Crystal Structure of an Ammonia-Permeable Aquaporin.
Aquaporins of the TIP subfamily (Tonoplast Intrinsic Proteins) have been suggested to facilitate permeation of water and ammonia across the vacuolar membrane of plants, allowing the vacuole to efficiently sequester ammonium ions and counteract cytosolic fluctuations of ammonia. Here, we report the structure determined at 1.18 Å resolution from twinned crystals of Arabidopsis thaliana aquaporin AtTIP2;1 and confirm water and ammonia permeability of the purified protein reconstituted in proteoliposomes as further substantiated by molecular dynamics simulations. The structure of AtTIP2;1 reveals an extended selectivity filter with the conserved arginine of the filter adopting a unique unpredicted position. The relatively wide pore and the polar nature of the selectivity filter clarify the ammonia permeability. By mutational studies, we show that the identified determinants in the extended selectivity filter region are sufficient to convert a strictly water-specific human aquaporin into an AtTIP2;1-like ammonia channel. A flexible histidine and a novel water-filled side pore are speculated to deprotonate ammonium ions, thereby possibly increasing permeation of ammonia. The molecular understanding of how aquaporins facilitate ammonia flux across membranes could potentially be used to modulate ammonia losses over the plasma membrane to the atmosphere, e.g., during photorespiration, and thereby to modify the nitrogen use efficiency of plants.


EANM/EARL harmonization strategies in PET quantification: from daily practice to multicentre oncological studies.
Quantitative positron emission tomography/computed tomography (PET/CT) can be used as diagnostic or prognostic tools (i.e. single measurement) or for therapy monitoring (i.e. longitudinal studies) in multicentre studies. Use of quantitative parameters, such as standardized uptake values (SUVs), metabolic active tumor volumes (MATVs) or total lesion glycolysis (TLG), in a multicenter setting requires that these parameters be comparable among patients and sites, regardless of the PET/CT system used. This review describes the motivations and the methodologies for quantitative PET/CT performance harmonization with emphasis on the EANM Research Ltd. (EARL) Fluorodeoxyglucose (FDG) PET/CT accreditation program, one of the international harmonization programs aiming at using FDG PET as a quantitative imaging biomarker. In addition, future accreditation initiatives will be discussed. The validation of the EARL accreditation program to harmonize SUVs and MATVs is described in a wide range of tumor types, with focus on therapy assessment using either the European Organization for Research and Treatment of Cancer (EORTC) criteria or PET Evaluation Response Criteria in Solid Tumors (PERCIST), as well as liver-based scales such as the Deauville score. Finally, also presented in this paper are the results from a survey across 51 EARL-accredited centers reporting how the program was implemented and its impact on daily routine and in clinical trials, harmonization of new metrics such as MATV and heterogeneity features.


An enlarged postcranial sample confirms Australopithecus afarensis dimorphism was similar to modern humans.
In a previous study, we introduced the template method as a means of enlarging the Australopithecus afarensis postcranial sample to more accurately estimate its skeletal dimorphism. Results indicated dimorphism to be largely comparable to that of Homo sapiens. Some have since argued that our results were biased by artificial homogeneity in our Au. afarensis sample. Here we report the results from inclusion of 12 additional, newly reported, specimens. The results are consistent with those of our original study and with the hypothesis that early hominid demographic success derived from a reproductive strategy involving male provisioning of pair-bonded females.


Small is challenging; distal femur fracture management in an elderly lady with achondroplastic dwarfism.
Achondroplasia is an autosomal dominant dwarfing condition that represents the most common form of skeletal dysplasia. The disease is caused by a mutation in the gene encoding fibroblast growth factor receptor 3 (FGFR3) found at the p16.3 locus on chromosome 4 which results in severe inhibition of subchondral bone growth. Anatomic variations, including articular and periarticular deformities classically seen with this condition, amplify the complexity of fracture fixation. Blood volume loss, age-related bone fragility, component selection and positioning pose serious orthopaedic challenges. Concomitant cardiovascular, respiratory and neurological comorbidities pose additional high-risk perioperative considerations. Despite an estimated prevalence of 1:25 000 in the general population, there is little literature concerning the operative and postoperative treatment challenges faced by orthopaedic surgeons dealing with fracture management on a patient with this condition. We present a case of an intercondylar femoral fracture in an elderly achondroplastic lady successfully treated with percutaneous screw fixation.


Equimolar doses of mannitol and hypertonic saline in the treatment of increased intracranial pressure.
To compare the effects of equimolar doses of 20% mannitol solution and of 7.45% hypertonic saline solution (HSS) in the treatment of patients with sustained elevated intracranial pressure (ICP). Parallel, randomized, controlled trial. Two intensive care units in a university hospital. A total of 20 stable patients with a sustained ICP of >20 mm Hg secondary to traumatic brain injury (n = 17) or stroke (n = 3). A single equimolar infusion (255 mOsm dose) of either 231 mL of 20% mannitol (mannitol group; n = 10 patients) or 100 mL of 7.45% hypertonic saline (HSS group; n = 10 patients) during 20 mins of administration. ICP, arterial blood pressure, cerebral perfusion pressure, blood flow velocities of middle cerebral artery using continuous transcranial Doppler, brain tissue oxygen tension, serum sodium and osmolality, and urine output during a study period of 120 mins. The two treatments equally and durably reduced ICP during the experiment. At 60 mins after the start of the infusion, ICP was reduced by 45% +/- 19% of baseline values (mean +/- sd) in the mannitol group vs. 35% +/- 14% of baseline values in the HSS group. Cerebral perfusion pressure and diastolic and mean blood flow velocities were durably increased in the mannitol group, resulting in lower values of pulsatility index at the different times of the experiment (p < .01 vs. HSS). No major changes in brain tissue oxygen tension were found after each treatment. Mannitol caused a significantly greater increase in urine output (p < .05) than HSS, although there was no difference in the vascular filling requirement between the two treatments. HSS caused a significant elevation of serum sodium and chloride at 120 mins after the start of the infusion (p < .01). A single equimolar infusion of 20% mannitol is as effective as 7.45% HSS in decreasing ICP in patients with brain injury. Mannitol exerts additional effects on brain circulation through a possible improvement in blood rheology. Pretreatment factors, such as serum sodium, systemic hemodynamics, and brain hemodynamics, thus should be considered when choosing between mannitol and HSS for patients with increased ICP.


Salicylate-induced degeneration of cochlea spiral ganglion neurons-apoptosis signaling.
Aspirin, whose active ingredient is sodium salicylate, is the most widely used drug worldwide, but it is not recommended for children because it may cause Reye's syndrome. High doses of salicylate also induce temporary hearing loss and tinnitus; while these disorders are believed to disappear when treatment is discontinued some data suggest that prolonged treatment may be neurotoxic. To investigate its ototoxicity, immature, postnatal day 3 rat cochlear organotypic cultures were treated with salicylate. Salicylate did not damage the sensory hair cells, but instead damaged the spiral ganglion neurons (SGN) and their peripheral fibers in a dose-dependent manner. The cross-sectional area of SGN decreased from 205 microm(2) in controls to 143, 116, and 91 microm(2) in cultures treated with 1, 3, or 5 mM salicylate, respectively. Morphological changes and caspase upregulation were indicative of caspase-mediated apoptosis. A quantitative RT-PCR apoptosis array identified a subset of genes up- or down regulated by salicylate. Eight genes showed a biologically relevant change (P<0.05, > or =2 fold change) after 3 h treatment with salicylate; seven genes (Tp53, Birc3, Tnfrsf5, Casp7, Nfkb1, Fas, Lta, Tnfsf10) were upregulated and one gene (Pycard) was downregulated. After 6 h treatment, only one gene (Nol3) was upregulated and two genes were downregulated (Cideb and Lhx4) while after 12 h treatment, two genes (Il10, Gadd45a) were upregulated and 4 (Prok2, Card10, Ltbr, Dapk1) were downregulated. High doses of salicylate in a physiologically relevant range can induce caspase-mediated cell death in immature SGN; changes in the expression of apoptotic genes particularly among members of the tumor necrosis factor (TNF) family appear to play an important role in the degeneration.


Vacuum-assisted closure in the treatment of early hip joint infections.
The aim of the present study was to evaluate the efficacy of the vacuum-assisted closure (V.A.C.) system in the treatment of early hip joint infections. 28 patients (11 m/17 f; mean age 71 y. [43-84]) with early hip joint infections have been treated by means of the V.A.C.-therapy. At least one surgical revision [1-7] has been unsuccessfully performed for infection treatment prior to V.A.C.-application. Pathogen organisms could have been isolated in 22/28 wounds. During revision, cup inlay and prosthesis head have been exchanged and 1-3 polyvinylalcohol sponges inserted into the wound cavity/ periprosthetically at an initial continuous pressure of 200 mm Hg. Postoperatively, a systemic antibiosis was given according to antibiogram. 48-72 h after surgery an alteration from haemorrhagic to serous fluid was observed in the V.A.C.-canister. Afterwards, the pressure was decreased to 150 mm Hg and remained at this level till sponge removal. After a mean period of 9 [3-16] days the inflammation parameters have been retrogressive and the sponges were removed. An infection eradication could be achieved in 26/28 cases. In the two remaining cases the infected prosthesis had to be explanted and a gentamicin-vancomycin-loaded spacer has been implanted, respectively. At a total mean follow-up of 36 [12-87] months no reinfection or infection persistence was observed. The V.A.C.-system can be a valuable contribution in the treatment of early joint infections when properly used. Indications should be early infections with well-maintained soft-tissues for retention of the negative atmospheric pressure.


