Secondary structure-induced aggregation by hydrogen peroxide: a stimuli-triggered open/close implementation by recombination.
The fabrication of reactive aggregation nanomaterials through assemblies in a facile and cost-effective manner is much desired but remains to be well explored. Here we show that exquisite and ultra-long (>2 μm) hybrid polymer nanorods (NRs) can be formed by a simple self-assembly of a phenylboronic acid modified genistein crosslinker (Ge-di(HMPBA-pin)) and d-α-tocopheryl polyethylene glycol 1000 (TPGS). The obtained NRs exhibit quantitative and sensitive colorimetric detection of H2O2 with a remarkable detection limit for different stromal materials. More significantly, the presence of H2O2 triggers a distinct morphological transformation of the polymer NR assembly into the secondary structure of micelles via the oxidative deboronation of boronate moieties in HMPBA-pin-SA. It spontaneously induces the aggregation of metal nanoparticles (Au NPs), metal nanorods (Au NRs), quantum dots (MoS2 QDs), metal ions (Cu2+), protein (ferritin) and tetraphenylethene (TPE) molecules, giving rise to a dramatic stimuli-triggered open/close switchable complexation and apparent colorimetric transitions in vitro. This study, for the first time, showcases the fascinating advantages of such unprecedented secondary structure-induced aggregation and uncovers the immense potential to design a plethora of other sensing systems by virtue of the alternate trigger-specific, sacrifice-aggregated building moieties.


The role of litigation in tobacco control.
There is now 50-years of experience in the United States litigating against the tobacco industry. As the base of evidence regarding health effects has evolved and new legal strategies have emerged, successive waves of litigation have occurred. The many failures by the first and second waves were followed by some notable successes in the third. Litigation by flight attendants and the states led to substantial settlements and some beneficial consequences for tobacco control. One of the most significant consequences of state litigation was access to the industry's documents, gained through the Minnesota settlement. These documents further empowered the tobacco control movement and strengthened the basis for legal action. The continuing litigation in the United States remains a threat to the industry, in spite of the mixed outcomes of recent cases.


A homozygous pathogenic missense variant broadens the phenotypic and mutational spectrum of CREB3L1-related osteogenesis imperfecta.
The cyclic adenosine monophosphate responsive element binding protein 3-like 1 (CREB3L1) gene codes for the endoplasmic reticulum stress transducer old astrocyte specifically induced substance (OASIS), which has an important role in osteoblast differentiation during bone development. Deficiency of OASIS is linked to a severe form of autosomal recessive osteogenesis imperfecta (OI), but only few patients have been reported. We identified the first homozygous pathogenic missense variant [p.(Ala304Val)] in a patient with lethal OI, which is located within the highly conserved basic leucine zipper domain, four amino acids upstream of the DNA binding domain. In vitro structural modeling and luciferase assays demonstrate that this missense variant affects a critical residue in this functional domain, thereby decreasing the type I collagen transcriptional binding ability. In addition, overexpression of the mutant OASIS protein leads to decreased transcription of the SEC23A and SEC24D genes, which code for components of the coat protein complex type II (COPII), and aberrant OASIS signaling also results in decreased protein levels of SEC24D. Our findings therefore provide additional proof of the potential involvement of the COPII secretory complex in the context of bone-associated disease.


The Arabidopsis IRX10 and IRX10-LIKE glycosyltransferases are critical for glucuronoxylan biosynthesis during secondary cell wall formation.
Arabidopsis IRX10 and IRX10-LIKE (IRX10-L) proteins are closely related members of the GT47 glycosyltransferase family. Single gene knock-outs of IRX10 or IRX10-L result in plants with either a weak or no mutant phenotype. However irx10 irx10-L double mutants are severely affected in their development, with a reduced rosette size and infrequent formation of a small infertile inflorescence. Plants homozygous for irx10 and heterozygous for irx10-L have an intermediate phenotype exhibiting a short inflorescence compared with the wild type, and an almost complete loss of fertility. Stem sections of the irx10 homozygous irx10-L heterozygous or irx10 irx10-L double mutants show decreased secondary cell-wall formation. NMR analysis shows that signals derived from the reducing end structure of glucuronoxylan were detected in the irx10 single mutant, and in the irx10 homozygous irx10-L heterozygous combination, but that the degree of polymerization of the xylan backbone was reduced compared with the wild type. Additionally, xylans from irx10 stem tissues have an almost complete loss of the GlcUA side chain, whereas the level of 4-O-Me-GlcUA was similar to that in wild type. Deletion of the predicted signal peptide from the N terminus of IRX10 or IRX10-L results in an inability to rescue the irx10 irx10-L double mutant phenotype. These findings demonstrate that IRX10 and IRX10-L perform a critical function in the synthesis of glucuronoxylan during secondary cell-wall formation, and that this activity is associated with the formation of the xylan backbone structure. This contrasts with the proposed function of the tobacco NpGUT1, which is closely related to the Arabidopsis IRX10 and IRX10-L proteins, in rhamnogalacturonan II biosynthesis.


Design and Analysis of a Single-Camera Omnistereo Sensor for Quadrotor Micro Aerial Vehicles (MAVs).
We describe the design and 3D sensing performance of an omnidirectional stereo (omnistereo) vision system applied to Micro Aerial Vehicles (MAVs). The proposed omnistereo sensor employs a monocular camera that is co-axially aligned with a pair of hyperboloidal mirrors (a vertically-folded catadioptric configuration). We show that this arrangement provides a compact solution for omnidirectional 3D perception while mounted on top of propeller-based MAVs (not capable of large payloads). The theoretical single viewpoint (SVP) constraint helps us derive analytical solutions for the sensor's projective geometry and generate SVP-compliant panoramic images to compute 3D information from stereo correspondences (in a truly synchronous fashion). We perform an extensive analysis on various system characteristics such as its size, catadioptric spatial resolution, field-of-view. In addition, we pose a probabilistic model for the uncertainty estimation of 3D information from triangulation of back-projected rays. We validate the projection error of the design using both synthetic and real-life images against ground-truth data. Qualitatively, we show 3D point clouds (dense and sparse) resulting out of a single image captured from a real-life experiment. We expect the reproducibility of our sensor as its model parameters can be optimized to satisfy other catadioptric-based omnistereo vision under different circumstances.


Monte Carlo simulations of weak polyelectrolyte microgels: pH-dependence of conformation and ionization.
In this study, we investigated the effect of pH on single weak acidic polyelectrolyte microgels under salt-free conditions with (i) varying microgel concentration, (ii) varying content of acidic groups and (iii) different crosslinking densities using Monte Carlo simulations under explicit consideration of the protonation/deprotonation reaction. We assessed both global properties, such as the degree of ionization, the degree of swelling and the counterion distribution, and local properties such as the radial network ionization profile and the ionization along the polymer chains as a function of pH. We found a pronounced suppression of the pH-dependent ionization of the microgels, as compared to the ideal titration behavior and a shift of the titration curve to a higher pH originating in the proximity of acidic groups in the microgel. In contrast to macroscopic gels, counterions can leave the microgel, resulting in an effective charge of the network, which hinders the ionization. A decreasing microgel concentration leads to an increased effective charge of the microgel and a more pronounced shift of the titration curve. The number of acidic groups showed only a weak effect on the ionization behavior of the microgels. For two different microgels with different crosslinking densities, similar scaling of the gel size was observed. A distinct transition from an uncharged and unswollen to a highly charged and expanded polymer network was observed for all investigated microgels. The degree of swelling mainly depends on the degree of ionization. An inhomogeneous distribution of the degree of ionization along the radial profile of the microgel was found.


Temporal phase-unwrapping algorithm for dynamic interference pattern analysis in interference-contrast microscopy.
A temporal phase-unwrapping algorithm has been developed for the analysis of dynamic interference patterns generated with interference-contrast microscopy in micromachined picoliter vials. These vials are etched in silicon dioxide, have a typical depth of 6 mum, and are filled with a liquid sample. In this kind of microscopy, fringe patterns are observed at the air-liquid interface. These fringe patterns are caused by interference between the directly reflected part of an incident plane wave and the part of that wave that is reflected on the bottom of the vial. The optical path difference (OPD) between both parts is proportional to the distance to the reflecting bottom of the vial. Evaporation decreases the OPD at the meniscus level and causes alternating constructive and destructive interference of the incident light, resulting in an interferogram. Imaging of the space-varying OPD yields a fringe pattern in which the isophotes correspond to isoheight curves of the meniscus. When the bottom is flat, the interference pattern allows for monitoring of the meniscus as a function of time during evaporation. However, when there are objects on the bottom of the vial, the heights of these objects are observed as phase jumps in the fringes proportional to their heights. First, we present a classical electromagnetic description of interference-contrast microscopy. Second, a temporal phase-unwrapping algorithm is described that retrieves the meniscus profile from the interference pattern. Finally, this algorithm is applied to measure height differences of objects on the bottom in other micromachined vials with a precision of ~5 nm.


Predicting cancer drug mechanisms of action using molecular network signatures.
Molecular signatures are a powerful approach to characterize novel small molecules and derivatized small molecule libraries. While new experimental techniques are being developed in diverse model systems, informatics approaches lag behind these exciting advances. We propose an analysis pipeline for signature based drug annotation. We develop an integrated strategy, utilizing supervised and unsupervised learning methodologies that are bridged by network based statistics. Using this approach we can: 1, predict new examples of drug mechanisms that we trained our model upon; 2, identify "New" mechanisms of action that do not belong to drug categories that our model was trained upon; and 3, update our training sets with these "New" mechanisms and accurately predict entirely distinct examples from these new categories. Thus, not only does our strategy provide statistical generalization but it also offers biological generalization. Additionally, we show that our approach is applicable to diverse types of data, and that distinct biological mechanisms characterize its resolution of categories across different data types. As particular examples, we find that our predictive resolution of drug mechanisms from mRNA expression studies relies upon the analog measurement of a cell stress-related transcriptional rheostat along with a transcriptional representation of cell cycle state; whereas, in contrast, drug mechanism resolution from functional RNAi studies rely upon more dichotomous (e.g., either enhances or inhibits) association with cell death states. We believe that our approach can facilitate molecular signature-based drug mechanism understanding from different technology platforms and across diverse biological phenomena.


