Optical side-band cooling of a low frequency optomechanical system.
For experimental investigations of macroscopic quantum superpositions and the possible role of gravitational effects on the reduction of the corresponding quantum wave function it is beneficial to consider large mass, low frequency optomechanical systems. We report optical side-band cooling from room temperature for a 1.5×10⁻¹⁰ kg (mode mass), low frequency side-band resolved optomechanical system based on a 5 cm long Fabry-Perot cavity. By using high-quality Bragg mirrors for both the stationary and the micromechanical mirror we are able to construct an optomechanical cavity with an optical linewidth of 23 kHz. This, together with a resonator frequency of 315 kHz, makes the system operate firmly in the side-band resolved regime. With the presented optomechanical system parameters cooling close to the ground state is possible. This brings us one step closer to creating and verifying macroscopic quantum superpositions.


An efficient, non-invasive approach for in-vivo sampling of hair follicles: design and applications in monitoring DNA damage and aging.
In accordance with the 3 Rs principle (to replace, reduce and refine) animal models in biomedical research, we have developed and applied a new approach for sampling and analyzing hair follicles in various experimental settings. This involves use of a convenient device for non-invasive collection of hair follicles and processing methods that provide sufficient amounts of biological material to replace stressful and painful biopsies. Moreover, the main components of hair follicles are live cells of epithelial origin, which are highly relevant for most types of malignant tumors, so they provide opportunities for studying aging-related pathologies including cancer. Here, we report the successful use of the method to obtain mouse hair follicular cells for genotyping, quantitative PCR, and quantitative immunofluorescence. We present proof of concept data demonstrating its utility for routine genotyping and monitoring changes in quality and expression levels of selected proteins in mice after gamma irradiation and during natural or experimentally induced aging. We also performed pilot translation of animal experiments to human hair follicles irradiated ex vivo. Our results highlight the value of hair follicles as biological material for convenient in vivo sampling and processing in both translational research and routine applications, with a broad range of ethical and logistic advantages over currently used biopsy-based approaches.


Detection of novel chlamydiae in cats with ocular disease.
To detect and characterize the full range of chlamydial infections in cats with ocular disease by use of polymerase chain reaction (PCR) assays, cytologic examination, immunohistochemical analysis, and evaluation of clinical information including status for feline herpesvirus-1 (FeHV-1). DNA extracted from 226 conjunctival samples obtained from cats with clinically diagnosed keratitis or conjunctivitis and 30 conjunctival samples from healthy cats. PCR assays for the 16S rRNA gene specific for the order Chlamydiales and a new Chlamydophila felis (formerly Chlamydia psittaci) species-specific 23S rRNA gene were performed. Seventy-four conjunctival samples were prepared with Romanowsky-type stain, grouped on the basis of inflammatory pattern, and screened for chlamydial inclusions by use of immunohistochemical analysis. Clinical information and FeHV-1 status were recorded. 26 (12%) specimens had positive results for the only known feline chlamydial pathogen, C felis. Surprisingly, an additional 88 (39%) were positive for non-C felis chlamydial DNA. Identification of non-C felis chlamydial DNA by direct sequencing revealed 16S rRNA gene sequences that were 99% homologous to the sequence for Neochlamydia hartmannellae, an amebic endosymbiont. Chlamydial prevalence was significantly higher in cats with ocular disease. Application of a broad-range detection method resulted in identification of a new agent associated with ocular disease in cats. Finding chlamydia-like agents such as N hartmannellae in coinfections with their obligate amebic host, Hartmannella vermiformis, raises questions about the potential role of these microorganisms in causation or exacerbation of ocular disease in cats.


Metabolic maturation of differentiating cardiosphere-derived cells.
Cardiosphere-derived cells (CDCs) can be expanded in vitro and induced to differentiate along the cardiac lineage. To recapitulate the phenotype of an adult cardiomyocyte, differentiating progenitors need to upregulate mitochondrial glucose and fatty acid oxidation. Here we cultured and differentiated CDCs using protocols aimed to maintain stemness or to promote differentiation, including triggering fatty acid oxidation using an agonist of peroxisome proliferator-activated receptor alpha (PPARα). Metabolic changes were characterised in undifferentiated CDCs and during differentiation towards a cardiac phenotype. CDCs from rat atria were expanded on fibronectin or collagen IV via cardiosphere formation. Differentiation was assessed using flow cytometry and qPCR and substrate metabolism was quantified using radiolabelled substrates. Collagen IV promoted proliferation of CDCs whereas fibronectin primed cells for differentiation towards a cardiac phenotype. In both populations, treatment with 5-Azacytidine induced a switch towards oxidative metabolism, as shown by changes in gene expression, decreased glycolytic flux and increased oxidation of glucose and palmitate. Addition of a PPARα agonist during differentiation increased both glucose and fatty acid oxidation and expression of cardiac genes. We conclude that oxidative metabolism and cell differentiation act in partnership with increases in one driving an increase in the other.


Mass spectrometric analysis of synaptosomal membrane preparations for the determination of brain receptors, transporters and channels.
The molecular composition of synaptic signal transduction machineries shapes synaptic neurotransmission. The repertoire of receptors, transporters and channels (RTCs) comprises major signaling events in the brain. RTCs are conventionally studied by candidate immunohistochemistry and biochemistry, which are low throughput with resolution greatly affected by available immunoreagents and membrane interference. Therefore, a comprehensive resource of synaptic brain RTCs is still lacking. In particular, studies on the detergent-soluble synaptosomal fraction, known to contain transporters and channels, are limited. We, therefore, performed sub-synaptosomal fractionation of rat cerebral cortex, followed by trypsin/chymotrypsin sequential digestion of a detergent-soluble synaptosomal fraction and a postsynaptic density preparation, stable-isotope tryptic peptide labeling and liquid chromatography mass spectrometry. Based on the current study, a total of 4784 synaptic proteins were submitted to the ProteomExchange database (PXD001948), including 274 receptors, 394 transporters/channels and 1377 transmembrane proteins. Function-based classification assigned 1781 proteins as probable drug targets with 834 directly linked to brain disorders. The analytical approach identified 499 RTCs that are not listed in the largest, curated database for synaptosomal proteins (SynProt). This is a threefold RTC increase over all other data collected to date. Taken together, we present a protein discovery resource that can serve as a benchmark for future molecular interrogation of synaptic connectivity.


The use of human skin fibroblasts to obtain potency estimates of drug binding to androgen receptors.
Although several drugs with antiandrogenic properties have been used to treat such conditions as prostatic carcinoma, precocious puberty, acne, and hirsutism, their relative strengths in human tissues are not known. Most of the compounds that are effective clinically in opposing androgen action interact with the androgen receptor in various assay systems. To determine in human cells the relative potencies of these agents as well as others with androgenic properties, we measured the abilities of various compounds to compete with [3H]dihydrotestosterone [( 3H]DHT) for androgen-binding sites in dispersed human genital skin fibroblasts at 22 degrees C. The concentrations of unlabeled DHT, methyltrienolone (a synthetic non- metabolizeable androgen), and testosterone required for 50% inhibition of [3H]DHT binding were similar, approximately 1 nM [0.87 +/- 0.12 (+/- SE), 1.18 +/- 0.18, and 1.01 +/- 0.20 nM, respectively]. The relative binding activities, defined by the ratio of the concentration of methyltrienolone to the concentration of competitor required for 50% displacement of [3H]DHT, were as follows: spironolactone greater than R2956 (a synthetic antiandrogen) greater than megestrol acetate greater than cyproterone acetate greater than estradiol greater than flutamide much greater than testolactone greater than cimetidine. Danazol, an androgen agonist that causes hirsutism, was nearly as effective as spironolactone in its ability to compete for the fibroblast androgen receptor, 50% inhibition of fibroblast [3H]DHT binding was achieved by 1.76 +/- 0.31 nM spironolactone and 2.85 +/- 0.50 nM danazol. Two other compounds that induce hirsutism, diphenylhydantoin and diazoxide, did not displace [3H]DHT. We conclude that 1) of the compounds tested, spironolactone, which is rapidly metabolized in vivo to a much less potent competitor, is the most potent antiandrogen in its ability to interact in vitro with human skin fibroblast androgen receptors; 2) estradiol is a relatively potent androgen receptor binder; and 3) this receptor assay, combined with metabolic clearance and pharmacokinetic considerations, should be useful in selecting drugs for androgen and antiandrogen therapy.


Clear Liquids versus Polyethylene Glycol Preparation for Video Capsule Endoscopy of the Small Bowel: A Randomized Controlled Trial.
An effective bowel cleanse can improve the imaging quality of video capsule endoscopy (VCE). We aimed to further investigate the optimal small bowel cleanse method by comparing the efficacy of 4 L of clear liquids, 2 L of polyethylene glycol (PEG), and 4 L of PEG on the image quality of VCE. A randomized controlled, non-inferiority trial was performed comparing 4 L of clear liquids (Group A), 2 L of PEG (Group B), and 4 L of PEG (Group C). The primary endpoint was image quality between the groups. The secondary endpoints included patient tolerability and side effects. Eighty-one patients were analyzed in group A, 84 patients were analyzed in group B, and 80 patients were analyzed in group C. Image quality scores revealed 4 L of clear liquids to be non-inferior to 2 L of PEG, and 2 L of PEG to be non-inferior to 4 L of PEG (p < 0.0167). Group A had a lower difficulty of completion rate than Group B and Group C and a lower rate of side effects when compared to Group C (p < 0.0167). Four liters of clear liquids should be considered a routine method for small bowel preparation prior to VCE.


Predictors of Overtesting in Pulmonary Embolism Diagnosis.
The benefits of computed tomography pulmonary angiography (CTPA) for pulmonary embolism (PE) diagnosis must be weighed against its risks, radiation-induced malignancy, and contrast-induced nephropathy. Appropriate use of CTPA can be assessed by monitoring yield, the percentage of tests positive for PE. We identify factors that are associated low CTPA yield, which may predict overtesting. This was a retrospective cohort study of six emergency departments between June 2014 and February 2017. The electronic health record was queried for CTPAs ordered for adult patients in the emergency department. We assessed the following patient factors: age, gender, body mass index, number of comorbidities, race, and ethnicity, provider factors: type (resident, fellow, attending, physician assistant) and environment factors: test time of day, season of visit, and crowdedness of the department. A total of 14,782 CTPAs were reviewed, of which 1366 were found to be positive for PE, resulting in an overall CTPA yield of 9.24%. Provider type was not associated with a difference in yield. Testing was less likely to be positive in younger patients, females, those with lower body mass indexes and those identifying as Asian or Hispanic. Testing was also less likely to be positive when ordered during the overnight shift and during the winter and spring seasons. Our study identified several patient and environmental factors associated with low CTPA yield suggesting potential targets for overtesting. Targeting education and clinical decision support to assist providers in these circumstances may meaningfully improve yields.