Prevalence and risk factors of anterior atlantoaxial subluxation in ankylosing spondylitis.
In ankylosing spondylitis (AS), the cervical spine, like other sections of the spine and sacroiliac joints, is vulnerable during the disease process. Atlantoaxial subluxation (AAS) has been studied in connection with AS, but its risk factors and progression have not been clarified. Therefore, this study assessed the prevalence and risk factors of AAS in patients with AS. A total of 819 patients with AS who fulfilled the modified New York criteria and were examined with a full-flexion lateral view of the cervical spine by radiograph were enrolled from an outpatient clinic. The medical records of the patients were retrospectively reviewed and the anterior atlantodental interval (AADI) in the lateral flexion view of the cervical spine radiograph was investigated by 2 experienced musculoskeletal radiologists. We defined the AAS as an AADI of > 3 mm, and progression of AADI as a progression rate > 0.5 mm/year. AAS was found in 14.1% (116/819) of patients. Progression of AADI occurred in 32.1% (26/81) patients with AAS and 5.0% (16/320) patients without AAS (p < 0.001). The development of AAS was significantly associated with elevated C-reactive protein [CRP; OR 2.19 (1.36-3.53)], peripheral arthritis [OR 2.05 (1.36-3.07)], use of anti-tumor necrosis factor antagonists because of failure of nonsteroidal antiinflammatory drugs/disease-modifying antirheumatic drugs [NSAID/DMARD; OR 2.28 (1.52-3.42)], and uveitis [OR 1.71 (1.13-2.59)]. These factors were adjusted for age, sex, and disease duration by logistic regression analysis. No clear association was found for HLA-B27, seropositivity, or smoking status with AAS. AAS is a frequent complication, and the progression of AADI was more rapid in cases with AAS. The presence of peripheral arthritis, or high disease activity with elevated CRP level or refractory to conventional NSAID/DMARD, independently increased the risk of AAS, suggesting that clinicians should focus on the detection and monitoring of AAS, especially in cases with associated risk factors.


The Clinical Relevance of the IBD-Associated Variation within the Risk Gene Locus Encoding Protein Tyrosine Phosphatase Non-Receptor Type 2 in Patients of the Swiss IBD Cohort.
The single nucleotide polymorphism (SNP) rs1893217 within the gene locus encoding protein tyrosine phosphatase non-receptor type 2 (PTPN2) results in a dysfunctional PTPN2 protein is associated with Crohn's disease (CD) and exists in perfect linkage disequilibrium with the CD- and ulcerative colitis (UC)-associated PTPN2 SNP rs2542151. We investigated associations of PTPN2 SNP rs1893217 and clinical characteristics of inflammatory bowel disease (IBD) patients. One thousand seventy three patients with CD and 734 patients with UC from the Swiss IBD Cohort Study (SIBDCS) were included. Epidemiologic, disease and treatment characteristics were analysed for an association with the presence of one of the rs1893217 genotypes 'homozygous wild-type' (TT), 'heterozygous' (CT) and 'homozygous variant' (CC). About 2.88% of IBD patients were identified with CC, 26.8% with CT and 70.4% with TT genotype. The CC-genotype was associated with the existence of gallstones in CD and pancolitis in UC patients. The presence of the C-allele (i.e. either CC or CT genotype) was associated with the onset of uveitis, but protected from aphthous oral ulcers in CD patients. UC patients carrying a C-allele were diagnosed at an older age but required intestinal surgery more often. The presence of the C-allele was associated with a successful treatment with anti-TNF antibodies in both CD and UC patients. IBD patients carrying the C-allele of PTPN2 SNP rs1893217 are at greater risk for developing a severe disease course but are more likely to respond to treatment with anti-TNF antibodies. These findings demonstrate a clinical relevance of this PTPN2 risk variant in IBD patients.


Depressed intracellular calcium transients and contraction in myocytes from hypertrophied and failing guinea pig hearts.
We investigated the basis for impaired left ventricular function of hearts in which hypertrophy was produced by gradual pressure overload. We measured myoplasmic free calcium concentration ([Ca2+]i) with fura-2 and sarcomere shortening in single myocytes isolated from control hearts and hypertrophied failing hearts. Diastolic [Ca2+]i was normal, but [Ca2+]i at the peak of contraction was depressed in myocytes from failing hypertrophied hearts. Increasing drive rate from 0.20 Hz to 5.00 Hz increased both diastolic and peak [Ca2+]i. Norepinephrine (3 x 10(-6) M) increased diastolic [Ca2+]i in all cells and tended to normalize peak [Ca2+]i in myocytes from hypertrophied failing hearts during 5.00 Hz drive. Depressed peak [Ca2+]i in the hypertrophied cells was paralleled by significant decreases in both the velocity and percent of sarcomere shortening, which were measured in cells not loaded with fura-2. Sarcomere length was correlated with estimates of [Ca2+]i in intact cells and with controlled levels of [Ca2+] in chemically "skinned" myocytes. A plot of sarcomere length against [Ca2+] gave a single continuous relationship that spanned resting and peak values at all drive rates in both the control and hypertrophied myocytes. Thus heart failure in this model is reflected in impaired myocyte contraction, which is closely related to reduced levels of [Ca2+]i during systole rather than to depressed myofilament sensitivity to Ca2+.


Mitochondrial genomics of gadine fishes: implications for taxonomy and biogeographic origins from whole-genome data sets.
Phylogenetic analysis of 13 substantially complete mitochondrial DNA genome sequences (14,036 bp) from 10 taxa of gadine codfishes and pollock provides highly corroborated resolution of outstanding questions on their biogeographic evolution. Of 6 resolvable nodes among species, 4 were supported by >95% of bootstrap replications in parsimony, distance, likelihood, and similarly high posterior probabilities in bayesian analyses, one by 85%-95% according to the method of analysis, and one by 99% by one method and a majority of the other two. The endemic Pacific species, walleye pollock (Theragra chalcogramma), is more closely related to the endemic Atlantic species, Atlantic cod (Gadus macrocephalus), than either is to a second Pacific endemic, Pacific cod (Gadus macrocephalus). The walleye pollock should thus be referred to the genus Gadus as originally described (Gadus chalcogrammus Pallas 1811). Arcto-Atlantic Greenland cod, previously regarded as a distinct species (G. ogac), are a genomically distinguishable subspecies within pan-Pacific G. macrocephalus. Of the 2 endemic Arctic Ocean genera, Polar cod (Boreogadus) as the outgroup to Arctic cod (Arctogadus) and Gadus sensu lato is more strongly supported than a pairing of Boreogadus and Arctogadus as sister taxa. Taking into consideration historical patterns of hydrogeography, we outline a hypothesis of the origin of the 2 endemic Pacific species as independent but simultaneous invasions through the Bering Strait from an Arcto-Atlantic ancestral lineage. In contrast to the genome data, the complete proteome sequence (3830 amino acids) resolved only 3 nodes with >95% confidence, and placed Alaska pollock outside the Gadus clade owing to reversal mutations in the ND5 locus that restore ancestral, non-Gadus, amino acid residues in that species.


Sonographic location of the radial nerve confirmed by nerve stimulation during axillary brachial plexus blockade.
: Visualization of the radial nerve can be a challenge during ultrasound-guided axillary blockade. The objective of this study was to plot the location and examine the sonographic appearance of the radial nerve in the axilla using both ultrasound visualization and nerve stimulator verification in patients undergoing axillary blockade. : Fifty-one patients requiring an axillary block for surgery were enrolled. Sonograms of the radial nerve at the point of best nerve stimulator response were recorded. The needle tip and center of the radial nerve were both plotted on a 2-dimensional scattergram using coordinates relative to the center of the axillary artery. A video tracing the path of the radial nerve was also recorded. : The radial nerve location was confirmed with both ultrasound imaging and nerve stimulation in 46 patients (90%). Five nerves (11%) were immediately deep to the axillary artery (ie, at a 6-o'clock position), 6 (13%) were cephalad, and the remaining 35 (76%) were caudad to the axillary artery. The 2-dimensional (2D) scattergrams reveal the variability in radial nerve location and clarity, of which 14 (30%) of the 46 visible nerves were difficult to trace precisely. A fascial plane between the medial and long heads of the triceps containing the radial nerve could be imaged clearly in 41 (91%) of the videos. : The radial nerve uncommonly lies immediately deep to the axillary artery. It is frequently located on the caudad side of the artery.


Thromboembolism with Janus Kinase (JAK) Inhibitors for Rheumatoid Arthritis: How Real is the Risk?
Two different Janus kinase (JAK) inhibitors-baricitinib and tofacitinib-are effective and licensed in active rheumatoid arthritis (RA). There have been recent concerns about potential thromboembolic risks with these drugs. Concerns about baricitinib focus on clinical trial findings. Using all publically available data, we estimate thromboembolic risks are approximately five events per 1000 patient years with 4 mg baricitinib daily. Concerns about tofacitinib have been raised by analyses of the Federal Drug Administration Adverse Event Reporting System (FAERs). These show some evidence of increased risks of pulmonary thrombosis, though not pulmonary embolism or venous thrombosis. Observational studies suggest in the general population and non-RA controls there are one to four thromboembolic events per 1000 patient years. In RA, thromboembolic risks increase to three to seven per 1000 patient years. The impact of biologics and disease-modifying anti-rheumatic drugs (DMARDs) on disease risk appears minimal, and the number of thromboembolic events is between four and eight per 1000 patient years. In the short term, full details of thromboembolic events in trials of JAK inhibitors need to be published. As the numbers of thromboembolic events will be small and patients enrolled in trials are not representative of all RA patients who may receive JAK inhibitors, this information is unlikely to provide definitive answers. Consequently, in the longer term, large observational studies are needed to accurately quantify thromboembolic risks attributable to JAK inhibitors and other drugs used to treat RA, and differentiate these from risks attributable to RA itself and its comorbidities.


MALDI-TOF and nESI Orbitrap MS/MS identify orthogonal parts of the phosphoproteome.
Phosphorylation is a reversible posttranslational protein modification which plays a pivotal role in intracellular signaling. Despite extensive efforts, phosphorylation site mapping of proteomes is still incomplete motivating the exploration of alternative methods that complement existing workflows. In this study, we compared tandem mass spectrometry (MS/MS) on matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) and nano-electrospray ionization (nESI) Orbitrap instruments with respect to their ability to identify phosphopeptides from complex proteome digests. Phosphopeptides were enriched from tryptic digests of cell lines using Fe-IMAC column chromatography and subjected to LC-MS/MS analysis. We found that the two analytical workflows exhibited considerable orthogonality. For instance, MALDI-TOF MS/MS favored the identification of phosphopeptides encompassing clear motif signatures for acidic residue directed kinases. The extent of orthogonality of the two LC-MS/MS systems was comparable to that of using alternative proteases such as Asp-N, Arg-C, chymotrypsin, Glu-C and Lys-C on just one LC-MS/MS instrument. Notably, MALDI-TOF MS/MS identified an unexpectedly high number and percentage of phosphotyrosine sites (∼20% of all sites), possibly as a direct consequence of more efficient ionization. The data clearly show that LC-MALDI MS/MS can be a useful complement to LC-nESI MS/MS for phosphoproteome mapping and particularly so for acidic and phosphotyrosine containing peptides.