[Crossed graft of embryonic testis between quail and chick embryos: contribution to the study of the mechanism of female bird embryo masculinization].
Quail, or chick embryonic testes grafted respectively in the extraembryonic coelom of chick or quail embryos induce both a Mullerian duct regression and a masculinization of the female host gonads up to the differentiation of two testes, in some cases. Such a result confirms the fact evidenced previously in other bird species (chick and duck) that the testis-inducer is interspecific. Quail cells are not observed in histological sections of embryonic gonads of testis-grafted chicks. This allows to discard a possible influence of cells migrating from the graft to the host. The grafted testes act then through substance/s secreted in the blood stream. Present and previous experimental data strongly suggest that the same substance, i.e. the so-called anti-Mullerian hormone, is responsible for both MD regression and gonadal sex reversal.


Presence of mother and unfamiliar female alters levels of testosterone, progesterone, cortisol, adrenocorticotropin, and behavior in maturing Guinea pigs.
Although the guinea pig is characterized by precocial physical development and minimal active maternal care, studies suggest the presence of the mother can influence neuroendocrine and behavioral activity of offspring even well beyond weaning. Previous results may have been influenced by the procedure of housing weaned subjects with the mother to within 2 days of testing. The present study examined approximately 40-day-old guinea pigs housed apart from the mother for 0 (not rehoused), 2, or 10 days. Rehousing without the mother led to elevations in plasma testosterone (measured in males), progesterone (measured in females), cortisol, and adrenocorticotropin (ACTH) (both measured in males and females). Offspring housed without the mother for 10 days had the highest progesterone, cortisol, and ACTH levels. Testosterone elevations were observed in 2-day-, but not 10-day-, rehoused animals. Regardless of rehousing condition, 60 min isolation in a novel test cage elevated progesterone, cortisol, and ACTH, and reduced testosterone. These effects were all moderated if the subject was tested with the mother or another female. Sexual behavior toward the mother was observed frequently, but only in males housed apart from her prior to testing. Overall, males and females that had been housed apart from the mother interacted with her as they would an unfamiliar female. Our results corroborate previous findings, suggest the effect of housing apart from the mother on male testosterone is transitory, and indicate that continuous housing with the mother past weaning suppresses circulating progesterone in females and cortisol and ACTH in both sexes.


Decoding post-stroke motor function from structural brain imaging.
Clinical research based on neuroimaging data has benefited from machine learning methods, which have the ability to provide individualized predictions and to account for the interaction among units of information in the brain. Application of machine learning in structural imaging to investigate diseases that involve brain injury presents an additional challenge, especially in conditions like stroke, due to the high variability across patients regarding characteristics of the lesions. Extracting data from anatomical images in a way that translates brain damage information into features to be used as input to learning algorithms is still an open question. One of the most common approaches to capture regional information from brain injury is to obtain the lesion load per region (i.e. the proportion of voxels in anatomical structures that are considered to be damaged). However, no systematic evaluation has yet been performed to compare this approach with using patterns of voxels (i.e. considering each voxel as a single feature). In this paper we compared both approaches applying Gaussian Process Regression to decode motor scores in 50 chronic stroke patients based solely on data derived from structural MRI. For both approaches we compared different ways to delimit anatomical areas: regions of interest from an anatomical atlas, the corticospinal tract, a mask obtained from fMRI analysis with a motor task in healthy controls and regions selected using lesion-symptom mapping. Our analysis showed that extracting features through patterns of voxels that represent lesion probability produced better results than quantifying the lesion load per region. In particular, from the different ways to delimit anatomical areas compared, the best performance was obtained with a combination of a range of cortical and subcortical motor areas as well as the corticospinal tract. These results will inform the appropriate methodology for predicting long term motor outcomes from early post-stroke structural brain imaging.


Liquid-crystal depolarizer consisting of randomly aligned hybrid orientation domains.
A novel depolarization method for linearly polarized incident light that uses a liquid-crystal (LC) cell with randomly aligned hybrid orientation domains is theoretically described by use of Mueller matrix calculations. The depolarization effect of the incident linear polarization is confirmed with Stokes parameter measurements. The unique optical properties of the fabricated LC depolarizer are revealed; that is, the intensity of the transmitted light is independent of the rotation of the analyzer. The degree of polarization becomes zero when the retardation of the LC depolarizer coincides with a half-wavelength.


Lactate is associated with mortality in very old intensive care patients suffering from COVID-19: results from an international observational study of 2860 patients.
Lactate is an established prognosticator in critical care. However, there still is insufficient evidence about its role in predicting outcome in COVID-19. This is of particular concern in older patients who have been mostly affected during the initial surge in 2020. This prospective international observation study (The COVIP study) recruited patients aged 70 years or older (ClinicalTrials.gov ID: NCT04321265) admitted to an intensive care unit (ICU) with COVID-19 disease from March 2020 to February 2021. In addition to serial lactate values (arterial blood gas analysis), we recorded several parameters, including SOFA score, ICU procedures, limitation of care, ICU- and 3-month mortality. A lactate concentration ≥ 2.0 mmol/L on the day of ICU admission (baseline) was defined as abnormal. The primary outcome was ICU-mortality. The secondary outcomes 30-day and 3-month mortality. In total, data from 2860 patients were analyzed. In most patients (68%), serum lactate was lower than 2 mmol/L. Elevated baseline serum lactate was associated with significantly higher ICU- and 3-month mortality (53% vs. 43%, and 71% vs. 57%, respectively, p < 0.001). In the multivariable analysis, the maximum lactate concentration on day 1 was independently associated with ICU mortality (aOR 1.06 95% CI 1.02-1.11; p = 0.007), 30-day mortality (aOR 1.07 95% CI 1.02-1.13; p = 0.005) and 3-month mortality (aOR 1.15 95% CI 1.08-1.24; p < 0.001) after adjustment for age, gender, SOFA score, and frailty. In 826 patients with baseline lactate ≥ 2 mmol/L sufficient data to calculate the difference between maximal levels on days 1 and 2 (∆ serum lactate) were available. A decreasing lactate concentration over time was inversely associated with ICU mortality after multivariate adjustment for SOFA score, age, Clinical Frailty Scale, and gender (aOR 0.60 95% CI 0.42-0.85; p = 0.004). In critically ill old intensive care patients suffering from COVID-19, lactate and its kinetics are valuable tools for outcome prediction. NCT04321265.


Impact of lifestyle in middle-aged women on mortality: evidence from the Royal College of General Practitioners' Oral Contraception Study.
Although many individuals have multiple lifestyle risk factors, few studies have investigated the impact of lifestyle risk factor combinations among women. To investigate the relationship between individual and combinations of lifestyle risk factors in middle-aged women with subsequent mortality, and to estimate the associated population attributable risks. Prospective cohort study. Royal College of General Practitioners' (RCGP) Oral Contraception Study, UK. In 1994-1995, women remaining under follow-up in the RCGP Oral Contraception Study were sent a lifestyle survey, from which modifiable risk factors were identified: pack-years smoked, physical inactivity, never drinking versus consuming at least 7 units of alcohol weekly, and being underweight, overweight, or obese. The cohort was followed to December 2006 or death. Population attributable risks were calculated. Of 10 059 women studied, 896 died. Pack-years smoked (11-20 years: adjusted hazard ratio [HR] = 1.82, 95% confidence interval [CI] = 1.46 to 2.27; >20 years: adjusted HR = 2.34, 95% CI = 2.00 to 2.74); never drinking alcohol (adjusted HR = 1.66, 95% CI = 1.34 to 2.05); being underweight (adjusted = HR 1.66, 95% CI = 1.03 to 2.68); and physical inactivity (<15 hours/week: adjusted HR = 1.73, 95% CI = 1.46 to 2.04) were significantly associated with mortality compared with their respective reference group. Women with multiple lifestyle risk factors had higher mortality risks than those reporting one factor. The population attributable risk of the combination of smoking, physical inactivity, body mass index outside normal range, and alcohol (never drinking or excess intake) was 59% (95% CI = 31% to 78%). Assuming a causal relationship between lifestyle and mortality, avoidance of four lifestyle risk factors would have prevented 60% of the deaths. The importance of avoiding smoking and undertaking physical inactivity during midlife should continue to be emphasised.


Rapid diversification of a species-rich genus of neotropical rain forest trees.
Species richness in the tropics has been attributed to the gradual accumulation of species over a long geological period in stable equatorial climates or, conversely, to speciation in response to late Tertiary geological events and unstable Pleistocene climates. DNA sequence data are consistent with recent diversification in Inga, a species-rich neotropical tree genus. We estimate that speciation was concentrated in the past 10 million years, with many species arising as recently as 2 million years ago. This coincides with the more recent major uplifts of the Andes, the bridging of the Isthmus of Panama, and Quaternary glacial cycles. Inga may be representative of other species-rich neotropical genera with rapid growth and reproduction, which contribute substantially to species numbers in the world's most diverse flora.


High-resolution cryo-electron microscopy structure of the Trypanosoma brucei ribosome.
Ribosomes, the protein factories of living cells, translate genetic information carried by messenger RNAs into proteins, and are thus involved in virtually all aspects of cellular development and maintenance. The few available structures of the eukaryotic ribosome reveal that it is more complex than its prokaryotic counterpart, owing mainly to the presence of eukaryote-specific ribosomal proteins and additional ribosomal RNA insertions, called expansion segments. The structures also differ among species, partly in the size and arrangement of these expansion segments. Such differences are extreme in kinetoplastids, unicellular eukaryotic parasites often infectious to humans. Here we present a high-resolution cryo-electron microscopy structure of the ribosome of Trypanosoma brucei, the parasite that is transmitted by the tsetse fly and that causes African sleeping sickness. The atomic model reveals the unique features of this ribosome, characterized mainly by the presence of unusually large expansion segments and ribosomal-protein extensions leading to the formation of four additional inter-subunit bridges. We also find additional rRNA insertions, including one large rRNA domain that is not found in other eukaryotes. Furthermore, the structure reveals the five cleavage sites of the kinetoplastid large ribosomal subunit (LSU) rRNA chain, which is known to be cleaved uniquely into six pieces, and suggests that the cleavage is important for the maintenance of the T. brucei ribosome in the observed structure. We discuss several possible implications of the large rRNA expansion segments for the translation-regulation process. The structure could serve as a basis for future experiments aimed at understanding the functional importance of these kinetoplastid-specific ribosomal features in protein-translation regulation, an essential step towards finding effective and safe kinetoplastid-specific drugs.


