bluuebunny/crossref_metadata_2025 huggingface dataset:
The migration of pesticide residues in groundwater at a bank filtration site (Krajkowo well field, Poland)
Authors: Dragon Krzysztof , Drozdzynski Dariusz , Gorski Jozef , Kruc Roksana
Published: October 2019

River bank filtration systems are widely used for water supply purposes. Using these systems, the movement of water over short distances between the river bottom and extraction wells can decrease the concentrations of some contaminants. Such systems are especially important for the removal of specific micro-pollutants that seasonally appear in river water. In this article, pesticides migration at the Krajkowo well field is analysed based on different water extraction schemes. The water is extracted by two groups of wells (one located 60–80 m from the Warta River, and the second located more than 400 m from the river) and by a horizontal well with radial drains located 5 m below the river bottom. Based on this scheme, the rate of pesticide residues removal was analysed in wells located at different distances from the river channel. The results of the three sampling campaigns conducted in summer and autumn 2017 and winter 2018 indicate the presence of pesticide compounds in the Warta River (max. total concentration of 0.171 μg/l). The pesticides were also present in the horizontal well (max. total concentration of 0.137 μg/l). Much smaller concentrations (max. 0.064 μg/l) were observed in vertical wells located 60–80 m from the river. Additionally, in the well located 250 m from the river, only two pesticide constituents were detected (at concentrations just above the detection limit), and in a well located 680 m from the river, the concentrations of pesticide residues were below the detection limit (excluding isoproturon, which was slightly above the detection limit). This research illustrates the effectiveness of pesticides removal by river bank filtration.
On the design of equity-oriented pharmaceutical copayments
Authors: González Paula
Published: October 2019

In this paper, we propose a new approach to address the problem of designing pharmaceutical copayments. The rationale for positive copayments in our setting lies in the presence of budgetary constraints and, hence, in the need to raise funds in order to finance the costs of the treatments. We use results from the literature on axiomatic bargaining with claims to incorporate criteria of distributive justice into the design of copayments. We find that if the government constrains patient rights to what is medically feasible, equity-based copayments vary from a percentage of the cost of the treatment, to a flat rate per prescription. If the government also takes into account the burden of disease experienced by patients, then copayments based on equity lead to a relation between copayments and clinical status that diverges from those proposals based on efficiency arguments. In particular, we show that equity-based copayments might be increasing in the health gains that the treatments provide to the patients. The reason is that these copayments try to avoid a “double jeopardy” problem, by ensuring that those patients with a large burden of disease do not face also an important monetary cost.
Investigation of dopant centres dominating the conduction process in the bulk of un-doped GaSb
Authors: Shura Megersa Wodajo
Published: October 2019

In this paper, first, the theoretical description of the effects of the dopant densities and the activation energies on the ionization densities, the chemical potentials corresponding to each dopant levels, the majority carrier densities and the Fermi-energy levels in one-acceptor-level system, highly compensated system and two-acceptor-level system are described in detail. Upon fitting the theoretical to the experimental results obtained by the temperature-dependent Hall effect measurements for three samples of un-doped GaSb, the dopant densities and the activation energies for a system with different dopants are investigated. The obtained results revealed that the dopant activation energy has less (no) effect on the Fermi-energy level and the majority carrier density in the highest temperature regimes. The doping density has also less (no) effect on the Fermi-energy level in the lowest temperature regimes. Finally, fitting of the theoretical to the experimental Hall effect measurements results confirmed the presence of three acceptor and one donor levels dominating the majority carrier densities at different temperature regions in all the samples of un-doped GaSb semiconductor.
Making urinary extracellular vesicles a clinically tractable source of biomarkers for inherited tubulopathies using a small volume precipitation method: proof of concept
Authors: Williams Timothy Lee , Bastos Carlos , Faria Nuno , Karet Frankl Fiona Eve
Published: October 2019

Biomarkers of inherited tubulopathies would be useful for clarifying diagnoses in patients where genetic screening is not readily available or where disease-attributable mutations are not found. Urinary extracellular vesicles (uEVs) obtained by ultracentrifugation can be used as a source of biomarkers for inherited tubulopathies such as Gitelman Syndrome (GS), however, ultracentrifugation requires costly equipment and is thus not usually accessible. In contrast, precipitation methods can extract uEVs using standard laboratory centrifuges, thus making uEVs extracted by this method clinically tractable as a source of biomarkers for GS and other inherited tubulopathies. Here we optimise a precipitation method for extracting urinary extracellular vesicles (uEVs) and provide proof of concept that these uEVs are a source of biomarkers using GS an exemplar tubulopathy. For method optimisation, uEVs were precipitated from fresh and frozen (for up to 6 years), small volume (1–2 mL) urine samples from healthy volunteers and GS patients. Nanoparticle tracking analysis was used to calculate the concentration of uEVs. Thiazide sensitive sodium-chloride cotransporter (NCC) content was determined by densitometry of Western blots. NCC content of uEVs was lower in GS patients (n = 11) than healthy volunteers (n = 12; P = 0.001). Three of four patients clinically suspected for GS, in whom only a singleSLC12A3mutation was identified, had lower uEV NCC content than all healthy volunteers tested. In the clinical setting, sufficient uEVs can be extracted from frozen, small volume urine samples using precipitation methods to distinguish patients with GS from healthy volunteers, and thus this source of uEVs could be utilised as an additional diagnostic test for GS and similar disorders.
Well-posedness issues on the periodic modified Kawahara equation
Authors: Kwak Chulkwang
Published: October 2019