Inflammatory profiles across the spectrum of disease reveal a distinct role for GM-CSF in severe COVID-19.
While it is now widely accepted that host inflammatory responses contribute to lung injury, the pathways that drive severity and distinguish coronavirus disease 2019 (COVID-19) from other viral lung diseases remain poorly characterized. We analyzed plasma samples from 471 hospitalized patients recruited through the prospective multicenter ISARIC4C study and 39 outpatients with mild disease, enabling extensive characterization of responses across a full spectrum of COVID-19 severity. Progressive elevation of levels of numerous inflammatory cytokines and chemokines (including IL-6, CXCL10, and GM-CSF) were associated with severity and accompanied by elevated markers of endothelial injury and thrombosis. Principal component and network analyses demonstrated central roles for IL-6 and GM-CSF in COVID-19 pathogenesis. Comparing these profiles to archived samples from patients with fatal influenza, IL-6 was equally elevated in both conditions whereas GM-CSF was prominent only in COVID-19. These findings further identify the key inflammatory, thrombotic, and vascular factors that characterize and distinguish severe and fatal COVID-19.


Advanced dementia: state of the art and priorities for the next decade.
Dementia is a leading cause of death in the United States. This article outlines the current understanding of advanced dementia and identifies research priorities for the next decade. Research over the past 25 years has largely focused on describing the experience of patients with advanced dementia. This work has delineated abundant opportunities for improvement, including greater recognition of advanced dementia as a terminal illness, better treatment of distressing symptoms, increased access to hospice and palliative care services, and less use of costly and aggressive treatments that may be of limited clinical benefit. Addressing those opportunities must be the overarching objective for the field in the coming decade. Priority areas include designing and testing interventions that promote high-quality, goal-directed care; health policy research to identify strategies that incentivize cost-effective and evidence-based care; implementation studies of promising interventions and policies; and further development of disease-specific outcome measures. There is great need and opportunity to improve outcomes, contain expenditures, reduce disparities, and better coordinate care for the millions of persons in the United States who have advanced dementia.


Ask versus tell: Potential confusion when child witnesses are questioned about conversations.
Children's potential confusion between "ask" and "tell" can lead to misunderstandings when child witnesses are asked to report prior conversations. The verbs distinguish both between interrogating and informing, and between requesting and commanding. Children's understanding was examined using both field (Study 1) and laboratory methods (Studies 2-4). Study 1 examined 100 5- to 12-year-olds' trial testimony in child sexual abuse cases, and found that potentially ambiguous use of ask and tell was common, typically found in yes-no questions that elicited unelaborated answers, and virtually never clarified by attorneys or child witnesses. Studies 2 to 4 examined 345 maltreated 6- to 11-year-olds' understanding of ask and tell. The results suggest that children initially comprehend telling as saying, and thus believed that asking is a form of telling. As such, they often endorsed asking as telling when asked yes-no questions, but distinguished between asking and telling when explicitly asked to choose. Their performance was impaired by movement between different use of the words. Child witnesses' characterization of their conversations can easily be misconstrued by the way in which they are questioned, leading questioners to misinterpret whether they were coached by disclosure recipients or coerced by abuse suspects. (PsycINFO Database Record


Ethnobotany of the Monpa ethnic group at Arunachal Pradesh, India.
The present paper documents the uses of plants in traditional herbal medicine for human and veterinary ailments, and those used for dietary supplements, religious purpose, local beverage, and plants used to poison fish and wild animals. Traditional botanical medicine is the primary mode of healthcare for most of the rural population in Arunachal Pradesh. Field research was conducted between April 2006 and March 2009 with randomly selected 124 key informants using semi-structured questionnaire. The data obtained was analyzed through informant consensus factor (FIC) to determine the homogeneity of informant's knowledge on medicinal plants. We documented 50 plants species belonging to 29 families used for treating 22 human and 4 veterinary ailments. Of the medicinal plants reported, the most common growth form was herbs (40%) followed by shrubs, trees, and climbers. Leaves were most frequently used plant parts. The consensus analysis revealed that the dermatological ailments have the highest FIC (0.56) and the gastro-intestinal diseases have FIC (0.43). FIC values indicated that there was high agreement in the use of plants in dermatological and gastro-intestinal ailments category among the users. Gymnocladus assamicus is a critically rare and endangered species used as disinfectant for cleaning wounds and parasites like leeches and lice on livestocks. Two plant species (Illicium griffithii and Rubia cordifolia) are commonly used for traditional dyeing of clothes and food items. Some of the edible plants recorded in this study were known for their treatment against high blood pressure (Clerodendron colebrookianum), diabetes mellitus (Momordica charantia), and intestinal parasitic worms like round and tape worms (Lindera neesiana, Solanum etiopicum, and Solanum indicum). The Monpas of Arunachal Pradesh have traditionally been using Daphne papyracea for preparing hand-made paper for painting and writing religious scripts in Buddhist monasteries. Three plant species (Derris scandens, Aesculus assamica, and Polygonum hydropiper) were frequently used to poison fish during the month of June-July every year and the underground tuber of Aconitum ferrox is widely used in arrow poisoning to kill ferocious animals like bear, wild pigs, gaur and deer. The most frequently cited plant species; Buddleja asiatica and Hedyotis scandens were used as common growth supplements during the preparation of fermentation starter cultures. The traditional pharmacopoeia of the Monpa ethnic group incorporates a myriad of diverse botanical flora. Traditional knowledge of the remedies is passed down through oral traditions without any written document. This traditional knowledge is however, currently threatened mainly due to acculturation and deforestation due to continuing traditional shifting cultivation. This study reveals that the rural populations in Arunachal Pradesh have a rich knowledge of forest-based natural resources and consumption of wild edible plants is still an integral part of their socio-cultural life. Findings of this documentation study can be used as an ethnopharmacological basis for selecting plants for future phytochemical and pharmaceutical studies.


Towards an open science publishing platform.
The way science and research is done is rapidly becoming more open and collaborative. The traditional way of publishing new findings in journals is becoming increasingly outdated and no longer serves the needs of much of science. Whilst preprints can bring significant benefits of removing delay and selection, they do not go far enough if simply implemented alongside the existing journal system. We propose that we need a new approach, an Open Science Platform, that takes the benefits of preprints but adds formal, invited, and transparent post-publication peer review. This bypasses the problems of the current journal system and, in doing so, moves the evaluation of research and researchers away from the journal-based Impact Factor and towards a fairer system of article-based qualitative and quantitative indicators. In the long term, it should be irrelevant where a researcher publishes their findings. What is important is that research is shared and made available without delay within a framework that encourages quality standards and requires all players in the research community to work as collaborators.


"Do-it-yourself": Vaccine rejection and complementary and alternative medicine (CAM).
In this article, we elucidate a symbiotic relationship between complementary and alternative medicine (CAM) and rejection of, or hesitancy towards, vaccination. In Fremantle, Western Australia, and Adelaide, South Australia, we conducted in-depth interviews from September 2013-December 2015 with 29 parents who had refused or delayed some or all of their children's vaccines. Our qualitative analysis found that for many, their do-it-yourself ethic and personal agency was enhanced by self-directed CAM use, alongside (sometimes informal) CAM practitioner instruction. Reifying 'the natural,' these parents eschewed vaccines as toxic and adulterating, and embraced CAM as a protective strategy for immune systems before, during and after illness. Users saw CAM as harm-free, and when it came to experiences that non-users might interpret as demonstrating CAM's ineffectiveness, they rationalised to the contrary. They also generally glossed over its profit motive. CAM emerged as part of an expert system countering Western medicine. CAM's faces were trusted and familiar, and its cottage capitalism appeared largely free from the taint of "Big Pharma." A few parents employed a scientific critique of CAM modalities - and a minority were dubious of its profit motive - but others rejected the epistemology underpinning biomedicine, framing CAM as a knowledge not poisoned by avarice; a wisdom whose very evidence-base (anecdote and history) was demeaned by an arrogant scientific process only permitting belief in that which could be quantified. However, all parents engaged with Western medicine for broken bones and, sometimes, medical diagnoses. Our analysis suggests that pro-vaccination health professionals, policymakers and information-providers seeking to address the role of CAM in vaccine rejection face significant challenges due to the epistemic basis of some parents' decisions. However, we make some suggestions for professional practice and policy to enhance trust in vaccination.


The first peripherally masked thiol dendrimers: a facile and highly efficient functionalization strategy of polyester dendrimers via one-pot xanthate deprotection/thiol-acrylate Michael addition reactions.
Introducing multiple reactive functional groups at the periphery of dendrimer materials presents considerable challenges if the functionality is able to self-react. An efficient and facile approach to introducing masked thiols at the surface of polyester dendrimers is presented. One-pot, deprotection/thiol-acrylate Michael addition from the xanthate-functional dendritic substrates (generation zero to two) has been achieved for the first time, with high efficiency demonstrated using three acrylates of varying chemistry and avoiding disulfide formation.


Maternal Tobacco and Alcohol Use in Relation to Child Malnutrition in Gauteng, South Africa: A Retrospective Analysis.
Tobacco and alcohol use among mothers is associated with numerous adverse consequences for affected offspring, including poor growth and development. This study determined the association between maternal tobacco and alcohol use, and malnutrition, among infants aged ≤ 12 months (n = 300), in selected health facilities situated in Gauteng, South Africa. Data on alcohol and tobacco use were collected using a validated questionnaire, in addition to mothers' socio-demographic characteristics and obstetric history. Stunting (low height/length-for-age), underweight (low weight-for-age) and thinness (low body mass index-for-age) were calculated using z-scores based on the World Health Organization child growth standards. The association of tobacco and alcohol use with stunting, underweight and thinness was analysed using logistic regression analysis. The results showed a mean age of 29 years (24.0; 35.0) for mothers and 7.6 ± 3 months for infants, and over half of the mothers were unemployed (63%). Approximately 18.7% of mothers had used tobacco and 3% had used alcohol during pregnancy. The prevalence of current tobacco and alcohol use among mothers were estimated at 14.3% and 49.7%, respectively, and almost three-quarters (67.3%) of them were still breastfeeding during the study period. Stunting (55%) was the most prevalent malnutrition indicator among infants, while underweight was 41.7%, and thinness was 22%. Current tobacco use was associated with increased odds of being thin [OR = 2.40, 95% CI: 1.09-5.45), and after adjusting for confounders, current alcohol use was associated with the likelihood of being underweight [AOR = 1.96, 95% CI: 1.06-3.63] among infants. Future prospective cohort studies that examine growth patterns among infants who are exposed to maternal tobacco and alcohol use from the intrauterine life to infancy are necessary to inform, partly, the public health programmes, to reduce malnutrition among children.