A crimp-like microarchitecture improves tissue production in fibrous ligament scaffolds in response to mechanical stimuli.
The aim of this study was to determine the influence of a crimp-like microarchitecture within electrospun polymer scaffolds on fibroblast extracellular matrix (ECM) production when cultured under dynamic conditions. Electrospun poly(L-lactide-co-D,L-lactide) scaffolds possessing a wave pattern similar to collagen crimp (amplitude: 5 μm and wavelength: 46 μm) were seeded with bovine fibroblasts and mechanically stimulated under dynamic uniaxial tension. The effect of strain amplitude (5%, 10% and 20%) was investigated in a short-term stimulation study. The 10% strain amplitude in the stimulated crimp-like fibre scaffold increased only collagen synthesis, while the 20% strain amplitude increased both collagen and sulphated proteoglycan synthesis compared to stimulated uncrimped (straight) fibre scaffolds and unloaded controls (crimp-like static fibre scaffolds). Alternatively, mechanical stimulation of fibroblasts seeded on uncrimped fibre scaffolds induced significant fibroblast proliferation compared to the stimulated crimp-like fibre scaffolds and no-load controls. Long-term, dynamic mechanical stimulation of fibroblasts seeded on crimp-like fibre scaffolds at 10% strain amplitude resulted in significantly up-regulated collagen accumulation and down-regulated sulphated proteoglycan accumulation. Additionally, the fibroblasts seeded on dynamically stimulated crimp-like fibre scaffolds appeared to form bundles that resembled fascicles, a characteristic hierarchical feature of the native ligament. Our findings demonstrate that fibroblasts seeded on crimp-like fibrous scaffolds respond more favourably (increased ECM synthesis and fascicle formation) to dynamic mechanical loading compared to those grown on scaffolds containing uncrimped (straight) fibres.


The role of actively released fibrin-conjugated VEGF for VEGF receptor 2 gene activation and the enhancement of angiogenesis.
A major challenge for therapeutic delivery of angiogenic agents such as vascular endothelial growth factor (VEGF) is to achieve sustained, low dose signaling leading to durable neovessel formation. To this end, we recently created a variant of VEGF(121), TG-VEGF(121) that directly binds to fibrin and gets released locally in proteolysis-triggered manner. Here we combined noninvasive biophotonic monitoring of VEGF receptor 2 gene activation in transgenic VEGFR2-luc mice and histomorphometry to compare endothelial activation and long-term neovascularization by actively released TG-VEGF(121)versus passively released, diffusible wild-type VEGF(121) in subcutaneous fibrin implants. Monitoring in real-time over 3 weeks of luciferase signal driven by the VEGFR2 promoter revealed endothelial activation in skin exposed to wild-type VEGF(121), but no detectable elevation over fibrin alone by TG-VEGF(121). Histology at 3 weeks, however, demonstrated that TG-VEGF(121) promoted vessel growth significantly more effectively and reliably than wild-type VEGF(121). The majority of vessels surviving to 3 weeks contained stabilizing smooth muscle cells. Yet, by 6 weeks, no extra vessels induced by exogenous VEGF were left. In conclusion, release of fibrin-conjugated variant TG-VEGF(121) elicited lower VEGFR2-luc activation than wild-type VEGF(121) yet significantly more vascularization. In the absence of true physiological demand, even stabilized vessels are ultimately regressed.


Hepatic differentiation from human mesenchymal stem cells on a novel nanofiber scaffold.
The emerging fields of tissue engineering and biomaterials have begun to provide potential treatment options for liver failure. The goal of the present study is to investigate the ability of a poly L-lactic acid (PLLA) nanofiber scaffold to support and enhance hepatic differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs). A scaffold composed of poly L-lactic acid and collagen was fabricated by the electrospinning technique. After characterizing isolated hMSCs, they were seeded onto PLLA nanofiber scaffolds and induced to differentiate into a hepatocyte lineage. The mRNA levels and protein expression of several important hepatic genes were determined using RT-PCR, immunocytochemistry and ELISA. Flow cytometry revealed that the isolated bone marrow-derived stem cells were positive for hMSC-specific markers CD73, CD44, CD105 and CD166 and negative for hematopoietic markers CD34 and CD45. The differentiation of these stem cells into adipocytes and osteoblasts demonstrated their multipotency. Scanning electron microscopy showed adherence of cells in the nanofiber scaffold during differentiation towards hepatocytes. Our results showed that expression levels of liver-specific markers such as albumin, α-fetoprotein, and cytokeratins 8 and 18 were higher in differentiated cells on the nanofibers than when cultured on plates. Importantly, liver functioning serum proteins, albumin and α-1 antitrypsin were secreted into the culture medium at higher levels by the differentiated cells on the nanofibers than on the plates, demonstrating that our nanofibrous scaffolds promoted and enhanced hepatic differentiation under our culture conditions. Our results show that the engineered PLLA nanofibrous scaffold is a conducive matrix for the differentiation of MSCs into functional hepatocyte-like cells. This represents the first step for the use of this nanofibrous scaffold for culture and differentiation of stem cells that may be employed for tissue engineering and cell-based therapy applications.


Serologic responses in indeterminate colitis patients before ileal pouch-anal anastomosis may determine those at risk for continuous pouch inflammation.
Although acute pouchitis after ileal pouch-anal anastomosis is common and easily treated, continuous pouch inflammation seen clinically as chronic, antibiotic-dependent pouchitis, and/or Crohn's disease remains a difficult management problem. Compared with ulcerative colitis, indeterminate colitis patients undergoing ileal pouch-anal anastomosis have a higher incidence of continuous pouch inflammation, which may represent persistent immune reactivity to microbial antigens. Antibody responses to three microbial antigens (oligomannan anti-Saccharomyces cerevisiae, outer membrane porin C of Escherichia coli, and an antigen (I2) from Pseudomonas flourescens) are more commonly seen in Crohn's disease, whereas antibodies to a cross-reactive antigen (perinuclear antineutrophil cytoplasmic antibodies) is more suggestive of ulcerative colitis. We examined whether preoperative serologic responses to these antigens were associated with Crohn's disease in indeterminate colitis patients after ileal pouch-anal anastomosis. Twenty-eight indeterminate colitis patients undergoing ileal pouch-anal anastomosis were prospectively assessed for the development of pouchitis or Crohn's disease. Serologic responses were determined by enzyme-linked immunosorbent assay and immunofluorescence. Patients were classified based on four predominant profiles of antibody expression. Antibody profiles were determined before knowledge of clinical outcome. Median follow-up was 38 (range, 3-75) months. Of 16 patients (61 percent) who developed pouch inflammation, 4 (25 percent) had acute pouchitis and 12 (75 percent) had continuous pouch inflammation (9 had chronic pouchitis, 3 had Crohn's disease). No preoperative clinical factor predicted the development of these pouch complications. Overall, 16 patients (57 percent) had a positive antibody reactivity profile. Serologic expression of any marker alone did not predict the development of continuous pouch inflammation. However, continuous pouch inflammation developed in 10 of 16 patients (63 percent) who had a positive antibody reactivity profile compared with only 2 of 12 patients (17 percent) who had a negative antibody reactivity profile (P = 0.015). Indeterminate colitis patients who have a positive antibody reactivity profile before ileal pouch-anal anastomosis have a significantly higher incidence of continuous pouch inflammation after surgery than those with a negative profile.


Chromosome structure and mitosis in the dinoflagellates: an ultrastructural approach to an evolutionary problem.
Chromosome structure and mitosis have been examined in three evolutionarily diverse members of the Pyrrophyta. Chromosome uncoiling, revealing the chromonema, has been correlated with the uptake of [3H]thymidine. In addition, chromosome uncoiling has been observed during gamete formation, gamete fusion, and in the nucleolar organizing region of the chromosomes suggesting that dinoflagellate chromosomes undergoing duplication, transcription or pairing have a morphology different from the characteristic tightly banded structure generally observed during most of interphase and mitosis. The dinoflagellate chromonema is composed of 2.5-nm fibers and 9.0-nm granules coiled into a helix around a central core of 9.0-nm fibers. Chromosome attachment to nuclear channels and kinetochore division and separation have been examined in several dinoflagellates. After evaluating many nuclear and cytoplasmic characteristics of the dinoflagellates it appears that this group of organisms are true eukaryotes which may be on the main line to the evolution of the mitotic spindle typical of higher plant and animals cells.U


Therapeutic hypothermia in stroke and traumatic brain injury.
Therapeutic hypothermia (TH) is considered to improve survival with favorable neurological outcome in the case of global cerebral ischemia after cardiac arrest and perinatal asphyxia. The efficacy of hypothermia in acute ischemic stroke (AIS) and traumatic brain injury (TBI), however, is not well studied. Induction of TH typically requires a multimodal approach, including the use of both pharmacological agents and physical techniques. To date, clinical outcomes for patients with either AIS or TBI who received TH have yielded conflicting results; thus, no adequate therapeutic consensus has been reached. Nevertheless, it seems that by determining optimal TH parameters and also appropriate applications, cooling therapy still has the potential to become a valuable neuroprotective intervention. Among the various methods for hypothermia induction, intravascular cooling (IVC) may have the most promise in the awake patient in terms of clinical outcomes. Currently, the IVC method has the capability of more rapid target temperature attainment and more precise control of temperature. However, this technique requires expertise in endovascular surgery that can preclude its application in the field and/or in most emergency settings. It is very likely that combining neuroprotective strategies will yield better outcomes than utilizing a single approach.