Aggregation and vesiculation of membrane proteins by curvature-mediated interactions.
Membrane remodelling plays an important role in cellular tasks such as endocytosis, vesiculation and protein sorting, and in the biogenesis of organelles such as the endoplasmic reticulum or the Golgi apparatus. It is well established that the remodelling process is aided by specialized proteins that can sense as well as create membrane curvature, and trigger tubulation when added to synthetic liposomes. Because the energy needed for such large-scale changes in membrane geometry significantly exceeds the binding energy between individual proteins and between protein and membrane, cooperative action is essential. It has recently been suggested that curvature-mediated attractive interactions could aid cooperation and complement the effects of specific binding events on membrane remodelling. But it is difficult to experimentally isolate curvature-mediated interactions from direct attractions between proteins. Moreover, approximate theories predict repulsion between isotropically curving proteins. Here we use coarse-grained membrane simulations to show that curvature-inducing model proteins adsorbed on lipid bilayer membranes can experience attractive interactions that arise purely as a result of membrane curvature. We find that once a minimal local bending is realized, the effect robustly drives protein cluster formation and subsequent transformation into vesicles with radii that correlate with the local curvature imprint. Owing to its universal nature, curvature-mediated attraction can operate even between proteins lacking any specific interactions, such as newly synthesized and still immature membrane proteins in the endoplasmic reticulum.


The ERICA Tool.
The ERICA Tool is a computerised, flexible software system that has a structure based upon the ERICA Integrated Approach to assessing the radiological risk to biota. The Tool guides the user through the assessment process, recording information and decisions and allowing the necessary calculations to be performed to estimate risks to selected animals and plants. Tier 1 assessments are media concentration based and use pre-calculated environmental media concentration limits to estimate risk quotients. Tier 2 calculates dose rates but allows the user to examine and edit most of the parameters used in the calculation including concentration ratios, distribution coefficients, percentage dry weight soil or sediment, dose conversion coefficients, radiation weighting factors and occupancy factors. Tier 3 offers the same flexibility as Tier 2 but allows the option to run the assessment probabilistically if the underling parameter probability distribution functions are defined. Results from the Tool can be put into context using incorporated data on dose-effects relationships and background dose rates.


S1P Stimulates Proliferation by Upregulating CTGF Expression through S1PR2-Mediated YAP Activation.
Dysregulation of the Hippo pathway in the liver results in overgrowth and eventually tumorigenesis. To date, several upstream mechanisms have been identified that affect the Hippo pathway, which ultimately regulate YAP, the major downstream effector of the pathway. However, upstream regulators of the Hippo pathway in the liver remain poorly defined. Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite that has been shown to stimulate hepatocellular carcinoma (HCC) cell proliferation, but whether the Hippo pathway is involved in S1P-stimulated HCC cell proliferation remains to be determined. Here it is demonstrated that S1P activates YAP and that the S1P receptor 2 (S1PR2/S1P2) mediates S1P-induced YAP activation in both human and mouse HCC cells. S1P promotes YAP-mediated upregulation of cysteine-rich protein 61 and connective tissue growth factor (CTGF), and stimulates HCC cell proliferation. By using siRNA-mediated knockdown approaches, only CTGF was required for S1P-stimulated cell proliferation. Of note, S1P activates YAP in a MST1/2-independent manner suggesting that the canonical Hippo kinase is not required for S1P-mediated proliferation in liver. The upregulation of CTGF and S1P2 were also observed in liver-specific YAP overexpression transgenic mouse hepatocytes. Moreover, YAP regulated liver differentiation-dependent gene expression by influencing the chromatin binding of HNF4α based on ChIP-seq analysis. Finally, results using gain- and loss-of-function approaches demonstrate that HNF4α negatively regulated S1P-induced CTGF expression.Implications: These findings reveal a role for S1P in stimulating HCC cell proliferation by upregulating CTGF expression through S1P2-mediated YAP activation. Mol Cancer Res; 16(10); 1543-55. ©2018 AACR.


A sham-controlled trial of acupressure on the quality of sleep and life in haemodialysis patients.
Sleep disorder in haemodialysis patients can lead to disturbance in their psychosocial function and interpersonal relations, and reduced quality of life. The aim of the present study was to investigate the effect of acupressure on the quality of sleep of haemodialysis patients. In a randomised controlled trial, 108 haemodialysis patients were randomly divided into three groups: true acupressure, placebo acupressure, and no treatment. The two acupressure groups received treatment three times a week for 4 weeks during dialysis. Routine care only was provided for the no treatment group. The main study outcome was sleep quality. The total Pittsburgh Sleep Quality Index score decreased significantly from 11.9±3.13 to 6.2±1.93 in the true acupressure group, from 11.3±3.69 to 10.6±3.82 in the sham acupressure group, and from 10.9±4.10 to 10.7±3.94 in the no treatment group. There was a significant difference between groups (p<0.001). Acupressure seems to have a positive effect on the sleep quality in haemodialysis patients. IRCT201106145864N2.


Management of actinic cheilitis using diclofenac 3% gel: a report of six cases.
Actinic cheilitis is a frequent manifestation of actinic dysplasia and requires early therapy to prevent its progression into invasive squamous cell carcinoma (SCC). Several therapies are used, ranging from unspecific lesion-adapted destructive techniques (i.e. laser) to ambitious surgical field-management (vermillionectomy). There is increasing awareness of the effectiveness of field adapted, non destructive therapies, such as photodynamic therapy or 5% imiquimod. Diclofenac 3% gel is used in the treatment of actinic keratosis (AK), but it has not been evaluated for the treatment of actinic cheilitis. This non-blinded, uncontrolled case series study evaluated the effects of diclofenac 3% gel in the treatment of actinic cheilitis. Six patients with histologically verified actinic cheilitis were treated with diclofenac 3% gel, twice daily for 6 weeks. Clinical assessment was performed 2-4 weeks after the end of treatment. Four out of six patients showed clinical clearing of actinic cheilitis 2-4 weeks after the end of treatment. Biopsies were taken from the treated areas at the final visit to verify clinical clearance. Side effects in most of the patients included mild erythema and mild to moderate swelling of the lips. Topical therapy with diclofenac 3% gel may be an efficient, cosmetically more appealing alternative treatment for actinic cheilitis than currently used destructive therapies. However, future studies and long-term follow-up of patients will be needed to compare its efficacy with established forms of therapy.


The NMDA receptor intracellular C-terminal domains reciprocally interact with allosteric modulators.
N-methyl-d-aspartate receptors (NMDARs) have multiple prominent roles in CNS function but their excessive or insufficient activity contributes to neuropathological/psychiatric disorders. Consequently, a variety of positive and negative allosteric modulators (PAMs and NAMs, respectively) have recently been developed. Although these modulators bind to extracellular domains, in the present report we find that the NMDAR's intracellular C-terminal domains (CTDs) significantly influence PAM/NAM activity. GluN2 CTD deletion robustly affected NAM and PAM activity with both enhancing and inhibiting effects that were compound-specific and NMDAR subunit-specific. In three cases, individual PAMs became NAMs at specific GluN2-truncated receptors. In contrast to GluN2, GluN1 CTD removal only reduced PAM activity of UBP684 and CIQ, and did not affect NAM activity. Consistent with these findings, agents altering phosphorylation state or intracellular calcium levels displayed receptor-specific and compound-specific effects on PAM activity. It is possible that the GluN2's M4 domain transmits intracellular modulatory signals from the CTD to the M1/M4 channel gating machinery and that this site is a point of convergence in the direct or indirect actions of several PAMs/NAMs thus rendering them sensitive to CTD status. Thus, allosteric modulators are likely to have a marked and varied sensitivity to post-translational modifications, protein-protein associations, and intracellular ions. The interaction between PAM activity and NMDAR CTDs appears reciprocal. GluN1 CTD-deletion eliminated UBP684, but not pregnenolone sulfate (PS), PAM activity. And, in the absence of agonists, UBP684, but not PS, was able to promote movement of fluorescently-tagged GluN1-CTDs. Thus, it may be possible to pharmacologically target NMDAR metabotropic activity in the absence of channel activation.


Deep brain stimulation plus best medical therapy versus best medical therapy alone for advanced Parkinson's disease (PD SURG trial): a randomised, open-label trial.
Surgical intervention for advanced Parkinson's disease is an option if medical therapy fails to control symptoms adequately. We aimed to assess whether surgery and best medical therapy improved self-reported quality of life more than best medical therapy alone in patients with advanced Parkinson's disease. The PD SURG trial is an ongoing randomised, open-label trial. At 13 neurosurgical centres in the UK, between November, 2000, and December, 2006, patients with Parkinson's disease that was not adequately controlled by medical therapy were randomly assigned by use of a computerised minimisation procedure to immediate surgery (lesioning or deep brain stimulation at the discretion of the local clinician) and best medical therapy or to best medical therapy alone. Patients were analysed in the treatment group to which they were randomised, irrespective of whether they received their allocated treatment. The primary endpoint was patient self-reported quality of life on the 39-item Parkinson's disease questionnaire (PDQ-39). Changes between baseline and 1 year were compared by use of t tests. This trial is registered with Current Controlled Trials, number ISRCTN34111222. 366 patients were randomly assigned to receive immediate surgery and best medical therapy (183) or best medical therapy alone (183). All patients who had surgery had deep brain stimulation. At 1 year, the mean improvement in PDQ-39 summary index score compared with baseline was 5.0 points in the surgery group and 0.3 points in the medical therapy group (difference -4.7, 95% CI -7.6 to -1.8; p=0.001); the difference in mean change in PDQ-39 score in the mobility domain between the surgery group and the best medical therapy group was -8.9 (95% CI -13.8 to -4.0; p=0.0004), in the activities of daily living domain was -12.4 (-17.3 to -7.5; p<0.0001), and in the bodily discomfort domain was -7.5 (-12.6 to -2.4; p=0.004). Differences between groups in all other domains of the PDQ-39 were not significant. 36 (19%) patients had serious surgery-related adverse events; there were no suicides but there was one procedure-related death. 20 patients in the surgery group and 13 in the best medical therapy group had serious adverse events related to Parkinson's disease and drug treatment. At 1 year, surgery and best medical therapy improved patient self-reported quality of life more than best medical therapy alone in patients with advanced Parkinson's disease. These differences are clinically meaningful, but surgery is not without risk and targeting of patients most likely to benefit might be warranted.


