Minimization of 1/fn phase noise in liquid crystal masks for reliable femtosecond pulse shaping.
Liquid-crystal spatial light modulators (LCM) are a common tool to tailor femtosecond laser pulses. The phase stability of 1 kHz, sub-20 fs visible shaped and unshaped pulses are investigated. Our results show that the spectral phase after the LCM varies from pulse to pulse leading to strong deviations from the predicted pulse shapes. This phase instability is generated only by LCM and is strongly temperature dependent. Based on the experimental data, a numerical model for the phase was developed that takes the temperature-dependent phase instability as well as pixel coupling across the LCM into account. Phase stability after the LCM can be improved by an order of magnitude by combining the control the temperature of the LCM and by using rapid-scan averaging. Reliable pulse shapes on a pulse-to-pulse basis are crucial, especially in coherent control experiments, where small differences between pulse shape are important.


Buparlisib plus fulvestrant versus placebo plus fulvestrant for postmenopausal, hormone receptor-positive, human epidermal growth factor receptor 2-negative, advanced breast cancer: Overall survival results from BELLE-2.
Buparlisib, a pan-phosphatidylinositol 3-kinase (PI3K) inhibitor, plus fulvestrant in the BELLE-2 study significantly improved progression-free survival (PFS) in patients with hormone receptor-positive, human epidermal growth factor receptor 2-negative advanced breast cancer. In this phase III study, patients were randomised 1:1 to buparlisib (100 mg/day; continuously in 28-day cycles) or placebo, plus fulvestrant (500 mg on cycle 1 day 15, and day 1 of subsequent cycles). Overall survival (OS) was assessed in the overall population and patients with known PI3K pathway status (both had shown significant PFS improvements). OS by PIK3CA status in circulating tumour DNA (ctDNA) was an exploratory end-point. A total of 2025 patients were screened for eligibility between 7th September 2012 and 10th September 2014, and 1178 received fulvestrant (500 mg) during a run-in phase; 31 discontinued. Of 1147 patients (median age 62 years), 98% had the Eastern Cooperative Oncology Group performance status ≤1, and 59% had visceral disease. Median follow-up from randomisation to data cut-off (23rd December 2016) was 37.6 months. Median OS trended in favour of the buparlisib arm in the overall population (33.2 versus 30.4 months; P = 0.045) and among patients with known PI3K pathway status (30.9 versus 28.9 months; P = 0.144); neither outcome was statistically significant. Median OS also trended in favour of buparlisib among patients with PIK3CA-mutant ctDNA (26.0 versus 24.8 months). Grade III/IV adverse events with ≥10% difference between the buparlisib versus placebo arms were elevated alanine aminotransferase (26% versus 1%), elevated aspartate aminotransferase (18% versus 3%) and hyperglycemia (15% versus <1%). OS results were in favour of buparlisib plus fulvestrant versus placebo plus fulvestrant; however, there is no statistical significance and more frequent grade III/IV adverse events were reported. Use of more selective PI3K inhibitors might provide the greatest clinical benefit and tolerable safety profile in this setting. Further evaluation of the predictive benefit of PIK3CA-mutant ctDNA is warranted. NCT01610284.


Myocardial perfusion imaging in patients with unprotected left main disease.
The management of patients with unprotected left main (LM) coronary artery disease remains challenging, with recent data casting a shadow of doubt on the safety of percutaneous coronary intervention. We aimed at describing the features of patients undergoing myocardial perfusion imaging (MPI) subsequently found to have LM disease. We queried our institutional database for subjects without prior revascularization or myocardial infarction (MI), who had undergone MPI followed by invasive coronary angiography within 6 months, comparing those with evidence of angiographically significant LM disease (i.e. diameter stenosis ≥50%) to those without significant coronary artery disease (CAD), or those with CAD not involving LM. Baseline, stress and imaging features were systematically collected and analyzed, and clinical outcomes (death, myocardial infarction, revascularization) sought. We included a total of 74 patients with LM disease, which were compared with 70 without CAD, and 920 with significant CAD not involving LM. MPI was remarkably safe in all subjects, and significant differences were found for several features, but particularly so for ST change, rate pressure product, and left ventricular ejection fraction (all P<0.05). Most patients with LM disease had moderate or severe ischemia, and the apical, lateral and inferior regions were the most sensitive ones. Clinical outcomes after an average of 35 months were worse in patients with LM disease than in subjects with significant CAD not involving LM, albeit non-significantly, possibly in light of the higher use of coronary artery bypass grafting. MPI is safe and informative in patients with LM disease, and multidimensional appraisal of MPI results may guide decision-making on top of providing prognostic detail and warranty period.


The gut microbiome as a driver of individual variation in cognition and functional behaviour.
Research into proximate and ultimate mechanisms of individual cognitive variation in animal populations is a rapidly growing field that incorporates physiological, behavioural and evolutionary investigations. Recent studies in humans and laboratory animals have shown that the enteric microbial community plays a central role in brain function and development. The 'gut-brain axis' represents a multi-directional signalling system that encompasses neurological, immunological and hormonal pathways. In particular it is tightly linked with the hypothalamic-pituitary-adrenal axis (HPA), a system that regulates stress hormone release and influences brain development and function. Experimental examination of the microbiome through manipulation of diet, infection, stress and exercise, suggests direct effects on cognition, including learning and memory. However, our understanding of these processes in natural populations is extremely limited. Here, we outline how recent advances in predominantly laboratory-based microbiome research can be applied to understanding individual differences in cognition. Experimental manipulation of the microbiome across natal and adult environments will help to unravel the interplay between cognitive variation and the gut microbial community. Focus on individual variation in the gut microbiome and cognition in natural populations will reveal new insight into the environmental and evolutionary constraints that drive individual cognitive variation.This article is part of the theme issue 'Causes and consequences of individual differences in cognitive abilities'.


The role of d-lactate in differential diagnosis of acute appendicitis.
Early diagnosis of acute appendicitis, known as the most frequent cause of acute surgical abdominal pathologies, dramatically decreases the related complications. D-lactate, produced by intestinal bacteria as a fermentation product, may be useful in diagnosing acute abdominal pathologies. The aim of this study was to investigate whether the presence of d-lactate would be a significant indicator in the early diagnosis of acute appendicitis. Eighty consecutive patients were prospectively included in this study. The patients were divided into four groups: acute appendicitis (group 1), perforated acute appendicitis (group 2), nonspecific abdominal pain (group 3), and acute abdomen other than acute appendicitis (group 4). For the control group, blood samples were taken in the same manner from 20 healthy subjects. There was no significant difference in blood d-lactate levels between the simple acute appendicitis and acute perforated appendicitis groups (p > .05). The blood d-lactate levels in groups 1 and 2 were significantly higher than those in groups 3 and 4, and the control group (p < .001). The reliability of d-lactate was determined as 97% sensitivity, 93% specificity, 90% positive predictive and 95% negative predictive values, and 95% accuracy. Based on findings in this study, blood d-lactate level may be a valuable diagnostic marker for the diagnosis of acute appendicitis.


Design of virtual libraries of umami-tasting molecules.
The introduction of molecular tools in food research offers the possibility to the food industry to benefit from the experience gained in the field by pharmaceutical companies. In this work we are showing how in silico virtual screening techniques based on molecular similarity were applied for identifying novel umami-tasting compounds. The results obtained suggest that 5'-ribonucleotides and monosodium glutamate might elicit the fifth basic taste via the same molecular mechanism. New algorithms were developed and used in this work, such as the dimension reduction of data sets by singular value decomposition and the introduction of the correlation dimension as a natural dimension of a chemical space. It is shown that the representations of molecular data sets in chemical spaces possess self-similar properties, characteristic of fractal objects.


Evaluation of assays for the detection of autoantibodies to the ribosomal P proteins.
Sera from systemic lupus erythematosus patients that had antibodies to the ribosomal P proteins were compared in several different assays. The enzyme-linked immunosorbent assay (ELISA) method was compared to the Western immunoblotting method using either affinity purified human or bovine ribosomal P proteins. All 30 normal sera had no significant reactivity with these antigens. The most sensitive test was the ELISA using the human P protein, where 31/32 patients were positive (97%). The assay with bovine proteins in ELISA yielded 28/32 (88%) positive results. Immunoblotting with either bovine or human P protein was equally effective with 30/32 (94%) positive. An ELISA incorporating human P proteins is a more sensitive assay for clinical diagnosis than an ELISA with the bovine protein. Immunoblotting is a sensitive method, but is less convenient and is not quantitative. The ELISA with the human protein appears to be the method of choice.


Synaptic basis of a sub-second representation of time in a neural circuit model.
Temporal sequences of neural activity are essential for driving well-timed behaviors, but the underlying cellular and circuit mechanisms remain elusive. We leveraged the well-defined architecture of the cerebellum, a brain region known to support temporally precise actions, to explore theoretically whether the experimentally observed diversity of short-term synaptic plasticity (STP) at the input layer could generate neural dynamics sufficient for sub-second temporal learning. A cerebellar circuit model equipped with dynamic synapses produced a diverse set of transient granule cell firing patterns that provided a temporal basis set for learning precisely timed pauses in Purkinje cell activity during simulated delay eyelid conditioning and Bayesian interval estimation. The learning performance across time intervals was influenced by the temporal bandwidth of the temporal basis, which was determined by the input layer synaptic properties. The ubiquity of STP throughout the brain positions it as a general, tunable cellular mechanism for sculpting neural dynamics and fine-tuning behavior.


Phase 1 single dose studies to optimize the pharmacokinetics of DG17, a novel HIV-protease inhibitor pro-drug, using sodium bicarbonate and ritonavir.
DG17 is an orally available prodrug of DG35 (a novel HIV protease inhibitor with variable pharmacokinetics). These studies aimed to optimize DG17 pharmacokinetics by gastric acid neutralization and ritonavir pharmacoenhancement. Both studies were conducted using a randomized, cross-over design in which 6 healthy individuals were administered a single dose of 100mg or 200mg DG17, half with the study intervention (sodium bicarbonate solution in the first study, low dose ritonavir in the second). After a one week washout period, each subject was then administered a second dose of DG17, with the study intervention only administered to the other half. Cmax and AUC increases with gastric acid neutralization were greatest in those with the lowest absorption of DG17 alone. All doses were subsequently given with sodium bicarbonate solution in the second study. Low-dose ritonavir co-administration with DG17 increased DG35 C(max) (median 1437 versus 100 ng/ml, p=0.028) and AUC (median 6975 versus 154ng/ml*hr, p=0.028) compared with DG17 without ritonavir. Plasma DG35 exceeded the IC(90) for HIV for > or = 12 hours following a single DG17/ritonavir dose. No significant adverse events occurred. Single dose DG17 is safe and best administered in a manner preventing gastric acid degradation and with low-dose ritonavir.