This paper is concerned with the Cauchy problem of the modified Kawahara equation (posed on

\mathbb{T}

), which is well-known as a model of capillary-gravity waves in an infinitely long canal over a flat bottom in a long wave regime [26]. We show in this paper some well-posedness results, mainly the
global well-posedness
in

L^{2}(\mathbb{T})

. The proof basically relies on the idea introduced in Takaoka-Tsutsumi's works [60,69], which weakens the non-trivial resonance in the cubic interactions (a kind of smoothing effect) for the local result, and the global well-posedness result immediately follows from

L^{2}

conservation law. An immediate application of Takaoka-Tsutsumi's idea is available only in

H^{s}(\mathbb{T})

,

s > 0

, due to the lack of

L^{4}

-Strichartz estimate for arbitrary

L^{2}

data, a slight modification, thus, is needed to attain the local well-posedness in

L^{2}(\mathbb{T})

. This is the first low regularity (global) well-posedness result for the periodic modified Kwahara equation, as far as we know. A direct interpolation argument ensures the
unconditional uniqueness
in

H^{s}(\mathbb{T})

,

s > \frac{1}{2}

, and as a byproduct, we show the weak ill-posedness below

H^{\frac{1}{2}}(\mathbb{T})

, in the sense that the flow map fails to be uniformly continuous.
Instabilities in non-ideal fluids
Authors: Robinet J.-C. , Gloerfelt X.
Published: October 2019

The recent study of Ren et al. (J. Fluid Mech., vol. 871, 2019, pp. 831–864) investigated the hydrodynamic linear stability of a compressible boundary layer over an insulated flat plate for a non-ideal gas (supercritical $\text{CO}_{2}$). In particular, the authors showed that in the transcritical regime (across the pseudo-critical line) the flow is strongly convectively unstable due to the co-existence of two unstable modes: Mode I, related to Tollmien–Schlichting instabilities and a new inviscid two-dimensional mode (Mode II) with a spatial growth rate one order of magnitude larger than Mode I for high Eckert numbers. In contrast to the transcritical regime, in the sub- and supercritical regimes, Mode II does not exist. Only Mode I drives the instabilities: viscous and two-dimensional for the subcritical regime and inflectional and three-dimensional for the supercritical regime.
The Chinese Unitary Multinational State: A Normative Reappraisal
Authors: El Kholi Hugo
Published: October 2019

This article provides a normative analysis of the rationale behind the creation of the Chinese unitary multinational state. Far from being irrelevant in the face of the well-known aversion of scientific socialism for normative thinking, such analysis illuminates the Chinese Communist Party’s long-standing commitment to the unitary multinational state as the best response to the national question. It shows that the Party’s leadership, in a conscious effort to adapt Stalin’s theory of nationality to Chinese revolutionary praxis, substantially revised the standard conception of national self-determination. Relying on its own materialist logic, it made participation in the struggle against alienation the normative basis to justify granting ethnic minorities special rights of regional autonomy under a common state. The creation of the Chinese unitary multinational state appears in this light as a normatively coherent attempt to guarantee that all ethnic groups that contributed to national liberation benefit equally from their imprescriptible right to collective self-determination.
The Politics of Emotions and Spectacles: The Case of the Turkish Language Olympiads
Authors: Aslan Senem
Published: October 2019

This article examines the Turkish Language Olympiads as a political-performative strategy that the Muslim nationalists used to communicate their ideology. I argue that to understand the rise of Muslim nationalism, we also need to understand how emotional appeal is created through spectacles like the Turkish Olympiads. The spectacle was effective in boosting people’s sense of national pride and self-confidence by resolving two important tensions of Kemalist nationalism. First, it addressed the tension between Westernization and nationalization. Depicting an image of Turkish national culture that is appreciated and imitated by foreigners, it contested the imitative, Westernist character of Kemalist nationalism. Second, recasting the outside world as friendly to Turks, even Turkophile, it challenged Kemalist nationalism’s emphasis on external threats. Turkish-speaking and -acting foreigners communicated a message of nationalist self-empowerment and confidence, calling into question people’s sense of fear and distrust of the outside world. How Muslim nationalism was promoted, particularly the performative-symbolic strategies that were used, are important to understand because of their emotional resonance and potential for mass mobilization.
Plasma polyphenols associated with lower high-sensitivity C-reactive protein concentrations: a cross-sectional study within the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort
Authors: Harms Laura M. , Scalbert Augustin , Zamora-Ros Raul , Rinaldi Sabina , Jenab Mazda , Murphy Neil , Achaintre David , Tjønneland Anne , Olsen Anja , Overvad Kim , Romana Mancini Francesca , Mahamat-Saleh Yahya , Boutron-Ruault Marie-Christine , Kühn Tilman , Katzke Verena , Trichopoulou Antonia , Martimianaki Georgia , Karakatsani Anna , Palli Domenico , Panico Salvatore , Sieri Sabina , Tumino Rosario , Sacerdote Carlotta , Bueno-de-Mesquita Bas , Vermeulen Roel C. H. , Weiderpass Elisabete , Nøst Therese Haugdahl , Lasheras Cristina , Rodríguez-Barranco Miguel , Huerta José María , Barricarte Aurelio , Dorronsoro Miren , Hultdin Johan , Schmidt Julie A. , Gunter Marc , Riboli Elio , Aleksandrova Krasimira
Published: October 2019