Diffusion weighted imaging evidence of extra-callosal pathways for interhemispheric communication after complete commissurotomy.
The integrity of white matter architecture in the human brain is related to cognitive processing abilities. The corpus callosum is the largest white matter bundle interconnecting the two cerebral hemispheres. "Split-brain" patients in whom all cortical commissures have been severed to alleviate intractable epilepsy demonstrate remarkably intact cognitive abilities despite the lack of this important interhemispheric pathway. While it has often been speculated that there are compensatory alterations in the remaining interhemispheric fibers in split-brain patients several years post-commissurotomy, this has never been directly shown. Here we examined extra-callosal pathways for interhemispheric communication in the brain of a patient who underwent complete cerebral commissurotomy using diffusion weighted imaging tractography. We found that compared with a healthy age-matched comparison group, the split-brain patient exhibited increased fractional anisotropy (FA) of the dorsal and ventral pontine decussations of the cortico-cerebellar interhemispheric pathways. Few differences were observed between the patient and the comparison group with respect to FA of other long-range intrahemispheric fibers. These results point to specific cerebellar anatomical substrates that may account for the spared interhemispheric coordination and intact cognitive abilities that have been extensively documented in this unique patient.


Abuse liability assessment of tobacco products including potential reduced exposure products.
The harm produced by tobacco products is a result of frequent use of a highly toxic product. Reducing the adverse public health impact of tobacco products might be most effectively achieved by reducing the likelihood of their use and the toxicity of the products. Products that retain some characteristics of cigarettes but have been altered with the intention of reducing toxicity have been referred to as modified risk tobacco products or potential reduced exposure products (MRTP/PREP). Evaluation of their content, emission, and toxicity is discussed in other articles in this special issue. Here, we discuss the methodology that has been used to examine the likelihood of abuse or addiction. Abuse liability assessment (ALA) methodology has been used by the Food and Drug Administration (FDA) and other drug regulatory agencies world-wide for decades to assess the risks posed by a wide variety of pharmacologically active substances. ALA is routinely required among other evaluations of safety during the pre-market assessment of new drugs, and is continually adapted to meet the challenges posed by new drug classes and drug formulations. In the 2009 law giving FDA regulation over tobacco products, FDA is now required to evaluate new tobacco products including MRTP/PREPs to determine their risk for abuse and toxicity at the population level. This article describes the traditional tools and methods of ALA that can be used to evaluate new tobacco and nicotine products including MRTP/PREPs. Such ALA data could contribute to the scientific foundation on which future public policy decisions are based.


Embryogenesis of the first circulating endothelial cells.
Prior to this study, the earliest appearance of circulating endothelial cells in warm-blooded animals was unknown. Time-lapse imaging of germ-line transformed Tie1-YFP reporter quail embryos combined with the endothelial marker antibody QH1 provides definitive evidence for the existence of circulating endothelial cells - from the very beginning of blood flow. Blood-smear counts of circulating cells from Tie1-YFP embryos showed that up to 30% of blood-borne cells are Tie1 positive; though cells expressing low levels of YFP were also positive for benzidine, a hemoglobin stain, suggesting that these cells were differentiating into erythroblasts. Electroporation-based time-lapse experiments, exclusively targeting the intra-embryonic mesoderm were combined with QH1 immunostaining. The latter antibody marks quail endothelial cells. Together the optical data provide conclusive evidence that endothelial cells can enter blood flow from vessels of the embryo proper, as well as from extra-embryonic areas. When Tie1-YFP positive cells and tissues are transplanted to wild type host embryos, fluorescent cells emigrate from such transplants and join host vessels; subsequently a few YFP cells are shed into circulation. These data establish that entering circulation is a commonplace activity of embryonic vascular endothelial cells. We conclude that in the class of vertebrates most closely related to mammals a normal component of primary vasculogenesis is production of endothelial cells that enter circulation from all vessels, both intra- and extra-embryonic.


Contextual and Developmental Differences in the Neural Architecture of Cognitive Control.
Because both development and context impact functional brain architecture, the neural connectivity signature of a cognitive or affective predisposition may similarly vary across different ages and circumstances. To test this hypothesis, we investigated the effects of age and cognitive versus social-affective context on the stable and time-varying neural architecture of inhibition, the putative core cognitive control component, in a subsample (N = 359, 22-36 years, 174 men) of the Human Connectome Project. Among younger individuals, a neural signature of superior inhibition emerged in both stable and dynamic connectivity analyses. Dynamically, a context-free signature emerged as stronger segregation of internal cognition (default mode) and environmentally driven control (salience, cingulo-opercular) systems. A dynamic social-affective context-specific signature was observed most clearly in the visual system. Stable connectivity analyses revealed both context-free (greater default mode segregation) and context-specific (greater frontoparietal segregation for higher cognitive load; greater attentional and environmentally driven control system segregation for greater reward value) signatures of inhibition. Superior inhibition in more mature adulthood was typified by reduced segregation in the default network with increasing reward value and increased ventral attention but reduced cingulo-opercular and subcortical system segregation with increasing cognitive load. Failure to evidence this neural profile after the age of 30 predicted poorer life functioning. Our results suggest that distinguishable neural mechanisms underlie individual differences in cognitive control during different young adult stages and across tasks, thereby underscoring the importance of better understanding the interplay among dispositional, developmental, and contextual factors in shaping adaptive versus maladaptive patterns of thought and behavior.SIGNIFICANCE STATEMENT The brain's functional architecture changes across different contexts and life stages. To test whether the neural signature of a trait similarly varies, we investigated cognitive versus social-affective context effects on the stable and time-varying neural architecture of inhibition during a period of neurobehavioral fine-tuning (age 22-36 years). Younger individuals with superior inhibition showed distinguishable context-free and context-specific neural profiles, evidenced in both static and dynamic connectivity analyses. More mature individuals with superior inhibition evidenced only context-specific profiles, revealed in the static connectivity patterns linked to increased reward or cognitive load. Delayed expression of this profile predicted poorer life functioning. Our results underscore the importance of understanding the interplay among dispositional, developmental, and contextual factors in shaping behavior.


LRP1 controls intracellular cholesterol storage and fatty acid synthesis through modulation of Wnt signaling.
The low-density lipoprotein receptor-related protein LRP1 is a cell surface receptor with functions in diverse physiological pathways, including lipid metabolism. Here we show that LRP1-deficient fibroblasts accumulate high levels of intracellular cholesterol and cholesteryl-ester when stimulated for adipocyte differentiation. We demonstrate that LRP1 stimulates a canonical Wnt5a signaling pathway that prevents cholesterol accumulation. Moreover, we show that LRP1 is required for lipolysis and stimulates fatty acid synthesis independently of the noradrenergic pathway, through inhibition of GSK3beta and its previously unknown target acetyl-CoA carboxylase (ACC). As a result of ACC inhibition, mature LRP1-deficient adipocytes of adult mice are hypotrophic, and lower uptake of fatty acids into adipose tissue leads to their redistribution to the liver. These results establish LRP1 as a novel integrator of adipogenic differentiation and fat storage signals.


Result-driven exploration of simulation parameter spaces for visual effects design.
Graphics artists commonly employ physically-based simulation for the generation of effects such as smoke, explosions, and similar phenomena. The task of finding the correct parameters for a desired result, however, is difficult and time-consuming as current tools provide little to no guidance. In this paper, we present a new approach for the visual exploration of such parameter spaces. Given a three-dimensional scene description, we utilize sampling and spatio-temporal clustering techniques to generate a concise overview of the achievable variations and their temporal evolution. Our visualization system then allows the user to explore the simulation space in a goal-oriented manner. Animation sequences with a set of desired characteristics can be composed using a novel search-by-example approach and interactive direct volume rendering is employed to provide instant visual feedback. A user study was performed to evaluate the applicability of our system in production use.


Metabolic constraint-based refinement of transcriptional regulatory networks.
There is a strong need for computational frameworks that integrate different biological processes and data-types to unravel cellular regulation. Current efforts to reconstruct transcriptional regulatory networks (TRNs) focus primarily on proximal data such as gene co-expression and transcription factor (TF) binding. While such approaches enable rapid reconstruction of TRNs, the overwhelming combinatorics of possible networks limits identification of mechanistic regulatory interactions. Utilizing growth phenotypes and systems-level constraints to inform regulatory network reconstruction is an unmet challenge. We present our approach Gene Expression and Metabolism Integrated for Network Inference (GEMINI) that links a compendium of candidate regulatory interactions with the metabolic network to predict their systems-level effect on growth phenotypes. We then compare predictions with experimental phenotype data to select phenotype-consistent regulatory interactions. GEMINI makes use of the observation that only a small fraction of regulatory network states are compatible with a viable metabolic network, and outputs a regulatory network that is simultaneously consistent with the input genome-scale metabolic network model, gene expression data, and TF knockout phenotypes. GEMINI preferentially recalls gold-standard interactions (p-value = 10(-172)), significantly better than using gene expression alone. We applied GEMINI to create an integrated metabolic-regulatory network model for Saccharomyces cerevisiae involving 25,000 regulatory interactions controlling 1597 metabolic reactions. The model quantitatively predicts TF knockout phenotypes in new conditions (p-value = 10(-14)) and revealed potential condition-specific regulatory mechanisms. Our results suggest that a metabolic constraint-based approach can be successfully used to help reconstruct TRNs from high-throughput data, and highlights the potential of using a biochemically-detailed mechanistic framework to integrate and reconcile inconsistencies across different data-types. The algorithm and associated data are available at https://sourceforge.net/projects/gemini-data/


Key issues in the clinical development and implementation of TB vaccines in South Africa.
Significant progress has been made in advancing the development pipeline for a new and more effective TB vaccine with some candidate vaccines now in late stage clinical evaluation. However, progress has been hampered by an incomplete understanding of the components of a protective immune response and limited animal models, rendering the field unable to reliably predict vaccine efficacy earlier in preclinical development, including by evaluation in animal models, and limiting the predictive utility of comparing immunogenic effects across vaccine candidates in phase I/II studies. Consequently, new candidate vaccines have to be evaluated for efficacy in large-scale phase II/III trials using clinical endpoints. Apart from the technical challenges of characterising TB incidence in target populations at high risk of acquiring TB disease and standardising case definitions in order to improve both the sensitivity and more importantly the specificity of trial endpoints, there is an urgency in expanding and supporting the considerable trial infrastructure that will be required to evaluate and ultimately license a new TB vaccine. In the longer term, implementation strategies are dependent on what policy makers most value. Economic analyses will be essential to guide policy and implementation. This paper outlines the gaps and challenges and identifies solutions for effectively developing and efficiently introducing a new TB vaccine.