Fruit Preference, Parasitism, and Offspring Fitness of Fopius arisanus (Hymenoptera: Braconidae) Exposed to Bactrocera dorsalis' (Diptera: Tephritidae) Infested Fruit Species.
Fopius arisanus (Sonan) (Hymenoptera: Braconidae) is a koinobiont solitary parasitoid of various fruit flies, particularly those in the genus Bactrocera. Researchers introduced F. arisanus into Africa for the biological control of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), a pest of a wide range of fruit trees and vegetables. However, the suitability of host fruit species as egg-laying substrates for parasitoid development remains poorly investigated in tropical Africa. The present study examines the preference and performance of F. arisanus on B. dorsalis reared on eleven fruit species through laboratory choice-test trials. We assessed the oviposition activity, parasitism rate, developmental time, and offspring fitness of F. arisanus on nine cultivated and two wild host fruits species. Oviposition attempts were higher on Psidium guajava (L.) (Myrtales: Myrtaceae) and Mangifera indica (L.) (Sapindales: Anacardiaceae) than on the other host fruits tested. The wasp parasitized host eggs in P. guajava in no-choice experiments. Psidium guajava, Irvingia wombulu (Vermoesen) (Malpighiales: Irvingiaceae), and Irvingia gabonensis (Aubry_Lecomte) Baill (Malpighiales: Irvingiaceae) were suitable for parasitism in choice tests. Of all host fruits tested, the body and hind tibia lengths of both parasitoid sexes emerging from M. indica were longer than on the others. The female ovipositor was long on Annona squamosa (L.) (Magnoliales: annonaceae) and short on Eribotrya japonica ([Thunb.] Lindl.; Rosales: Rosaceae). We obtained the longest preimaginal developmental time for both sexes on E. japonica and the shortest for females and males on Carica papaya (L.) (Brassicales: Caricaceae). These results demonstrate the ability of some tested fruit species to serve for the permanent establishment of F. arisanus in the field.


VE-cadherin enables trophoblast endovascular invasion and spiral artery remodeling during placental development.
During formation of the mammalian placenta, trophoblasts invade the maternal decidua and remodel spiral arteries to bring maternal blood into the placenta. This process, known as endovascular invasion, is thought to involve the adoption of functional characteristics of vascular endothelial cells (ECs) by trophoblasts. The genetic and molecular basis of endovascular invasion remains poorly defined, however, and whether trophoblasts utilize specialized endothelial proteins in an analogous manner to create vascular channels remains untested. Vascular endothelial (VE-)cadherin is a homotypic adhesion protein that is expressed selectively by ECs in which it enables formation of tight vessels and regulation of EC junctions. VE-cadherin is also expressed in invasive trophoblasts and is a prime candidate for a molecular mechanism of endovascular invasion by those cells. Here, we show that VE-cadherin is required for trophoblast migration and endovascular invasion into the maternal decidua in the mouse. VE-cadherin deficiency results in loss of spiral artery remodeling that leads to decreased flow of maternal blood into the placenta, fetal growth restriction, and death. These studies identify a non-endothelial role for VE-cadherin in trophoblasts during placental development and suggest that endothelial proteins may play functionally unique roles in trophoblasts that do not simply mimic those in ECs.


Mouse malaria nephropathy.
Mice were infected with 1X 107 Plasmodium berghei Yoelii parasites intraperitoneally. Circulating parasite, malaria antibody and C3 concentrations were measures: parasitaemia and hypocomplementaemia were transient, but the antibody response was persistent. Animals were sacrificed at intervals and their kidneys examined: a glomerulonephritis associates with predominantly mesangial deposits of C3, IgG1, IgM and some IgA always developed after 7 days and persisted for up to 6 mth. Malaria antigen and antibody were demonstrated within the glomeruli. Microscopic haematuria occurred with proteinuria but without marked deterioration in renal function. Strains producing high and low affinity antibody were equally susceptible to the disease. Treatment with glucocorticoid, immunosuppressive, platelet function inhibiting and/or anticoagulant drugs, or indomethacin from the 1st day of infection failed to prevent development of the disease or to lead to its early cure. Eradication of the infection within its first 3 days prevented glomerular deposition of antibody and complement, and infection with a smaller antigen load followed by later treatment also produced subsequent cure.


Embryogenesis - the humble beginnings of plant life.
Each plant starts life from the zygote formed by the fusion of an egg and a sperm cell. The zygote gives rise to a multicellular embryo that displays a basic plant body organization and is surrounded by nutritive endosperm and maternal tissue. How the body organization is generated had already been studied before the genome sequence of Arabidopsis thaliana was completed 10 years ago, but several regulatory mechanisms of embryo development have since been discovered or analysed in more detail. Although this progress did not strictly depend on the availability of the genome sequence itself, several advances were considerably facilitated. In this review, we mainly address early embryo development, highlighting general mechanisms and crucial regulators, including phytohormones, that are involved in patterning the embryo and were mainly analysed in the post-genome decade. We also highlight some unsolved problems, provide a brief outlook on the future of Arabidopsis embryo research, and discuss how the knowledge gained from Arabidopsis could be translated to crop species.


Pulmonary disease in selected protozoal infections.
Protozoa of the phylum Apicomplexa, including the genera Plasmodium, Babesia, Toxoplasmosis, and Cryptosporidium, are a group of closely related organisms that seldom cause pulmonary disease. All but Babesia are members of the subclass Coccidiasina ("Coccidians"). (The fungal organism Coccidioides immitis, a frequent cause of pulmonary disease in endemic areas, was first believed by Rixsford and Gilchrist to be a member of the subclass Coccidiasina; hence its name, meaning coccidioidal-like). Some species, such as Toxoplasma gondii, occasionally cause pulmonary disease by directly infecting lung parenchyma, whereas others, such as Cryptosporidium and Microsporidium, can be occasionally visualized in respiratory secretions or lung biopsy specimens, but their role in causing respiratory disease in human beings is more questionable. In contrast, pulmonary disease associated with infections caused by Malaria and Babesia is often the result of a systemic inflammatory response.


[Approaches to surgical treatment of snoring and sleep apnea].
Examination and treatment data on 186 patients suffering from snoring and sleep apnea and surgical methods to manage these pathological conditions are reviewed. The stages of the surgical treatment of snoring and sleep apnea are the following: correction of nasal breathing, surgical intervention on the palatine using different tools (laser, scalpel, loop, electric knife). Effectiveness of the treatment methods, complications, postoperative developments are compared.


Changes in intracellular Ca2+ concentration induced by L-type Ca2+ channel current in guinea pig gastric myocytes.
We investigated the relationship between voltage-operated Ca2+ channel current and the corresponding intracellular Ca2+ concentration ([Ca2+]i) change (Ca2+ transient) in guinea pig gastric myocytes. Fluorescence microspectroscopy was combined with conventional whole cell patch-clamp technique, and fura 2 (80 microM) was added to CsCl-rich pipette solution. Step depolarization to 0 mV induced inward Ca2+ current (ICa) and concomitantly raised [Ca2+]i. Both responses were suppressed by nicardipine, an L-type Ca2+ channel blocker, and the voltage dependence of Ca2+ transient was similar to the current-voltage relation of ICa. When pulse duration was increased by up to 900 ms, peak Ca2+ transient increased and reached a steady state when stimulation was for longer. The calculated fast Ca2+ buffering capacity (B value), determined as the ratio of the time integral of ICa divided by the amplitude of Ca2+ transient, was not significantly increased after depletion of Ca2+ stores by the cyclic application of caffeine (10 mM) in the presence of ryanodine (4 microM). The addition of cyclopiazonic acid (CPA, 10 microM), a sarco(endo)plasmic reticulum Ca(2+)-ATPase inhibitor, decreased B value by approximately 20% in a reversible manner. When KCl pipette solution was used, Ca(2+)-activated K+ current [IK(Ca)] was also recorded during step depolarization. CPA sensitively suppressed the initial peak and oscillations of IK(Ca) with irregular effects on Ca2+ transients. The above results suggest that, in guinea pig gastric myocyte, Ca2+ transient is tightly coupled to ICa during depolarization, and global [Ca2+]i is not significantly affected by Ca(2+)-induced Ca2+ release from sarcoplasmic reticulum during depolarization.


Factor structure of the NIH Toolbox Cognition Battery in individuals with acquired brain injury.
The National Institutes of Health Toolbox Cognition Battery (NIHTB-CB) measures reading, vocabulary, episodic memory, working memory, executive functioning, and processing speed. While previous research has validated the factor structure in healthy adults, the factor structure has not been examined in adults with neurological impairments. Thus, this study evaluated the NIHTB-CB factor structure in individuals with acquired brain injury. A sample of 392 individuals (ages 18-84) with acquired brain injury (n = 182 TBI, n = 210 stroke) completed the NIHTB-CB along with neuropsychological tests as part of a larger, multisite research project. Confirmatory factor analyses supported a 5-factor solution that included reading, vocabulary, episodic memory, working memory, and processing speed/executive functioning. This structure generally held in TBI and stroke subsamples as well as in subsamples of those with severe TBI and stroke injuries. The factor structure of the NIHTB-CB is similar in adults with acquired brain injury to adults from the general population. We discuss the implications of these findings for clinical practice and clinical research. (PsycINFO Database Record


Peroxidase activity of the coronene bisimide supramolecular architecture and its applications in colorimetric sensing of H2O2 and glucose.
A new water soluble coronene bisimide derivative (CTDI) was designed and synthesized. CTDI self-assembled in an aqueous solution and formed supramolecular nanofibers through π-π stacking and hydrophobic interactions. The nanofibers exhibit distinct peroxidase-like catalytic activity, and can catalyze the redox reaction of 3,3,5,5-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide. Clear assay solution color changes were observed. The peroxidase-like catalytic property was utilized for the sensitive detection of H2O2 and glucose. The assay shows excellent sensitivity, and 1 μM of glucose could be easily detected. Glucose detection in dilute human blood samples was also demonstrated, and the results were verified using a commercial glucose meter. Our method is simple, convenient, sensitive and selective, and could facilitate the sensing of glucose in relation to biological and biomedical research applications.