Effect of electrical stimulation on acupuncture points in diabetic patients with gastric dysrhythmia: a pilot study.
Abnormal gastric slow-wave frequencies have been observed in diabetic gastroparesis and are associated with impaired antral motor activity. In this study, we aimed at evaluating the effect of acupuncture on gastric slow waves in diabetic patients with symptoms suggesting gastric motor dysfunction. Fifteen patients with type II diabetes who had had dyspeptic symptoms for more than 3 months were enrolled. Two acupuncture needles were inserted into the subjects' legs at the Zusanli points, and electrical stimulation (2-Hz pulses) was delivered for 30 min. Cutaneous electrogastrography was performed for 30 min at baseline, for 30 min during acupuncture, and for an additional 30 min after acupuncture. Serum gastrin, motilin, and human pancreatic polypeptide levels were also measured. There was a significant increase in the percentages of normal frequency during and after acupuncture (baseline vs. acupuncture and after acupuncture 21.99 +/- 19.38% vs. 45.93 +/- 19.72 and 48.92 +/- 19.56%; p < 0.01). In addition, the percentage of tachygastric frequency was decreased significantly during and after acupuncture. The dominant frequency was also changed significantly. There was an increase of serum human pancreatic polypeptide during acupuncture (baseline vs. acupuncture 56.96 +/- 27.64 vs. 73.11 +/- 22.37 pmol/l; p < 0.05). The results of this study revealed that electrical stimulation at the Zusanli points could increase the percentage of normal electrogastrography frequency and decrease the percentage of tachygastric frequency in diabetic patients. The data indicate that acupuncture may enhance the regularity of gastric myoelectrical activity in diabetic patients.


Surgical transvenous embolization of a cortically draining carotid cavernous fistula via a vein of the sylvian fissure.
Percutaneous transvenous embolization is one of the most effective treatments of intracranial dural arteriovenous fistulas (AVFs) involving the dural sinuses. We present a unique case of surgical transvenous embolization in a 48-year-old man with a dural AVF of the cavernous sinus who presented with intracerebral hematoma. The dural AVF drained only into the vein of the sylvian fissure on angiography. Transvenous embolization via the vein of the sylvian fissure during craniotomy obliterated the AVF completely.


Comparative maps of motion and assembly of filamentous actin and myosin II in migrating cells.
To understand the mechanism of cell migration, one needs to know how the parts of the motile machinery of the cell are assembled and how they move with respect to each other. Actin and myosin II are thought to be the major structural and force-generating components of this machinery (Mitchison and Cramer, 1996; Parent, 2004). The movement of myosin II along actin filaments is thought to generate contractile force contributing to cell translocation, but the relative motion of the two proteins has not been investigated. We use fluorescence speckle and conventional fluorescence microscopy, image analysis, and computer tracking techniques to generate comparative velocity and assembly maps of actin and myosin II over the entire cell in a simple model system of persistently migrating fish epidermal keratocytes. The results demonstrate contrasting polarized assembly patterns of the two components, indicate force generation at the lamellipodium-cell body transition zone, and suggest a mechanism of anisotropic network contraction via sliding of myosin II assemblies along divergent actin filaments.


Current concepts review: intra-articular fractures of the calcaneus.
1) Intra-articular fractures of the calcaneus are associated with severe long-term consequences for function and pain. The condition of the soft tissues is of paramount importance when determining the method of treatment,the timing of surgery, and the post-injury rehabilitation.2) Intra-articular fractures are difficult to fully evaluate with plain radiographs. Computed tomography may assist in assessing the fracture pattern and planning for surgery. The likelihood of a good to excellent outcome is increased when an anatomic reduction is obtained.3) The outcome after operative management is difficult to characterize and appears to be influenced by factors related to the fracture, the patient, and the experience of the institution where the patient obtains treatment.All these factors should be factors in the decision to operate, but no single factor reliably determines the most appropriate treatment.4) Open reduction and internal fixation through an extensile approach achieves acceptable results in carefully selected patients. The use of a limited exposure with minimally invasive techniques may decrease the incidence of wound complications. However, this option is technically demanding and the quality of the reduction achieved may be more difficult to obtain and determine intraoperatively. Open fractures should be promptly debrided. The choice of fixation after reduction is based on the surgeons assessment of the soft tissue and the risk of infection.5) Post-traumatic arthritis of the subtalar joint is a common complication. Successful salvage can be achieved with a subtalar arthrodesis. However, these results may be influenced by the institution at which the initial management was rendered.


Genome-wide association study for agronomic and physiological traits in spring wheat evaluated in a range of heat prone environments.
We identified 27 stable loci associated with agronomic traits in spring wheat using genome-wide association analysis, some of which confirmed previously reported studies. GWAS peaks identified in regions where no QTL for grain yield per se has been mapped to date, provide new opportunities for gene discovery and creation of new cultivars with desirable alleles for improving yield and yield stability in wheat. We undertook large-scale genetic analysis to determine marker-trait associations (MTAs) underlying agronomic and physiological performance in spring wheat using genome-wide association studies (GWAS). Field trials were conducted at seven sites in three countries (Sudan, Egypt, and Syria) over 2-3 years in each country. Twenty-five agronomic and physiological traits were measured on 188 wheat genotypes. After correcting for population structure and relatedness, a total of 245 MTAs distributed over 66 loci were associated with agronomic traits in individual and mean performance across environments respectively; some of which confirmed previously reported loci. Of these, 27 loci were significantly associated with days to heading, thousand kernel weight, grain yield, spike length, and leaf rolling for mean performance across environments. Despite strong QTL by environment interactions, eight of the loci on chromosomes 1A, 1D, 5A, 5D, 6B, 7A, and 7B had pleiotropic effects on days to heading and yield components (TKW, SM-2, and SNS). The winter-type alleles at the homoeologous VRN1 loci significantly increased days to heading and grain yield in optimal environments, but decreased grain yield in heat prone environments. Top 20 high-yielding genotypes, ranked by additive main effects and multiplicative interaction (AMMI), had low kinship relationship and possessed 4-5 favorable alleles for GY MTAs except two genotypes, Shadi-4 and Qafzah-11/Bashiq-1-2. This indicated different yield stability mechanisms due to potentially favorable rare alleles that are uncharacterized. Our results will enable wheat breeders to effectively introgress several desirable alleles into locally adapted germplasm in developing wheat varieties with high yield stability and enhanced heat tolerance.


ERdj3 regulates BiP occupancy in living cells.
Co-chaperones regulate chaperone activities and are likely to impact a protein-folding environment as much as the chaperone itself. As co-chaperones are expressed substoichiometrically, the ability of co-chaperones to encounter a chaperone is crucial for chaperone activity. ERdj3, an abundant soluble endoplasmic reticulum (ER) co-chaperone of the Hsp70 BiP, stimulates the ATPase activity of BiP to increase BiP's affinity for client (or substrate) proteins. We investigated ERdj3 availability, how ERdj3 levels impact BiP availability, and the significance of J proteins for regulating BiP binding of clients in living cells. FRAP analysis revealed that overexpressed ERdj3-sfGFP dramatically decreases BiP-GFP mobility in a client-dependent manner. By contrast, ERdj3-GFP mobility remains low regardless of client protein levels. Native gels and co-immunoprecipitations established that ERdj3 associates with a large complex including Sec61α. Translocon binding probably ensures rapid encounters between emerging nascent peptides and stimulates BiP activity in the crucial early stages of secretory protein folding. Importantly, mutant BiP exhibited significantly increased mobility when it could not interact with any ERdjs. Thus, ERdjs appear to play the dual roles of increasing BiP affinity for clients and regulating delivery of clients to BiP. Our data suggest that BiP engagement of clients is enhanced in ER subdomains enriched in ERdj proteins.


Volunteering by older adults and risk of mortality: a meta-analysis.
Organizational volunteering has been touted as an effective strategy for older adults to help themselves while helping others. Extending previous reviews, we carried out a meta-analysis of the relation between organizational volunteering by late-middle-aged and older adults (minimum age = 55 years old) and risk of mortality. We focused on unadjusted effect sizes (i.e., bivariate relations), adjusted effect sizes (i.e., controlling for other variables such as health), and interaction effect sizes (e.g., the joint effect of volunteering and religiosity). For unadjusted effect sizes, on average, volunteering reduced mortality risk by 47%, with a 95% confidence interval ranging from 38% to 55%. For adjusted effect sizes, on average, volunteering reduced mortality risk by 24%, with a 95% confidence interval ranging from 16% to 31%. For interaction effect sizes, we found preliminary support that as public religiosity increases, the inverse relation between volunteering and mortality risk becomes stronger. The discussion identifies several unresolved issues and directions for future research.


Infectivity reacquisition by Trypanosoma brucei brucei cultivated with tsetse salivary glands.
Reacquisition of infectivity four mice was observed in cultures of Trypanosoma brucei brucei grown in the presence of tests fly salivary gland explants in a medium based on the amino acid composition of Glossina hemolymph and containing fetal bovine serum. High infection rates were obtained in mice inoculated with about 1.5 X 10(8) organisms. Infectivity reacquisition was correlated with invasion of the salivary glands by the parasites. Few small trypanosomes with subterminal kinetoplasts (metacyclic-like forms) were found in the infective inoculums. The parasitemias in mice consisted of pleomorphic cultivable trypanosomes. Cultures initiated by these organisms and then placed with the head-salivary gland preparations became infective for mice.


Latino parents' perceptions of the eating and physical activity experiences of their pre-school children at home and at family child-care homes: a qualitative study.
Research indicates that healthful eating and physical activity (PA) practices implemented in child-care settings can have a positive effect on children's healthful behaviours in this setting, and this effect on healthful behaviours may possibly transfer to the home environment. While more research is needed to examine whether behaviours learned in family child-care homes (FCCH) transfer, the potential for transferability is especially important given that Latino children's home environment has been characterized by obesogenic parenting practices. We aimed to examine Latino parents' perceptions of their pre-school children's eating and PA experiences at home and at FCCH. Qualitative study. Six focus groups were conducted in Spanish (n 36). Transcripts were analysed using thematic analysis to identify key concepts and themes. Analyses revealed that Latino parents perceive their children have healthier eating and PA experiences at FCCH than at home. Parents attributed this to FCCH providers providing an environment conducive to healthful eating and PA due to providers having more knowledge and skills, time and resources, and being required to follow rules and regulations set by the state that promote healthful eating and PA. Understanding parental perceptions, attitudes and practices related to establishing and maintaining an environment conducive to children's healthful eating and PA at home and at the FCCH is essential for the design of successful interventions to promote children's healthful behaviours in these two settings. Given that parents perceive their children as having more healthful behaviours while at FCCH, interventions that address both settings jointly may be most effective than those addressing only one environment by itself.