The work ability index and functional capacity among older workers.
Decreases in functional ability due to aging can impair work capacity and productivity among older workers. This study compares the sociodemographics, health conditions, and physical functioning abilities of young and old workers as well as correlates of physical functioning capacity with the work ability index (WAI). This exploratory, cross-sectional study examined employees of a higher education institution (HEI) and those of a metallurgical industry. Older workers (50 years old or above) were matched for gender and occupation type with younger workers (less than 50 years old). The following evaluations were applied: the multidimensional assessment questionnaire (which included sociodemographic, clinical, health perception, and physical health indices), the WAI, and a battery of physical functional tests. Diseases and regularly used medications were more common among the group of aging workers. The WAI did not differ between groups (p=0.237). Both groups showed similar physical functional capacity performances with regard to walking speed, muscle strength, and lower limb physical functioning. Aging workers showed a poorer performance on a test of right-leg support (p=0.004). The WAI was moderately correlated with the sit-to-stand test among older female workers (r=0.573, p=0.051). Unfavorable general health conditions did not affect the assessment of work ability or most of the tests of physical functional capacity in the aging group.


Management of Spinal Implants in Acute Pediatric Surgical Site Infections: A Multicenter Study.
A retrospective review of patients who underwent posterior spinal fusion (PSF) and returned within 90 days with an acute infection. The study motive is to identify and understand the risk factors associated with failure of retaining spinal implants and failure to treat acute infection. The natural history of early surgical site infection (SSI) (less than 3 months) after PSF is not known and removing the implants early after PSF risks pseudarthrosis and deformity progression. Patients ranging from 1999 to 2011 with surgical site infections (SSIs) who required irrigation and debridement within 3 months of PSF were identified from 4 institutions. Univariable and multivariable regression analysis were used to identify risk factors associated with failure of acute infection treatment. Eighty-two patients (59 female, 23 male) with a mean age of 13.6 years were identified. Median follow-up after initial surgery was 33 months (range: 12-112 months). Sixty-two (76%) were treated successfully with acute treatment and did not return with recurrent infection (cleared infection, group C); 20 (24%) returned later with chronic infection (recurrent infection, group R). Multivariable analysis indicated that patients with stainless steel implants (OR = 6.4, 95% CI = 1.7-32.1; p = .009) and older subjects (OR = 1.3, 95% CI = 1.0-1.6; p = .03) were more likely to present with recurrent infection. There was no difference between the groups with regard to the initial time of presentation post fusion, proportion of non-idiopathic diagnosis, rate of positive cultures, culture species, presence of fusion to pelvis, and time on antibiotic treatment. Seventy-six percent of patients presenting with an SSI less than 3 months after PSF did not require implant removal to clear their infection. Early postoperative SSIs can be treated with retention or implant exchange. Older patients and patients with stainless steel instrumentation are more likely to present with a late recurrent infection compared to other metals. Level III.


Dysesthetic pain in patients with syringomyelia.
Dysesthethic pain is a common complaint of patients with syringomyelia, traumatic paraplegia, and various myelopathic conditions. Because cavitary lesions of the spinal cord can be defined with good resolution by magnetic resonance imaging, syringomyelia provides a potential model for examining anatomic correlates of central pain. In this study, a syndrome of segmental dysesthesias, characterized by burning pain, hyperesthesia, and a variable incidence of trophic changes, was described by 51 of 137 patients (37%) with syringomyelia at the time of clinical presentation. Complete magnetic resonance scans, including axial images, demonstrated extension of the syrinx into the dorsolateral quadrant of the spinal cord on the same side and at the level of pain in 43 of 51 patients (84%). Surgical treatment of syringomyelia resulted in the relief or improvement of dysesthetic pain in 22 of 37 patients (59%), but 15 patients (41%) reported no improvement or an intensification of pain despite collapse of the syrinx. Postoperative dysesthetic pain was often a disabling complaint that responded poorly to medical therapy, including analgesics, sedatives, antiepileptics, antispasmodics, and anti-inflammatory agents. In most cases, there was a gradual improvement of symptoms, although six patients continued to complain of pain 24 to 74 months postoperatively. Prompt but transient relief was achieved in two of two patients with regional sympathetic blocks, and prolonged relief was achieved in one patient by stellate ganglionectomy. We conclude that painful dysesthesias can be caused by a disturbance of pain-modulating centers in the dorsolateral quadrant of the spinal cord and have certain causalgia-like features that respond in an unpredictable way to surgical collapse of the syrinx.


Home care needs of extremely obese elderly European women.
To examine the health and needs of extremely obese women aged over 65 years receiving home care in Europe. A cross-sectional assessment study based on the Aged in Home Care (AdHOC) project recruited 2974 women aged 65 or over who were receiving home care at 11 sites in European countries. Extreme obesity was defined as 'Obesity of such a degree as to interfere with normal activities, including respiration'. Resident Assessment Instrument for Home Care (RAI-HC version 2.0); Activity of Daily Living Scale; Instrumental Activity of Daily Living Scale; the Minimum Data Set Cognitive Performance Scale; and a health profile. One hundred and twenty women (4.0%) were extremely obese. They were younger than their thinner counterparts, with a median age of 78.3 versus 83.3 years, and they more often had multiple health complaints and needed more help with mobility outside the home. The extremely obese had received home care longer than the non-extremely obese (median 28.7 versus 36.6 months). Extremely obese women also needed more help with personal care than the other group and, due to lower age, they were less cognitively impaired. Extreme obesity is a problem that increasingly affects home care of elderly women.


Interictal pattern-induced visual discomfort and ictal photophobia in episodic migraineurs: an association of interictal and ictal photophobia.
Pattern-induced visual discomfort and photophobia are frequently observed symptoms in migraineurs. The presumed pathophysiologic mechanisms of pattern glare and photophobia seem to overlap anatomically within the central nervous system. To assess the relationship between interictal pattern-induced visual discomfort and ictal photophobia in episodic migraineurs. We compared pattern-induced visual discomfort among 3 groups: controls, migraineurs without ictal photophobia (MwoP), and migraineurs with ictal photophobia (MwP). Photophobia was assessed with a validated photophobia questionnaire. Visual discomfort tests were performed using 3 striped patterns with different spatial frequencies. After viewing the patterns for 10 seconds, subjects were asked to report the severity of visual discomfort using 4 scales (none, mild, moderate, and severe) and using a 0-10 visual analog scale (VAS). We compared the proportion of subjects choosing moderate-to-severe discomfort and the median values of VAS scores for each pattern among the 3 groups. We enrolled 35 controls, 18 MwoP, and 44 MwP, and there were no significant differences in clinical features among the 3 groups. MwP reported a significantly higher proportion of moderate-to-severe discomfort and higher median VAS scores than the controls and MwoP did. The intensity of discomfort increased with higher frequency of visual stimuli. We conclude that MwP experienced more severe pattern-induced visual discomfort as compared with the controls and MwoP.


Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA.
A quantitative molecular technique was developed for rapid analysis of microbial community diversity in various environments. The technique employed PCR in which one of the two primers used was fluorescently labeled at the 5' end and was used to amplify a selected region of bacterial genes encoding 16S rRNA from total community DNA. The PCR product was digested with restriction enzymes, and the fluorescently labeled terminal restriction fragment was precisely measured by using an automated DNA sequencer. Computer-simulated analysis of terminal restriction fragment length polymorphisms (T-RFLP) for 1,002 eubacterial sequences showed that with proper selection of PCR primers and restriction enzymes, 686 sequences could be PCR amplified and classified into 233 unique terminal restriction fragment lengths or "ribotypes." Using T-RFLP, we were able to distinguish all bacterial strains in a model bacterial community, and the pattern was consistent with the predicted outcome. Analysis of complex bacterial communities with T-RFLP revealed high species diversity in activated sludge, bioreactor sludge, aquifer sand, and termite guts; as many as 72 unique ribotypes were found in these communities, with 36 ribotypes observed in the termite guts. The community T-RFLP patterns were numerically analyzed and hierarchically clustered. The pattern derived from termite guts was found to be distinctly different from the patterns derived from the other three communities. Overall, our results demonstrated that T-RFLP is a powerful tool for assessing the diversity of complex bacterial communities and for rapidly comparing the community structure and diversity of different ecosystems.


The balance between stress and personal capital during pregnancy and the relationship with adverse obstetric outcomes: findings from the 2007 Los Angeles Mommy and Baby (LAMB) study.
Stress during pregnancy is a salient risk factor for adverse obstetric outcomes. Personal capital during pregnancy, defined as internal and social resources that help women cope with or decrease their exposure to stress, may reduce the risk of poor obstetric outcomes. Using data from the 2007 Los Angeles Mommy and Baby study (N = 3,353), we examined the relationships between the balance of stress and personal capital during pregnancy, or the stress-to-capital ratio (SCR), and adverse obstetric outcomes (i.e., pregnancy complications, preterm birth (PTB), low birth weight (LBW), and small for gestational age (SGA)). Women with a higher SCR (i.e., greater stress relative to personal capital during pregnancy) were significantly more likely to experience at least one pregnancy complication, PTB, and lower gestational age, but not LBW or SGA. Accounting for pregnancy complications completely mediated the association between the SCR and PTB. Our findings indicate that experiencing greater stress relative to personal capital during pregnancy is associated with an increased risk for pregnancy complications, PTB, and lower gestational age and that pregnancy complications may be a mechanism by which the SCR is related to adverse obstetric outcomes.