Caregivers' descriptions of patients with advanced breast cancer in connection with supervision sessions in a surgical ward.
The purpose of this study was to describe the content of caregivers' (health care professionals') presentation of care situations as told at supervision sessions. For 1 year, 21 caregivers in a surgical ward of a county hospital in the middle of Sweden participated in a clinical supervision session 2 hours every third week. The participants in the supervision sessions were divided so as to form three mixed groups composed of registered nurses, practical nurses, physiotherapists, and physicians. The purpose of the supervision sessions was to give caregivers the opportunity to reflect on different care situations in a way that contributed to the development of patient care. The 38 supervision sessions were tape-recorded and transcribed verbatim. The transcripts were analyzed using a qualitative content analysis. The caregivers described difficult care situations focusing on the feelings of patients, relatives, and caregivers, with an emphasis on the caregivers' being dominant. Difficult care situations were described as giving caregivers feelings of discomfort, powerlessness, and reduced self-esteem. These feelings were described as arising in connection with caring for women with advanced breast cancer and other seriously ill patients in an organization lacking clear goals and rules. This study found that supervision sessions offering an opportunity to reflect on the difficult care situations are important for caregivers. These sessions seem therefore to be of vital importance for the future development of cancer care on the surgical ward.


Integrating conflict analysis and consensus reaching in a decision support system for water resource management.
The importance of shared decision processes in water management derives from the awareness of the inadequacy of traditional--i.e. engineering--approaches in dealing with complex and ill-structured problems. It is becoming increasingly obvious that traditional problem solving and decision support techniques, based on optimisation and factual knowledge, have to be combined with stakeholder based policy design and implementation. The aim of our research is the definition of an integrated decision support system for consensus achievement (IDSS-C) able to support a participative decision-making process in all its phases: problem definition and structuring, identification of the possible alternatives, formulation of participants' judgments, and consensus achievement. Furthermore, the IDSS-C aims at structuring, i.e. systematising the knowledge which has emerged during the participative process in order to make it comprehensible for the decision-makers and functional for the decision process. Problem structuring methods (PSM) and multi-group evaluation methods (MEM) have been integrated in the IDSS-C. PSM are used to support the stakeholders in providing their perspective of the problem and to elicit their interests and preferences, while MEM are used to define not only the degree of consensus for each alternative, highlighting those where the agreement is high, but also the consensus label for each alternative and the behaviour of individuals during the participative decision-making. The IDSS-C is applied experimentally to a decision process regarding the use of treated wastewater for agricultural irrigation in the Apulia Region (southern Italy).


PI 3-kinase signalling in platelets: the significance of synergistic, autocrine stimulation.
Phosphoinositide 3-kinases (PI 3Ks) play a key role in regulation of intracellular signalling and cellular function, including cell proliferation, apoptosis, chemotaxis, membrane trafficking and platelet activation. The PI 3Ks are grouped into three classes on the basis on their structure and in vitro substrate specificity. Class I are activated by a variety of agonists which mediate their effect through tyrosine kinase-linked or G-protein-linked receptors. In vivo class I PI 3Ks seem to preferentially phosphorylate the D3 hydroxyls of the inositol moiety of PtdIns(4,5)P2 to produce PtdIns(3,4,5)P3. However, class II PI 3Ks preferentially phosphorylate the D3 hydroxyl of PtdIns and PtdIns(4)P to produce PtdIns(3)P and PtdIns(3,4)P2, respectively. The late accumulation of PtdIns(3,4)P2 has been suggested to play an important role in irreversible platelet aggregation. In human platelets the class II PI 3K isoform HsC2-PI 3K is activated in an integrin alpha IIb beta 3 + fibrinogen-dependent manner. Class III PI 3Ks phosphorylate PtdIns to produce PtdIns(3)P, which play a crucial role in vesicular trafficking. Recent work has suggested that crosstalk between individual receptors and their downstream signal pathways play a central role in PI 3K signalling responses. In this review, we will concentrate on recent advances regarding the regulation of platelet PI 3Ks.


[Properties of conductivity channels induced in phospholipid bilayer membranes by alpha-staphylotoxin].
Interaction between alpha-straphylotoxin (alpha-T) and synthetic phospholipid membranes was shown to be accompanied by the formation of conductivity channels of different amplitude. Dependence of channel conductivity on salt concentration, medium pH, as well as selectivity for different cations and anions was determined. Current--voltage characteristics of BPhM modified by alpha-T were measured. The conductivity fluctuations observed are suggested to be concerned with the initiation of phospholipid domens (with changed mobility of their fatty acid chains) in the bilayer structure.


[Evaluation of clinical competence of family medicine trainer].
We show the first experience of the application of an objective and structured clinical evaluation (OSCE) procedure to family medicine trainers, that has been carried out in Andalucia. The objective is to use a competence evaluation instrument that, in the short term, will be used not only for trainer accreditation but also for other public sanitary professionals. Tutors of family and community medicine residents. Observational descriptive. Educational unity of family medicine. The competencial components to be assessed are the following: anamnesis, physical exploration, communication, technical skill, management, family attention y preventive activities. The clinical situations were selected using the following priority criteria: prevalence, clinical gravity, prevention and early diagnosis importance, case complexity, doctor's capacity of evaluation and simplicity. Thirteen family medicine trainers took part in the OSCE. Their average age was 42.8 +/- 3.6 years. The test had an overall reliability coefficient (Cronbach's alpha) of 0.73. The overall mean score of the participants was 73 +/- 6.2. The best results about the competencial components were family attention, communication and technical skill. The OSCE can be a convenient tool for family medical trainer evaluation, helping to orientate their education in the weak points and, in the near future, it can also be used as an instrument do accredit family medicine trainers.


Increased colonic inflammatory injury and formation of aberrant crypt foci in Nrf2-deficient mice upon dextran sulfate treatment.
Chronic inflammation has been associated with increased risk of developing cancer. The transcription factor NF-E2-related factor 2 (Nrf2) controls the expression of numerous antioxidative enzymes that have been shown to attenuate acute inflammation. The present study investigated the role of Nrf2 genotype in modulating inflammation-promoted colorectal tumorigenesis. Nrf2 wild-type (WT) and Nrf2-deficient (N0) mice were administered a single dose of azoxymethane followed by a 1-week dose of drinking water with or without 1% dextran sulfate sodium (DSS). Aberrant crypt foci were counted 3 weeks after the cessation of DSS treatment. DSS treatment significantly increased numbers of aberrant crypt foci in N0 mice, but not WT mice. The extent of inflammation over the course of DSS treatment was analyzed in both genotypes. Histological analysis of colon sections revealed that N0 mice had markedly increased inflammation and mucosal damage when compared to WT mice beginning on Day 6 of DSS treatment. Although similar levels of inflammatory and oxidative damage biomarkers were evident in colons from WT and N0 mice at the start of DSS treatment, increased colonic proinflammatory cytokine mRNA transcript levels, myeloperoxidase activity and 3-nitrotyrosine immunoreactivity were observed on Day 6 of DSS treatment in N0 mice, but not WT mice. Additionally, DSS treatment resulted in increased lipid peroxidation and loss of aconitase activity in N0 mice, but not WT mice, reflecting increased oxidative damage in colons from N0 mice. Taken together, these results clearly illustrate the role of Nrf2 in regulating an adaptive response that protects against early-phase inflammation-mediated tumorigenesis.


Application of a ketogenic diet in children with autistic behavior: pilot study.
A pilot prospective follow-up study of the role of the ketogenic diet was carried out on 30 children, aged between 4 and 10 years, with autistic behavior. The diet was applied for 6 months, with continuous administration for 4 weeks, interrupted by 2-week diet-free intervals. Seven patients could not tolerate the diet, whereas five other patients adhered to the diet for 1 to 2 months and then discontinued it. Of the remaining group who adhered to the diet, 18 of 30 children (60%), improvement was recorded in several parameters and in accordance with the Childhood Autism Rating Scale. Significant improvement (> 12 units of the Childhood Autism Rating Scale) was recorded in two patients (pre-Scale: 35.00 +/- 1.41[mean +/- SD]), average improvement (> 8-12 units) in eight patients (pre-Scale: 41.88 +/- 3.14[mean +/- SD]), and minor improvement (2-8 units) in eight patients (pre-Scale: 45.25 +/- 2.76 [mean +/- SD]). Although these data are very preliminary, there is some evidence that the ketogenic diet may be used in autistic behavior as an additional or alternative therapy.


Synthesis and first evaluation of [¹⁸F]fluorocyano- and [¹⁸F]fluoronitro-quinoxalinedione as putative AMPA receptor antagonists.
Derivatives of quinoxalinedione (QX) were chosen as chemical lead for the development of new radioligands of the AMPA receptor, since there are several examples of QX-derivatives with high affinity. The radiosyntheses of the new compounds 6-[(18)F]fluoro-7-nitro-QX([(18)F]FNQX) and 7-[(18)F]fluoro-6-cyano-QX ([(18)F]FCQX) with radiochemical yields of 8±2 and 3± 2%, respectively, as well as the evaluation of their binding properties to the AMPA-receptor were performed. A comparison of the Ki-values of the new QX-derivatives FCQX and FNQX with mono-substituted cyanoand nitro-QX shows negligibly small differences of affinity (within the range of 1.4 to 5 µM), but exhibits a tenfold lower affinity than derivatives with two electron withdrawing groups like the 7-cyano-6-nitro-compound CNQX and the 6,7- dinitro compound DNQX. Thus, with respect to the low affinity and a high non-specific binding with in vitro and ex vivo autoradiographic studies, the new compounds do not lend themselves for in vivo imaging.