Survey of nutrition knowledge of Canadian physicians.
Previous reports have indicated that physicians generally have little training in nutrition and a poor knowledge of the subject. A survey was carried out to determine the nutrition knowledge of physicians working in general practice. A questionnaire with multiple-choice questions was mailed to 248 physicians working in Alberta, Canada, mainly in Edmonton and Calgary. Non-respondents received a second questionnaire and a phone call. Completed questionnaires were received from 36.1% (84 of 233 eligible physicians). The average correct response was 63.1%. The results indicate that physicians are generally aware of information which has been publicized in the medical press: which nutrients are antioxidants; the nutrient associated with the prevention of neural tube defects (folate); the preventive action of fruit and vegetables against cancer; the energy value of fat (9 kcals/g); and the recommended fat intake (under 30% of energy). By contrast they have a poor knowledge of other important topics in nutrition: the typical salt intake of Canadians; the association between excess protein intake and calcium loss; the type of dietary fiber helpful in lowering the blood cholesterol level (soluble fiber); and the nutrient which helps prevent thrombosis (omega-3 fat). These results support other data that physicians need more training in nutrition.


Girls' and boys' trajectories of appearance anxiety from age 10 to 15 years are associated with earlier maturation and appearance-related teasing.
Adolescents' appearance-related concerns can provoke increasing emotional, social, and eating-related problems. The aims of this five-wave (2.5-year), multiple-informant longitudinal study were to (a) examine growth trajectories of appearance anxiety symptoms and appearance esteem, (b) identify whether trajectories differed by gender, and (c) examine several launching factors including parent-reported physical maturation, peer-rated physical appearance, body mass index, and appearance teasing by parents and peers. Participants were 387 adolescents (44% boys) aged 10 to 13 years at the first assessment. Steep growth in appearance anxiety symptoms was found for both girls and boys, but there was no average change in appearance esteem. Girls had more elevated appearance anxiety symptoms and lower appearance esteem than boys, girls' body mass index was associated with symptoms, and earlier physical maturation and teasing about appearance, alone and in combination, were associated with growth in appearance anxiety symptoms for girls and boys. Earlier maturing boys who were highly teased by parents, but even more so when teased by peers, were at utmost risk for elevated appearance anxiety symptoms and increasing symptoms over time. In contrast, all girls exhibited elevated or increasing appearance anxiety symptoms across time, with the exception of girls with the latest maturation who also reported little teasing about their appearance.


Why time poverty matters for individuals, organisations and nations.
Over the last two decades, global wealth has risen. Yet material affluence has not translated into time affluence. Most people report feeling persistently 'time poor'-like they have too many things to do and not enough time to do them. Time poverty is linked to lower well-being, physical health and productivity. Individuals, organisations and policymakers often overlook the pernicious effects of time poverty. Billions of dollars are spent each year to alleviate material poverty, while time poverty is often ignored or exacerbated. In this Perspective, we discuss the societal, organisational, institutional and psychological factors that explain why time poverty is often under appreciated. We argue that scientists, policymakers and organisational leaders should devote more attention and resources toward understanding and reducing time poverty to promote psychological and economic well-being.


Contrasting Rates of LINE-1 Amplification among New World Primates of the Atelidae Family.
LINE-1 (L1) retrotransposons constitute the dominant category of transposons in mammalian genomes. L1 elements are active in the vast majority of mammals, and only a few cases of L1 extinction have been documented. The only possible case of extinction in primates was suggested for South American spider monkeys. However, these previous studies were based on a single species. We revisited this question with a larger phylogenetic sample, covering all 4 genera of Atelidae and 3 species of spider monkeys. We used an enrichment method to clone recently inserted L1 elements and performed an evolutionary analysis of the sequences. We were able to identify young L1 elements in all taxa, suggesting that L1 is probably still active in all Atelidae examined. However, we also detected considerable variations in the proportion of recent elements indicating that the rate of L1 amplification varies among Atelidae by a 3-fold factor. The extent of L1 amplification in Atelidae remains overall lower than in other New World monkeys. Multiple factors can affect the amplification of L1, such as the demography of the host and the control of transposition. These factors are discussed in the context of host life history.


Concentration- and schedule-dependent effects of chemotherapy on the angiogenic potential and drug sensitivity of vascular endothelial cells.
The anti-angiogenic activity of chemotherapy is both dose- and schedule-dependent. While conventional maximum tolerated dose (MTD) chemotherapy exerts only mild and reversible anti-angiogenic effects, low-dose metronomic (LDM) chemotherapy was developed to specifically target tumour angiogenesis. However, the long-term effects of either MTD or LDM chemotherapy on vascular endothelial cells have never been investigated. Here, we demonstrated that repeated exposure to MTD and LDM chemotherapy differentially impact on the angiogenic potential and chemosensitivity of immortalized endothelial cells. Repeated MTD vinblastine treatment of vascular endothelial cells led to an increased proliferation rate and resistance to paclitaxel. In contrast, repeated LDM treatment with vinblastine or etoposide impaired the angiogenic potential of endothelial cells and increased their chemosensitivity. This effect was associated with a significant decrease in βII- and βIII-tubulin expression. Functional analysis using siRNA showed that silencing the expression of βIII-tubulin in endothelial cells significantly decreased their capacity to form vascular structures and increased their sensitivity to the anti-angiogenic and vascular-disrupting effects of chemotherapy, whereas silencing βII-tubulin expression had no effect. Collectively our results show that LDM chemotherapy impairs the angiogenic potential of endothelial cells while increasing their chemosensitivity-an effect at least in part mediated by the down-regulation of βIII-tubulin expression. Furthermore, our study suggests that βIII-tubulin represents an attractive therapeutic target to increase the anti-angiogenic effects of chemotherapy and overall anti-tumour efficacy.


Differential protein immunoexpression profiles in appendiceal mucinous neoplasms: a special reference to classification and predictive factors.
Appendiceal mucinous neoplasms have been the focus of considerable debate in recent years. We histologically classified 70 appendiceal mucinous neoplasms into three categories: 32 mucinous adenoma, 23 mucinous neoplasm of uncertain malignant potential, and 15 mucinous adenocarcinomas. Immunohistochemistry was performed for 24 proteins in different functional categories, specifically, oncogenic proteins (bcl-2, beta-catenin, CEA, C-erbB2, c-kit, Cox-2, Cyclin D1, EGFR, Ki-67, NF-kappaB, VEGF), tumor suppressors (E-cadherin, FHIT, hMLH1, p53, p63, smad4), cell-cycle regulators (p21, p27, p16), and mucin proteins (MUC1, MUC2, MUC5AC, MUC6). Our data showed that 9 out of the 24 proteins were more frequently altered in the mucinous adenocarcinoma group than in the mucinous adenoma group (P<0.05), including beta-catenin (13% in mucinous adenoma vs 60% in mucinous adenocarcinoma), CyclinD1 (44 vs 87%), Ki-67 (high labeling index: 31 vs 67%), NF-kappaB (19 vs 60%), VEGF (16 vs 87%), E-cadherin (0 vs 47%), p53 (6 vs 40%), MUC2 (9 vs 67%), and MUC5AC (3 vs 40%). The distinct immunoexpression profile of mucinous neoplasm of uncertain malignant potential was placed between those of mucinous adenoma and mucinous adenocarcinoma (P<0.05). Moreover, the mucinous adenoma, mucinous neoplasm of uncertain malignant potential, and mucinous adenocarcinoma categories displayed differences in terms of the number of altered markers among the nine proteins (P<0.05; mean 1.4 vs 2.6 vs 5.5, respectively). In mucinous adenocarcinoma, the p53 status was related to disease-free survival and overall survival of patients (P<0.05, both). NF-kappaB status and the number of altered protein markers made statistically marginal impacts on disease-free survival; also beta-catenin loss, on overall survival of patients. In conclusion, protein immunoexpression profiles may facilitate the classification of appendiceal mucinous neoplasms. In our study, the three tumor categories of mucinous adenoma, mucinous neoplasm of uncertain malignant potential, and mucinous adenocarcinoma exhibited distinct immunoexpression profiles. Five and more altered protein markers, p53 overexpression, NF-kappaB positivity, and beta-catenin loss were predictive factors of adverse clinical outcomes in appendiceal mucinous adenocarcinomas.


Genetic risk factors for cerebral small-vessel disease in hypertensive patients from a genetically isolated population.
Asymptomatic cerebral lesions on MRI such as white matter lesions (WML), lacunes and microbleeds are commonly seen in older people. We examined the role of a series of candidate genes involved in blood pressure regulation and amyloid metabolism. The study was embedded in a family-based cohort sampled from a Dutch genetically isolated population. We selected individuals between 55 and 75 years of age with hypertension (N=129). Volumes of WML and presence of lacunes and microbleeds were assessed with MRI. We studied three genes involved in blood pressure regulation (angiotensin, angiotensin II type 1 receptor, α-adducin) and two genes involved in the amyloid pathway (apolipoprotein E (APOE) and sortilin-related receptor gene (SORL1)). All participants had WML (median volume, 3.1 ml; interquartile range, 1.5-6.5 ml); lacunar infarcts were present in 15.5% and microbleeds in 23.3%. Homozygosity for the APOE ε4 allele was associated with lacunes (OR, 4.8; 95% CI, 1.2 to 19.3). Individuals carrying two copies of the variant allele of four single nucleotide polymorphism (SNPs) located at the 3'-end of SORL1 (rs1699102, rs3824968, rs2282649, rs1010159) had significantly more often microbleeds (highest OR, 6.87; 95% CI, 1.78 to 26.44). The association of SORL1 with microbleeds suggests that the amyloid cascade is involved in the aetiology of microbleeds in populations with hypertension.


Predictors of caregiver burden and care costs for older persons with dementia in Taiwan.
ABSTRACTObjectives:As a degenerative disease, the progression of dementia needs continued care provision and poses both psychological and financial burden for family caregivers of persons with dementia (PWD). This study seeks to compare predictors of care costs and caregiver burden, and to identify modifiable factors that could alleviate the burden faced by dementia caregivers. This study interviewed 231 PWD-caregiver dyads in a dementia clinic at a teaching hospital in southern Taiwan in 2013. A follow-up study was conducted a year later, and 167 dyads completed the second interview. Data collected included PWD characteristics, caregiver characteristics, relationship to PWD, and social support to caregivers. Caregiver burden was measured with the Zarit Burden Interview instrument. The association between each predictor variable and cost of care and caregiver burden scores was examined using linear mixed models. Predictors of care costs were found to be different from predictors of caregiver burden: functional declines measured by Katz's activities of daily living (ADL) scale were associated with total cost as compared to behavioral disturbance measured by Neuropsychiatric Inventory (NPI), which showed no impact on care costs. However, NPI was a significant predictor of caregiver burden. Caregivers who were better-off financially also reported significantly lower caregiver burden. Since predictors of care costs were different from the predictors of caregiver burden, providing training to caregivers in addressing PWD's behavioral disturbance and proving financial assistance to low income caregivers could be effective in reducing caregiver burden.