Experimental studies have reported on the anti-inflammatory properties of polyphenols. However, results from epidemiological investigations have been inconsistent and especially studies using biomarkers for assessment of polyphenol intake have been scant. We aimed to characterise the association between plasma concentrations of thirty-five polyphenol compounds and low-grade systemic inflammation state as measured by high-sensitivity C-reactive protein (hsCRP). A cross-sectional data analysis was performed based on 315 participants in the European Prospective Investigation into Cancer and Nutrition cohort with available measurements of plasma polyphenols and hsCRP. In logistic regression analysis, the OR and 95 % CI of elevated serum hsCRP (>3 mg/l) were calculated within quartiles and per standard deviation higher level of plasma polyphenol concentrations. In a multivariable-adjusted model, the sum of plasma concentrations of all polyphenols measured (per standard deviation) was associated with 29 (95 % CI 50, 1) % lower odds of elevated hsCRP. In the class of flavonoids, daidzein was inversely associated with elevated hsCRP (OR 0·66, 95 % CI 0·46, 0·96). Among phenolic acids, statistically significant associations were observed for 3,5-dihydroxyphenylpropionic acid (OR 0·58, 95 % CI 0·39, 0·86), 3,4-dihydroxyphenylpropionic acid (OR 0·63, 95 % CI 0·46, 0·87), ferulic acid (OR 0·65, 95 % CI 0·44, 0·96) and caffeic acid (OR 0·69, 95 % CI 0·51, 0·93). The odds of elevated hsCRP were significantly reduced for hydroxytyrosol (OR 0·67, 95 % CI 0·48, 0·93). The present study showed that polyphenol biomarkers are associated with lower odds of elevated hsCRP. Whether diet rich in bioactive polyphenol compounds could be an effective strategy to prevent or modulate deleterious health effects of inflammation should be addressed by further well-powered longitudinal studies.
Impact of Ramadan diurnal intermittent fasting on the metabolic syndrome components in healthy, non-athletic Muslim people aged over 15 years: a systematic review and meta-analysis
Authors: Faris ‘Mo’ez Al-Islam’ E. , Jahrami Haitham A. , Alsibai Joud , Obaideen Asma A.
Published: October 2019

Studies on the impact of Ramadan diurnal intermittent fasting (RDIF) on the metabolic syndrome (MetS) components among healthy Muslims observing Ramadan month have yielded contradictory results. This comprehensive meta-analysis aimed to obtain a more stable estimate of the effect size of fasting during Ramadan on the MetS components, examine variability among studies, assess the generalisability of reported results and perform subgroup analyses for associated factors. We searched the CINAHL, Cochrane, EBSCOhost, Google Scholar, ProQuest Medical, PubMed/MEDLINE, ScienceDirect, Scopus and Web of Science databases for relevant studies published from 1950 to March 2019. The MetS components analysed were: waist circumference (WC), systolic blood pressure (SBP), fasting plasma/serum glucose (FG), TAG, and HDL-cholesterol. We identified eighty-five studies (4326 participants in total) that were conducted in twenty-three countries between 1982 and 2019. RDIF-induced effect sizes for the MetS components were: small reductions in WC (no. of studies K = 24, N 1557, Hedges’ g = −0·312, 95 % CI −0·387, −0·236), SBP (K = 22, N 1172, Hedges’ g = −0·239, 95 % CI −0·372, −0·106), FG (K = 51, N 2318, Hedges’ g = −0·101, 95 % CI −0·260, 0·004) and TAG (K = 63, N 2862, Hedges’ g = −0·088, 95 % CI −0·171, −0·004) and a small increase in HDL-cholesterol (K = 57, N 2771, Hedges’ g = 0·150, 95 % CI 0·064, 0·236). We concluded that among healthy people, RDIF shows small improvement in the five MetS components: WC, SBP, TAG, FG and HDL.
A new species ofHedruris(Nematoda: Hedruridae) from freshwater turtles, its life cycle and biogeographic distribution of the genus
Authors: Palumbo E. , Servián A. , Sánchez R. , Diaz J.I.
Published: October 2019

We describeHedruris dratinin. sp. (Nematoda, Hedruridae) fromHydromedusa tectiferaandPhrynops hilariiin Argentina based on morphological and molecular characters. Also, we provide information about its life cycle. The new species differs from other species of the genus by possessing the excretory pore, nerve ring and deirids at equal distance from the anterior end. Additionally,H. dratinin. sp. has mammilated eggs and males possess nine pairs of caudal papillae. The subadults and adults ofH. dratinin. sp andH. orestiaewere characterized by sequencing the small subunit ribosomal DNA (18S). We present for the first time a life cycle of a species ofHedruristhat includes an amphipod as intermediate host and a reptile as definitive host. Furthermore, we analysed the host and geographic distribution of allHedrurisspecies. Although the genus has a cosmopolitan distribution and parasitizes a great host diversity, the majority of species have a Gondwanian distribution, with amphibians being the preferred hosts.
Power in the metropolis: the impact of economic and demographic growth on the Antwerp City Council (1400–1550)
Authors: Everaert Janna
Published: October 2019