Transformation of tooth type induced by inhibition of BMP signaling.
Mammalian dentitions are highly patterned, with different types of teeth positioned in different regions of the jaws. BMP4 is an early oral epithelial protein signal that directs odontogenic gene expression in mesenchyme cells of the developing mandibular arch. BMP4 was shown to inhibit expression of the homeobox gene Barx-1 and to restrict expression to the proximal, presumptive molar mesenchyme of mouse embryos at embryonic day 10. The inhibition of BMP signaling early in mandible development by the action of exogenous Noggin protein resulted in ectopic Barx-1 expression in the distal, presumptive incisor mesenchyme and a transformation of tooth identity from incisor to molar.


RIG-I-Like Receptor LGP2 Is Required for Tumor Control by Radiotherapy.
Dendritic cells (DC) play an essential role in innate immunity and radiation-elicited immune responses. LGP2 is a RIG-I-like receptor involved in cytoplasmic RNA recognition and antiviral responses. Although LGP2 has also been linked to cell survival of both tumor cells and T cells, the role of LGP2 in mediating DC function and antitumor immunity elicited by radiotherapy remains unclear. Here, we report that tumor DCs are linked to the clinical outcome of patients with breast cancer who received radiotherapy, and the presence of DC correlates with gene expression of LGP2 in the tumor microenvironment. In preclinical models, host LGP2 was essential for optimal antitumor control by ionizing radiation (IR). The absence of LGP2 in DC dampened type I IFN production and the priming capacity of DC. In the absence of LGP2, MDA5-mediated activation of type I IFN signaling was abrogated. The MDA5/LGP2 agonist high molecular weight poly I:C improved the antitumor effect of IR. This study reveals a previously undefined role of LGP2 in host immunity and provides a new strategy to improve the efficacy of radiotherapy. SIGNIFICANCE: These findings reveal an essential role of LGP2 in promoting antitumor immunity after radiotherapy and provide a new strategy to enhance radiotherapy.


Contingent monetary reinforcement of smoking reductions, with and without transdermal nicotine, in outpatients with schizophrenia.
This study was conducted to examine the effects of contingent monetary reinforcement (CM) for smoking reduction, with and without transdermal nicotine, on cigarette smoking in individuals with schizophrenia. Fourteen outpatients participated in each of 3 conditions: (a) CM combined with 21 mg transdermal nicotine, (b) CM combined with placebo patch, and (c) noncontingent reinforcement combined with placebo patch. Each condition lasted 5 days. Carbon monoxide levels were measured 3 times daily, and nicotine withdrawal symptoms were measured once daily in each condition. Results indicated that CM reduced smoking but that 21 mg transdermal nicotine did not enhance that effect. These results offer further evidence supporting the efficacy of CM for reducing smoking among people with schizophrenia, but higher doses of nicotine replacement therapy, or another pharmacotherapy, may be needed to enhance that effect.


Socialising Health Burden Through Different Network Topologies: A Simulation Study.
An aging population and the expectation of premium quality health services combined with the increasing economic burden of the healthcare system requires a paradigm shift toward patient oriented healthcare. The guardian angel theory described by Szolovits [1] explores the notion of enlisting patients as primary providers of information and motivation to patients with similar clinical history through social connections. In this study, an agent based model was developed to simulate to explore how individuals are affected through their levels of intrinsic positivity. Ring, point-to-point (paired buddy), and random networks were modelled, with individuals able to send messages to each other given their levels of variables positivity and motivation. Of the 3 modelled networks it is apparent that the ring network provides the most equal, collective improvement in positivity and motivation for all users. Further study into other network topologies should be undertaken in the future.


A functional ryanodine-sensitive intracellular Ca2+ store is present in vascular endothelial cells.
The presence of the ryanodine receptor was recently demonstrated in vascular and endocardial endothelium, but its function has not been established. We investigated whether functional ryanodine-sensitive Ca2+ stores are present in cultured endothelial cells from rat aorta (RAECs), human aorta (HAECs), human umbilical vein (HUVECs), and bovine pulmonary artery (BPAECs) and what role these may play in intracellular Ca2+ regulation. Under resting conditions, HAECs, BPAECs, and HUVECs demonstrated a slow increase in intracellular Ca2+ (indexed by indo 1 fluorescence) on exposure to 5 mumol/L ryanodine, whereas RAECs did not. However, after an initial bradykinin exposure in RAECs, ryanodine markedly blunted the rapid increase in Ca2+ on a second exposure to bradykinin. In HUVECs, ryanodine in buffer with 1.5 mmol/L Ca2+ did not inhibit the agonist-sensitive Ca2+ increase, whereas it blunted the rapid increase in Ca2+ on histamine exposure in buffer with 5 mmol/L Ca2+, suggesting that increasing [Ca2+] enhances the binding of ryanodine to its receptor. Thus, functional ryanodine-sensitive Ca2+ stores are present in vascular endothelial cells. These appear to be involved in regulation of Ca2+ storage and release from agonist-sensitive intracellular compartments.


Final year student nurses' experiences of wound care: an evaluation.
This article reports on research to explore if pre-registration nursing students felt prepared to manage patients' skin integrity effectively on registration. Final year nursing students completing adult, child and mental health fields were invited to complete questionnaires to investigate the amount of teaching sessions delivered in university in relation to managing skin integrity during their 3-year training programme, discover if pre-registration nursing students received supplementary management of skin integrity teaching in the clinical areas, explore which member of staff in the clinical areas supported the students' learning in the area of skin integrity. Data was collected on 217 final year students (196 females and 21 males) at two higher education institutions in the north of England. The majority of respondents (n = 146; 68%) reported receiving less than 10 hours formal teaching at university on the subject of skin integrity over their 3-year courses. Of those registered on degree courses, 134 students (71%) reported receiving less than 10 hours formal teaching over their 3-year courses, compared with only 12 students (46%) registered on diploma courses. Some 198 (99%) of respondents reported that their clinical teaching was undertaken by registered nurses all or some of the time. Other health professionals were reported to provide substantially less clinical teaching; with the next largest contribution reported to be provided by specialist nurses, who provided all clinical teaching to 36 respondents (19%) and some clinical teaching to 115 respondents (59%). Some 149 respondents (70%) reported that the teaching they received had developed their knowledge and skills to maintain skin integrity for all patients. Respondents claimed that teaching received had developed their knowledge and skills, reporting an average of 16.9 hours spent in directed study; whereas those who did not claim that teaching they had received had developed their knowledge and skills reported an average of 7.6 hours spent in directed study. The results of this study suggest that diplomate nurses are likely to feel more confident and competent than their graduate counterparts, despite spending the same amount of time with mentors and their peers.


Lack of action of exogenously administered T3 on the fetal rat brain despite expression of the monocarboxylate transporter 8.
Mutations of the monocarboxylate transporter 8 gene (MCT8, SLC16A2) cause the Allan-Herndon-Dudley syndrome, an X-linked syndrome of severe intellectual deficit and neurological impairment. Mct8 transports thyroid hormones (T4 and T3), and the Allan-Herndon-Dudley syndrome is likely caused by lack of T3 transport to neurons during critical periods of fetal brain development. To evaluate the role of Mct8 in thyroid hormone action in the fetal brain we administered T4 or T3 to thyroidectomized pregnant dams treated with methyl-mercapto-imidazol to produce maternal and fetal hypothyroidism. Gene expression was then measured in the fetal cerebral cortex. T4 increased Camk4, Sema3c, and Slc7a3 expression, but T3 was without effect. To investigate the cause for the lack of T3 action we analyzed the expression of organic anion transport polypeptide (Oatp14, Slco1c1), a T4 transporter, and Mct8 (Slc16a2), a T4 and T3 transporter, by confocal microscopy. Both proteins were present in the brain capillaries forming the blood-brain barrier and in the epithelial cells of the choroid plexus forming the blood-cerebrospinal fluid barrier. It is concluded that T4 from the maternal compartment influences gene expression in the fetal cerebral cortex, possibly after transport via organic anion transporter polypeptide and/or Mct8, and conversion to T3 in the astrocytes. On the other hand, T3 does not reach the target neurons despite the presence of Mct8. The data indicate that T4, through local deiodination, provides most T3 in the fetal rat brain. The role of Mct8 as a T3 transporter in the fetal rat brain is therefore uncertain.


Management of Type B Aortic Dissections: Treatment of Acute Dissections and Acute Complications from Chronic Dissections.
Aortic dissection is one of the most catastrophic vascular challenges faced by interventionalists, often misdiagnosed and frequently associated with devastating consequences. Stanford type B aortic dissections affects the descending thoracic aorta without any involvement of the ascending aorta. Dissections associated with refractory pain, rapid aneurysm formation, malperfusion syndromes, rupture, or impending rupture are categorized as complicated and distinct from initially uncomplicated type B dissections without the aforementioned complications. Traditionally, uncomplicated type B dissection is managed medically whereas complicated type B dissection requires intervention to limit mortality. Over the last two decades, thoracic endovascular aortic repair (TEVAR) has emerged as an alternative to open surgical repair for treatment of complicated type B dissection. Various societal guidelines now unanimously propose TEVAR as the treatment of choice for management of complicated disease based on its relatively low morbidity and mortality compared to open surgical outcomes. As the consensually acknowledged standard of care for management of patients with complicated lesions, TEVAR is currently being considered for select patients with initially uncomplicated dissection to mitigate the risks of mid and late-term disease progression and aortic-related mortality. This article describes the decision-making treatment algorithm for management of type B aortic dissection. It also provides a comprehensive review of the indications and procedural recommendations for performing TEVAR based on current evidence in the literature. Additionally, the article guides the readers through step-by-step practical considerations, from choosing the optimal graft to insuring its ideal deployment in type B dissection, as well as providing advice on how to handle a variety of procedure-related complications.