Persistent chemopreventive effect of S-adenosyl-L-methionine on the development of liver putative preneoplastic lesions induced by thiobenzamide in diethylnitrosamine-initiated rats.
S-Adenosyl-L-methionine (SAM) is a strong chemopreventive agent of rat liver carcinogenesis. Examination was made to determine whether inhibition by SAM of the development of preneoplastic liver lesions persists to SAM withdrawal in diethylnitrosamine-initiated F344 rats promoted with thiobenzamide (TB). The rats were subjected, 2 weeks after initiation, to 5 weeks feeding with a 0.1% TB diet followed by a TB-free diet for 6 weeks and then a second TB treatment for 3 weeks. SAM (384 micromol/kg/day) was injected i.m. during the first TB cycle (treatment A) or for 6 weeks after the first TB cycle (treatment B). Many gamma-glutamyltranspeptidase (GGT)-positive lesions developed in initiated rats after the first TB cycle. They decreased in number after TB withdrawal, while partial recovery of lesion number and a great increase in volume occurred after the second TB cycle. Liver ornithine decarboxylase (ODC) activity and c-myc and c-Ha-ras mRNAs increased during the TB cycles and returned to normal liver values after TB withdrawal. Number and size of GGT-positive lesions, DNA synthesis of GGT-positive cells, liver ODC activity and c-myc and c-Ha-ras mRNA levels decreased as a consequence of SAM treatment A. The recovery of these parameters, induced by a second TB cycle in rats not treated with SAM, was prevented by SAM treatment B. These results suggest that SAM causes a persistent decrease in growth capacity of preneoplastic liver lesions in rats subjected to a diethylnitrosamine/TB protocol.


Intraoperative ultrasonography (IOUS)-guided vs conventional laparoscopic nephrectomy: a randomised control trial.
To assess the efficacy of routine use of intraoperative ultrasonography (IOUS) in improving perioperative outcomes in patients undergoing IOUS-guided laparoscopic nephrectomy (IOUS-LN) and conventional laparoscopic nephrectomy (C-LN). This was a parallel-arm, single-blinded, randomised controlled trial (CTRI/2021/12/038906). All patients undergoing LN, either for benign or malignant causes, were included. Patients undergoing partial/cytoreductive nephrectomy, with venous thrombus were excluded. In the study arm, IOUS-guided renal vascular assessment was performed after colon mobilisation and a standard LN was performed in the control arm. The primary outcome was intraoperative duration. The secondary outcomes were blood loss, need for open conversion, blood transfusion, perioperative complications, duration of Intensive Care Unit (ICU) stay and length of hospitalisation (LOH). The patients were followed for 3 months after surgery. A total of 104 patients were included, with 52 in each arm. Demographic characteristics were comparable in both arms. A significant reduction in the operative duration (mean [sd] 181.69 [40.8] vs 199.7 [41.8] min, P = 0.02) was seen in the IOUS-LN group. The difference in blood loss showed no significant difference when compared between both groups (median [interquartile range] 84.55 [74-105.5] vs 99.95 [78.5-111] mL, P = 0.08). On subgroup analysis, the reduction in the operative duration was significant in patients who underwent laparoscopic simple nephrectomy (LSN; mean [sd] 194.4 [42.5] vs 221.2 [36.4] min, P = 0.01), whereas comparable operative durations were seen in patients undergoing laparoscopic radical nephrectomy (LRN; mean [sd] 168.96 [35.3] vs 178.3 [35.9] min, P = 0.34). Similar conversion rates were seen in both groups (P = 0.98) along with blood transfusions (P = 0.78). The LOH, ICU stay, and complications were similar in both groups. Significantly less blood loss (P = 0.03) was noted with IOUS in patients undergoing LSN. IOUS did not influence any outcomes in patients undergoing LRN. Intraoperative ultrasonography significantly reduced the operative duration in LN, but with no significant reduction in the volume of blood loss. Significant reduction in intraoperative duration and blood loss was seen in patients who underwent LSN on subgroup analysis.


A developmental defect in Plasmodium falciparum male gametogenesis.
Asexually replicating populations of Plasmodium parasites, including those from cloned lines, generate both male and female gametes to complete the malaria life cycle through the mosquito. The generation of these sexual forms begins with the induction of gametocytes from haploid asexual stage parasites in the blood of the vertebrate host. The molecular processes that govern the differentiation and development of the sexual forms are largely unknown. Here we describe a defect that affects the development of competent male gametocytes from a mutant clone of P. falciparum (Dd2). Comparison of the Dd2 clone to the predecessor clone from which it was derived (W2'82) shows that the defect is a mutation that arose during the long-term cultivation of asexual stages in vitro. Light and electron microscopic images, and indirect immunofluorescence assays with male-specific anti-alpha-tubulin II antibodies, indicate a global disruption of male development at the gametocyte level with at least a 70-90% reduction in the proportion of mature male gametocytes by the Dd2 clone relative to W2'82. A high prevalence of abnormal gametocyte forms, frequently containing multiple and unusually large vacuoles, is associated with the defect. The reduced production of mature male gametocytes may reflect a problem in processes that commit a gametocyte to male development or a progressive attrition of viable male gametocytes during maturation. The defect is genetically linked to an almost complete absence of male gamete production and of infectivity to mosquitoes. This is the first sex-specific developmental mutation identified and characterized in Plasmodium.


Search for quantitative trait loci affecting growth and carcass traits in a cross population of beef and dairy cattle.
A genome scan to detect QTL influencing growth and carcass-related traits was conducted in a Charolais x Holstein crossbred cattle population. Phenotypic measurements related to growth and carcass traits were made on the 235 second-generation crossbred males of this herd (F2 and reciprocal backcrosses), which were born in 4 consecutive annual cohorts. Traits measured in vivo were related to birth dimensions, growth rates, and ultrasound measurements of fat and muscle depth. The animals were slaughtered near a target BW of 550 kg, and a wide range of postmortem traits were measured: visual assessment of carcass conformation and carcass fatness, estimated subcutaneous fat percentage, weights of kidney knob and channel fat, and weights of carcass components after commercial and full-tissue dissections. The whole population, including grandparents, parents, and the crossbred bulls, was genotyped initially for 139 genome-wide microsatellite markers. Twenty-six additional markers were subsequently analyzed to increase marker density on some of the chromosomes where QTL had been initially identified. The linear regression analyses based on the 165 markers revealed a total of 51 significant QTL at the suggestive level, 21 of which were highly significant (F-value >or=9; based on the genome-wide thresholds obtained in the initial scan). A large proportion of the highly significant associations were found on chromosomes 5 and 6. The most highly significant QTL was localized between markers DIK1054 and DIK082 on chromosome 6 and explained about 20% of the phenotypic variance for the total bone proportion estimated after the commercial dissection. In the adjacent marker interval on this chromosome, 2 other highly significant QTL were found that explain about 30% of the phenotypic variance for birth dimension traits (BW and body length at birth). On chromosome 5, the most significant association influenced the lean:bone ratio at the forerib joint and was flanked by markers DIK4782 and BR2936. Other highly significant associations were detected on chromosomes 10 (estimated subcutaneous fat percentage), 11 (total saleable meat proportion), 16 (prehousing growth rate), and 22 (bone proportion at the leg joint). These results provide a useful starting point for the identification of the genes associated with traits of direct interest to the beef industry, using fine mapping or positional candidate gene approaches.


International guidelines for prevention and management of post-operative chronic pain following inguinal hernia surgery.
To provide uniform terminology and definition of post-herniorrhaphy groin chronic pain. To give guidelines to the scientific community concerning the prevention and the treatment of chronic groin and testicular pain. A group of nine experts in hernia surgery was created in 2007. The group set up six clinical questions and continued to work on the answers, according to evidence-based literature. In 2008, an International Consensus Conference was held in Rome with the working group, with an audience of 200 participants, with a view to reaching a consensus for each question. A consensus was reached regarding a definition of chronic groin pain. The recommendation was to identify and preserve all three inguinal nerves during open inguinal hernia repair to reduce the risk of chronic groin pain. Likewise, elective resection of a suspected injured nerve was recommended. There was no recommendation for a procedure on the resected nerve ending and no recommendation for using glue during hernia repair. Surgical treatment (including all three nerves) should be suggested for patients who do not respond to no-surgery pain-management treatment; it is advisable to wait at least 1 year from the previous herniorraphy. The consensus reached on some open questions in the field of post-herniorrhaphy chronic pain may help to better analyze and compare studies, avoid sending erroneous messages to the scientific community, and provide some guidelines for the prevention and treatment of post-herniorraphy chronic pain.


Cancer diagnosis and prognosis decoded by blood-based circulating microRNA signatures.
In the recent years, circulating microRNAs (miRNAs) have garnered a lot of attention and interest in the field of disease biomarkers. With characteristics such as high stability, low cost, possibility of repeated sampling and minimal invasiveness, circulating miRNAs are ideal for development into diagnostic tests. There have been many studies reported on the potential of circulating miRNAs as early detection, prognostic, and predictive biomarkers in cancer. Here, we have reviewed the application of plasma and serum miRNAs as biomarkers for cancer focusing on epithelial carcinomas [prostate, breast, lung, colorectal, and gastric cancer (GC)] and hematological malignancies (leukemia and lymphoma). We have also addressed the common challenges that need to be overcome to achieve a successful bench to bedside transition.


Atomic Layer Deposition-A Versatile Toolbox for Designing/Engineering Electrodes for Advanced Supercapacitors.
Atomic layer deposition (ALD) has become the most widely used thin-film deposition technique in various fields due to its unique advantages, such as self-terminating growth, precise thickness control, and excellent deposition quality. In the energy storage domain, ALD has shown great potential for supercapacitors (SCs) by enabling the construction and surface engineering of novel electrode materials. This review aims to present a comprehensive outlook on the development, achievements, and design of advanced electrodes involving the application of ALD for realizing high-performance SCs to date, as organized in several sections of this paper. Specifically, this review focuses on understanding the influence of ALD parameters on the electrochemical performance and discusses the ALD of nanostructured electrochemically active electrode materials on various templates for SCs. It examines the influence of ALD parameters on electrochemical performance and highlights ALD's role in passivating electrodes and creating 3D nanoarchitectures. The relationship between synthesis procedures and SC properties is analyzed to guide future research in preparing materials for various applications. Finally, it is concluded by suggesting the directions and scope of future research and development to further leverage the unique advantages of ALD for fabricating new materials and harness the unexplored opportunities in the fabrication of advanced-generation SCs.