Current historiography endorses a narrative that the political elite of pre-industrial gateway cities became more ‘open’ in the wake of efflorescence and that their city councils became populated with merchants. Yet, according to the existing literature, Antwerp challenges this narrative, as the influx of merchants was very limited during late fifteenth and early sixteenth centuries when Antwerp transformed from a medium-sized Brabantine city into the leading economic centre in western Europe. Moreover, scholars disagree on whether the economic expansion had any impact at all on the composition and profile of Antwerp's political elite. By analysing the social composition of the city council and how this evolved from the beginning of Antwerp's commercial expansion around 1400 until its apogee around 1550, I revisit the question whether Antwerp constitutes an exception to the established pattern of elite formation in gateway cities and, if so, why.
Pain drawings, interpreter support and clinical findings among immigrant patients on sick leave in Swedish primary health care
Authors: Löfvander Monica
Published: October 2019

Aim:

To evaluate the spread of pain and its correlates among immigrant patients on sick leave.

Background:

Backache, outspread pain and sick-leave questions are problematic to handle primary health care, especially in multicultural settings.

Methods:

Two hundred and thirty-five patients 20–45 years on paid sick leave (59% women, 93% foreign-born, mostly non-Europeans). Many had little formal education. One-third had professional interpreter support. The patients pointed out on their bodies where they felt pain. This information was transferred on a pain drawing [pain drawing fields (PDFs) 0–18] by a doctor. Major depression and psychosocial stressors were assessed usingDiagnostic and Statistical Manual of Mental Disorders. Nociceptive locations for pain were established (pain-sites 0–18). Dependent variable was the number of PDFs. Independent variables were social data, sick leave, interpreter, depression, stress levels and number of pain sites. Calculations were done using descriptive methods and multi-variable linear regression in full models, by gender.

Findings:

Many patients had depression (51% women versus 32% men). A majority were exposed to psychosocial stressors. Women had more PDFs, in median 5 [inter-quartile ranges (IQR) 4–8] versus men 3 (IQR 2–5), and also more pain sites, in median 3 (IQR 2–5) versus men in median 2 (IQR 1–3). For men, the regression calculations revealed that numbers of PDFs associated only with increasing numbers of pain sites (B 0.871P< 0.001). For women, this association was weaker (B 0.364,P< 0.001), with significant values also for age (B 0.103) and sick leave > one year (B 0.767,P= 0.010), and a negative predicting value for interpreter support (B −1.198,P< 0.043). To conclude, PDFs associated often with somatic findings but varied much among the women. This implies potential problems regarding cause, function and sick leave questions. However, support by professional interpreters may facilitate a shared understanding with immigrant women having long-standing pain.
Inter-professional education and primary care: EFPC position paper
Authors: Miller Robin , Scherpbier Nynke , van Amsterdam Loes , Guedes Virgínia , Pype Peter
Published: October 2019

Inter-professional education (IPE) can support professionals in developing their ability to work collaboratively. This position paper from the European Forum for Primary Care considers the design and implementation of IPE within primary care. This paper is based on workshops and is an evidence review of good practice. Enablers of IPE programmes are involving patients in the design and delivery, providing a holistic focus, focussing on practical actions, deploying multi-modal learning formats and activities, including more than two professions, evaluating formative and summative aspects, and encouraging team-based working. Guidance for the successful implementation of IPE is set out with examples from qualifying and continuing professional development programmes.
Loneliness among cancer caregivers: A narrative review
Authors: Gray Tamryn F. , Azizoddin Desiree R. , Nersesian Paula V.
Published: October 2019

Objective

Providing care to a loved one with cancer places demands on caregivers that result in changes to their daily routines and disruptions to their social relationships that then contribute to loneliness. Though caregivers’ psychosocial challenges have been well studied, loneliness — a determinant of health — has not been well studied in this population. This narrative review sought to describe the current evidence on loneliness among caregivers of cancer patients. We aimed to (1) define loneliness, (2) describe its prevalence, (3) describe the association between loneliness and health outcomes, (4) describe risks and consequences of loneliness among cancer caregivers, (5) identify ways to assess loneliness, and (6) recommend strategies to mitigate loneliness in this unique population.

Method

We used evidence from articles listed in PubMed, PsycINFO, and Cumulative Index to Nursing and Allied Health Literature (CINAHL) databases, book chapters, and reports. Articles were reviewed for the following inclusion criteria: (1) published in English, (2) caregivers of cancer patients, (3) loneliness as a study variable, and (4) peer-reviewed with no restriction on the timeframe of publication. Caregivers were defined as relatives, friends, or partners who provide most of the care and support for someone with cancer.

Results

Eighteen studies met inclusion criteria and were included in the analysis. Caregivers’ experiences of loneliness can contribute to negative effects on one's social, emotional, and physical well-being. Social support interventions may not be sufficient to address this problem. Existing recommendations to mitigate loneliness include cognitive and psychological reframing, one-on-one and group therapy, befriending, resilience training, and technology-based interventions.

