing (n = 759) and from a Ugandan study cohort (n = 349). Amplicons spanning HIV reverse transcriptase codons 90 to 234 were sequenced with both MiSeq sequencing and conventional Sanger sequencing methods. Sequences were evaluated for nucleotide concordance between methods, using coverage and mixture parameters for quality control. Consensus sequences were also analyzed for disparities in the identification of drug resistance mutations. Sanger and MiSeq sequencing was successful for 881 samples (80%) and 892 samples (81%), respectively, with 832 samples having results from both methods. Most failures were for samples with viral loads of <3.0 log10 HIV RNA copies/ml. Overall, 99.3% nucleotide concordance between methods was observed. MiSeq sequencing achieved 97.4% sensitivity and 99.3% specificity in detecting resistance mutations identified by Sanger sequencing. Findings suggest that the Illumina MiSeq platform can yield high-quality data with a high-multiplex "wide" sequencing approach. This strategy can be used for multiple HIV subtypes, demonstrating the potential for widespread individual testing and annual population surveillance in resource-limited settings.