The Alexander Project 1998-2000: susceptibility of pathogens isolated from community-acquired respiratory tract infection to commonly used antimicrobial agents.
The Alexander Project is a continuing surveillance study, begun in 1992, examining the susceptibility of pathogens involved in adult community-acquired respiratory tract infections (CARTI) to a range of antimicrobial agents. This study tested the susceptibility of isolates of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis collected between 1998 and 2000 to 23 antimicrobials. Minimum inhibitory concentrations of agents were determined using the broth microdilution method and interpreted according to NCCLS and pharmacokinetic/pharmacodynamic (PK/PD) breakpoints. In total, 8882 isolates of S. pneumoniae, 8523 isolates of H. influenzae and 874 isolates of M. catarrhalis were collected during 1998-2000 from centres in 26 countries. The world-wide prevalence of penicillin resistance (penicillin MICs > or = 2 mg/l) in isolates of S. pneumoniae was 18.2% over the study period, and the prevalence of macrolide resistance (erythromycin MICs > or = 1 mg/l) in this pathogen was 24.6%. Over the study period, macrolide resistance exceeded penicillin resistance in 19 of the 26 countries included in the study. Of the non-fluoroquinolone agents, the only oral agents to which over 90% of S. pneumoniae isolates were susceptible at both NCCLS and PK/PD breakpoints were amoxicillin (95.1%) and co-amoxiclav (95.5-97.9%). The prevalence of fluoroquinolone-resistant S. pneumoniae (ofloxacin MICs > or = 8 mg/l) was 1.1%. Gemifloxacin was the most potent fluoroquinolone tested against S. pneumoniae (99.9% susceptible). In isolates of H. influenzae, beta-lactamase production was 16.9%, whereas the prevalence of beta-lactamase-negative, ampicillin-resistant strains was low (0.2%). beta-Lactamase production in M. catarrhalis world-wide remained high over the period studied (92.1%). Using PK/PD breakpoints, the most active non-fluoroquinolone agents against H. influenzae were ceftriaxone (100% susceptible), cefixime (99.8%) and co-amoxiclav (98.1-99.6%). Co-amoxiclav, cefdinir and cefixime (100%) were the most active beta-lactams against M. catarrhalis. Both H. influenzae and M. catarrhalis were highly susceptible to the fluoroquinolones. These data demonstrate the continued evolution of and geographical variation in bacterial resistance and highlight the need for appropriate prescribing of antimicrobials in CARTI, using agents with adequate activity, based on local susceptibility profiles and PK/PD parameters.


The endoplasmic reticulum: Homeostasis and crosstalk in retinal health and disease.
The endoplasmic reticulum (ER) is the largest intracellular organelle carrying out a broad range of important cellular functions including protein biosynthesis, folding, and trafficking, lipid and sterol biosynthesis, carbohydrate metabolism, and calcium storage and gated release. In addition, the ER makes close contact with multiple intracellular organelles such as mitochondria and the plasma membrane to actively regulate the biogenesis, remodeling, and function of these organelles. Therefore, maintaining a homeostatic and functional ER is critical for the survival and function of cells. This vital process is implemented through well-orchestrated signaling pathways of the unfolded protein response (UPR). The UPR is activated when misfolded or unfolded proteins accumulate in the ER, a condition known as ER stress, and functions to restore ER homeostasis thus promoting cell survival. However, prolonged activation or dysregulation of the UPR can lead to cell death and other detrimental events such as inflammation and oxidative stress; these processes are implicated in the pathogenesis of many human diseases including retinal disorders. In this review manuscript, we discuss the unique features of the ER and ER stress signaling in the retina and retinal neurons and describe recent advances in the research to uncover the role of ER stress signaling in neurodegenerative retinal diseases including age-related macular degeneration, inherited retinal degeneration, achromatopsia and cone diseases, and diabetic retinopathy. In some chapters, we highlight the complex interactions between the ER and other intracellular organelles focusing on mitochondria and illustrate how ER stress signaling regulates common cellular stress pathways such as autophagy. We also touch upon the integrated stress response in retinal degeneration and diabetic retinopathy. Finally, we provide an update on the current development of pharmacological agents targeting the UPR response and discuss some unresolved questions and knowledge gaps to be addressed by future research.


Computational Alanine Scanning and Structural Analysis of the SARS-CoV-2 Spike Protein/Angiotensin-Converting Enzyme 2 Complex.
The recent emergence of the pathogen severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent for the coronavirus disease 2019 (COVID-19), is causing a global pandemic that poses enormous challenges to global public health and economies. SARS-CoV-2 host cell entry is mediated by the interaction of the viral transmembrane spike glycoprotein (S-protein) with the angiotensin-converting enzyme 2 gene (ACE2), an essential counter-regulatory carboxypeptidase of the renin-angiotensin hormone system that is a critical regulator of blood volume, systemic vascular resistance, and thus cardiovascular homeostasis. Accordingly, this work reports an atomistic-based, reliable in silico structural and energetic framework of the interactions between the receptor-binding domain of the SARS-CoV-2 S-protein and its host cellular receptor ACE2 that provides qualitative and quantitative insights into the main molecular determinants in virus/receptor recognition. In particular, residues D38, K31, E37, K353, and Y41 on ACE2 and Q498, T500, and R403 on the SARS-CoV-2 S-protein receptor-binding domain are determined as true hot spots, contributing to shaping and determining the stability of the relevant protein-protein interface. Overall, these results could be used to estimate the binding affinity of the viral protein to different allelic variants of ACE2 receptors discovered in COVID-19 patients and for the effective structure-based design and development of neutralizing antibodies, vaccines, and protein/protein inhibitors against this terrible new coronavirus.


Effectiveness of Primary Health Care Services in Addressing Mental Health Needs of Minority Refugee Population in New Zealand.
Many people are displaced from their country of origin and become refugees, mostly due to armed conflicts, political violence and human rights abuse. Refugees have complex mental, physical, and social health problems related to their traumatic background and the experiences they have endured during their refugee journey. The aim of this qualitative exploratory study was to examine the effectiveness of primary health care services in addressing mental health needs of Bhutanese refugee women resettled in New Zealand. This study included focus group discussion with Bhutanese women and men followed by interviews with health service providers. The findings of this study highlighted inadequacies and constraints in addressing Bhutanese refugee women's mental health needs in New Zealand and provided evidence for recommendations to address these inadequacies.


Ribosome Stoichiometry: From Form to Function.
The existence of eukaryotic ribosomes with distinct ribosomal protein (RP) stoichiometry and regulatory roles in protein synthesis has been speculated for over 60 years. Recent advances in mass spectrometry (MS) and high-throughput analysis have begun to identify and characterize distinct ribosome stoichiometry in yeast and mammalian systems. In addition to RP stoichiometry, ribosomes host a vast array of protein modifications, effectively expanding the number of human RPs from 80 to many thousands of distinct proteoforms. Is it possible that these proteoforms combine to function as a 'ribosome code' to tune protein synthesis? We outline the specific benefits that translational regulation by specialized ribosomes can offer and discuss the means and methodologies available to correlate and characterize RP stoichiometry with function. We highlight previous research with a focus on formulating hypotheses that can guide future experiments and crack the ribosome code.


Modeling photoionization of aqueous DNA and its components.
Radiation damage to DNA is usually considered in terms of UVA and UVB radiation. These ultraviolet rays, which are part of the solar spectrum, can indeed cause chemical lesions in DNA, triggered by photoexcitation particularly in the UVB range. Damage can, however, be also caused by higher energy radiation, which can ionize directly the DNA or its immediate surroundings, leading to indirect damage. Thanks to absorption in the atmosphere, the intensity of such ionizing radiation is negligible in the solar spectrum at the surface of Earth. Nevertheless, such an ionizing scenario can become dangerously plausible for astronauts or flight personnel, as well as for persons present at nuclear power plant accidents. On the beneficial side, ionizing radiation is employed as means for destroying the DNA of cancer cells during radiation therapy. Quantitative information about ionization of DNA and its components is important not only for DNA radiation damage, but also for understanding redox properties of DNA in redox sensing or labeling, as well as charge migration along the double helix in nanoelectronics applications. Until recently, the vast majority of experimental and computational data on DNA ionization was pertinent to its components in the gas phase, which is far from its native aqueous environment. The situation has, however, changed for the better due to the advent of photoelectron spectroscopy in liquid microjets and its most recent application to photoionization of aqueous nucleosides, nucleotides, and larger DNA fragments. Here, we present a consistent and efficient computational methodology, which allows to accurately evaluate ionization energies and model photoelectron spectra of aqueous DNA and its individual components. After careful benchmarking, the method based on density functional theory and its time-dependent variant with properly chosen hybrid functionals and polarizable continuum solvent model provides ionization energies with accuracy of 0.2-0.3 eV, allowing for faithful modeling and interpretation of DNA photoionization. The key finding is that the aqueous medium is remarkably efficient in screening the interactions within DNA such that, unlike in the gas phase, ionization of a base, nucleoside, or nucleotide depends only very weakly on the particular DNA context. An exception is the electronic interaction between neighboring bases which can lead to sequence-specific effects, such as a partial delocalization of the cationic hole upon ionization enabled by presence of adjacent bases of the same type.


Production of chimeras between the Chinese soft-shelled turtle and Peking duck through transfer of early blastoderm cells.
Chimeras are useful models for studies of developmental biology and cell differentiation. Intraspecies and interspecies germline chimeras have been produced in previous studies, but the feasibility of producing chimeras between animals of two different classes remains unclear. To address this issue, we attempted to produce chimeras between the Chinese soft-shelled turtle and the Peking duck by transferring stage X blastoderm cells to recipient embryos. We then examined the survival and development of the PKH26-labeled donor cells in the heterologous embryos. At early embryonic stages, both turtle and duck donor cells that were labeled with PKH26 were readily observed in the brain, neural tube, heart and gonads of the respective recipient embryos. Movement of turtle donor-derived cells was observed in the duck host embryos after 48 h of incubation. Although none of the hatchlings presented a chimeric phenotype, duck donor-derived cells were detected in a variety of organs in the hatchling turtles, particularly in the gonads. Moreover, in the hatched turtles, mRNA expression of tissue-specific duck genes MEF2a and MEF2c was detected in many tissues, including the muscle, heart, small and large intestines, stomach and kidney. Similarly, SPAG6 mRNA was detected in a subset of turtle tissues, including the gonad and the small and large intestines. These results suggest that duck donor-derived cells can survive and differentiate in recipient turtles; however, no turtle-derived cells were detected in the hatched ducks. Our findings indicate that chimeras can be produced between animals of two different classes.