Neurohospitalists: an emerging subspecialty.
Akin to the rapid growth in hospitalist medicine seen in the prior decade, there has been a recent explosion in the need for neurohospitalists. Factors driving this demand include nationally mandated quality and safety measures, the increasing complexity and age of the hospitalized patient, and diminished training in diagnosis and management of neurological illnesses for internal medicine residents. The role of the neurohospitalist is varied and may include not only providing neurological care to hospitalized patients but also serving as a leader in an institution's push to meet quality and safety measures. Close collaboration with intensivists, vascular neurologists, and outpatient neurologists is both a challenge and essential for successful delivery of care both during hospitalization and after discharge. Future challenges facing neurohospitalists include defining its relationship to other fields, instituting a pathway for certification, and conducting research to guide the development of evidence-based practice and quality measures.


The neprilysin pathway in heart failure: a review and guide on the use of sacubitril/valsartan.
Inhibition of neurohumoural pathways such as the renin angiotensin aldosterone and sympathetic nervous systems is central to the understanding and treatment of heart failure (HF). Conversely, until recently, potentially beneficial augmentation of neurohumoural systems such as the natriuretic peptides has had limited therapeutic success. Administration of synthetic natriuretic peptides has not improved outcomes in acute HF but modulation of the natriuretic system through inhibition of the enzyme that degrades natriuretic (and other vasoactive) peptides, neprilysin, has proven to be successful. After initial failures with neprilysin inhibition alone or dual neprilysin-angiotensin converting enzyme (ACE) inhibition, the Prospective comparison of angiotensin receptor neprilysin inhibitor (ARNI) with ACEI to Determine Impact on Global Mortality and morbidity in Heart Failure trial (PARADIGM-HF) trial demonstrated that morbidity and mortality can be improved with the angiotensin receptor blocker neprilysin inhibitor sacubitril/valsartan (formerly LCZ696). In comparison to the ACE inhibitor enalapril, sacubitril/valsartan reduced the occurrence of the primary end point (cardiovascular death or hospitalisation for HF) by 20% with a 16% reduction in all-cause mortality. These findings suggest that sacubitril/valsartan should replace an ACE inhibitor or angiotensin receptor blocker as the foundation of treatment of symptomatic patients (NYHA II-IV) with HF and a reduced ejection fraction. This review will explore the background to neprilysin inhibition in HF, the results of the PARADIGM-HF trial and offer guidance on how to use sacubitril/valsartan in clinical practice.


Mutations within the catalytic motif of DNA adenine methyltransferase (Dam) of Aeromonas hydrophila cause the virulence of the Dam-overproducing strain to revert to that of the wild-type phenotype.
In this study, we demonstrated that the methyltransferase activity associated with Dam was essential for attenuation of Aeromonas hydrophila virulence. We mutated aspartic acid and tyrosine residues to alanine within the conserved DPPY catalytic motif of Dam and transformed the pBAD/damD/A, pBAD/damY/A, and pBAD/damAhSSU (with the native dam gene) recombinant plasmids into the Escherichia coli GM33 (dam-deficient) strain. Genomic DNA (gDNA) isolated from either of the E. coli GM33 strains harboring the pBAD vector with the mutated dam gene was resistant to DpnI digestion and sensitive to DpnII restriction endonuclease cutting. These findings were contrary to those with the gDNA of E. coli GM33 strain containing the pBAD/damAhSSU plasmid, indicating nonmethylation of E. coli gDNA with mutated Dam. Overproduction of mutated Dam in A. hydrophila resulted in bacterial motility, hemolytic and cytotoxic activities associated with the cytotoxic enterotoxin (Act), and protease activity similar to that of the wild-type (WT) bacterium, which harbored the pBAD vector and served as a control strain. On the contrary, overproduction of native Dam resulted in decreased bacterial motility, increased Act-associated biological effects, and increased protease activity. Lactone production, an indicator of quorum sensing, was increased when the native dam gene was overexpressed, with its levels returning to that of the control strain when the dam gene was mutated. These effects of Dam appeared to be mediated through a regulatory glucose-inhibited division A protein. Infection of mice with the mutated Dam-overproducing strains resulted in mortality rates similar to those for the control strain, with 100% of the animals dying within 2 to 3 days with two 50% lethal doses (LD50s) of the WT bacterium. Importantly, immunization of mice with a native-Dam-overproducing strain at the same LD50 did not result in any lethality and provided protection to animals after subsequent challenge with a lethal dose of the control strain.


Rapid identification of the three homoeologues of the wheat dwarfing gene Rht using a novel PCR-based screen of three-dimensional BAC pools.
A high-throughput two-step PCR strategy for the identification of selected genes from a BAC library derived from hexaploid wheat (16,974 Mbp) is described. The screen is based on the pooling of DNA from BAC clones into 675 "superpools" arrayed in a three-dimensional configuration. Each BAC clone is represented in three superpools to allow the identification of candidate 384-well plates of clones after the first round of PCR; identification is facilitated by an associated Perl script. A second round of PCR detects the specific BAC clone within the candidate plate that corresponds to the gene of interest. Thus, a single copy of the target gene can be identified from the library of over 700,000 clones (approximately 5 genome equivalents) by assaying only three 384-well plates. The pooling strategy was validated by screening the library with primers specific for the reduced height (Rht-1a) gene. Using relatively stringent selection criteria, 13 Rht-containing clones were identified from 17 candidate plates, and sequence analysis of the amplified products showed that all three Rht homoeologues were represented. Furthermore, the method confirmed the estimated coverage of the BAC library. Thus, this methodology allows the rapid and cost-effective identification of genes, and their homoeologues, from large-insert libraries of complex genomes such as hexaploid wheat.


Surgicric 2: A comparative bench study with two established emergency cricothyroidotomy techniques in a porcine model.
'Can't Intubate, Can't Oxygenate' is a rare but life threatening event. Anaesthetists must be trained and have appropriate equipment available for this. The ideal equipment is a topic of ongoing debate. To date cricothyroidotomy training for anaesthetists has concentrated on cannula techniques. However cases reported to the NAP4 audit illustrated that they were associated with a high failure rate. A recent editorial by Kristensen and colleagues suggested all anaesthetists must master a surgical technique. The surgical technique for cricothyroidotomy has been endorsed as the primary technique by the recent Difficult Airway Society 2015 guidelines. We conducted a bench study comparing the updated Surgicric 2 device with a scalpel-bougie-tube surgical technique, and the Melker seldinger technique, using a porcine model. Twenty six senior anaesthetists (ST5+) participated. The primary outcome was insertion time. Secondary outcomes included success rate, ease of use, device preference and tracheal trauma. There was a significant difference (P<0.001) in the overall comparisons of the insertion times. The surgical technique had the fastest median time of 62 s. The surgical and Surgicric techniques were significantly faster to perform than the Melker (both P<0.001). The surgical technique had a success rate of 85% at first attempt, and 100% within two attempts, whereas the others had failed attempts. The surgical technique was ranked first by 50% participants and had the lowest grade of posterior tracheal wall trauma, significantly less than the Surgicric 2 (P=0.002). This study supports training in and the use of surgical cricothyroidotomy by anaesthetists.


Decolorization of synthetic textile dyes by lignin peroxidase of Phanerochaete chrysosporium.
Neem hull waste (containing a high amount of lignin and other phenolic compounds) was used for lignin peroxidase production by Phanerochaete chrysosporum under solid-state fermentation conditions. Maximum decolorization achieved by partially purified lignin peroxidase was 80% for Porocion Brilliant Blue HGR, 83 for Ranocid Fast Blue, 70 for Acid Red 119 and 61 for Navidol Fast Black MSRL. The effects of different concentrations of veratryl alcohol, hydrogen peroxide, enzyme and dye on the efficiency of decolorization have been investigated. Maximum decolorization efficiency was observed at 0.2 and 0.4 mmol/L hydrogen peroxide, 2.5 mmol/L veratryl alcohol and pH 5.0 after a 1-h reaction, using 50 ppm of dyes and 9.96 mkat/L of enzyme.


Escherichia coli DNA polymerase III is responsible for the high level of spontaneous mutations in mutT strains.
Reactive oxygen species induce oxidative damage in DNA precursors, i.e. dNTPs, leading to point mutations upon incorporation. Escherichia coli mutT strains, deficient in the activity hydrolysing 8-oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP), display more than a 100-fold higher spontaneous mutation frequency over the wild-type strain. 8-oxo-dGTP induces A to C transversions when misincorporated opposite template A. Here, we report that DNA pol III incorporates 8-oxo-dGTP ≈ 20 times more efficiently opposite template A compared with template C. Single, double or triple deletions of pol I, pol II, pol IV or pol V had modest effects on the mutT mutator phenotype. Only the deletion of all four polymerases led to a 70% reduction of the mutator phenotype. While pol III may account for nearly all 8-oxo-dGTP incorporation opposite template A, it only extends ≈ 30% of them, the remaining 70% being extended by the combined action of pol I, pol II, pol IV or pol V. The unique property of pol III, a C-family DNA polymerase present only in eubacteria, to preferentially incorporate 8-oxo-dGTP opposite template A during replication might explain the high spontaneous mutation frequency in E. coli mutT compared with the mammalian counterparts lacking the 8-oxo-dGTP hydrolysing activities.


The relationships between impact location and post-impact ball speed, bat torsion, and ball direction in cricket batting.
Three-dimensional kinematic data of bat and ball were recorded for 239 individual shots performed by twenty batsmen ranging from club to international standard. The impact location of the ball on the bat face was determined and assessed against the resultant instantaneous post-impact ball speed and measures of post-impact bat torsion and ball direction. Significant negative linear relationships were found between post-impact ball speed and the absolute distance of impact from the midline medio-laterally and sweetspot longitudinally. Significant cubic relationships were found between the distance of impact from the midline of the bat medio-laterally and both a measure of bat torsion and the post-impact ball direction. A "sweet region" on the bat face was identified whereby impacts within 2 cm of the sweetspot in the medio-lateral direction, and 4.5 cm in the longitudinal direction, caused reductions in ball speed of less than 6% from the optimal value, and deviations in ball direction of less than 10° from the intended target. This study provides a greater understanding of the margin for error afforded to batsmen, allowing researchers to assess shot success in more detail, and highlights the importance of players generating consistently central impact locations when hitting for optimal performance.