Thymol as an Adjuvant to Restore Antibiotic Efficacy and Reduce Antimicrobial Resistance and Virulence Gene Expression in Enterotoxigenic Escherichia coli Strains.
The continuous spread of antimicrobial resistance is endangering the efficient control of enterotoxigenic Escherichia coli (ETEC), which is mainly responsible for post-weaning diarrhea onset in piglets. Thymol, the key constituent of thyme essential oil, is already used in animal nutrition for its antimicrobial action. The aim of this study was to investigate the potential adjuvant effect of thymol to re-establish antibiotic efficacy against highly resistant ETEC field strains. Secondly, we evaluated the modulation of virulence and antibiotic resistance genes. Thymol showed the capacity to control ETEC growth and, when combined with ineffective antibiotics, it increased their antimicrobial power. In particular, it showed significant effects when blended with colistin and tetracycline, suggesting that the adjuvant effects rely on the presence of complementary mechanisms of action between molecules, or the absence of resistance mechanisms that inactivate antibiotics and target sites. Furthermore, our findings demonstrate that, when added to antibiotics, thymol can help to further downregulate several virulence and antibiotic resistance genes, offering new insights on the potential mechanisms of action. Therefore, in a one-health approach, our study supports the beneficial effects of combining thymol with antibiotics to restore their efficacy, together with the possibility of targeting gene expression as a pioneering approach to manage ETEC pathogenicity.


Genome-wide profiling of alternative splicing in Alzheimer's disease.
Alternative splicing is a highly regulated process which generates transcriptome and proteome diversity through the skipping or inclusion of exons within gene loci. Identification of aberrant alternative splicing associated with human diseases has become feasible with the development of new genomic technologies and powerful bioinformatics. We have previously reported genome-wide gene alterations in the neocortex of a well-characterized cohort of Alzheimer's disease (AD) patients and matched elderly controls using a commercial exon microarray platform [1]. Here, we provide detailed description of analyses aimed at identifying differential alternative splicing events associated with AD.


Co-producing Human and Animal Experimental Subjects: Exploring the Views of UK COVID-19 Vaccine Trial Participants on Animal Testing.
Preclinical (animal) testing and human testing of drugs and vaccines are rarely considered by social scientists side by side. Where this is done, it is typically for theoretically exploring the ethics of the two situations to compare relative treatment. In contrast, we empirically explore how human clinical trial participants understand the role of animal test subjects in vaccine development. Furthermore, social science research has only concentrated on broad public opinion and the views of patients about animal research, whereas we explore the views of a public group particularly implicated in pharmaceutical development: experimental subjects. We surveyed and interviewed COVID-19 vaccine trial participants in Oxford, UK, on their views about taking part in a vaccine trial and the role of animals in trials. We found that trial participants mirrored assumptions about legitimate reasons for animal testing embedded in regulation and provided insight into (i) the nuances of public opinion on animal research; (ii) the co-production of human and animal experimental subjects; (iii) how vaccine and medicine testing, and the motivations and demographics of clinical trial participants, change in an outbreak; and (iv) what public involvement can offer to science.


Erythrocyte Partitioning Profile of Isosteviol in Human and Rat Blood.
Isosteviol is a synthetic derivative of steviol glycosides with promising pharmacological properties and might find future use as a cardioprotective agent. A simple LC-MS/MS technique was developed and validated for the bioanalysis of isosteviol in plasma and erythrocytes. This method was subsequently utilized for the in vitro assessment of isosteviol's partitioning into blood compartments of humans and rats. Fresh blood samples from healthy humans and Wistar rats were equilibrated with 1, 10, and 30 µM isosteviol at 37 °C in a shaking dry-bath. The levels of isosteviol in plasma and erythrocytes partitions were determined in these samples, after separation, at intervals over a 60-minute period. The data derived was used to estimate erythrocyte-to-plasma and blood-to-plasma coefficients. Mean erythrocyte-to-plasma partition coefficients (SD) after 60 minutes of equilibration were observed to be 0.039 (0.002) and 0.040 (0.003) in humans and rats, respectively. Derived values for the blood-to-plasma ratio (SD) were 0.576 (0.001) in humans and 0.543 (0.007) in rats, whereas plasma component binding was estimated to be more than 96%. The findings suggest that isosteviol preferentially partitions into plasma compartments in humans and rats. The significance of this profile for the efficacy, tissue uptake, and retention of isosteviol will have to be further studied.


Identifying and sequencing a Mycobacterium sp. strain F4 as a potential bioremediation agent for quinclorac.
Quinclorac is a widely used herbicide in rice filed. Unfortunately, quinclorac residues are phytotoxic to many crops/vegetables. The degradation of quinclorac in nature is very slow. On the other hand, degradation of quinclorac using bacteria can be an effective and efficient method to reduce its contamination. In this study, we isolated a quinclorac bioremediation bacterium strain F4 from quinclorac contaminated soils. Based on morphological characteristics and 16S rRNA gene sequence analysis, we identified strain F4 as Mycobacterium sp. We investigated the effects of temperature, pH, inoculation size and initial quinclorac concentration on growth and degrading efficiency of F4 and determined the optimal quinclorac degrading condition of F4. Under optimal degrading conditions, F4 degraded 97.38% of quinclorac from an initial concentration of 50 mg/L in seven days. Our indoor pot experiment demonstrated that the degradation products were non-phytotoxic to tobacco. After analyzing the quinclorac degradation products of F4, we proposed that F4 could employ two pathways to degrade quinclorac: one is through methylation, the other is through dechlorination. Furthermore, we reconstructed the whole genome of F4 through single molecular sequencing and de novo assembly. We identified 77 methyltransferases and eight dehalogenases in the F4 genome to support our hypothesized degradation path.


Smoking and perceived stress in relation to short salivary telomere length among caregivers of children with disabilities.
Telomere length (TL), the length of repeated DNA sequence that forms protective caps at the end of chromosomes, has emerged as a novel biomarker of cell aging and oxidative stress. There is increasing research exploring the associations of smoking and perceived stress with TL, and the results are inconsistent. This study aimed to examine whether smoking and perceived stress were associated with shortened salivary TL among primary caregivers of children with disabilities. Using a quantitative polymerase chain reaction method, salivary TL was assessed among 89 caregivers aged 19-69 years (87% were women) who took care of disabled children in the Patagonia Region, Chile. Interviewer-administered questionnaires were used to collect information on sociodemographic and lifestyle factors. The 14-item Perceived Stress Scale was used to assess perceived stress. Mean relative TL was 0.92 (standard error = 0.03). Smokers had age-adjusted mean TL that was 0.07 units lower (β = -0.07, standard error = 0.03; p = 0.012) than non-smokers. Smokers were 2.17 times more likely to have shorter TL ( < 0.73, the lowest quartile of TL) than non-smokers (odds ratio = 3.17; 95% confidence interval = 1.05-9.52) with adjustment for age and perceived stress. Caregivers with higher perceived stress were 2.13 times more likely to have shorter TL (odds ratio = 3.13; 95% confidence interval = 1.03-9.55) than caregivers with lower perceived stress after adjustment for age and smoking. This study provides the first evidence of strong associations between smoking and perceived stress and shortened salivary TL among caregivers of children with disabilities. Larger studies with detailed information on smoking status are warranted to confirm our findings.


Long-term Lifestyle-modification Programs for Overweight and Obesity Management in the Arab States: Systematic Review and Meta-analysis.
Obesity is a growing health problem worldwide. It is associated with serious medical and psychosocial comorbidities that increase the risk of mortality. However, strong evidence confirms lifestyle-modification programs as the cornerstone treatment for excess weight and obesity. The aim of this systematic review was to assess the effectiveness of the lifestyle-modification programs for weight management delivered in Arabic-speaking countries. The PubMed database was searched, and studies conducted in humans were identified and screened as per the Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) guidelines. Data were collated using meta-analysis and a narrative approach. Of the 1057 articles retrieved, six studies, conducted in four Arab countries, comprising a total of 444 adolescent and adult participants of both genders with overweight and obesity, met the inclusion criteria and were reviewed. Most studies that assessed weight loss at 6-month follow-up showed no significant reduction in body weight. Meta-analysis confirmed that the lifestyle-modification programs delivered were no more effective than other treatments. Only one article reported significant weight-loss maintenance after 12 months of follow-up. However this was a prospective non-controlled study in which the weight loss maintained (=4%) did not conform to the standard for clinical significance (>10%). Lifestyle-modification programs for weight management delivered in Arabic-speaking countries seem lacking in effectiveness due to methodological weaknesses in program adaptation, a lack of expert clinical supervision before and during implementation, and the presence of barriers to lifestyle modification, especially for women. Future studies should bear these features in mind.


Sequence and phylogenetic analyses of novel totivirus-like double-stranded RNAs from field-collected powdery mildew fungi.
The identification of mycoviruses contributes greatly to understanding of the diversity and evolutionary aspects of viruses. Powdery mildew fungi are important and widely studied obligate phytopathogenic agents, but there has been no report on mycoviruses infecting these fungi. In this study, we used a deep sequencing approach to analyze the double-stranded RNA (dsRNA) segments isolated from field-collected samples of powdery mildew fungus-infected red clover plants in Japan. Database searches identified the presence of at least ten totivirus (genus Totivirus)-like sequences, termed red clover powdery mildew-associated totiviruses (RPaTVs). The majority of these sequences shared moderate amino acid sequence identity with each other (<44%) and with other known totiviruses (<59%). Nine of these identified sequences (RPaTV1a, 1b and 2-8) resembled the genome of the prototype totivirus, Saccharomyces cerevisiae virus-L-A (ScV-L-A) in that they contained two overlapping open reading frames (ORFs) encoding a putative coat protein (CP) and an RNA dependent RNA polymerase (RdRp), while one sequence (RPaTV9) showed similarity to another totivirus, Ustilago maydis virus H1 (UmV-H1) that encodes a single polyprotein (CP-RdRp fusion). Similar to yeast totiviruses, each ScV-L-A-like RPaTV contains a -1 ribosomal frameshift site downstream of a predicted pseudoknot structure in the overlapping region of these ORFs, suggesting that the RdRp is translated as a CP-RdRp fusion. Moreover, several ScV-L-A-like sequences were also found by searches of the transcriptome shotgun assembly (TSA) libraries from rust fungi, plants and insects. Phylogenetic analyses show that nine ScV-L-A-like RPaTVs along with ScV-L-A-like sequences derived from TSA libraries are clustered with most established members of the genus Totivirus, while one RPaTV forms a new distinct clade with UmV-H1, possibly establishing an additional genus in the family. Taken together, our results indicate the presence of diverse, novel totiviruses in the powdery mildew fungus populations infecting red clover plants in the field.