Capillary isoelectric focusing coupled offline to matrix assisted laser desorption/ionization mass spectrometry.
This work presents several critical details for making cIEF-MALDI-MS a robust technique which will allow for more routine application and aid in automation. This includes emphasis on the hardware necessary for syringe pump mobilization and proper protocol for preventing disruption from gas bubbles. Following these guidelines, excellent elution time reproducibility is demonstrated for six pI markers (RSD <5%). Additionally, the pI markers are used to calibrate the pH gradient and determine experimental pIs of proteins detected offline by mass spectrometry. This was demonstrated using a standard protein mixture of myoglobin and two forms of beta-lactoglobulin. Experimental determination of protein pIs and molecular weights were found to be in agreement with literature values. The technical details discussed provide a sound foundation for applying the offline coupling of MALDI-MS with cIEF.


Functional analysis of the BMP9 response of ALK1 mutants from HHT2 patients: a diagnostic tool for novel ACVRL1 mutations.
Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant genetically inheritable vascular dysplasia caused by mutations in genes encoding receptors of the transforming growth factor-beta (TGF-beta) family: ENG, encoding endoglin (HHT1), and ACVRL1, encoding activin receptor-like kinase-1 (ALK1; HHT2). Our recent discovery of bone morphogenetic protein 9 (BMP9) as the specific ligand for ALK1 allowed us to reevaluate the functional significance of ACVRL1 mutations. We generated 19 ALK1 mutants reproducing HHT2 mutations (4 were novel mutations) found throughout the protein. We show that all ALK1 mutant proteins were expressed by transfected cells; most of them were present at the cell surface and retained their ability to bind BMP9 (except for the extracellular mutants). However, most were defective in BMP9 signaling. None of the ALK1 mutants had a dominant negative effect on wild-type ALK1 activity. These data demonstrate that mutations of ACVRL1 fit with a functional haploinsufficiency model affecting BMP9 signaling. Our study also identified 4 ACVRL1 mutations (D179A, R386C, R454W, and A482V) that did not alter the BMP9 responses that are polymorphisms and 2 novel mutations that are pathogenic (L381P and I485F). This demonstrates that the analysis of BMP9 responses can be used as a diagnostic tool by geneticists confronted with novel or conflicting ACVRL1 mutations.


Digestive symptoms of COVID-19 and expression of ACE2 in digestive tract organs.
SARS-CoV-2 has resulted in numerous cases of Coronavirus Disease 2019 (COVID-19) worldwide. In addition to fever and respiratory symptoms, digestive symptoms also are observed in some patients with COVID-19. Angiotensin-converting enzyme 2 (ACE2) was reported to be the receptor for SARS-CoV-2. The aim of this study was to comprehensively investigate the digestive symptoms that occur in COVID-19 patients, and the potential pathogenic route of the SARS-CoV-2 infection in digestive tract organs (from the oral cavity to the gastrointestinal tract). We investigated the digestive symptoms of 48 patients with COVID-19 and explored ACE2 expression in digestive tract and lung cancers, based on a series of bulk and single-cell RNA sequencing data obtained from public databases. We found that 25% (12/48) of the patients with COVID-19 suffered from digestive symptoms, among which pharyngalgia (7/48) was the most common manifestation, followed by diarrhea (3/48), anorexia (3/48), and nausea (1/48). The bulk tissue RNA sequencing analysis indicated that digestive tract organs had higher ACE2 expression levels compared to the lung, and the expression of ACE2 in the lung increased with age. Single-cell RNA-Seq results showed that the ACE2-positive-cell ratio in digestive tract organs was significantly higher compared to the lung. ACE2 expression was higher in tumor cells compared to normal control (NC) tissues. While in gastric tissues, ACE2 expression gradually increased from chronic gastritis to metaplasia, to early cancer. Our data might provide a theoretical basis for screening the SARS-CoV-2 susceptible population and for the clinical classification of treatment of patients with COVID-19.


Chrysin (5,7-dihydroxyflavone) exerts anxiolytic-like effects through GABAA receptors in a surgical menopause model in rats.
The present study investigated the effects of the flavonoid chrysin (5,7-dihydroxyflavone) on anxiety-like behavior in rats in a model of surgical menopause and evaluated the participation of γ-aminobutyric acid-A (GABAA) receptors in these actions. At 12 weeks post-ovariectomy, the effects of different doses of chrysin (0.5, 1, 2, and 4 mg/kg) were evaluated in the elevated plus maze, light/dark test, and locomotor activity test, and comparisons were made with the clinically effective anxiolytic diazepam. The participation of GABAA receptors in the actions of chrysin was explored by pretreating the rats with the noncompetitive GABAA chloride ion channel antagonist picrotoxin (1 mg/kg). The results showed that chrysin (2 and 4 mg/kg) reduced anxiety-like behavior in both the elevated plus maze and light/dark test, and these effects were similar to diazepam. Pretreatment with picrotoxin had no effects on its own but prevented the anxiolytic-like effects of chrysin in both tests. Chrysin also increased rearing and grooming, without significantly altering the number of crossings in the locomotor activity test; these effects were also similar to diazepam. In conclusion, the flavonoid chrysin produced anxiolytic-like effects through actions on GABAA receptors in a model of surgical menopause in rats. These findings support the hypothesis that this flavonoid could be a future natural alternative for ameliorating symptoms of anxiety after surgical menopause in women.


How Business Cycles Affect the Healthcare Sector: A Cross-country Investigation.
The long-term relationship between the general economy and healthcare expenditures has been extensively researched, to explain differences in healthcare spending between countries, but the midterm (i.e., business cycle) perspective has been overlooked. This study explores business cycle sensitivity in both public and private parts of the healthcare sector across 32 countries. Responses to the business cycle vary notably, both across spending sources and across countries. Whereas in some countries, consumers and/or governments cut back, in others, private and/or public healthcare buyers tend to spend more. We also assess long-term consequences of business cycle sensitivity and show that public cost cutting during economic downturns deflates the mortality rates, whereas private cut backs increase the long-term growth in total healthcare expenditures. Finally, multiple factors help explain variability in cyclical sensitivity. Private cost cuts during economic downturns are smaller in countries with a predominantly publicly funded healthcare system and more preventive public activities. Public cut backs during contractions are smaller in countries that rely more on tax-based resources rather than social health insurances. Copyright © 2015 John Wiley & Sons, Ltd.


Immunisation of two rodent species with new live-attenuated mutants of Yersinia pestis CO92 induces protective long-term humoral- and cell-mediated immunity against pneumonic plague.
We showed recently that the live-attenuated Δlpp ΔmsbB Δail and Δlpp ΔmsbB::ailL2 mutants of Yersinia pestis CO92 provided short-term protection to mice against developing subsequent lethal pneumonic plague. These mutants were either deleted for genes encoding Braun lipoprotein (Lpp), an acetyltransferase (MsbB) and the attachment invasion locus (Ail) (Δlpp ΔmsbB Δail) or contained a modified version of the ail gene with diminished virulence (Δlpp ΔmsbB::ailL2). Here, long-term immune responses were first examined after intramuscular immunisation of mice with the above-mentioned mutants, as well as the newly constructed Δlpp ΔmsbB Δpla mutant, deleted for the plasminogen-activator protease (pla) gene instead of ail. Y. pestis-specific IgG levels peaked between day 35 and 56 in the mutant-immunised mice and were sustained until the last tested day 112. Splenic memory B cells peaked earlier (day 42) before declining in the Δlpp ΔmsbB::ailL2 mutant-immunised mice while being sustained for 63 days in the Δlpp ΔmsbB Δail and Δlpp ΔmsbB Δpla mutant-immunised mice. Splenic CD4+ T cells increased in all immunised mice by day 42 with differential cytokine production among the immunised groups. On day 120, immunised mice were exposed intranasally to wild-type (WT) CO92, and 80-100% survived pneumonic challenge. Mice immunised with the above-mentioned three mutants had increased innate as well as CD4+ responses immediately after WT CO92 exposure, and coupled with sustained antibody production, indicated the role of both arms of the immune response in protection. Likewise, rats vaccinated with either Δlpp ΔmsbB Δail or the Δlpp ΔmsbB Δpla mutant also developed long-term humoral and cell-mediated immune responses to provide 100% protection against developing pneumonic plague. On the basis of the attenuated phenotype, the Δlpp ΔmsbB Δail mutant was recently excluded from the Centers for Disease Control and Prevention select agent list.


The role of parental rearing practices and family demographics on oral health-related quality of life in children.
To evaluate the direct and indirect (via oral health-related behaviour) effects of parental rearing practices on children's Oral Health-Related Quality of Life (OHRQoL) within a family-focused, comprehensive predictive model. Participants were 11- to 14-year-old children and their parents living in Telangana State, India (N = 1130). Children were clinically assessed for dental caries, gingivitis, oral hygiene status, fluorosis, and malocclusion, and completed a self-administered questionnaire on oral health-related behaviour and OHRQoL. Parents answered questions related to their socioeconomic status (SES), family circumstances, parent's perceptions of child's OHRQoL, and child rearing practices. Structural equation modelling was used to evaluate the pathways through which parenting practices were associated with children's OHRQoL. Parents with higher positive (β = -0.106) and lower power assertion rearing practices (β = 0.103) had children with better OHRQoL. Parental rearing practices did not have any effect on children's oral hygiene behaviour. Children who had malocclusion (β = 0.076) and fluorosis (β = 0.38) had lower OHRQoL. Family SES had a significant effect on children's oral hygiene behaviour and oral hygiene status with children of higher SES demonstrating better oral hygiene behaviour and status. Children living in single-parent families reported poorer oral hygiene behaviour (β = -0.048) than those living in other types of families. Parental rearing practices had direct effects on OHRQoL. However, the hypothesised indirect effects of these practices on OHRQoL via poor oral health behaviour were not supported.


Myelo-optic neuropathy caused by aconitine in rabbit model.
A single dose of aconitine (0.6 mg/kg), the principal constituent of aconite, was administered intraperitoneally in order to evaluate its toxic effects on the visual system of a rabbit model. The typical pattern in which the alteration in the visual evoked potential occurred after an aconitine injection consisted of a delay in the onset and peak latency, and a reduction in the amplitude. Histopathologically, there was damage to the myelin sheath of the visual pathway, spinal cord and peripheral nerves. These findings suggest that a toxic dose of aconitine may cause myelo-optic neuropathy in rabbits.