Significance of results

Limited attention to loneliness in cancer caregivers poses a twofold problem that impacts patient and caregiver outcomes. Interventions are critically needed to address loneliness as a determinant of health in caregivers, given their pivotal role in providing care and impacting health outcomes for people with cancer.
Quality improvement in hospitals in the Russian Federation, 2000–2016: a systematic review
Authors: Vlassov Vasiliy V. , Bates Katie , McKee Martin
Published: October 2019

We reviewed published evidence on quality improvement in hospitals in the Russian Federation since 2000. We used three data sources: MEDLINE, ‘Rossiiskaia Meditsina’ (Central Scientific Medical Library), and elibrary.ru using specific search terms. No language or study design restrictions were imposed. In total, 1717 articles were identified; 51 met the inclusion criteria and were thematically analysed. Russian legislation, government acts and grey literature were sourced to contextualise identified themes. Since 2010, the Federal Ministry of Health has increasingly sought to improve quality of care, providing additional resources and new initiatives across the health system. These include clinical practice guidelines, pay for performance schemes, electronic medical records, more specialist care, paraclinical care, and quality control systems. Quality of care, increasingly a concern of the Russian government, is said to be improving. Yet most initiatives have rarely been evaluated. This reflects the limited capacity for health services research in Russia. It seems likely that the full potential for improvements in quality of care in Russia is still to be realised.
Red Skies, Black Holes, Green Lakes, and Other Colorful Things
Authors:   AGU
Published: October 2019

What Earth and space science stories are we recommending this week?
CtBP—a targetable dependency for tumor-initiating cell activity and metastasis in pancreatic adenocarcinoma
Authors: Chawla Ayesha T. , Chougoni Kranthi Kumar , Joshi Prashant J. , Cororaton Agnes D. , Memari Patrick , Stansfield John C. , Park Haemin , Seth Rashmi , Szomju Barbara , Sima Adam P. , Idowu Michael O. , Ellis Keith C. , Grossman Steven R.
Published: October 2019

Ctbp2 is a uniquely targetable oncogenic transcriptional coregulator, exhibiting overexpression in most common solid tumors, and critical to the tumor-initiating cell (TIC) transcriptional program. In the “CKP” mouse pancreatic ductal adenocarcinoma (PDAC) model driven by mutant K-Ras, Ctbp2 haploinsufficiency prolonged survival, abrogated peritoneal metastasis, and caused dramatic downregulation of c-Myc, a known critical dependency for TIC activity and tumor progression in PDAC. A small-molecule inhibitor of CtBP2, 4-chloro-hydroxyimino phenylpyruvate (4-Cl-HIPP) phenocopied Ctbp2 deletion, decreasing tumor burden similarly to gemcitabine, and the combination of 4-Cl-HIPP and gemcitabine further synergistically suppressed tumor growth. Pharmacodynamic monitoring revealed that the 4-Cl-HIPP/gemcitabine combination induced robust and synergistic tumor apoptosis and marked downregulation of the TIC marker CD133 in CKP PDAC tumors. Collectively, our data demonstrate that targeting CtBP represents a fruitful avenue for development of highly active agents in PDAC that cooperate with standard therapy to limit both primary and metastatic tumor burden.
Engaging with research impact assessment for an environmental science case study
Authors: Fryirs Kirstie A. , Brierley Gary J. , Dixon Thom
Published: October 2019

Impact assessment is embedded in many national and international research rating systems. Most applications use the Research Impact Pathway to track inputs, activities, outputs and outcomes of an invention or initiative to assess impact beyond scholarly contributions to an academic research field (i.e., benefits to environment, society, economy and culture). Existing approaches emphasise easy to attribute ‘hard’ impacts, and fail to include a range of ‘soft’ impacts that are less easy to attribute, yet are often a dominant part of the impact mix. Here, we develop an inclusive 3-part impact mapping approach. We demonstrate its application using an environmental initiative.
In vivo non-invasive monitoring of dystrophin correction in a new Duchenne muscular dystrophy reporter mouse
Authors: Amoasii Leonela , Li Hui , Zhang Yu , Min Yi-Li , Sanchez-Ortiz Efrain , Shelton John M. , Long Chengzu , Mireault Alex A. , Bhattacharyya Samadrita , McAnally John R. , Bassel-Duby Rhonda , Olson Eric N.
Published: October 2019

Duchenne muscular dystrophy (DMD) is a fatal genetic disorder caused by mutations in the dystrophin gene. To enable the non-invasive analysis of DMD gene correction strategies in vivo, we introduced a luciferase reporter in-frame with the C-terminus of the dystrophin gene in mice. Expression of this reporter mimics endogenous dystrophin expression and DMD mutations that disrupt the dystrophin open reading frame extinguish luciferase expression. We evaluated the correction of the dystrophin reading frame coupled to luciferase in mice lacking exon 50, a common mutational hotspot, after delivery of CRISPR/Cas9 gene editing machinery with adeno-associated virus. Bioluminescence monitoring revealed efficient and rapid restoration of dystrophin protein expression in affected skeletal muscles and the heart. Our results provide a sensitive non-invasive means of monitoring dystrophin correction in mouse models of DMD and offer a platform for testing different strategies for amelioration of DMD pathogenesis.
Atomic-level insight into mRNA processing bodies by combining solid and solution-state NMR spectroscopy
Authors: Damman Reinier , Schütz Stefan , Luo Yanzhang , Weingarth Markus , Sprangers Remco , Baldus Marc
Published: October 2019