Is bad living better than good death? Impact of demographic and cultural factors on health state preference.
The aim of this study was to examine the impact of demographic and cultural factors on health preferences among Chinese general population. The Chinese EQ-5D-5L valuation study was conducted between December 2012 and January 2013. A total of 1296 participants were recruited from the general public at Beijing, Chengdu, Guiyang, Nanjing, and Shenyang. Each participant was interviewed to measure preferences for ten EQ-5D-5L health states using composite time trade-off and seven pairs of states using discrete choice experiment (data were not included in this study). At the end of the interview, each participant was also asked to provide their demographic information and answers to two questions about their attitudes towards whether bad living is better than good death (LBD) and whether they believe in an afterlife. Generalized linear model and random effects logistic models were used to examine the impact of demographic and cultural factors on health preferences. Participants who had serious illness experience received college or higher education, or agree with LBD were more likely to value health states positively and have a narrower score range. Participants at Beijing were more likely to be non-traders, value health states positively, less likely to reach the lowest possible score, and have narrower score range compared with all other four cities after controlling for all other demographic and culture factors. Health state preference is significantly affected by factors beyond demographics. These factors should be considered in achieving a representative sample in valuation studies in China.


Molecular Similarity for Drug Discovery, Target Prediction and Chemical Space Visualization.
Similar drug molecules often have similar properties and activities. Therefore, quantifying molecular similarity is central to drug discovery and optimization. Here I review computational methods using molecular similarity measures developed in my group within the interdisciplinary network NCCR TransCure investigating the physiology, structural biology and pharmacology of ion channels and membrane transporters. We designed a 3D molecular shape and pharmacophore comparison algorithm to optimize weak and unselective inhibitors by scaffold hopping and discovered potent and selective inhibitors of the ion channels TRPV6 and TRPM4, of endocannabinoid membrane transport, and of the divalent metal transporters DMT1 and ZIP8. We predicted off-target effects by combining molecular similarity searches from different molecular fingerprints against target annotated compounds from the ChEMBL database. Finally, we created interactive chemical space maps reflecting molecular similarities to facilitate the selection of screening compounds and the analysis of screening results. These different tools are available online at https://gdb.unibe.ch/tools/.


Using the assessment of motor and process skills to compare occupational performance between clinic and home settings.
The study described in this article examined the effect of home versus clinic settings on the instrumental activities of daily living (IADL) performance of older adults. Twenty older adults living in the community were evaluated in their homes and in an occupational therapy clinic with the Assessment of Motor and Process Skills (AMPS). The motor and process ability measures were compared between the two settings with many-faceted Rasch analysis. The subjects' motor ability measures tended to remain stable from clinic to home settings. The process ability measures tended not to remain stable from clinic to home settings, because 10 of the 20 subjects performed significantly better in their homes. These findings support the idea that process skill abilities are affected by the environment to a greater degree than are motor skills abilities and that for persons living in the community, the familiar home environment tends to support IADL performance. If an occupational therapist wants to know how a person performs IADLs, the therapist should evaluate that person's performance in the environment in which the client will be functioning.


'Candidatus Xenohaliotis californiensis', a newly described pathogen of abalone, Haliotis spp., along the west coast of North America.
Withering syndrome is a fatal disease of wild and cultured abalone, Haliotis spp., that inhabit the west coast of North America. The aetiological agent of withering syndrome has recently been identified as a member of the family Rickettsiaceae in the order Rickettsiales. Using a combination of morphological, serological, life history and genomic (16S rDNA) characterization, we have identified this bacterium as a unique taxon and propose the provisional status of 'Candidatus Xenohaliotis californiensis'. The Gram-negative, obligate intracellular pleomorphic bacterium is found within membrane-bound vacuoles in the cytoplasm of abalone gastrointestinal epithelial cells. The bacterium is not cultivable on synthetic media or in fish cell lines (e.g. CHSE-214) and may be controlled by tetracyclines (oxytetracycline) but not by chloramphenicol, clarithromycin or sarafloxicin. Phylogenetic analysis based on the 16S rDNA of 'Candidatus Xenohaliotis californiensis' places it in the alpha-subclass of the class Proteobacteria but not to the four recognized subtaxa of the alpha-Proteobacteria (alpha-1, alpha-2, alpha-3 and alpha-4). The bacterium can be detected in tissue squashes stained with propidium iodide, microscopic examination of stained tissue sections, PCR or in situ hybridization. 'Candidatus Xenohaliotis californiensis' can be differentiated from other closely related alpha-Proteobacteria by its unique 16S rDNA sequence.


Determination of host proteins composing the microenvironment of coronavirus replicase complexes by proximity-labeling.
Positive-sense RNA viruses hijack intracellular membranes that provide niches for viral RNA synthesis and a platform for interactions with host proteins. However, little is known about host factors at the interface between replicase complexes and the host cytoplasm. We engineered a biotin ligase into a coronaviral replication/transcription complex (RTC) and identified >500 host proteins constituting the RTC microenvironment. siRNA-silencing of each RTC-proximal host factor demonstrated importance of vesicular trafficking pathways, ubiquitin-dependent and autophagy-related processes, and translation initiation factors. Notably, detection of translation initiation factors at the RTC was instrumental to visualize and demonstrate active translation proximal to replication complexes of several coronaviruses. Collectively, we establish a spatial link between viral RNA synthesis and diverse host factors of unprecedented breadth. Our data may serve as a paradigm for other positive-strand RNA viruses and provide a starting point for a comprehensive analysis of critical virus-host interactions that represent targets for therapeutic intervention.


Inhibitors of histone deacetylases suppress cisplatin-induced p53 activation and apoptosis in renal tubular cells.
Inhibitors of histone deacetylases, including suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA), are emerging anticancer agents. In the current study, we examined the cytoprotective effects of these agents. Cisplatin induced 40-50% apoptosis in rat kidney proximal tubular cells in 18 h, which was suppressed to 20-30% by 1-5 microM SAHA or 0.1 microM TSA. Consistently, SAHA partially prevented cisplatin-induced caspase activation. The cytoprotective effects of SAHA and TSA were associated with long-term cell survival. During cisplatin treatment, Bax translocated to mitochondria, leading to cytochrome c release. Both Bax translocation and cytochrome c release were ameliorated by SAHA. Mechanistically, SAHA inhibited and TSA delayed p53 phosphorylation, acetylation, and activation during cisplatin incubation. At the upstream signaling level, SAHA blocked cisplatin-induced phosphorylation of Chk2, a key DNA damage response kinase. Interestingly, in HCT116 colon cancer cells, SAHA suppressed cisplatin-induced p53 activation, but enhanced apoptosis. The results suggest that inhibitors of histone deacetylases can protect against cisplatin nephrotoxicity by attenuating DNA damage response and associated p53 activation.


Guanine nucleotides induce tyrosine phosphorylation and activation of the respiratory burst in neutrophils.
Activation of the NADPH oxidase was examined in electrically permeabilized human neutrophils exposed to non-hydrolysable guanine nucleotides. Guanosine 5'-[gamma-thio]triphosphate (GTP[S]) induced a marked increase in the rate of O2 consumption, which was partially resistant to staurosporine, an inhibitor of protein kinase C, under conditions where the response to diacylglycerol was virtually abolished. The respiratory burst elicited by GTP[S] was dependent on the presence of ATP and Mg2+, suggesting involvement of phosphorylation reactions. Accordingly, phosphoprotein formation was greatly stimulated by the guanine nucleotide. The polypeptide phosphorylation pattern induced by GTP[S] was similar to, but not identical with, that observed with diacylglycerol, indicating the activation of kinases other than protein kinase C by the guanine nucleotide. The possible involvement of tyrosine kinases was assessed by immunoblotting using anti-phosphotyrosine antibodies. Treatment of electroporated cells with GTP[S] stimulated the accumulation of tyrosine-phosphorylated proteins. This effect was not induced by diacylglycerol, indicating that tyrosine phosphorylation is not secondary to stimulation of protein kinase C. The results indicate that, in neutrophils, activated G-proteins can stimulate tyrosine kinase and/or inhibit tyrosine phosphatase activity. Changes in the amounts of tyrosine-phosphorylated proteins may signal activation of the respiratory burst.


Relation of obesity and gender to left ventricular hypertrophy in normotensive and hypertensive adults.
Although it is recognized that both hypertension and obesity are associated with increased left ventricular mass, the relative impacts of obesity, arterial hypertension, and gender on the prevalence of ventricular hypertrophy remain uncertain. Accordingly, echocardiographic left ventricular mass normalized for height to the power of the allometric or growth relation between ventricular mass and height was compared in 164 normotensive subjects (85 men [24 obese] and 79 women [28 obese], aged 45 +/- 12 years) and 475 hypertensive patients (325 men [126 obese] and 150 women [85 obese], aged 54 +/- 10 years) from an adult employed population. Gender-specific upper normal limits were used to identify ventricular hypertrophy. Left ventricular mass/height 2.7 was higher in obese than normal-weight normotensive subjects (P < .004) independently of the level of blood pressure and identified a higher prevalence of hypertrophy (mainly eccentric) in obese than in normal-weight normotensive subjects (14% versus 5%, P < .04), a difference that was not detected by left ventricular mass/body surface area. Left ventricular mass/height identified hypertrophy in 52% of obese and 30% of normal-weight hypertensive patients (P < .0001) because of higher prevalences in obese than normal-weight patients of both eccentric (34% versus 20%) and concentric ventricular hypertrophy (18% versus 10%). The increase in left ventricular mass was independent of blood pressure values in obese normotensive women (but not men), and the prevalence of supranormal left ventricular mass/height 2.7 was even higher in hypertensive obese women (58%) than men (49%).(ABSTRACT TRUNCATED AT 250 WORDS)


The socioeconomic cost of chronic rhinosinusitis study.
Chronic rhinosinusitis (CRS) is highly prevalent, affecting 11% of the population. Studies evaluating the socio-economic impact of CRS are mostly limited to the US population. Currently there is no study that has evaluated the socio-economic costs of CRS in the UK. A case-control study of patients with CRS and healthy controls was conducted to investigate the wider socio-economic impact of the disease. Data on demographic and socioeconomic characteristics, out-of-pocket expenditure (OOPE), health resource utilisation, productivity losses and health-related quality of life (HRQoL) via the EQ-5D and SNOT-22 instruments, were collected from questionnaires. A total of 139 CRS participants and 67 control participants completed the questionnaires. The average total OOPE per patient extrapolated to a 12-month period was £304.84. Other important findings include significantly higher reported primary care interactions (4.14 vs. 1.16) as well as secondary care interactions (2.61 vs 0.4) in CRS group as compared to controls over three-months. The average total missed workdays was estimated to be 18.7 per patient per year. The estimated incremental healthcare cost of CRS per year is £16.8 billion or £2.8 billion per million inhabitants. Factors predictive of a higher OOPE include higher household occupancy and income and these accounted for only 9.7% of the total variance in total OOPEs. Other socioeconomic, demographic and HRQoL variables were not found to be predictive factors of OOPE. This study showed that CRS has a significant wider economic burden beyond the immediate direct healthcare costs. CRS participants had a high level of healthcare service use, OOPE and productivity loss. Results from this study will add to the existing limited data both for the UK and abroad and emphasises the need for effective treatments for these patients to reduce the disease impact.