The origins of premating reproductive isolation: testing hypotheses in the grasshopper Chorthippus parallelus.
There are many proposed routes for the origin of premating reproductive isolation, but few systematic studies aimed at testing their relative importance. Accumulated information about the biogeographical history of the European meadow grasshopper, Chorthippus parallelus, has allowed us to make a planned series of comparisons among populations aimed at distinguishing the contributions of some of these hypotheses. We have compared the effects on assortative mating of long-term isolation in glacial refugia, founder events during postglacial colonization, and sympatry with a closely related species. A likelihood-based analysis allowed us to separate effects of variation in male and female mating propensity among populations from variation in mate choice leading to assortative mating. All three effects contributed significantly to the overall variation in mating pattern in a set of 21 pairwise comparisons among seven populations. Male cuticular composition, but not other candidate signals, was significantly associated with the level of assortative mating. Of the hypotheses for the origin of reproductive isolation, only the predictions of the founder hypothesis explained a significant amount of the variation in assortative mating. This does not rule out the possiblity that there may be some other explanation. Having established the pattern of divergence, it is possible to generate hypotheses that explain our results at least as well as the founder hypothesis. However, because many such post hoc hypotheses are possible, they cannot be tested with this dataset. On this basis, our results favor the hypothesis that some aspect of the colonization process tends to accelerate divergence in mating signals leading to premating reproductive isolation. This could be accomplished through any one of several mechanisms. Colonization involves many bottlenecks as new populations are established at the edge of the range by long-distance migrants. Genetic effects may be important, but these bottlenecks may also alter the conditions under which mates are found and chosen, as suggested by Kaneshiro. At the same time, the colonizing populations may encounter novel environmental challenges.


Negative controls: a tool for detecting confounding and bias in observational studies.
Noncausal associations between exposures and outcomes are a threat to validity of causal inference in observational studies. Many techniques have been developed for study design and analysis to identify and eliminate such errors. Such problems are not expected to compromise experimental studies, where careful standardization of conditions (for laboratory work) and randomization (for population studies) should, if applied properly, eliminate most such noncausal associations. We argue, however, that a routine precaution taken in the design of biologic laboratory experiments--the use of "negative controls"--is designed to detect both suspected and unsuspected sources of spurious causal inference. In epidemiology, analogous negative controls help to identify and resolve confounding as well as other sources of error, including recall bias or analytic flaws. We distinguish 2 types of negative controls (exposure controls and outcome controls), describe examples of each type from the epidemiologic literature, and identify the conditions for the use of such negative controls to detect confounding. We conclude that negative controls should be more commonly employed in observational studies, and that additional work is needed to specify the conditions under which negative controls will be sensitive detectors of other sources of error in observational studies.


Physical activity and global genomic DNA methylation in a cancer-free population.
Changes in DNA methylation may represent an intermediate step between the environment and human diseases. Little is known on whether behavioral risk factors may modify gene expression through DNA methylation. To assess whether DNA methylation is associated with different levels of physical activity, we measured global genomic DNA methylation using bisulfite converted DNA and real time PCR (MethyLight) for LINE-1 in peripheral blood of 161 participants aged 45-75 years enrolled in the North Texas Healthy Heart Study and levels of physical activity using an accelerometer (Actigraph GT1M Monitor). We found that individuals with physical activity 26-30 min/day had a significantly higher level of global genomic DNA methylation compared to those with physical activity ≤ 10 min/day (β=2.52, 95%CI: 0.70, 4.35) However, the association was attenuated and became statistically insignificant after multivariate adjustment (β=1.24, 95%CI:-0.93, 3.40). There were some suggestions of a positive association between physical activity and global genomic DNA methylation in non-Hispanics (β=1.50, 95%CI: -0.08, 3.08) that warrants further investigation.


Management of isolated yew berry toxicity with sodium bicarbonate: a case report in treatment efficacy.
Taxus species are known to be toxic and may result in significant dysrhythmias. Treatment of taxus induced cardiac dysrhythmias is based largely on case reports. We describe a case of a 24-year-old male with Taxus cuspidate (yew berry) toxicity initially treated with amiodarone bolus and infusion and subsequently managed with sodium bicarbonate boluses and continuous infusion. The patient was found at home by his parents with witnessed "seizure-like"activity 2 hours after reportedly chewing and swallowing 168 yew seeds. The initial prehospital rhythm strip demonstrated ventricular tachycardia (VT); the patient was hypotensive with fluctuating levels of alertness. Prehospital cardioversion was attempted without success. Staff at the local presenting emergency department (ED) consulted toxicology for management of the presumed yew berry ingestion, complicated by cardiac dysrhythmias and mental status change with seizure. Amiodarone 300-mg IV and diazepam 5-mg IV were given. Cardioversion was attempted 4 times without change in the wide complex tachycardia, presumed to be VT, at a rate of 166. An amiodarone drip at 1 mg/min was initiated. The patient was transferred to an intensive care unit (ICU) at a regional toxicology center. On arrival to the toxicology center the patient was alert and verbally appropriate without complaints. Initial heart rate was 76 and regular with premature ventricular contractions (PVCs). A wide complex tachycardia associated with hypotension recurred; however, normal mental status was maintained. A bolus of 100 mEq of sodium bicarbonate (NaHCO3) was given intravenously followed by sodium bicarbonate infusion at 37.5 mEq/hr. The amiodarone drip was discontinued. Subsequent electrocardiograms (EKG's) revealed a prolonged, but steadily narrowing QRS complex. Ultimately, the QRS complex closed to 92 ms, with a rate of 94, PR 154 and a QT/QTc of 390/487. This case describes successful treatment of an isolated Taxus cuspidate (yew berry) ingestion with significant toxicity initially with amiodarone bolus and infusion. Due to lack of significant change in telemetry recordings with amiodarone, treatment with sodium bicarbonate bolus and infusion was initiated. While the QRS narrowed significantly temporally related to the bicarbonate, it is difficult to determine if correction of the cardiac dysrhythmias was solely due to the sodium bicarbonate, or the synergism of sodium bicarbonate and amiodarone, or possibly spontaneous improvement due to taxine clearance. One should use caution while drawing conclusions from a single case; however, based on the clinical improvement of this patient, both with EKG recordings and vital signs, this report would suggest that isolated Taxus cuspidate ingestion from yew berry plants can be treated with sodium bicarbonate.


Understanding the Role of Social Factors in Farmworker Housing and Health.
Differences in social advantage significantly influence health conditions and life expectancy within any population. Such factors reproduce historic class, race, and ethnic disparities in community success. Few populations in the United States face more social and economic disadvantage than farmworkers, and farmworker housing has significant potential to ameliorate or amplify the health impact of those disadvantages. Drawing on the limited direct research on farmworkers, and on additional research about poor, isolated, and immigrant societies, we propose four mechanisms through which housing can be expected to affect farmworker health: quality of social capital within farmworker communities, stress effects of poor housing situations, effects of housing on social support for healthy behaviors, and interactions among these factors, especially effects on children that can last for generations. Policy and planning definitions of "adequate" farmworker housing should take a more holistic view of housing needs to support specific social and community benefits in design decisions.


A neuroimmunometabolic view on the cephalic phase of insulin release.
The cephalic phase of insulin secretion (CPIS) plays a crucial role in glucose homeostasis. However, the neural basis of CPIS and its overall relevance to metabolic health are poorly understood. Here, we preview the findings of Wiedemann et al. (2022) that address the role of IL-1β in the integration of neuro-mediated insulin release following cephalic stimulation and CPIS dysregulation in obesity.


First Report of Biscogniauxia mediterranea on English Ash in Italy.
In July 2010, extensive decline of English ash (Fraxinus excelsior L.) was observed near Chiusi della Verna, Tuscany, in central Italy (altitude 956 m a.s.l.; lat. 43° 41' 54″ N, long. 11° 56' 9″E). The symptoms on the tree trunks and leaves included lengthwise bark cracks, detached bark, withering of the crown where the bark was detached, and extensive microphyllia. In September 2010, perithecial stromata were observed in the parts of the tree that had lost their bark. They were applanate and black and fruiting bodies were recognized as Biscogniauxia mediterranea (De Not.) Kuntze (basionym Sphaeria mediterranea (De Not.), Ascomycotina, Xylaraceae), a fungus causing charcoal canker that attacks oaks, Acer spp., Castanea sativa Miller, Fagus sylvatica L., and Platanus acerifolia Willd. The biometric characteristics of 30 stromata were examined. The stromata were slightly convex, ellipsoid, and elongate, 7.2 to 20.5 × 3.5 to 4.2 cm. The perithecia were ovoid to tubular, 0.74 to 0.80 × 0.12 to 0.15 mm; the asci were short and stipitate, 7.9 to 10.0 × 120.2 to 170.4 μm. The ascospores were ovoid, brownish-black, with narrowed and roundish ends, 6.9 to 9.1 × 14.6 to 20.0 μm. Colonies grown on PDA at 25°C for 5 days were grey viewed from the top and black viewed from the dish underside. A comparison with the data in the literature (1) confirmed the macro- and microscopic identification. Traditional identification was further confirmed by sequence information from the rDNA ITS region. A BLAST search of the ITS sequence of our B. mediterranea isolate (GenBank Accession No. JX262798) revealed an exact match (100%) with several reference sequences of the fungus present in the database, mainly from oak hosts. Four branches each of five English ash trees growing in a natural environment were inoculated at the trunk junction with a 10-5 ml ascospore suspension. Control consisted of a branch per tree inoculated with an identical volume of sterile water only. After 25 days, the bark became detached and after a further 15 days the typical black stromata appeared. The pathogen was reisolated from the lesions, confirming Koch's postulates. No symptoms were observed on control branches, which presented healed wounds. B. mediterranea was first detected in 1986 on Quercus cerris L. and Q. pubescens Willd. in the Circeo natural park in central Italy (altitude 50 m a.s.l; lat. 41° 27' 55″44 N, long. 12° 53' 53″52 E). From there it spread north to other parts of the country. All oak species in Italy appear to be susceptible. Its northward expansion is likely associated with the high temperature and water stress that have been affecting the Italian peninsula for the last few years (4). Most recently, B. mediterranea has also been reported on Q. cerris in the Karst region of Slovenia (lat. 45° 43' 03″7 N, long. 13° 45' 20″ 4 E). This confirms its current spread to the more northerly territories, most likely because of ongoing changes in the climate that are creating optimal conditions for its survival in areas that were previously unsuitable to it. References: (1) P. Capretti and L. Mugnai, Inform. Fitopatol. 37:39, 1987. (2) D. Jurc and N. Ogris. Plant Pathol. 55:299, 2006. (3) A. Ragazzi. Micologia Italiana 1:29, 2009. (4) A. Vannini and R. Valentini. Tree Physiol. 14:129, 1994.