Liquid–liquid phase separation is increasingly recognized as a process involved in cellular organization. Thus far, a detailed structural characterization of this intrinsically heterogeneous process has been challenging. Here we combine solid- and solution-state NMR spectroscopy to obtain atomic-level insights into the assembly and maturation of cytoplasmic processing bodies that contain mRNA as well as enzymes involved in mRNA degradation. In detail, we have studied the enhancer of decapping 3 (Edc3) protein that is a central hub for processing body formation in yeast. Our results reveal that Edc3 domains exhibit diverse levels of structural organization and dynamics after liquid–liquid phase separation. In addition, we find that interactions between the different Edc3 domains and between Edc3 and RNA in solution are largely preserved in the condensed protein state, allowing processing bodies to rapidly form and dissociate upon small alterations in the cellular environment.
3D extracellular matrix microenvironment in bioengineered tissue models of primary pediatric and adult brain tumors
Authors: Sood Disha , Tang-Schomer Min , Pouli Dimitra , Mizzoni Craig , Raia Nicole , Tai Albert , Arkun Knarik , Wu Julian , Black Lauren D. , Scheffler Bjorn , Georgakoudi Irene , Steindler Dennis A. , Kaplan David L.
Published: October 2019

Dynamic alterations in the unique brain extracellular matrix (ECM) are involved in malignant brain tumors. Yet studies of brain ECM roles in tumor cell behavior have been difficult due to lack of access to the human brain. We present a tunable 3D bioengineered brain tissue platform by integrating microenvironmental cues of native brain-derived ECMs and live imaging to systematically evaluate patient-derived brain tumor responses. Using pediatric ependymoma and adult glioblastoma as examples, the 3D brain ECM-containing microenvironment with a balance of cell-cell and cell-matrix interactions supports distinctive phenotypes associated with tumor type-specific and ECM-dependent patterns in the tumor cells’ transcriptomic and release profiles. Label-free metabolic imaging of the composite model structure identifies metabolically distinct sub-populations within a tumor type and captures extracellular lipid-containing droplets with potential implications in drug response. The versatile bioengineered 3D tumor tissue system sets the stage for mechanistic studies deciphering microenvironmental role in brain tumor progression.
Aldehyde-alcohol dehydrogenase forms a high-order spirosome architecture critical for its activity
Authors: Kim Gijeong , Azmi Liyana , Jang Seongmin , Jung Taeyang , Hebert Hans , Roe Andrew J. , Byron Olwyn , Song Ji-Joon
Published: October 2019

Aldehyde-alcohol dehydrogenase (AdhE) is a key enzyme in bacterial fermentation, converting acetyl-CoA to ethanol, via two consecutive catalytic reactions. Here, we present a 3.5 Å resolution cryo-EM structure of full-length AdhE revealing a high-order spirosome architecture. The structure shows that the aldehyde dehydrogenase (ALDH) and alcohol dehydrogenase (ADH) active sites reside at the outer surface and the inner surface of the spirosome respectively, thus topologically separating these two activities. Furthermore, mutations disrupting the helical structure abrogate enzymatic activity, implying that formation of the spirosome structure is critical for AdhE activity. In addition, we show that this spirosome structure undergoes conformational change in the presence of cofactors. This work presents the atomic resolution structure of AdhE and suggests that the high-order helical structure regulates its enzymatic activity.
Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
Authors: Rimbault Charlotte , Maruthi Kashyap , Breillat Christelle , Genuer Camille , Crespillo Sara , Puente-Muñoz Virginia , Chamma Ingrid , Gauthereau Isabel , Antoine Ségolène , Thibaut Coraline , Tai Fabienne Wong Jun , Dartigues Benjamin , Grillo-Bosch Dolors , Claverol Stéphane , Poujol Christel , Choquet Daniel , Mackereth Cameron D. , Sainlos Matthieu
Published: October 2019

Designing highly specific modulators of protein-protein interactions (PPIs) is especially challenging in the context of multiple paralogs and conserved interaction surfaces. In this case, direct generation of selective and competitive inhibitors is hindered by high similarity within the evolutionary-related protein interfaces. We report here a strategy that uses a semi-rational approach to separate the modulator design into two functional parts. We first achieve specificity toward a region outside of the interface by using phage display selection coupled with molecular and cellular validation. Highly selective competition is then generated by appending the more degenerate interaction peptide to contact the target interface. We apply this approach to specifically bind a single PDZ domain within the postsynaptic protein PSD-95 over highly similar PDZ domains in PSD-93, SAP-97 and SAP-102. Our work provides a paralog-selective and domain specific inhibitor of PSD-95, and describes a method to efficiently target other conserved PPI modules.
Interplay and Targetome of the Two Conserved Cyanobacterial sRNAs Yfr1 and Yfr2 in Prochlorococcus MED4
Authors: Lambrecht S. Joke , Kanesaki Yu , Fuss Janina , Huettel Bruno , Reinhardt Richard , Steglich Claudia
Published: October 2019