Fatigue in osteoarthritis: a qualitative study.
Fatigue is recognized as a disabling symptom in many chronic conditions including rheumatic disorders such as rheumatoid arthritis (RA) and lupus. Fatigue in osteoarthritis (OA) is not routinely evaluated and has only been considered in a very limited number of studies. To date, these studies have focused primarily on patients with OA under rheumatological care, which represent the minority of people living with OA. The purpose of this study was to increase our understanding of the fatigue experience in community dwelling people with OA. In 2004, 8 focus groups were conducted with 28 men and 18 women (mean age 72.3) with symptomatic hip or knee OA recruited from a population-based cohort. Participants completed a self-administered questionnaire, which included demographics, measures of OA severity (WOMAC), depression (CES-D) and fatigue (FACIT). Sessions were audio taped and transcribed verbatim. Two researchers independently reviewed the transcripts to identify themes. Findings were compared and consensus reached. Mean pain, disability, depression and fatigue scores were 8.7/20, 27.8/68, 15.4/60, and 30.9/52, respectively. Participants described their fatigue as exhaustion, being tired and "coming up against a brick wall". Participants generally perceived fatigue as different from sleepiness and distinguished physical from mental fatigue. Factors believed to increase fatigue included OA pain and pain medications, aging, various types of weather and poor sleep. Mental health was identified as both affecting fatigue and being affected by fatigue. Participants described fatigue as impacting physical function, and their ability to participate in social activities and to do household chores. Rest, exercise, and avoiding or getting assistance with activities were cited as ways of coping. Participants generally did not discuss their fatigue with anyone except their spouses. Participants with OA described experiencing notable amounts of fatigue and indicated that it had a substantial impact on their lives. Further research is required to better understand the role of fatigue in OA in order to identify strategies to reduce its impact.


Clinical and genetic heterogeneity of hypochondroplasia.
Hypochondroplasia (HCH) is an autosomal dominant condition characterised by short stature, micromelia, and lumbar lordosis. In a series of 29 HCH probands (13 sporadic cases, 16 familial cases), we tested their DNA for the N540K recurrent mutation previously described in the proximal tyrosine kinase domain of the FGFR3 gene on chromosome 4p16.3, and we detected this mutation in 21/29 HCH patients. Interestingly, three familial cases were clearly unlinked to chromosome 4p16.3. Reviewing the clinical and radiological manifestations of the disease a posteriori, we observed that the N540K mutation was associated with relative macrocrania with a high and large forehead and short hands. By contrast, in the three pedigrees inconsistent with linkage to chromosome 4p16.3, the clinical phenotype was milder, macrocephaly and shortening of the long bones was less obvious, the hands were normal, and no metaphyseal flaring was noted. This study supports the view that HCH is a clinically and genetically heterogeneous condition.


Influence of water stress on endogenous hormone contents and cell damage of maize seedlings.
Phytohormones play critical roles in regulating plant responses to stress. We investigated the effects of water stress induced by adding 12% (w/v) polyethylene glycol to the root medium on the levels of abscisic acid (ABA), indole-3-acid (IAA), zeatin (ZT), and gibberellin(3) (GA(3)) in maize leaves. The results suggested that water stress had significant effects on the four hormone levels. There was a transient increase in the IAA content during the initial stage of adaptation to water stress in maize leaves, but it dropped sharply thereafter in response to water stress. ABA content increased dramatically in maize leaves after 24 h of exposure to water stress, and then the high levels of ABA were maintained to the end. The contents of ZT and GA(3) rapidly declined in maize leaves subjected to water stress. The effects of water stress on chlorophyll content, electrolyte leakage and malondialdehyde levels in maize leaves were also studied. The variation of cell damage was negatively correlated with ZT and GA(3) levels in maize leaves under water stress. Thus, we explored the roles of ZT and GA(3) on the growth of maize seedlings under water stress by exogenous application. It is possible that both ZT and GA(3) were effective in protecting maize seedlings from water stress, which would be of great importance for the improvement of drought tolerance in maize by genetic manipulation.


The adult lifespan of the female honey bee (Apis mellifera): Metabolic rate, AGE pigment and the effect of dietary fatty acids.
Female honey bees can be queens or workers and although genetically identical, workers have an adult lifespan of weeks while queens can live for years. The mechanisms underlying this extraordinary difference remain unknown. This study examines three potential explanations of the queen-worker lifespan difference. Metabolic rates were similar in age-matched queens and workers and thus are not an explanation. The accumulation of fluorescent AGE pigment has been successfully used as a good measure of cellular senescence in many species. Unlike other animals, AGE pigment level reduced during adult life of queens and workers. This unusual finding suggests female honey bees can either modify, or remove from their body, AGE pigment. Another queen-worker difference is that, as adults, workers eat pollen but queens do not. Pollen is a source of polyunsaturated fatty acids. Its consumption explains the queen-worker difference in membrane fat composition of female adult honey bees which has previously been suggested as a cause of the lifespan difference. We were able to produce "queen-worker" membrane differences in workers by manipulation of diet that did not change worker lifespan and we can, thus, also rule out pollen consumption by workers as an explanation of the dramatic queen-worker lifespan difference.


Place field repetition and spatial learning in a multicompartment environment.
Recent studies have shown that place cells in the hippocampus possess firing fields that repeat in physically similar, parallel environments. These results imply that it should be difficult for animals to distinguish parallel environments at a behavioral level. To test this, we trained rats on a novel odor-location task in an environment with four parallel compartments which had previously been shown to yield place field repetition. A second group of animals was trained on the same task, but with the compartments arranged in different directions, an arrangement we hypothesised would yield less place field repetition. Learning of the odor-location task in the parallel compartments was significantly impaired relative to learning in the radially arranged compartments. Fewer animals acquired the full discrimination in the parallel compartments compared to those trained in the radial compartments, and the former also required many more sessions to reach criterion compared to the latter. To confirm that the arrangement of compartments yielded differences in place cell repetition, in a separate group of animals we recorded from CA1 place cells in both environments. We found that CA1 place cells exhibited repeated fields across four parallel local compartments, but did not do so when the same compartments were arranged radially. To confirm that the differences in place field repetition across the parallel and radial compartments depended on their angular arrangement, and not incidental differences in access to an extra-maze visual landmark, we repeated the recordings in a second set of rats in the absence of the orientation landmark. We found, once again, that place fields showed repetition in parallel compartments, and did not do so in radially arranged compartments. Thus place field repetition, or lack thereof, in these compartments was not dependent on extra-maze cues. Together, these results imply that place field repetition constrains spatial learning.


Associations between school meal-induced dietary changes and metabolic syndrome markers in 8-11-year-old Danish children.
We recently showed that provision of Nordic school meals rich in fish, vegetables and potatoes and with reduced intakes of fat improved blood pressure, insulin resistance assessed by the homeostatic model (HOMA-IR), and plasma triacylglycerol despite increasing waist circumference in Danish 8-11-year-olds. This study explored whether intake or biomarkers of key dietary components in the schools meals were associated with these metabolic syndrome (MetS) markers during the 6-month intervention. Data from 7-day dietary records and measurements of whole-blood docosahexaenoic acid (DHA, 22:6n-3), blood pressure, fasting blood MetS markers, waist circumference and android/total fat mass assessed by dual-energy X-ray absorptiometry collected at baseline, 3 and 6 months from 523 children were analyzed in linear mixed-effects models adjusted for puberty, growth and fasting. After adjustment for multiple testing, whole-blood DHA was negatively associated with HOMA-IR (P < 0.001) and triacylglycerol (P < 0.0001). Potato intake was positively associated with waist circumference (P < 0.01), but not with android/total fat mass (P = 0.94). Intakes of whole-grain as well as dietary fiber, protein and fat were not associated with any of the MetS markers. DHA in whole-blood, an indicator of DHA and fish intake, seemed to be the main diet-related predictor of the beneficial effects of the school meals on MetS markers. Increased potato intake was associated with increased waist circumference, but this may not only be due to an increase in abdominal fat, as no association was seen with fat distribution.


Effectiveness of the course of medical ethics for undergraduate medical students.
Judgment This study was done in order to evaluate the effectiveness of the revisions made in the course of medical ethics for undergraduate medical students. Medical Students of Tehran University of Medical Sciences who took the course of medical ethics in a semester before the implementation of the revision and those who took the course after the implementation of the revision at the beginning and at the end of course responded to two questionnaires (one for evaluating knowledge and the other for assessing their moral judgment). Response rate was between 70 to 93.1 percent. Students' knowledge was significantly higher in the semester after the course revision (mean ± SD: 6.12 ± 1.3) in comparison with the semester before the reform (mean ± SD: 3.63 ± 1.7) (P=0.001). Students' knowledge after taking this course showed an increase of about 60% when compared with their knowledge level before starting the course (P=0.001). There was no significant difference in the level of moral judgment before and after taking the revised course of medical ethics while moral judgment level of students in two semester[before (21.21 ± 4.0) and after 15.25 ± 2.87) reform] were significantly different (P=0.02). The revisions made in the course of medical ethics for medical students were effective in improving students' knowledge but could not improve their moral judgment. This could be due to the short length of this course and also the small sample size in this study. We suggest that this study should be repeated with larger sample size and also with other methods of a course evaluation.


Airborne Optical and Thermal Remote Sensing for Wildfire Detection and Monitoring.
For decades detection and monitoring of forest and other wildland fires has relied heavily on aircraft (and satellites). Technical advances and improved affordability of both sensors and sensor platforms promise to revolutionize the way aircraft detect, monitor and help suppress wildfires. Sensor systems like hyperspectral cameras, image intensifiers and thermal cameras that have previously been limited in use due to cost or technology considerations are now becoming widely available and affordable. Similarly, new airborne sensor platforms, particularly small, unmanned aircraft or drones, are enabling new applications for airborne fire sensing. In this review we outline the state of the art in direct, semi-automated and automated fire detection from both manned and unmanned aerial platforms. We discuss the operational constraints and opportunities provided by these sensor systems including a discussion of the objective evaluation of these systems in a realistic context.