Self-assembly of cytotoxic peptide amphiphiles into supramolecular membranes for cancer therapy.
Peptide amphiphiles (PAs) provide a versatile platform for the design of complex and functional material constructs for biomedical applications. The hierarchical self-assembly of PAs with biopolymers is used to create robust hybrid membranes with molecular order on the micron scale. Fabrication of membranes by assembling hyaluronic acid with positively charged PA nanostructures containing anti-cancer PAs bearing a (KLAKLAK)(2) peptide sequence is reported here. Changes in membrane microstructure as the positively charged PA nanostructures vary from cylindrical nanofibers to spherical aggregates are characterized. Results indicate that formation of highly aligned fibrous membranes requires a threshold concentration of nanofibers in solution. Additionally, variation of PA nanostructure morphology from spherical aggregates to cylindrical nanofibers allows membranes to act either as reservoirs for sustained release of cytotoxicity upon enzymatic degradation or as membranes with surface-bound cytotoxicity, respectively. Thus, the self-assembly processes of these PA-biopolymer membranes can be potentially used to design delivery platforms for anti-cancer therapeutics.


Translesion synthesis in Escherichia coli: lessons from the NarI mutation hot spot.
Duplication of DNA containing damaged bases is a challenge to DNA polymerases that normally replicate with high speed, high accuracy and high processivity undamaged templates only. When a replicative DNA polymerase encounters a chemically altered base that it is unable to copy, a process called translesion synthesis (TLS) takes place during which the replicative polymerase is transiently replaced by a so-called specialized or lesion bypass polymerase. In addition to the central players that are the replicative and translesion DNA polymerases, TLS pathways involve accessory factors such as the general replication processivity factor (i.e. the beta-clamp in prokaryotes and PCNA in eukaryotes). In Escherichia coli, besides the beta-clamp, RecA plays a fundamental role as a co-factor of Pol V the major bypass polymerase in this organism. An integrated view of TLS pathways necessarily requires both genetic and biochemical studies. In this review we will attempt to summarize the insights into TLS gained over the last 25 years by studying a frameshift mutation hot spot, the NarI site. This site was initially discovered by serendipity when establishing a forward mutation spectrum induced by a chemical hepatocarcinogen, N-2-acetylaminofluorene (AAF). Indeed, this chemical carcinogen covalently binds to DNA forming adducts with guanine residues. When bound to G* in the NarI site, 5'-GGCG*CC-, AAF induces the loss of the G*pC dinucleotide at a frequency that is approximately 10(7)-fold higher than the spontaneous frequency. In vivo studies showed that the NarI mutation hot spot is neither restricted to the NarI sequence itself, nor to the carcinogen AAF. Instead, the hot spot requires a sequence containing at least two GpC repeats and any of a family of aromatic amides and nitro aromatic compounds that form a large class of human carcinogens. Genetic analysis initially revealed that the NarI frameshift pathway is SOS dependent but umuDC (i.e. Pol V) independent. More recently, DNA Pol II was identified as the enzyme responsible of this frameshift pathway. Concurrently the AAF adduct in the NarI site can be bypassed in an error-free way by Pol V. The NarI site thus offers a unique possibility to study the interplay between two specialized DNA polymerases, Pol II and Pol V, that can both extend replication intermediates formed when the replicative Pol III dissociates in the vicinity of the damage. Full reconstitution of the two pathways led us to highlight a key feature for TLS pathways, namely that it is critical the specialized DNA polymerase synthesizes, during the course of a single binding event, a patch of DNA synthesis (TLS patch) that is long enough as to "hide the lesion induced distortion" from the proofreading activity upon reloading of the replicative DNA polymerase (or any exonuclease that may get access to the primer when the specialized DNA polymerase detaches). The beta-clamp, to which all DNA polymerases bind, plays a critical role in allowing the specialized DNA polymerases to synthesize TLS patches that are long enough to resist such "external proofreading" activities.


Ecotoxicity of pentachlorophenol in contaminated soil as affected by soil type.
Four uncontaminated soils were chosen with a wide range of pH, organic carbon, and clay content to allow us to determine the properties that were most influential on pentachlorophenol (PCP) toxicity. The soils were contaminated in the laboratory at concentrations of 50 and 100 mg/kg and target organisms were exposed to the contaminated soil. Germination and emergence of lettuce seedlings was found to be dependent upon PCP concentration and soil type, and responses were highly correlated to extractable concentrations. Earthworms were sensitive to PCP, regardless of soil properties, and mortality was observed in most samples at the 100 mg/kg concentration. Toxic responses by the worms were not strongly related to soil properties or extractable concentrations. The importance of soil chemical and physical properties on toxicity and bioavailability depends upon the target organism. In the case of lettuce seedlings, PCP is acquired through the aqueous phase; therefore, the chemical interaction between PCP and soil controls toxicity. Since earthworms ingest soil and potentially can change the chemical environment of exposure, the impact of soil properties on PCP toxicity is less apparent.


Treatment of chronic hepatitis B infection: an update of Swedish recommendations.
The main goal for treatment of chronic hepatitis B is to prevent complications such as liver cirrhosis or hepatocellular carcinoma. Knowledge from population studies of the long-term risk of chronic HBV infection, as well as the recent introduction of pegylated interferon and additional nucleoside analogues has changed the therapeutic situation. Recently, a Swedish expert panel convened to update the national recommendations for treatment. The panel recommends treatment for patients with active HBV infection causing protracted liver inflammation or significant liver fibrosis, verified by liver histology. In general, pegylated interferon alpha-2a is recommended as first-line treatment, in particular for HBeAg-positive patients with HBV genotypes A or B. Among nucleoside analogues, entecavir is the first choice and adefovir or tenofovir can be used as alternatives. Lamivudine monotherapy is not recommended due to the high risk of resistance development. Combinations of nucleoside analogues such as tenofovir and lamivudine or emtricitabine are alternatives for patients with non-response or infection with resistant variants, or as first choice for patients with advanced liver disease. Nucleoside analogue treatment should be monitored to detect primary non-response and virological breakthrough. Special recommendations are given for HBV/HIV coinfected patients, immunosuppressed patients, children, and for treatment before and after liver transplantation. The present guideline is translated from Swedish, where it is published on the MPA and RAV websites (www.mpa.se and www.rav.nu.se) including 7 separate papers based on thorough literature search. The complete reference list can be received from the Medical Products Agency upon request.


Maternal obesity and offspring cognition: the role of inflammation.
High pre-pregnancy body mass index (ppBMI) has been linked to neurodevelopmental impairments in childhood. However, very few studies have investigated mechanisms in human cohorts. Among 1361 mother-child pairs in Project Viva, we examined associations of ppBMI categories with the Peabody Picture Vocabulary Test III [PPVT] and Wide Range Assessment of Visual Motor Abilities [WRAVMA] in early childhood (median 3.2y); and with the Kaufman Brief Intelligence test (KBIT) and WRAVMA in mid-childhood (7.7y). We further examined the role of maternal inflammation in these associations using the following measures from the 2nd trimester of pregnancy: plasma C-reactive protein (CRP), dietary inflammatory index (DII), and plasma omega-6 (n-6): n-3 fatty acid ratio. Children of mothers with prenatal obesity (ppBMI ≥30 kg/m2) had WRAVMA scores that were 2.1 points lower (95% CI: -3.9, -0.2) in early childhood than children of normal weight mothers (ppBMI 18.5-<25 kg/m2), in a covariate adjusted model. This association was attenuated when we additionally adjusted for maternal CRP (β -1.8 points; 95% CI: -3.8, 0.2) but not for other inflammatory markers. PpBMI was not associated with other cognitive outcomes. Maternal inflammation may modestly mediate the association between maternal obesity and offspring visual motor abilities.


Visual impairment is associated with physical and mental comorbidities in older adults: a cross-sectional study.
Visual impairment is common in older people and the presence of additional health conditions can compromise health and rehabilitation outcomes. A small number of studies have suggested that comorbities are common in visual impairment; however, those studies have relied on self-report and have assessed a relatively limited number of comorbid conditions. We conducted a cross-sectional analysis of a dataset of 291,169 registered patients (65-years-old and over) within 314 primary care practices in Scotland, UK. Visual impairment was identified using Read Code ever recorded for blindness and/or low vision (within electronic medical records). Prevalence, odds ratios (from prevalence rates standardised by stratifying individuals by age groups (65 to 69 years; 70 to 74; 75 to 79; 80 to 84; and 85 and over), gender and deprivation quintiles) and 95% confidence intervals (95% CI) of 37 individual chronic physical/mental health conditions and total number of conditions were calculated and compared for those with visual impairment to those without. Twenty seven of the 29 physical health conditions and all eight mental health conditions were significantly more likely to be recorded for individuals with visual impairment compared to individuals without visual impairment, after standardising for age, gender and social deprivation. Individuals with visual impairment were also significantly more likely to have more comorbidities (for example, five or more conditions (odds ratio (OR) 2.05 95% CI 1.94 to 2.18)). Patients aged 65 years and older with visual impairment have a broad range of physical and mental health comorbidities compared to those of the same age without visual impairment, and are more likely to have multiple comorbidities. This has important implications for clinical practice and for the future design of integrated services to meet the complex needs of patients with visual impairment, for example, embedding depression and hearing screening within eye care services.