Involving elderly research participants in the co-design of a future multi-generational cohort study.
It has been proposed that the existing ASPirin in Reducing Events in the Elderly Extension observational cohort study (ASPREE-XT) would provide a platform for a future multigenerational research study (MGRS). An advert was sent to 14,268 participants (aged 74 years and older, from Australia, and located in both metropolitan and rural locations) to invite them to share views and preferences about being involved in the co-design of a future MGRS, as their preferences were not known. The objective of this article is to report as a case study the process of involving study participants and how this impacted the co-design of a proposed multi-generational research study, using a novel standardised reporting tool. We used participatory action research to involve elderly research participants in the co-design of a proposed multi-generational cohort study between 2017 and 2019 using newsletters, telephone interviews and an in-person workshop. We used the novel 'Standardised Data on Initiatives Alpha Version 0.1' (STARDIT 0.1) to plan and report how participant involvement activities positively impacted the study design. Fifty-nine ASPREE-XT participants were interviewed by telephone and 18 participants attended a face-to-face event. Involving participants positively impacted the proposed study design by improving the research objectives, developing protocols, influencing funding decisions and improving ethics applications. Learning points included the importance of maintaining the ideals of ASPREE-XT (respect, quality and transparency); research participants' preference for the option of receiving results (including genetic results); participants' need for involvement in decisions about recruitment, data access, governance and other ethical issues; and the preference for different communication methods, including both face-to-face and online methods. Data from the process indicated it was highly valued by all stakeholders, including research participants, study staff and lead investigators. Involvement of participants was described by a lead study investigator of ASPREE-XT as "enormously helpful". This case study demonstrates that including participants in the design of a research study positively impacted the study design, participants and researchers. Using a standardised reporting tool to describe the methods and impacts provides a way for learning from this case study to inform future research studies planning to involve people.


Isolation and characterization of nucleotide-binding site resistance gene homologues in common bean (Phaseolus vulgaris).
Common bean production is constrained by many fungal, viral, and bacterial pathogens. Thus, the identification of resistance (R) genes is an important focal point of common bean research. The main goal of our study was to identify resistance gene homologues (RGH) in the crop, using degenerate primers designed from conserved sequences in the nucleotide-binding site (NBS) domains of R-genes from the model legume Medicago truncatula. Total DNA of the Andean common bean genotype G19833 was used for amplification of over 500 primer combinations. Sequencing of amplicons showed that 403 cloned fragments had uninterrupted open reading frames and were considered representative of functional RGH genes. The sequences were grouped at two levels of nucleotide identity (90 and 80%) and representative sequences of each group were used for phylogenetic analyses. The RGH sequence diversity of common bean was divided into TIR and non-TIR families, each with different clusters. The TIR sequences grouped into 14 clades while non-TIR sequences grouped into seven clades. Pairwise comparisons showed purifying selection, although some sequences may have been the result of diversifying selection. Knowledge about RGH genes in common bean can allow the design of molecular markers for pyramiding of resistance genes against various pathogens.


Novel fluorogenic substrates for assaying retroviral proteases by resonance energy transfer.
The 11-kD protease (PR) encoded by the human immunodeficiency virus 1 (HIV-1) is essential for the correct processing of viral polyproteins and the maturation of infectious virus, and is therefore a target for the design of selective acquired immunodeficiency syndrome (AIDS) therapeutics. To facilitate the identification of novel inhibitors of HIV-1 PR, as well as to permit detailed studies on the enzymology and inhibition of this enzyme, a continuous assay for its activity was developed that was based on intramolecular fluorescence resonance energy transfer (RET). The assay used the quenched fluorogenic substrate 4-(4-dimethylaminophenylazo)benzoic acid (DABCYL)--Ser Gln Asn Tyr Pro Ile Val Gln--5-[(2-aminoethyl)amino]naphthalene-1 sulfonic acid (EDANS), whose peptide sequence is derived from a natural processing site for HIV-1 PR. Incubation of recombinant HIV-1 PR with the fluorogenic substrate resulted in specific cleavage at the Tyr-Pro bond and a time-dependent increase in fluorescence intensity that was linearly related to the extent of substrate hydrolysis. An internally quenched fluorogenic substrate was also designed that was selectively cleaved by the related PR from avian myeloblastosis virus (AMV). The fluorescence quantum yields of the HIV-1 PR and AMV PR substrates in the RET assay increased by 40.0- and 34.4-fold, respectively, per mole of substrate cleaved. Because of its simplicity, rapidity, and precision in the determination of reaction rates required for kinetic analysis, this method offers many advantages over the commonly used high-performance liquid chromatography- or electrophoresis-based assays for peptide substrate hydrolysis by retroviral PRs.


Wildlife ecotoxicology of pesticides: can we track effects to the population level and beyond?
During the past 50 years, the human population has more than doubled and global agricultural production has similarly risen. However, the productive arable area has increased by just 10%; thus the increased use of pesticides has been a consequence of the demands of human population growth, and its impact has reached global significance. Although we often know a pesticide's mode of action in the target species, we still largely do not understand the full impact of unintended side effects on wildlife, particularly at higher levels of biological organization: populations, communities, and ecosystems. In these times of regional and global species declines, we are challenged with the task of causally linking knowledge about the molecular actions of pesticides to their possible interference with biological processes, in order to develop reliable predictions about the consequences of pesticide use, and misuse, in a rapidly changing world.


In vitro study of doxorubicin-induced oxidative stress in spermatogonia and immature Sertoli cells.
Pediatric chemotherapy treatments can impair long-term male fertility. Unfortunately, no fertility preservation solution is available for pre-pubertal boys. Studies suggest that doxorubicin, used against pediatric cancers, induces oxidative stress in the testis. However, the targeted testicular cell types remain unknown. The goal of this study was to determine whether doxorubicin can induce oxidative stress in rat spermatogonia (GC-6Spg) and immature Sertoli (Ser-W3) cell lines, and to assess their protection by antioxidants. Using the MTT assay, we have shown that doxorubicin induces a time- and dose-dependent cytotoxicity in these two cell lines, Ser-W3 being more sensitive than GC-6Spg. After 3 h of treatment, reactive oxygen species and nuclear 8-oxo-deoxyguanosine increase in Ser-W3, but not in GC-6Spg. Moreover, after 6 h of treatment, intracellular reduced glutathione levels decrease significantly in Ser-W3 cells. These results show that doxorubicin induces oxidative stress in the Ser-W3 cell line. However, a depletion in glutathione does not affect their survival, and supplementation only offers a weak protection after exposure to doxorubicin, suggesting that the glutathione system is not essential for Ser-W3 cell line's defense against doxorubicin. On the other hand, among four antioxidants selected from the literature, none reduces the cytotoxicity of doxorubicin in Ser-W3 cells. Together, our data suggest that oxidative stress may not be a major pathway for doxorubicin's cytotoxicity in GC-6Spg and Ser-W3 lines. This study provides new insights in the mechanisms by which chemotherapies affect the pre-pubertal testis, with the long-term goal to help improve the quality of life of pediatric cancer survivors.


Orbit-infiltrating mast cells, monocytes, and macrophages produce PDGF isoforms that orchestrate orbital fibroblast activation in Graves' ophthalmopathy.
Platelet-derived growth factors (PDGF) are regulators of fibroblast activity that may be involved in the pathophysiology of Graves' ophthalmopathy (GO). We unraveled the expression and origin of PDGF family members in GO orbital tissue and investigated the effect of PDGF isoforms on IL-6 and hyaluronan production and proliferation by orbital fibroblasts. PDGF-A, PDGF-B, PDGF-C, PDGF-D, PDGF-Rα, and PDGF-Rβ expression was determined by real-time quantitative PCR and PDGF-A and PDGF-B protein expression was determined by Western blot in orbital tissues. Orbital tissues were immunohistochemically stained for PDGF-A and PDGF-B expression, together with stainings for T cells, monocytes, B cells, macrophages, and mast cells. Effects of PDGF-AA, PDGF-AB, and PDGF-BB on orbital fibroblast proliferation and IL-6 and hyaluronan production were examined. Finally, effects of PDGF-BB- and PDGF-AA-neutralizing antibodies on IL-6 and hyaluronan production in GO whole orbital tissue cultures were tested. GO orbital tissue showed increased PDGF-A and PDGF-B mRNA and protein levels. Increased numbers of PDGF-A- and PDGF-B-positive monocytes, macrophages, and mast cells were present in GO orbital tissue. PDGF-BB stimulated proliferation and hyaluronan and IL-6 production by orbital fibroblasts the most, followed by PDGF-AB and PDGF-AA. Finally, in particular imatinib mesylate and PDGF-BB-neutralizing antibodies reduced IL-6 and hyaluronan production by whole orbital tissue cultures from GO patients. In GO, mast cells, monocytes, and macrophages may activate orbital fibroblasts via secretion of especially PDGF-AB and PDGF-BB. Preclinical studies with whole orbital tissue cultures show that blocking PDGF-B chain containing isoforms can be a promising treatment for GO.