The sRNA Yfr1 and members of the Yfr2 sRNA family are almost universally present within cyanobacteria. The conserved motifs of these sRNAs are nearly complementary to each other, suggesting their ability to participate in crosstalk. The conserved motif of Yfr1 is shared by members of the Yfr10 sRNA family, members of which are otherwise less conserved in sequence, structure, and synteny compared to Yfr1. The different structural properties enable the discrimination of unique targets of Yfr1 and Yfr10. Unlike most studied regulatory sRNAs, Yfr1 gene expression only slightly changes under the tested stress conditions and is present at high levels at all times. In contrast, cellular levels of Yfr10 increase during the course of acclimation to darkness, and levels of Yfr2 increase when cells are shifted to high light or nitrogen limitation conditions. In this study, we investigated the targetomes of Yfr2, Yfr1, and Yfr10 in Prochlorococcus MED4, establishing CRAFD-Seq as a new method for identifying direct targets of these sRNAs that is applicable to all bacteria, including those that are not amenable to genetic modification. The results suggest that these sRNAs are integrated within a regulatory network of unprecedented complexity in the adjustment of carbon and nitrogen-related primary metabolism.
Imaging Rheumatoid Arthritis in Mice Using Combined Near Infrared and 19F Magnetic Resonance Modalities
Authors: Vu-Quang Hieu , Vinding Mads Sloth , Jakobsen Maria , Song Ping , Dagnaes-Hansen Frederik , Nielsen Niels Chr. , Kjems Jørgen
Published: October 2019

Rheumatoid arthritis (RA) is an autoimmune disease that causes pain and tissue destruction in people worldwide. An accurate diagnosis is paramount in order to develop an effective treatment plan. This study demonstrates that combining near infrared (NIR) imaging and 19F MRI with the injection of labelled nanoparticles provides high diagnostic specificity for RA. The nanoparticles were made from poly(ethylene glycol)-block-poly(lactic-co-glycolic acid) (NP) or PLGA-PEG-Folate (Folate-NP), loaded with perfluorooctyl bromide (PFOB) and indocyanine green (ICG) and evaluated in vitro and in a collagen-induced arthritic (CIA) mouse model. The different particles had a similar size and a spherical shape according to dynamic light scattering (DLS) and transmission electron microscopy (TEM). Based on flow cytometry and 19F MRI analysis, Folate-NP yielded a higher uptake than NP in activated macrophages in vitro. The potential RA-targeting ability of the particles was studied in CIA mice using NIR and 19F MRI analysis. Both NP and Folate-NP accumulated in the RA tissues, where they were visible in NIR and 19F MRI for up to 24 hours. The presence of folate as a targeting ligand significantly improved the NIR signal from inflamed tissue at the early time point (2 hours), but not at later time points. Overall, these results suggest that our nanoparticles can be applied for combined NIR and 19F MRI imaging for improved RA diagnosis.
Early skeletal muscle pathology and disease progress in the dy3K/dy3K mouse model of congenital muscular dystrophy with laminin α2 chain-deficiency
Authors: Gawlik Kinga I. , Körner Zandra , Oliveira Bruno M. , Durbeej Madeleine
Published: October 2019

Deficiency of laminin α2 chain leads to a severe form of congenital muscular dystrophy (LAMA2-CMD), and dystrophic symptoms progress rapidly in early childhood. Currently, there is no treatment for this detrimental disorder. Development of therapies is largely hindered by lack of understanding of mechanisms involved in the disease initiation and progress, both in patients but also in mouse models that are commonly used in the preclinical setup. Here, we unveil the first pathogenic events and characterise the disease development in a mouse model for LAMA2-CMD (dy3K/dy3K), by analysing muscles at perinatal, neonatal and postnatal stages. We found that apoptotic muscle fibres were present as early as postnatal day 1. Other typical dystrophic hallmarks (muscle degeneration, inflammation, and extensive production of the extracellular matrix proteins) were clearly evident already at postnatal day 4, and the highest degree of muscle deterioration was reached by day 7. Interestingly, the severe phenotype of limb muscles partially recovered on days 14 and 21, despite worsening of the general condition of the dy3K/dy3K mouse by that age. We found that masticatory muscles were severely affected in dy3K/dy3K mice and this may be an underlying cause of their malnutrition, which contributes to death around day 21. We also showed that several signalling pathways were affected already in 1-day-old dy3K/dy3K muscle. Therapeutic tests in the dy3K/dy3K mouse model should therefore be initiated shortly after birth, but should also take into account timing and correlation between regenerative and pathogenic events.
Digitizable therapeutics for decentralized mitigation of global pandemics
Authors: Hacohen Adar , Cohen Reuven , Efroni Sol , Barzel Baruch , Bachelet Ido
Published: October 2019

When confronted with a globally spreading epidemic, we seek efficient strategies for drug dissemination, creating a competition between supply and demand at a global scale. Propagating along similar networks, e.g., air-transportation, the spreading dynamics of the supply vs. the demand are, however, fundamentally different, with the pathogens driven by contagion dynamics, and the drugs by commodity flow. We show that these different dynamics lead to intrinsically distinct spreading patterns: while viruses spread homogeneously across all destinations, creating a concurrent global demand, commodity flow unavoidably leads to a highly uneven spread, in which selected nodes are rapidly supplied, while the majority remains deprived. Consequently, even under ideal conditions of extreme production and shipping capacities, due to the inherent heterogeneity of network-based commodity flow, efficient mitigation becomes practically unattainable, as homogeneous demand is met by highly heterogeneous supply. Therefore, we propose here a decentralized mitigation strategy, based on local production and dissemination of therapeutics, that, in effect, bypasses the existing distribution networks. Such decentralization is enabled thanks to the recent development of digitizable therapeutics, based on, e.g., short DNA sequences or printable chemical compounds, that can be distributed as digital sequence files and synthesized on location via DNA/3D printing technology. We test our decentralized mitigation under extremely challenging conditions, such as suppressed local production rates or low therapeutic efficacy, and find that thanks to its homogeneous nature, it consistently outperforms the centralized alternative, saving many more lives with significantly less resources.
An Internally Quenched Fluorescent Peptide Substrate for Protealysin
Authors: Karaseva Maria A. , Chukhontseva Ksenia N. , Lemeskina Irina S. , Pridatchenko Marina L. , Kostrov Sergey V. , Demidyuk Ilya V.
Published: October 2019