A study on the distribution of methylchloroform and n-octane in the mouse during and after inhalation.
The distribution of methylchloroform and n-octane, respectively, in the blood, liver, kidney, and brain of mice was studied at different inspired air concentrations and after different exposure times. The air concentration varied between 10 and 10,000 ppm; and the exposure time, between 0.5 and 24 h. The resulting solvent concentrations in kidney and brain were about the same, but the liver concentrations were usually somewhat higher for both solvents. There was a linear dependence between inspired air concentration and tissue concentrations at fixed exposure times. A correlation between blood and organ concentrations was observed in animals exposed at different inhalation air concentrations but not in animals exposed only at one fixed concentration. The ratios between the concentrations of the solvents in the organs and blood were higher for n-octane than for methylchloroform. The ratios increased as the exposure concentration increased for all organs studied in the case of n-octane but only for the liver in the case of methylchloroform. When the exposure dose, i.e., inspired air concentration X time, was generated in different ways, a high concentration during a short exposure resulted in a ten times higher organ concentration than a low concentration during a long exposure. The liver, kidney, and brain concentrations generally did not differ more than twice between methylchloroform and n-octane after exposure of the same concentration and duration. The blood concentration, however, was much less in n-octane exposed animals than in methylchloroform exposed ones. A pharmacokinetic model with both uptake and elimination of the first order fitted the empirical data better for methylchloroform than a model with zero order uptake and first order elimination. Postexposure concentrations of methylchloroform were linear in a semilog graph. A one-compartment pharmacokinetic model was in accordance with the experimental data for methylchloroform. For n-octane, however, at least a two-compartment model must be assumed.


Movement activity based classification of animal behaviour with an application to data from cheetah (Acinonyx jubatus).
We propose a new method, based on machine learning techniques, for the analysis of a combination of continuous data from dataloggers and a sampling of contemporaneous behaviour observations. This data combination provides an opportunity for biologists to study behaviour at a previously unknown level of detail and accuracy; however, continuously recorded data are of little use unless the resulting large volumes of raw data can be reliably translated into actual behaviour. We address this problem by applying a Support Vector Machine and a Hidden-Markov Model that allows us to classify an animal's behaviour using a small set of field observations to calibrate continuously recorded activity data. Such classified data can be applied quantitatively to the behaviour of animals over extended periods and at times during which observation is difficult or impossible. We demonstrate the usefulness of the method by applying it to data from six cheetah (Acinonyx jubatus) in the Okavango Delta, Botswana. Cumulative activity data scores were recorded every five minutes by accelerometers embedded in GPS radio-collars for around one year on average. Direct behaviour sampling of each of the six cheetah were collected in the field for comparatively short periods. Using this approach we are able to classify each five minute activity score into a set of three key behaviour (feeding, mobile and stationary), creating a continuous behavioural sequence for the entire period for which the collars were deployed. Evaluation of our classifier with cross-validation shows the accuracy to be 83%-94%, but that the accuracy for individual classes is reduced with decreasing sample size of direct observations. We demonstrate how these processed data can be used to study behaviour identifying seasonal and gender differences in daily activity and feeding times. Results given here are unlike any that could be obtained using traditional approaches in both accuracy and detail.


Role of aldehyde dehydrogenase isozymes in the defense of rat lens and human lens epithelial cells against oxidative stress.
4-Hydroxynonenal (HNE), a metastable lipid peroxidation product, is highly toxic to various cell types if not detoxified. Because of its constant exposure to light, the ocular lens continuously generates reactive oxygen species which, under conditions of oxidative stress, may lead to excessive lipid peroxidation and consequent formation of lipid-derived aldehydes (LDAs) such as HNE. The contribution of various isozymes of aldehyde dehydrogenase (ALDH) to the oxidation of LDAs has never been systematically investigated in the lens. The present study was undertaken to ascertain the role of ALDH1A1 and -3A1 in HNE metabolism and HNE-induced toxicity in cultured human lens epithelial cells (HLECs) and in rat and mouse lenses. The metabolism of 3H-HNE was studied in ALDH3A1-knockout mouse lens and in HLECs transfected with ALDH1A1- or -3A1-specific antisense RNA and short interfering (Si)RNA. Appropriate controls were used, including wild-type mouse lens, scrambled oligonucleotides, and a transfection reagent. Transfected HLECs were exposed to oxidative stress (Fenton reaction) or HNE (30 microM) for 3 hours. Toxicity parameters, such as cell viability, apoptosis, and protein-HNE adducts and oxidation of exogenously added 3H-HNE were measured. Rat lenses were transfected with the SiRNA specific to ALDH1A1, and oxidation of 3H-HNE and the susceptibility of the transfected lenses to oxidation-induced opacification were measured. Rat lenses transfected with ALDH1A1-specific SiRNA, or cultured in the presence of the ALDH inhibitor cyanamide/disulfiram and subjected to oxidative stress displayed accelerated loss of transparency and a diminished capacity to oxidize HNE. Similarly, inhibition of ALDH1A1 in HLECs by ALDH1A1-specific antisense RNA or SiRNA was associated with decreased oxidation of 3H-HNE and increased susceptibility of the cells to oxidative damage, including apoptosis. Furthermore, 3H-HNE metabolism and HNE-induced toxicity were not affected in ALDH3A1-specific SiRNA- or antisense RNA-treated rat lenses, HLECs, or ALDH3A1-null mouse lenses. The results suggest that, under oxidative stress, HNE produced in the lens epithelium can cause toxicity and thus contribute to oxidation-induced cataractogenesis. Furthermore, the studies indicate that ALDH1A1 is a critical isozyme for maintaining clarity in human, rat, and mouse lenses.


Identification of KIAA1018/FAN1, a DNA repair nuclease recruited to DNA damage by monoubiquitinated FANCD2.
DNA interstrand crosslinks (ICLs) are highly toxic because they block the progression of replisomes. The Fanconi Anemia (FA) proteins, encoded by genes that are mutated in FA, are important for repair of ICLs. The FA core complex catalyzes the monoubiquitination of FANCD2, and this event is essential for several steps of ICL repair. However, how monoubiquitination of FANCD2 promotes ICL repair at the molecular level is unknown. Here, we describe a highly conserved protein, KIAA1018/MTMR15/FAN1, that interacts with, and is recruited to sites of DNA damage by, the monoubiquitinated form of FANCD2. FAN1 exhibits endonuclease activity toward 5' flaps and has 5' exonuclease activity, and these activities are mediated by an ancient VRR_nuc domain. Depletion of FAN1 from human cells causes hypersensitivity to ICLs, defects in ICL repair, and genome instability. These data at least partly explain how ubiquitination of FANCD2 promotes DNA repair.


Two interacting binding sites for quinacrine derivatives in the active site of trypanothione reductase: a template for drug design.
Trypanothione reductase is a key enzyme in the trypanothione-based redox metabolism of pathogenic trypanosomes. Because this system is absent in humans, being replaced with glutathione and glutathione reductase, it offers a target for selective inhibition. The rational design of potent inhibitors requires accurate structures of enzyme-inhibitor complexes, but this is lacking for trypanothione reductase. We therefore used quinacrine mustard, an alkylating derivative of the competitive inhibitor quinacrine, to probe the active site of this dimeric flavoprotein. Quinacrine mustard irreversibly inactivates Trypanosoma cruzi trypanothione reductase, but not human glutathione reductase, in a time-dependent manner with a stoichiometry of two inhibitors bound per monomer. The rate of inactivation is dependent upon the oxidation state of trypanothione reductase, with the NADPH-reduced form being inactivated significantly faster than the oxidized form. Inactivation is slowed by clomipramine and a melarsen oxide-trypanothione adduct (both are competitive inhibitors) but accelerated by quinacrine. The structure of the trypanothione reductase-quinacrine mustard adduct was determined to 2.7 A, revealing two molecules of inhibitor bound in the trypanothione-binding site. The acridine moieties interact with each other through pi-stacking effects, and one acridine interacts in a similar fashion with a tryptophan residue. These interactions provide a molecular explanation for the differing effects of clomipramine and quinacrine on inactivation by quinacrine mustard. Synergism with quinacrine occurs as a result of these planar acridines being able to stack together in the active site cleft, thereby gaining an increased number of binding interactions, whereas antagonism occurs with nonplanar molecules, such as clomipramine, where stacking is not possible.


Essential role of the C-terminal helical domain in active site formation of selenoprotein MsrA from Clostridium oremlandii.
We previously determined the crystal structures of 1-Cys type selenoprotein MsrA from Clostridium oremlandii (CoMsrA). The overall structure of CoMsrA is unusual, consisting of two domains, the N-terminal catalytic domain and the C-terminal distinct helical domain which is absent from other known MsrA structures. Deletion of the helical domain almost completely abolishes the catalytic activity of CoMsrA. In this study, we determined the crystal structure of the helical domain-deleted (ΔH-domain) form of CoMsrA at a resolution of 1.76 Å. The monomer structure is composed of the central rolled mixed β-sheet surrounded by α-helices. However, there are significant conformational changes in the N- and C-termini and loop regions of the ΔH-domain protein relative to the catalytic domain structure of full-length CoMsrA. The active site structure in the ΔH-domain protein completely collapses, thereby causing loss of catalytic activity of the protein. Interestingly, dimer structures are observed in the crystal formed by N-terminus swapping between two molecules. The ΔH-domain protein primarily exists as a dimer in solution, whereas the full-length CoMsrA exists as a monomer. Collectively, this study provides insight into the structural basis of the essential role of the helical domain of CoMsrA in its catalysis.


Prescribing in pregnancy and during breast feeding: using principles in clinical practice.
Prescribing in pregnancy often causes uncertainty and anxiety for the clinician and may lead to the omission of necessary treatment. Many drugs have inadequate data to assure safety, and therefore the clinician is left with a dilemma as to where the balance of risks and benefits lie with respect to the mother and her fetus. Understanding under what circumstances women can be prescribed medication and using principles of prescribing in pregnancy to further clarify the potential risks will aid good clinical decision-making. An appreciation of the available resources and the conviction to find the best available evidence will best serve the patient and her fetus. Teratogenicity refers to the potential for a drug to cause fetal malformations and affects the embryo 3-8 weeks after conception. Teratogenic drugs are associated with an increased risk of malformations, but the majority of babies are born with no abnormalities. In addition, approximately 2-3% of infants are born with major malformations with no association with maternal medication, and this and other confounding factors need to be addressed during counselling of a woman. Fetotoxicity refers to the functional changes that can occur to the fetus as a result of medication in the second and third trimesters. These effects are more subtle and more difficult to assess and therefore there are fewer data to support or refute these types of associations. For the majority of drugs, the neonatal dose from breast feeding is a fraction of the dose exposure in utero.