Comparison between transcriptomic responses to short-term stress exposures of a common Holarctic and endemic Lake Baikal amphipods.
Lake Baikal is one of the oldest freshwater lakes and has constituted a stable environment for millions of years, in stark contrast to small, transient bodies of water in its immediate vicinity. A highly diverse endemic endemic amphipod fauna is found in one, but not the other habitat. We ask here whether differences in stress response can explain the immiscibility barrier between Lake Baikal and non-Baikal faunas. To this end, we conducted exposure experiments to increased temperature and the toxic heavy metal cadmium as stressors. Here we obtained high-quality de novo transcriptome assemblies, covering mutiple conditions, of three amphipod species, and compared their transcriptomic stress responses. Two of these species, Eulimnogammarus verrucosus and E. cyaneus, are endemic to Lake Baikal, while the Holarctic Gammarus lacustris is a potential invader. Both Baikal species possess intact stress response systems and respond to elevated temperature with relatively similar changes in their expression profiles. G. lacustris reacts less strongly to the same stressors, possibly because its transcriptome is already perturbed by acclimation conditions.


Effectiveness and safety of oral anticoagulants in elderly patients with atrial fibrillation.
To assess the risk of stroke/systemic embolism (SE) and major bleeding associated with the use of oral anticoagulants in elderly patients with atrial fibrillation (AF) in a real-world population. We identified all anticoagulant-naive initiators of warfarin, dabigatran, rivaroxaban and apixaban for the indication AF in Norway between January 2013 and December 2017. Multivariate competing risk regression was used to calculate subhazard ratios (SHRs) describing associations between non-vitamin K antagonist oral anticoagulants (NOACs) compared with warfarin for risk of stroke/SE and major bleeding. Among 30 401 patients ≥75 years identified (median age 82 years, 53% women, mean CHA2DS2-VaSc score 4.5), 3857 initiated dabigatran, 6108 rivaroxaban, 13 786 apixaban and 6650 warfarin. Reduced dose was initiated in 11 559 (49%) of the NOAC-treated patients. For stroke, the SHRs for standard dose NOAC against warfarin were 0.80 (95% CI 0.57 to 1.13) for dabigatran; 1.07 (95% CI 0.89 to 1.30) for rivaroxaban and 0.95 (95% CI 0.78 to 1.15) for apixaban. For major bleeding, the SHRs against warfarin were 0.75 (95% CI 0.52 to 1.08) for dabigatran; 0.96 (95% CI 0.78 to 1.16) for rivaroxaban and 0.74 (95% CI 0.60 to 0.91) for apixaban. Comparing reduced doses of NOACs with warfarin yielded similar results. Sensitivity analyses were in accordance with the main results. In this nationwide cohort study of patients ≥75 years initiating oral anticoagulation for AF, standard and reduced dose NOACs were associated with similar risks of stroke/SE as warfarin and lower or similar risks of bleeding. The NOACs seem to be a safe option also in elderly patients.


The harvest plot: a method for synthesising evidence about the differential effects of interventions.
One attraction of meta-analysis is the forest plot, a compact overview of the essential data included in a systematic review and the overall 'result'. However, meta-analysis is not always suitable for synthesising evidence about the effects of interventions which may influence the wider determinants of health. As part of a systematic review of the effects of population-level tobacco control interventions on social inequalities in smoking, we designed a novel approach to synthesis intended to bring aspects of the graphical directness of a forest plot to bear on the problem of synthesising evidence from a complex and diverse group of studies. We coded the included studies (n = 85) on two methodological dimensions (suitability of study design and quality of execution) and extracted data on effects stratified by up to six different dimensions of inequality (income, occupation, education, gender, race or ethnicity, and age), distinguishing between 'hard' (behavioural) and 'intermediate' (process or attitudinal) outcomes. Adopting a hypothesis-testing approach, we then assessed which of three competing hypotheses (positive social gradient, negative social gradient, or no gradient) was best supported by each study for each dimension of inequality. We plotted the results on a matrix ('harvest plot') for each category of intervention, weighting studies by the methodological criteria and distributing them between the competing hypotheses. These matrices formed part of the analytical process and helped to encapsulate the output, for example by drawing attention to the finding that increasing the price of tobacco products may be more effective in discouraging smoking among people with lower incomes and in lower occupational groups. The harvest plot is a novel and useful method for synthesising evidence about the differential effects of population-level interventions. It contributes to the challenge of making best use of all available evidence by incorporating all relevant data. The visual display assists both the process of synthesis and the assimilation of the findings. The method is suitable for adaptation to a variety of questions in evidence synthesis and may be particularly useful for systematic reviews addressing the broader type of research question which may be most relevant to policymakers.


Structural and functional association of androgen receptor with telomeres in prostate cancer cells.
Telomeres protect the ends of linear chromosomes from being recognized as damaged DNA, and telomere stability is required for genome stability. Here we demonstrate that telomere stability in androgen receptor (AR)-positive LNCaP human prostate cancer cells is dependent on AR and androgen, as AR inactivation by AR antagonist bicalutamide (Casodex), AR-knockdown, or androgen-depletion caused telomere dysfunction, and the effect of androgen-depletion or Casodex was blocked by the addition of androgen. Notably, neither actinomycin D nor cycloheximide blocked the DNA damage response to Casodex, indicating that the role of AR in telomere stability is independent of its role in transcription. We also demonstrate that AR is a component of telomeres, as AR-bound chromatin contains telomeric DNA, and telomeric chromatin contains AR. Importantly, AR inactivation by Casodex caused telomere aberrations, including multiple abnormal telomere signals, remindful of a fragile telomere phenotype that has been described previously to result from defective telomere DNA replication. We suggest that AR plays an important role in telomere stability and replication of telomere DNA in prostate cancer cells, and that AR inactivation-mediated telomere dysfunction may contribute to genomic instability and progression of prostate cancer cells.


The recovery of human polymorphonuclear leucocytes from sublytic complement attack is mediated by changes in intracellular free calcium.
Using polymorphonuclear leucocyte-erythrocyte ghost hybrids entrapping the calcium-activated photoprotein obelin, we have demonstrated that sublytic amounts of the complement membrane attack complex induce a rapid but transient increase in intracellular free calcium ion concentration ([Ca2+]i). This increase in [Ca2+]i occurs prior to, and is required for, rapid removal of membrane attack complexes from the cell surface. The increase in [Ca2+]i is not only due to increased influx from outside the cell, but also results from mobilization of intracellular stores. The possible mechanism of mobilization of calcium, and the importance of an increase in [Ca2+]i as a mediator of recovery processes in nucleated cells, are discussed.


Evidence of Extended Thermo-Stability of Typhoid Polysaccharide Conjugate Vaccines.
Typhoid conjugate vaccines (TCV) are effective in preventing enteric fever caused by Salmonella enterica serovar Typhi in Southeast Asia and Africa. To facilitate vaccination with the Vi capsular polysaccharide-tetanus toxoid conjugate vaccine, Typbar TCV, and allow it to be transported and stored outside a cold chain just prior to administration, an extended controlled-temperature conditions (ECTC) study was performed to confirm the quality of the vaccine at 40 °C for 3 days at the end of its shelf-life (36 months at 2-8 °C). Studies performed in parallel by the vaccine manufacturer, Bharat Biotech International Limited, and an independent national control laboratory (NIBSC) monitored its stability-indicating parameters: O-acetylation of the Vi polysaccharide, integrity of the polysaccharide-protein conjugate, and its molecular size and pH. ECTC samples stored at 40 °C and 45 °C in comparison with control samples stored at 4 °C and 55 or 56 °C, were shown to have stable O-acetylation and pH; only very slight increases in the percentage of free saccharide and corresponding decreases in molecular size were observed. The deoxycholate method for precipitating conjugated polysaccharide was very sensitive to small incremental increases in percentage of free saccharide, in line with storage temperature and duration. This extended ECTC study demonstrated minimal structural changes to the Vi polysaccharide and conjugate vaccine and a stable formulation following extended exposure to elevated temperatures for the desired durations. This outcome supports the manufacturer's ECTC claim for the vaccine to be allowed to be taken outside the cold chain before its administration.


Analysis of Patterns of Physical Activity and Sedentary Behavior in an Urban Slum Setting in Nairobi, Kenya.
Insufficient physical activity and sedentary behavior are key risk factors for the emergence of noncommunicable diseases in the sub-Saharan African setting. Given the limited evidence base, research is required to understand the trends. This study describes the patterns of physical activity and sedentary behavior in a large sample of urban slum residents in Nairobi, Kenya. We used data collected from 5190 study participants as part of cardiovascular disease risk assessment. Data were collected about work-, transport-, and recreation-related physical activity as well as sitting and sleeping time. Using time spent on each type of physical activity and respective metabolic equivalents (METs), patterns of physical activity and associated factors were evaluated using descriptive statistics, Pearson correlations, and logistic regression. Nearly 50% of the study population was involved in work-related physical activities, whereas only 6.3% was involved in recreation-related physical activities. Involvement in physical activities decreased with age, and 17.4% had <600 MET-minutes per week. Higher sitting time was associated with insufficient physical activity. There were substantial gender differences in the time spent for physical activity. Given the positive relationship between insufficient physical activity and sedentary behavior, complementary interventions that improve physical activity and at the same time reduce sitting time are needed.


Beyond the landscape: Resistance modelling infers physical and behavioural gene flow barriers to a mobile carnivore across a metropolitan area.
Urbanization affects key aspects of wildlife ecology. Dispersal in urban wildlife species may be impacted by geographical barriers but also by a species' inherent behavioural variability. There are no functional connectivity analyses using continuous individual-based sampling across an urban-rural continuum that would allow a thorough assessment of the relative importance of physical and behavioural dispersal barriers. We used 16 microsatellite loci to genotype 374 red foxes (Vulpes vulpes) from the city of Berlin and surrounding rural regions in Brandenburg in order to study genetic structure and dispersal behaviour of a mobile carnivore across the urban-rural landscape. We assessed functional connectivity by applying an individual-based landscape genetic optimization procedure. Three commonly used genetic distance measures yielded different model selection results, with only the results of an eigenvector-based multivariate analysis reasonably explaining genetic differentiation patterns. Genetic clustering methods and landscape resistance modelling supported the presence of an urban population with reduced dispersal across the city border. Artificial structures (railways, motorways) served as main dispersal corridors within the cityscape, yet urban foxes avoided densely built-up areas. We show that despite their ubiquitous presence in urban areas, their mobility and behavioural plasticity, foxes were affected in their dispersal by anthropogenic presence. Distinguishing between man-made structures and sites of human activity, rather than between natural and artificial structures, is thus essential for better understanding urban fox dispersal. This differentiation may also help to understand dispersal of other urban wildlife and to predict how behaviour can shape population genetic structure beyond physical barriers.