Resources for the Comprehensive Discovery of Functional RNA Elements.
Transcriptome-wide maps of RNA binding protein (RBP)-RNA interactions by immunoprecipitation (IP)-based methods such as RNA IP (RIP) and crosslinking and IP (CLIP) are key starting points for evaluating the molecular roles of the thousands of human RBPs. A significant bottleneck to the application of these methods in diverse cell lines, tissues, and developmental stages is the availability of validated IP-quality antibodies. Using IP followed by immunoblot assays, we have developed a validated repository of 438 commercially available antibodies that interrogate 365 unique RBPs. In parallel, 362 short-hairpin RNA (shRNA) constructs against 276 unique RBPs were also used to confirm specificity of these antibodies. These antibodies can characterize subcellular RBP localization. With the burgeoning interest in the roles of RBPs in cancer, neurobiology, and development, these resources are invaluable to the broad scientific community. Detailed information about these resources is publicly available at the ENCODE portal (https://www.encodeproject.org/).


Morphological and Whole-Word Semantic Processing Are Distinct: Event Related Potentials Evidence From Spoken Word Recognition in Chinese.
Behavioral and imaging studies in alphabetic languages have shown that morphological processing is a discrete and independent element of lexical access. However, there is no explicit marker of morphological structure in Chinese complex words, such that the extent to which morpheme meaning is related to word meaning is unknown. Event-related potentials (ERPs) were used in the present study to investigate the dissociation of morphemic and whole-word meaning in an auditory-auditory primed lexical decision task. All the prime and target words are compounds consisting of two Chinese morphemes. The relationship between morpheme and whole-word meaning was manipulated while controlling the phonology and orthography of the first syllable in each prime-target pair. A clear dissociation was found between morphemic and whole-word meaning on N400 amplitude and topography. Specifically, sharing a morpheme produced a larger N400 in the anterior-central electrode sites, while sharing whole-word meaning produced a smaller N400 in central-posterior electrode sites. In addition, the morphological N400 effect was negatively correlated with the participants' reading ability, with better readers needing less orthographic information to distinguish different morphemes in compound words. These findings indicate that morphological and whole-word meaning are dissociated in spoken Chinese compound word recognition and that even in the spoken language modality, good readers are better able to access the meaning of individual morphemes in Chinese compound word processing.


RNA Binding Protein HuR Promotes Autophagosome Formation by Regulating Expression of Autophagy-Related Proteins 5, 12, and 16 in Human Hepatocellular Carcinoma Cells.
Autophagy is a process of lysosomal self-degradation of cellular components by forming autophagosomes. Autophagosome formation is an essential process in autophagy and is fine-tuned by various autophagy-related gene (ATG) products, including ATG5, ATG12, and ATG16. Although several reports have shown that numerous factors affect multiple levels of gene regulation to orchestrate cellular autophagy, the detailed mechanism of autophagosome formation still needs further investigation. In this study, we demonstrate that the RNA binding protein HuR (human antigen R) performs an essential function in autophagosome formation. We observe that HuR silencing leads to inhibition of autophagosome formation and autophagic flux in liver cells. Ribonucleoprotein immunoprecipitation (RIP) assay allows the identification of ATG5, ATG12, and ATG16 mRNAs as the direct targets of HuR. We further show that HuR mediates the translation of ATG5, ATG12, and ATG16 mRNAs by binding to their 3' untranslated regions (UTRs). In addition, we show that HuR expression positively correlates with the levels of ATG5 and ATG12 in hepatocellular carcinoma (HCC) cells. Collectively, our results suggest that HuR functions as a pivotal regulator of autophagosome formation by enhancing the translation of ATG5, ATG12, and ATG16 mRNAs and that augmented expression of HuR and ATGs may participate in the malfunction of autophagy in HCC cells.


Mechanisms of mutational robustness in transcriptional regulation.
Robustness is the invariance of a phenotype in the face of environmental or genetic change. The phenotypes produced by transcriptional regulatory circuits are gene expression patterns that are to some extent robust to mutations. Here we review several causes of this robustness. They include robustness of individual transcription factor binding sites, homotypic clusters of such sites, redundant enhancers, transcription factors, redundant transcription factors, and the wiring of transcriptional regulatory circuits. Such robustness can either be an adaptation by itself, a byproduct of other adaptations, or the result of biophysical principles and non-adaptive forces of genome evolution. The potential consequences of such robustness include complex regulatory network topologies that arise through neutral evolution, as well as cryptic variation, i.e., genotypic divergence without phenotypic divergence. On the longest evolutionary timescales, the robustness of transcriptional regulation has helped shape life as we know it, by facilitating evolutionary innovations that helped organisms such as flowering plants and vertebrates diversify.


Tumor necrosis factor receptor p55 mediates deletion of peripheral cytotoxic T lymphocytes in vivo.
Cellular death of activated lymphocytes down-regulates immune responses and is involved in maintaining self tolerance. Signals associated with ligation of the membrane molecule Fas lead to lymphocyte apoptosis, but additional, Fas-independent mechanisms have been postulated. Here, we show a marked expansion and prolonged persistence of functional activated cytotoxic T cells in mice lacking the tumor necrosis factor (TNF) receptor p55. In the absence of this receptor, peripheral lymphocyte apoptosis was significantly reduced in vivo. The prolonged thymocyte survival was associated with functional anergy, since the T cells no longer proliferated in vitro when stimulated with peptide antigen. However, specific cytotoxic effector function was easily detected in vitro. We conclude that the TNF receptor p55 is involved in peripheral T cell deletion in vivo.


Extraordinarily high spectral sensitivity in refractive index sensors using multiple optical modes.
The extraordinary spectral sensitivity of surface plasmon resonance (SPR) sensors is commonly attributed to the modal overlap or unique dispersion of surface plasmons. In contrast to this belief, we show that such high sensitivity is due to the multi-mode nature of the sensing scheme. This concept of multi-mode sensing can be applied to dielectric systems as well in order to achieve similar extraordinary spectral sensitivity. We also show that there is a fundamental constraint between the spectral sensitivity and quality factor in such multi-mode sensing approach.


A light-gated potassium channel for sustained neuronal inhibition.
Currently available inhibitory optogenetic tools provide short and transient silencing of neurons, but they cannot provide long-lasting inhibition because of the requirement for high light intensities. Here we present an optimized blue-light-sensitive synthetic potassium channel, BLINK2, which showed good expression in neurons in three species. The channel is activated by illumination with low doses of blue light, and in our experiments it remained active over (tens of) minutes in the dark after the illumination was stopped. This activation caused long periods of inhibition of neuronal firing in ex vivo recordings of mouse neurons and impaired motor neuron response in zebrafish in vivo. As a proof-of-concept application, we demonstrated that in a freely moving rat model of neuropathic pain, the activation of a small number of BLINK2 channels caused a long-lasting (>30 min) reduction in pain sensation.


Sex chromosome-specific regulation in the Drosophila male germline but little evidence for chromosomal dosage compensation or meiotic inactivation.
The evolution of heteromorphic sex chromosomes (e.g., XY in males or ZW in females) has repeatedly elicited the evolution of two kinds of chromosome-specific regulation: dosage compensation--the equalization of X chromosome gene expression in males and females--and meiotic sex chromosome inactivation (MSCI)--the transcriptional silencing and heterochromatinization of the X during meiosis in the male (or Z in the female) germline. How the X chromosome is regulated in the Drosophila melanogaster male germline is unclear. Here we report three new findings concerning gene expression from the X in Drosophila testes. First, X chromosome-wide dosage compensation appears to be absent from most of the Drosophila male germline. Second, microarray analysis provides no evidence for X chromosome-specific inactivation during meiosis. Third, we confirm the previous discovery that the expression of transgene reporters driven by autosomal spermatogenesis-specific promoters is strongly reduced when inserted on the X chromosome versus the autosomes; but we show that this chromosomal difference in expression is established in premeiotic cells and persists in meiotic cells. The magnitude of the X-autosome difference in transgene expression cannot be explained by the absence of dosage compensation, suggesting that a previously unrecognized mechanism limits expression from the X during spermatogenesis in Drosophila. These findings help to resolve several previously conflicting reports and have implications for patterns of genome evolution and speciation in Drosophila.


Endometriosis - The challenge for occupational life of diagnosed women: A review of quantitative studies.
The aim of the article was to summarize the results offered in the research on occupational functioning of women with endometriosis. We followed PRISMA guidelines. Database search was done in November 2017 using EBSCO. In the review were included the articles clearly referring to the relationships between endometriosis and work or any aspect of work (e.g., work productivity, work ability, absenteeism). Eight papers were included in the final review. Seven studies were cross-sectional, 1 retrospective. The majority of researchers used standardized research measures, such as Work Ability Index, Work Productivity and Activity Impairment Questionnaire or Health Related Productivity Questionnaire. Only in 2 studies the reference group was considered, one of them consisted of healthy women. The results clearly indicate that endometriosis is a disease that causes major disturbance in occupational functioning of the suffering women. In the future the researchers should search for individual and organizational correlates of the improvement of occupational functioning in this group of patients. The methodology of the existing studies has been evaluated and the methodological cues for future research has been given. Med Pr 2018;69(6):663-671.


Persistent infection of MDCK cells by influenza C virus: initiation and characterization.
Persistent influenza C virus infection was readily initiated in Madin-Darby canine kidney (MDCK) cells at low m.o.i. and has been maintained for over 1 year. The persistently infected (p.i.) cultures were characterized by the following properties: virus infection was limited to a minority of cells, small amounts of infectious virus were produced together with low levels of interferon (IFN) and the cultures were resistant to superinfection by homologous virus and vesicular stomatitis virus, but not by influenza A and B viruses. These properties fluctuated cyclically with passage of the p.i. culture. When p.i. cultures were cured by cultivation in the presence of antiserum, the cultures lost their IFN-producing activity and became as susceptible to homologous virus as normal MDCK cell culture. The results suggest that persistent influenza C virus infection may be regulated by endogenously produced IFN. Under the condition of high m.o.i. a persistent influenza C virus infection could not be initiated in MDCK cells due to the development of cytopathic effects.