Apolipoprotein D in the aging brain and in Alzheimer's dementia.
Apolipoprotein D (apoD) levels were examined in the temporal cortex as well as an assessment of the location of apoD positive cells within the brain by immunohistochemical and biochemical methods in young control (YC), aged control (AC), and Alzheimer's demented (AD) probands. Scattered apoD positive astrocytes and oligodendrocytes were found throughout the white matter by immunohistochemistry. ApoD immunoreactivity was also observed in the cerebellar oligodendrocytes of the YC group. There was faint positive apoD staining in scattered cortical astrocytes and a few neurons in the same group. In contrast, some of the AC and all of the AD probands had intense and frequent apoD immunostained cortical astrocytes and pyramidal neurons. The cortical senile plaques and neurofibrillary tangles were apoD immunonegative. No quantitative differences were found between the cortical apoD levels in the AC and AD groups, determined by immunoblotting. ApoD detected in the brain tissue was different in molecular weight (29 kDal) from that seen in CSF or in the serum (32 kDal). Our results indicate apoD is present in the human brain, especially in glial cells, and has increased abundance in the elderly and AD subjects.


Molecular Pathways: Targeting ATR in Cancer Therapy.
The human ATR gene encodes a kinase that is activated by DNA damage and replication stress as a central transducer of a checkpoint signaling pathway. Once activated, ATR phosphorylates multiple substrates, including the kinase Chk1, to regulate cell-cycle progression, replication fork stability, and DNA repair. These events promote cell survival during replication stress and in cells with DNA damage. Accordingly, there has been the tantalizing possibility that ATR inhibitors would be therapeutically useful, especially if they were more effective in tumor versus normal cells. Indeed, multiple studies have demonstrated that alterations that promote tumorigenesis, such as defects in the ATM-p53 pathway, constitutive oncogene activation, and acquisition of the alternative lengthening of telomeres pathway, render tumor cells sensitive to ATR inhibitor monotherapy and/or increase the synergy between ATR inhibitors and genotoxic chemotherapies. Now, nearly two decades after the discovery of ATR, two highly selective and potent ATR inhibitors, AZD6738 and VX-970, are in early-phase clinical trials either as monotherapies or paired with a variety of genotoxic chemotherapies. These trials will generate important insights into the effects of ATR inhibition in humans and the potential role of inhibiting this kinase in the treatment of human malignancies.


Solution of the comparator theory of associative learning.
We derive an analytical solution of the comparator theory of associative learning, as formalized by Stout and Miller (2007). The solution enables us to calculate exactly the predicted responding to stimuli in any experimental design and for any choice of model parameters. We illustrate its utility by calculating the predictions of comparator theory in some paradigmatic designs: acquisition of conditioned responses, compound conditioning, blocking, unovershadowing, and backward blocking. We consider several versions of the theory: first-order comparator theory (close to the original ideas of Miller & Matzel, 1988), second-order comparator theory (Denniston, Savastano, & Miller, 2001), and sometimes-competing retrieval (Stout & Miller, 2007). We show that all versions of comparator theory make a number of surprising predictions, some of which appear hard to reconcile with empirical data. Our solution paves the way for a fuller understanding of the theory and for its empirical evaluation.


The role of the NFAT signaling pathway in retinal neovascularization.
The purpose of the present study was to investigate the role of nuclear factor of activated T cells (NFAT), a transcription factor downstream of VEGF, in angiogenic cell behaviors of human retinal microvascular endothelial cells (HRMEC), and to assess the efficacy of NFAT signaling inhibitors in a rat model of oxygen-induced retinopathy (OIR). Human retinal microvascular endothelial cells were treated with VEGF in the presence or absence of the NFAT inhibitor of NFAT-calcineurin association-6 (INCA-6), and NFAT translocation was evaluated using immunocytochemistry (ICC). Human retinal microvascular endothelial cells were treated with increasing doses of INCA-6, and cell proliferation and tube formation were assessed. Rats subjected to OIR were administered increasing doses of INCA-6 or the CN inhibitor FK-506, and the retinal neovascular area was measured. Nuclear factor of activated T-cells c1 was translocated to the nucleus of HRMEC treated with VEGF, and INCA-6 treatment blocked translocation. Inhibitor of NFAT-calcineurin association-6inhibited HRMEC proliferation and tube formation in a dose-dependent manner. Both INCA-6 and FK-506 treatment significantly reduced pathologic neovascularization in OIR. This investigation has demonstrated that in HRMEC, NFATc1 is activated downstream of VEGF signaling and NFAT signaling plays a key role in angiogenic cell behaviors. In addition, NFAT inhibition is shown to be highly efficacious in an OIR model. These findings indicate that the NFAT signaling pathway may serve as a suitable therapeutic target for the treatment of neovascular eye disease.


Bicycle riding and erectile dysfunction: a review.
For many years, reports in the literature have implicated bicycle riding as causing increased risk of erectile dysfunction (ED). Perineal compression during cycling has been associated with the development of sexual complications. To review current literature on the rationale for ED from bicycle riding and outcome of bicycle riding on erectile function and to present available research on preventative measures specifically regarding bicycle riding. A systematic comprehensive literature review. There is a significant relationship between cycling-induced perineal compression leading to vascular, endothelial, and neurogenic dysfunction in men and the development of ED. Research on female bicyclists is very limited but indicates the same impairment as in male bicyclists. Preventative measures including use of a properly fitted bicycle, a riding style with a suitable seat position and an appropriate bicycle seat can help prevent impairment of erectile function. There is a need for further research on safe bicycle and bicycle seat design and investigations that address the underlying mechanisms leading to cycling-related sexual dysfunction in both male and female bicyclists.


Brief report: Response inhibition and processing speed in children with motor difficulties and developmental coordination disorder.
A previous study reported that children with poor motor skills, classified as having motor difficulties (MD) or Developmental Coordination Disorder (DCD), produced more errors in a motor response inhibition task compared to typically developing (TD) children but did not differ in verbal inhibition errors. The present study investigated whether these groups differed in the length of time they took to respond in order to achieve these levels of accuracy, and whether any differences in response speed could be explained by generally slow information processing in children with poor motor skills. Timing data from the Verbal Inhibition Motor Inhibition test were analyzed to identify differences in performance between the groups on verbal and motor inhibition, as well as on processing speed measures from standardized batteries. Although children with MD and DCD produced more errors in the motor inhibition task than TD children, the current analyses found that they did not take longer to complete the task. Children with DCD were slower at inhibiting verbal responses than TD children, while the MD group seemed to perform at an intermediate level between the other groups in terms of verbal inhibition speed. Slow processing speed did not account for these group differences. Results extended previous research into response inhibition in children with poor motor skills by explicitly comparing motor and verbal responses, and suggesting that slow performance, even when accurate, may be attributable to an inefficient way of inhibiting responses, rather than slow information processing speed per se.


Pharmacogenetics of weight gain following switch from efavirenz- to integrase inhibitor-containing regimens.
Excessive weight gain affects some persons with HIV after switching to integrase strand transfer inhibitor (INSTI)-containing antiretroviral therapy (ART). We studied associations between CYP2B6 genotype and weight gain after ART switch among ACTG A5001 and A5322 participants. Eligible participants switched from efavirenz- to INSTI-containing ART, had genotype data, and had weight data at least once from 4 weeks to 2 years post-switch. Multivariable linear mixed effects models adjusted for race/ethnicity, CD4, age, BMI and INSTI type assessed relationships between CYP2B6 genotype and estimated differences in weight change. A total of 159 eligible participants switched ART from 2007 to 2019, of whom 138 had plasma HIV-1 RNA < 200 copies/mL (65 CYP2B6 normal, 56 intermediate, 17 poor metabolizers). Among participants with switch HIV-1 RNA < 200 copies/mL, weight increased in all 3 CYP2B6 groups. The rate of weight gain was greater in CYP2B6 poor than in CYP2B6 normal metabolizers overall, and within 9 subgroups (male, female, White, Black, Hispanic, dolutegravir, elvitegravir, raltegravir, and TDF in the pre-switch regimen); only in Hispanic and elvitegravir subgroups were these associations statistically significant (P < 0.05). Compared to normal metabolizers, CYP2B6 intermediate status was not consistently associated with weight gain. CYP2B6 poor metabolizer genotype was associated with greater weight gain after switch from efavirenz- to INSTI-containing ART, but results were inconsistent. Weight gain in this setting is likely complex and multifactorial.


Influence of intrinsic nerves on electromyogram of cat colon in vitro.
The electromyogram of the circular muscle layer of the cat colon was studied in vitro in superfused strips of muscle. Records exhibited electrical slow waves and migrating spike bursts, as described previously. Both the neurotoxin, tetrodotoxin, and the local anesthetic lidocaine, (P less than 0.05) prolonged the duration of migrating spike bursts, but migrating spike bursts were not affected by the adrenergic alpha-antagonist, phenoxybenzamine, nor by the adrenergic beta-antagonist, propranolol. Also, physostigmine and atropine did not affect them. Large concentrations of catecholamines did abolish them. This suggests that the migrating spike burst represents periodic release of the muscle from the tonic influence of nonadrenergic inhibitory nerves in the intramural plexuses. Slow-wave frequency and the congruence of slow waves were not affected (P greater than 0.05) by the antagonists listed above, nor by cholinergic and adrenergic agonists. This suggests that the slow waves are not importantly controlled by intrinsic nerves.


Plant extracts, spices, and essential oils inactivate Escherichia coli O157:H7 and reduce formation of potentially carcinogenic heterocyclic amines in cooked beef patties.
Meats need to be heated to inactivate foodborne pathogens such as Escherichia coli O157:H7. High-temperature treatment used to prepare well-done meats increases the formation of carcinogenic heterocyclic amines (HCAs). We evaluated the ability of plant extracts, spices, and essential oils to simultaneously inactivate E. coli O157:H7 and suppress HCA formation in heated hamburger patties. Ground beef with added antimicrobials was inoculated with E. coli O157:H7 (10(7) CFU/g). Patties were cooked to reach 45 °C at the geometric center, flipped, and cooked for 5 min. Samples were then taken for microbiological and mass spectrometry analysis of HCAs. Some compounds were inhibitory only against E. coli or HCA formation, while some others inhibited both. Addition of 5% olive or apple skin extracts reduced E. coli O157:H7 populations to below the detection limit and by 1.6 log CFU/g, respectively. Similarly, 1% lemongrass oil reduced E. coli O157:H7 to below detection limits, while clove bud oil reduced the pathogen by 1.6 log CFU/g. The major heterocyclic amines 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were concurrently reduced with the addition of olive extract by 79.5% and 84.3% and with apple extract by 76.1% and 82.1%, respectively. Similar results were observed with clove bud oil: MeIQx and PhIP were reduced by 35% and 52.1%, respectively. Addition of onion powder decreased formation of PhIP by 94.3%. These results suggest that edible natural plant compounds have the potential to prevent foodborne infections as well as carcinogenesis in humans consuming heat-processed meat products.