Protealysin, a metalloprotease of Serratia proteamaculans, is the prototype of a subgroup of the M4 peptidase family. Protealysin-like proteases (PLPs) are widely spread in bacteria but also occur in fungi and certain archaea. The interest in PLPs is primarily due to their putative involvement in the bacterial pathogenesis in animals and plants. Studying PLPs requires an efficient quantitative assay for their activity; however, no such assay has been reported so far. Here, we used the autoprocessing site sequence of the protealysin precursor to construct an internally quenched fluorescent peptide substrate 2-aminobenzoyl-L-arginyl-L-seryl-L-valyl-L-isoleucyl-L-(ε-2,4-dinitrophenyl)lysine. Protealysin and thermolysin, the prototype of the M4 family, proved to hydrolyze only the Ser-Val bond of the substrate. The substrate exhibited a KM = 35 ± 4 μM and kcat = 21 ± 1 s−1 for protealysin as well as a KM = 33 ± 8 μM and kcat = 7 ± 1 s−1 for thermolysin at 37 °C. Comparison of the effect of different enzymes (thermolysin, trypsin, chymotrypsin, savinase, and pronase E) on the substrate has demonstrated that it is not strictly specific for protealysin; however, this enzyme has higher molar activity even compared to the closely related thermolysin. Thus, the proposed substrate can be advantageous for quantitative studies of protealysin as well as for activity assays of other M4 peptidases.
Interactions of a medicinal climber Tinospora cordifolia with supportive interspecific plants trigger the modulation in its secondary metabolic profiles
Authors: Sharma Bhawana , Yadav Aarti , Dabur Rajesh
Published: October 2019

Tinospora cordifolia (TC) is scientifically proven immunomodulatory drug being used for centuries. Ancient literature reported that inter-specific interactions change medicinal properties of TC. Thus, the current study is aimed to understand the influence of interspecific biotic interactions on chemo-profiles of TC. To explore it, TC samples collected from six co-occurring plants, i.e. Azarditchita indica, Acacia nilotica, Albezia lebbeck, Ficus benghalensis, Tamarandus indica and Acacia leucophloea were analyzed by HPLC-ESI-QTOF-MS. Mass data were subjected to multivariate analysis. Support vector machines (SVMs) was found to be best classifier (r2 < 0.93). Data analysis showed the specific compounds in all TC due to inter-specific interactions. Data were further analyzed with SNK post-hoc test followed by permutative (n = 50) Bonferroni FDR multiple testing correction. The compound without any missing values reduced the number of variables to 133 (p < 0.01). Statistical analysis revealed that TC having interactions with A.lebbeck and A. nilotica formed the most distant groups. However, TC co-occurred with A. indica showed the highest number of up-regulated metabolites, including jatrorrhizine, chrysin, peonidin, 6-methylcoumarin and some terpenoids. Some metabolites, including jatrorrhizine and magnoflorine were quantified to confirm the accuracy of qualitative analysis. Results demonstrated the influence of inter-specific biotic interactions on TC chemo-profiles, hence its medicinal properties.
Sex differences in risk factors for stroke in patients with hypertension and hyperhomocysteinemia
Authors: Pang Hui , Fu Qiang , Cao Qiumei , Hao Lin , Zong Zhenkun
Published: October 2019

Data on the sex-specific differences in risk of stroke among patients with H-type hypertension are limited. We aimed to analyze interactions between sex and other risk factors on stroke, including the sex-methylenetetrahydrofolate reductase (MTHFR) interaction. A retrospective analysis of baseline data from 2040 patients with hypertension and hyperhomocysteinemia (HHcy) included demographic characteristics, biomarkers, history of chronic diseases and lifestyle factors. Polymerase chain reaction-restriction fragment length polymorphism method was used to investigate the C677T polymorphism of MTHFR gene. We examined independent effects and interactions between sex and stratified factors on the risk of stroke by logistic regression model. A total of 1412 patients suffered stroke, and the prevalence of stroke was 70.65% in men and 66.53% in women. Both men and women had independent risk factors for stroke, including diabetes mellitus, atrial fibrillation, smoking, increased level of systolic blood pressure (SBP) and plasma total homocysteine (tHcy), as well as the decreased level of high-density lipoprotein cholesterol. Diastolic blood pressure (DBP) -specific risk of stroke was unique to men. Interactions between sex and other risk factors on stroke risk were statistically significant: age, fasting plasma glucose (FPG), SBP, DBP, triglycerides (TG) and tHcy. Furthermore, tHcy interacted with age, SBP and DBP in men, and age, SBP, DBP, FPG, and TG in women to modulate the risk of stroke. Although TT genotype did not have an independent effect on stroke, it could interact with sex and FPG, TG and SBP to increase stroke. In conclusion, sex-specific differences are useful to stratify the risk of stroke and assist clinicians in the decision to select a reasonable therapeutic option for high-risk patients.