Factors Influencing the Adjuvant Therapy Decision: Results of a Real-World Multicenter Data Analysis of 904 Melanoma Patients.
Adjuvant treatment of melanoma patients with immune-checkpoint inhibition (ICI) and targeted therapy (TT) significantly improved recurrence-free survival. This study investigates the real-world situation of 904 patients from 13 German skin cancer centers with an indication for adjuvant treatment since the approval of adjuvant ICI and TT. From adjusted log-binomial regression models, we estimated relative risks for associations between various influence factors and treatment decisions (adjuvant therapy yes/no, TT vs. ICI in BRAF mutant patients). Of these patients, 76.9% (95% CI 74-80) opted for a systemic adjuvant treatment. The probability of starting an adjuvant treatment was 26% lower in patients >65 years (RR 0.74, 95% CI 68-80). The most common reasons against adjuvant treatment given by patients were age (29.4%, 95% CI 24-38), and fear of adverse events (21.1%, 95% CI 16-28) and impaired quality of life (11.9%, 95% CI 7-16). Of all BRAF-mutated patients who opted for adjuvant treatment, 52.9% (95% CI 47-59) decided for ICI. Treatment decision for TT or ICI was barely associated with age, gender and tumor stage, but with comorbidities and affiliated center. Shortly after their approval, adjuvant treatments have been well accepted by physicians and patients. Age plays a decisive role in the decision for adjuvant treatment, while pre-existing autoimmune disease and regional differences influence the choice between TT or ICI.


Integrating multiple fitting regression and Bayes decision for cancer diagnosis with transcriptomic data from tumor-educated blood platelets.
The application of machine learning in cancer diagnostics has shown great promise and is of importance in clinic settings. Here we consider applying machine learning methods to transcriptomic data derived from tumor-educated platelets (TEPs) from individuals with different types of cancer. We aim to define a reliability measure for diagnostic purposes to increase the potential for facilitating personalized treatments. To this end, we present a novel classification method called MFRB (for Multiple Fitting Regression and Bayes decision), which integrates the process of multiple fitting regression (MFR) with Bayes decision theory. MFR is first used to map multidimensional features of the transcriptomic data into a one-dimensional feature. The probability density function of each class in the mapped space is then adjusted using the Gaussian probability density function. Finally, the Bayes decision theory is used to build a probabilistic classifier with the estimated probability density functions. The output of MFRB can be used to determine which class a sample belongs to, as well as to assign a reliability measure for a given class. The classical support vector machine (SVM) and probabilistic SVM (PSVM) are used to evaluate the performance of the proposed method with simulated and real TEP datasets. Our results indicate that the proposed MFRB method achieves the best performance compared to SVM and PSVM, mainly due to its strong generalization ability for limited, imbalanced, and noisy data.


HapFIC: An Adaptive Force/Position Controller for Safe Environment Interaction in Articulated Systems.
Haptic interaction is essential for the dynamic dexterity of animals, which seamlessly switch from an impedance to an admittance behaviour using the force feedback from their proprioception. However, this ability is extremely challenging to reproduce in robots, especially when dealing with complex interaction dynamics, distributed contacts, and contact switching. Current model-based controllers require accurate interaction modelling to account for contacts and stabilise the interaction. In this manuscript, we propose an adaptive force/position controller that exploits the fractal impedance controller's passivity and non-linearity to execute a finite search algorithm using the force feedback signal from the sensor at the end-effector. The method is computationally inexpensive, opening the possibility to deal with distributed contacts in the future. We evaluated the architecture in physics simulation and showed that the controller can robustly control the interaction with objects of different dynamics without violating the maximum allowable target forces or causing numerical instability even for very rigid objects. The proposed controller can also autonomously deal with contact switching and may find application in multiple fields such as legged locomotion, rehabilitation and assistive robotics.


Highly corrected submicrometer grid patterning on curved surfaces.
A compact holographic projector system was built and tested. This projection system offers a practical approach for making a highly corrected mesh or grid pattern on curved surfaces. The pattern can range in size from multimicrometer to submicrometer dimensions and be recorded in either positive or negative photoresist. Standing-wave interference patterns in the form of a diverging close-packed lattice of either hexagonal or square rodlike intensity maxima extending outward from a point or a locus of points are produced by multiple-beam holography that involves the combination of a holographic diffraction grating and a hypercomatic focusing objective.


Physical Activity and Sex Hormone-Binding Globulin in Older Adults.
Besides its known function as a transport protein for testosterone and other steroid hormones, sex hormone-binding globulin (SHBG) is a biomarker associated with many adverse health effects. The aim of this study was to investigate the association of physical activity with SHBG serum levels in older adults. The physical activity and SHBG values for 1,259 older adults (43.4% female; 56.6% male) with a mean age of 75.6 ± 6.5 years were included in the analysis. The average daily walking duration was 104.2 ± 40.4 (mean ± SD) min. A positive dose-response relationship of daily walking duration with quartiles of SHBG was seen after adjustment for age, sex, history of cardiovascular diseases, diabetes, smoking, γ-glutamyl transferase, and C-reactive protein (p for trend = .010). However, this relationship disappeared after adjustment for body mass index (p for trend = .977). Body mass index seems to be an important determinant of SHBG and a possible confounding factor in the relationship of physical activity and SHBG.


The Rising Challenge of Training Physician-Scientists: Recommendations From a Canadian National Consensus Conference.
Physician-scientists are individuals who actively participate in patient care, have undergone additional research training, and devote the majority of their time to research. Physician-scientists are traditionally the primary catalysts in bridging the translational gap-that is, the failure to link fundamental new knowledge in the pathobiology of disease with advances in health care and health policy in a timely manner. However, there has been a shift away from training physician-scientists, and financial support for the physician-scientist is diminishing globally, causing the translational gap to grow. Given its socialized health care system and cultural and geographic diversity, Canada can serve as a unique case study in understanding how to address this phenomenon as a national priority. To this end, a Canadian national consensus conference was convened to develop recommendations for training programs and early-career supports for physician-scientists. Five recommendations were generated: (1) Establish an independent, national council whose mandate is to provide pan-Canadian oversight of physician-scientist training programs; (2) develop capacity for funding and mentorship support for physician-scientists; (3) develop coherent networks across a broad range of clinician-scientists, including physician-scientists, to reflect the unique cultural and geographic diversity of Canada and to reflect the interdisciplinarity of health research; (4) ensure that medical school curricula integrate, as a core curriculum feature, an understanding of the scientific basis of health care, including research methodologies; and (5) ensure that the funding of the physician-scientist trainee is viewed as portable and distinct from the operational funding provided to the training program itself.


Unique clinical characteristics, autoantibodies and medication use in Native American patients with systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with varied morbidity and mortality. We assessed clinical presentations, autoantibody specificities and therapeutic interventions in Native American (NA) patients with SLE. Patients with SLE meeting 1997 American College of Rheumatology classification criteria (n=3148) were enrolled between 1992 and 2010 in the multiethnic, cross-sectional Lupus Family Registry and Repository. Clinical, demographic and therapeutic information were extracted from medical records using a standardised form and formalised training. Autoantibodies were assessed by indirect immunofluorescence (antinuclear antibodies (ANA) and antidouble-stranded DNA), precipitin (ENA) and ELISA (IgG and IgM anticardiolipins). NA patients met SLE classification at a younger age (29.89±12.3 years) than European Americans (EA; 32.02±12.87, P=0.0157) and a similar age to African-Americans (AAs) and Hispanics (HIS). More NA patients had concurrent rheumatic diseases or symptoms, such as Raynaud's phenomenon, interstitial lung disease, Sjӧgren's syndrome and systemic sclerosis. Compared with EAs, NAs were more likely to have high-titre ANA (≥1:3240; P<0.0001) and had more SLE-associated autoantibodies. Autoantibodies with unknown specificities were more common in NAs (41%) compared with other racial/ethnic groups in this collection (AA: 24%, P=0.0006; EA: 17%, P<0.0001; HIS: 23%, P=0.0050). Fewer NA patients used hydroxychloroquine (68%) compared with others (AA: 74%, P=0.0308; EA: 79%, P=0.0001, HIS: 77%, P=0.0173); this was influenced by lower hydroxychloroquine use in NA patients from Latin America (32%). NA patients had higher rates of methotrexate use (28%) compared with AA (18%, P=0.0006) and HIS patients (14%, P=0.0003), higher azathioprine use (38%) compared with EA patients (30%, P=0.0105) and higher mycophenolate mofetil use (26%) compared with EA (17%, P=0.0012) and HIS patients (11%, P<0.0001). NA patients are diagnosed with SLE earlier in life and present worse concurrent rheumatic disease symptoms than EA patients. NA patients also are more likely to have expanded autoantibody profiles and precipitins of unknown specificities.


The Prospective Urban Rural Epidemiology (PURE) study: PURE Turkey.
The Prospective Urban Rural Epidemiology (PURE) study is a prospective cohort study that collects data on social, environmental, and individual risk factors and chronic diseases among residents of 25 countries in the range of 35 to 70 years of age, living in rural and urban areas. The PURE study is directed by the Population Health Research Institute of McMaster University in Canada. In Turkey, the study is conducted by the Metabolic Syndrome Society. In Turkey, the study is being conducted in 8 cities. The initial fieldwork began in 2008. Questionnaires were completed, and anthropometric measurements, blood and urine samples, handgrip strength evaluations, electrocardiogram readings, and spirometer and body composition measurements were obtained. Each year, participants were followed up via telephone. Every third year, questionnaires, field measurements, and biological data sampling were repeated. PURE Turkey has 4056 participants (female: 60.7%, male: 39.3%; mean age: 50±9.1 years). Among them, 43.9% had metabolic syndrome and 52.8% were obese. The prevalence of hypertension was 41.1% and proportion of controlled hypertension was 34%. A total of 2098 (51.7%) of the participants had a total cholesterol of ≥200 mg/dL or were using a lipid lowering agent. In patients with diabetes, 79.8% had low-density lipoprotein cholesterol levels ≥100 mg/dL. Although a dramatic change was not observed in those parameters in the follow-up years, the prevalence of diabetes mellitus increased from 13.7% in 2008 to 21% in 2015. The baseline and follow-up data of the PURE study were analyzed with the other countries participating in the study and reported for international publication. The PURE study is a large, ongoing, prospective epidemiological study that is investigating the "causes of the causes" of noncommunicable diseases in the world. In addition to revealing the health status of nations, the study also has the potential to affect health politics.