Effects of epidermal growth factor on the tyrosine phosphorylation of mitogen-activated protein kinases in monolayer cultures of porcine granulosa cells.
We have examined porcine granulosa cells (pGCs) for the presence of immunodetectable mitogen-activated protein (MAP) kinases (extracellular signal-regulated kinases, ERK) and have further studied the effects of epidermal growth factor (EGF) on the activation of these kinases. Cell lysates prepared from untreated monolayer cultures of pGCs were subjected to Western immunoblotting analysis using monoclonal antibodies to ERK1, ERK2 and pan-specific ERK. MAP kinases were detected having mol wts of 87K (ERK87), 54K (ERK54), 44K (ERK1), and 42K (ERK2). Treatment of pGCs with increasing concentrations (1-10 ng/ml) of EGF for 10 min resulted in electrophoretic mobility shifts of ERK1 and ERK2 suggesting hyperphosphorylation. Immunoprecipitation with an antiphosphotyrosine antibody (PY20), followed by Western analysis using pan-ERK, revealed a marked concentration-dependent increase in tyrosine phosphorylation of ERK2 in response to EGF treatment. The mobility shift and tyrosine phosphorylation of ERK2 was observed as early as 1 min after treatment with 10 ng/ml EGF. In-gel myelin basic protein (MBP) kinase assays revealed significant MBP kinase activity associated with ERK1 and ERK2 in total cell lysates and ERK2 in PY20 immunoprecipitates. Although ERK1 displayed a moderate mobility shift in response to EGF, tyrosine phosphorylation of this MAP kinase was not appreciably increased by EGF. Furthermore, PY20 immunoprecipitates demonstrated minimal MBP kinase associated with ERK1 in response to EGF treatment. Electrophoretic migration, tyrosine phosphorylation, and MBP kinase activity of the ERK54 and ERK87 was not effected regardless of EGF concentration or duration of treatment. These data demonstrate for the first time that pGCs contain immunodetectable MAP kinases. EGF, in a concentration- and time-dependent manner, increases tyrosine phosphorylation and MBP kinase activity (i.e. activation) of ERK2, and to a lesser degree ERK1, suggesting that the activation of MAP kinase may mediate the mitogenic action of EGF in pGCs.


Randomized Clinical Trial of First-Line Genome Sequencing in Pediatric White Matter Disorders.
Genome sequencing (GS) is promising for unsolved leukodystrophies, but its efficacy has not been prospectively studied. A prospective time-delayed crossover design trial of GS to assess the efficacy of GS as a first-line diagnostic tool for genetic white matter disorders took place between December 1, 2015 and September 27, 2017. Patients were randomized to receive GS immediately with concurrent standard of care (SoC) testing, or to receive SoC testing for 4 months followed by GS. Thirty-four individuals were assessed at interim review. The genetic origin of 2 patient's leukoencephalopathy was resolved before randomization. Nine patients were stratified to the immediate intervention group and 23 patients to the delayed-GS arm. The efficacy of GS was significant relative to SoC in the immediate (5/9 [56%] vs 0/9 [0%]; Wild-Seber, p < 0.005) and delayed (control) arms (14/23 [61%] vs 5/23 [22%]; Wild-Seber, p < 0.005). The time to diagnosis was significantly shorter in the immediate-GS group (log-rank test, p = 0.04). The overall diagnostic efficacy of combined GS and SoC approaches was 26 of 34 (76.5%, 95% confidence interval = 58.8-89.3%) in <4 months, greater than historical norms of <50% over 5 years. Owing to loss of clinical equipoise, the trial design was altered to a single-arm observational study. In this study, first-line GS provided earlier and greater diagnostic efficacy in white matter disorders. We provide an evidence-based diagnostic testing algorithm to enable appropriate clinical GS utilization in this population. ANN NEUROL 2020;88:264-273.


Interaction of pyridine derivatives with sepiolite.
In this study, the adsorption behavior of pyridine derivatives, i.e., 2-aminopyridine and 2,2'-bipyridyl, onto sepiolite, a natural clay mineral, has been investigated by bottle adsorption and IR spectroscopic techniques. The results indicate that 2-aminopyridine and 2,2'-bipyridyl molecules adsorb onto sepiolite through hydrogen bonding of the amino groups to the water molecules in the octahedral sheet and to the surface hydroxyls (Si-OH) in the tetrahedral sheet. These findings reveal that pyridine molecules not only adsorb onto the external surface of sepiolite but are also incorporated in its channels and tunnels with adsorption taking place at corners and/or edges, depending on the position of water molecules. A structural model is proposed to account for the orientation of pyridine derivatives in the sepiolite matrix.


ERS statement on chronic thromboembolic pulmonary hypertension.
Chronic thromboembolic pulmonary hypertension (CTEPH) is a rare complication of acute pulmonary embolism, either symptomatic or not. The occlusion of proximal pulmonary arteries by fibrotic intravascular material, in combination with a secondary microvasculopathy of vessels <500 µm, leads to increased pulmonary vascular resistance and progressive right heart failure. The mechanism responsible for the transformation of red clots into fibrotic material remnants has not yet been elucidated. In patients with pulmonary hypertension, the diagnosis is suspected when a ventilation/perfusion lung scan shows mismatched perfusion defects, and confirmed by right heart catheterisation and vascular imaging. Today, in addition to lifelong anticoagulation, treatment modalities include surgery, angioplasty and medical treatment according to the localisation and characteristics of the lesions.This statement outlines a review of the literature and current practice concerning diagnosis and management of CTEPH. It covers the definitions, diagnosis, epidemiology, follow-up after acute pulmonary embolism, pathophysiology, treatment by pulmonary endarterectomy, balloon pulmonary angioplasty, drugs and their combination, rehabilitation and new lines of research in CTEPH.It represents the first collaboration of the European Respiratory Society, the International CTEPH Association and the European Reference Network-Lung in the pulmonary hypertension domain. The statement summarises current knowledge, but does not make formal recommendations for clinical practice.


Aminopeptidases of malaria parasites: new targets for chemotherapy.
Novel targets for new drug development are urgently required to combat malaria, a disease that puts half of the world's population at risk. One group of enzymes identified within the genome of the most lethal of the causative agents of malaria, Plasmodium falciparum, that may have the potential to become new targets for antimalarial drug development are the aminopeptidases. These enzymes catalyse the cleavage of the N-terminal amino acids from proteins and peptides. P. falciparum appears to encode for at least nine aminopeptidases, two neutral aminopeptidases, one aspartyl aminopeptidase, one aminopeptidase P, one prolyl aminopeptidase and four methionine aminopeptidases. Recent advances in our understanding of these genes and their protein products are outlined in this review, including their potential for antimalarial drug development.


The Treasure Vault Can be Opened: Large-Scale Genome Skimming Works Well Using Herbarium and Silica Gel Dried Material.
Genome skimming has the potential for generating large data sets for DNA barcoding and wider biodiversity genomic studies, particularly via the assembly and annotation of full chloroplast (cpDNA) and nuclear ribosomal DNA (nrDNA) sequences. We compare the success of genome skims of 2051 herbarium specimens from Norway/Polar regions with 4604 freshly collected, silica gel dried specimens mainly from the European Alps and the Carpathians. Overall, we were able to assemble the full chloroplast genome for 67% of the samples and the full nrDNA cluster for 86%. Average insert length, cover and full cpDNA and rDNA assembly were considerably higher for silica gel dried than herbarium-preserved material. However, complete plastid genomes were still assembled for 54% of herbarium samples compared to 70% of silica dried samples. Moreover, there was comparable recovery of coding genes from both tissue sources (121 for silica gel dried and 118 for herbarium material) and only minor differences in assembly success of standard barcodes between silica dried (89% ITS2, 96% matK and rbcL) and herbarium material (87% ITS2, 98% matK and rbcL). The success rate was > 90% for all three markers in 1034 of 1036 genera in 160 families, and only Boraginaceae worked poorly, with 7 genera failing. Our study shows that large-scale genome skims are feasible and work well across most of the land plant families and genera we tested, independently of material type. It is therefore an efficient method for increasing the availability of plant biodiversity genomic data to support a multitude of downstream applications.


Older parents enjoy better filial piety and care from daughters than sons in China.
This study analyzes the unique datasets of the Chinese Longitudinal Healthy Longevity Survey using logistic regression and controlling for various covariates. Our analyses clearly demonstrate that disabled older parents are more satisfied with care provided by daughters than sons and that older parents enjoy greater filial piety from and better relationships with daughters than sons. The daughter-advantages of enjoying greater filial piety from and better relationships with children are stronger among the oldest-old aged 80+ than the young-old aged 65-79, and surprisingly more profound in rural areas than urban areas, while son-preference is much more prevalent among rural residents. We also discuss why China's rigorous fertility policy until October-2015 and much less-developed pension system in rural areas substantially contribute to sustaining traditional son-preference and a high sex ratio at birth (SRB) when fertility is low. We recommend China take integrative public health policy actions of informing the public that having daughter(s) is beneficial for old age care, developing the rural pension system and implementing the universal two-child policy as soon as possible. We believe that these policy actions would help to reduce son-preference, bring down the high SRB, and enable more future elderly parents to enjoy better care from their children and healthier lives.


Grape stalks as substrate for white rot fungi, lignocellulolytic enzyme production and dye decolorization.
The aim of this work was to evaluate the potential of grape stalks, an agroindustrial waste, for growth and lignocellulolytic enzyme production via solid-state fermentation, using the following three white rot fungi: Trametes trogii, Stereum hirsutum and Coriolus antarcticus. The decolorization of several dyes by the above mentioned cultures was also investigated. Similar values of dry weight loss of the substrate were measured after 60 days (33-43 %). C. antarcticus produced the highest laccase and Mn-peroxidase activities (33.0 and 1.6 U/g dry solid). The maximum endoglucanase production was measured in S. hirsutum cultures (10.4 U/g), while the endoxylanase peak corresponded to T. trogii (14.6 U/g). The C. antarcticus/grape stalk system seems potentially competitive in bioremediation of textile processing effluents, attaining percentages of decolorization of 93, 86, 82, 82, 77, and 58% for indigo carmine, malachite green, azure B, remazol brilliant blue R, crystal violet and xylidine, respectively, in 5 h.