bluuebunny/crossref_metadata_2025 huggingface dataset:
Soluble Fibrin Monomer Complexes Demonstrated by Agarose Gel Filtration and by Adsorption on Insolubilized Fibrinogen
Authors: von Hugo R , Hafter R , Stemberger A , Graeff H
Published: July 2018

Incubation of fibrinogen with small amounts of thrombin resulted in the occurrence of soluble fibrin monomer complexes. These complexes consisted predominantly of a derivative with a higher molecular weight than that of fibrinogen. It was characterized by its relative electrophoretic mobility in 5% PAA gel (0.28 × 10-5 cm2/V × sec) and its elution position prior to the fibrinogen peak following gel filtration. Using adsorption chromatography on insolubilized fibrinogen the derivative dissociated at a ratio of almost 1 : 1 into one part which was adsorbed and into fibrinogen which was not adsorbed. The part which was adsorbed seemed to be the thrombin mediated fibrin monomer. This study confirms the concept that dissociable dimeric fibrinogen-fibrin monomer complexes occur after limited action of thrombin on fibrinogen.
The Filtragometer: A New Device for Measuring Platelet Aggregation in Venous Blood of Man
Authors: Hornstra G , ten Hoor F
Published: July 2018

A new device for the direct assessment of spontaneous platelet aggregation in human venous blood is described: the Filtragometer. The principle of the method is based on measurement of the pressure difference across a filter with pores of 20 μπι diameter through which blood from a forearm vein is drawn. Platelet aggregates, obstructing the filter, cause a change in the pressure difference which is proportional to the degree of platelet aggregation. The measurement takes only a short time and a small amount (5-10 ml) of blood.

Platelet aggregation as measured with the Filtragometer depends on the type of anticoagulant used. The Filtragometer response decreases on inhibition of platelet stickiness in vitro by prostaglandin E1 and in vivo by aspirin ingestion. Moreover it appeared to be higher in a group with a high thrombosis tendency than in a group less susceptible to fatal arterial thrombosis.

The Filtragometer seems especially useful in monitoring the results of diet and/or drug therapy.
Maternal Serum FDP and Circulating Fetal Red Cells throughout Pregnancy. A Longitudinal Study
Authors: Hoq M. S , Pepper J , Prescott R. J , Robertson J , Cash J. D
Published: July 2018

The presence of fetal red cells in the maternal circulation and the serum FDP content of 73 women was followed serially throughout a normal pregnancy. There was a significant increase in the mean serum FDP levels in late pregnancy, which was due to episodic elevations occurring in approximately 65% of women. These transient elevations appeared to increase in frequency and severity as pregnancy progressed. There was also an increase in the number of occasions fetal red cells were detected in the maternal circulation in the later months of pregnancy, but this phenomenon did not appear to be associated in any way to the serum FDP elevations. It is concluded that the episodic rises in serum FDP occurring in normal pregnancy are not related to episodes of occult disseminated intravascular coagulation secondary to placental haemorrhage as was previously hypothesized. Their etiology and clinical significance remain unknown.
Adverse Effect of Heparin in Thrombin-Antithrombin III Interaction
Authors: Marciniak Ewa
Published: July 2018

Thrombin, while reacting in the presence of heparin, impairs the inhibitory capacity of antithrombin III so that subsequent inhibition of thrombin or factor Xa is decreased or abolished. This adverse effect of heparin has been observed directly with at least 1.5 Iowa units of thrombin per each unit of purified human antithrombin III participating in the reaction. The inhibitory capacity was then totally destroyed and some residual thrombin remained in the active form. With a lower enzyme/inhibitor ratio inactivation of thrombin in the presence of heparin was fast and complete, however, a significant decrease of inhibitory capacity below that found in reaction without heparin, has been established by measuring the residual antithrombin III activity. In defibrinated human plasma at least 2 units of thrombin per each antithrombin III unit were required to demonstrate directly the adverse effect of heparin, but a fast depletion of inhibitory capacity has been also observed after repeated additions of small thrombin portions into plasma heparinized in vitro or in vivo. Portions of enzyme initially added disappeared with great velocity; subsequent portions, however, accumulated building up a high thrombin level not seen in the absence of heparin. The accumulation of residual enzyme was more extensive in plasma containing about 1 heparin unit per ml than anticoagulant at lower concentrations and was particularly noticeable in antithrombin III deficient plasma. These results may have some bearings on the approach to heparin therapy in the event when thrombin continuously generates or when a marked deficiency of antithrombin III exists.
Einfluß von Zytostatik auf das Gerinnungssystem
Authors: Fellin F. M. , Barsigian C. , Martinez J. , Base W.
Published: July 2018

Zytostatika werden sehr viel in der Behandlung von Malignomen verwendet. Blutungskomplikationen und thrombotische Ereignisse treten bei Patienten mit Karzinomen entweder durch die Krebserkrankung selbst oder aufgrund der Behandlung der Krebserkrankung auf. Die Mechanismen für die thrombotischen und hämorrhagischen Syndrome sind bisher noch unzureichend untersucht. Diese Arbeit gibt eine Übersicht der wesentlichen thrombotischen und hämorrhagischen Zustände, die sich aus der Anwendung antineoplastischer Substanzen ergeben, und versucht eine Darstellung des gegenwärtigen Wissensstandes der Pathophysiologie dieser Zustände.
Wrist Swelling in Kienböck's Disease
Authors: Ichinose Hirofusa , Nakao Etsuhiro , Shinohara Takaaki , Tatebe Masahiro , Yamada Harumoto , Nakamura Ryogo
Published: July 2018

Background and Purpose Wrist swelling is a frequent clinical manifestation of Kienböck's disease, but no study has reported the site and pathology of wrist swelling in this disease. The aim of this study is to elucidate the site and pathology of wrist swelling in Kienböck's disease.

Materials and Methods Dorsal and palmar soft tissue thicknesses of the wrist were measured on standard lateral radiographs of the wrist in 26 patients with Kienböck's disease and 30 subjects without intra-articular lesion. Axial magnetic resonance imaging (MRI) views were examined to detect the site of swelling. The dorsal capsular ligament in three patients with Kienböck's disease underwent histological examination.

Results Radiographic study confirmed dorsal wrist swelling in 24 of 26 (92%) patients examined compared with the contralateral unaffected wrists. MRI demonstrated thickening of the dorsal capsular ligament and extensor layer with synovial proliferation. Histological examination revealed nonspecific chronic inflammation.

Conclusion Dorsal wrist swelling in Kienböck's disease is a common manifestation and constitutes a part of pathology of Kienböck's disease, although further study is required to clarify the relation between wrist swelling and etiology of Kienböck's disease.

Level of Evidence This is a Level III study.
Short-Term Results after Total Trapeziectomy with a Poly-L/D-Lactide Spacer
Authors: Kestens Bart , Van Laere Sven , Goubau Jean , Goorens Chul , Van Royen Kjell
Published: July 2018

Background Proximal migration of the first metacarpal can be seen after total trapeziectomy and various techniques have been described to prevent this subsidence.

Purpose We hypothesized the insertion of a poly-L/D-lactide spacer to prevent proximal migration of the first metacarpal without the need of an additional ligament reconstruction, allowing early mobilization and less demanding rehabilitation.

Patients and Methods Ten thumbs were treated with a total trapeziectomy and insertion of a poly-L/D-lactide scaffold. Clinical and radiological evaluation was performed after 6 months and 1 year. Patient satisfaction, pain, Quick Disabilities of the Arm, Shoulder, and Hand (QuickDASH) score, mobility of the thumb, and strength were assessed.

Results Pain according to the visual analog scale decreased (p = 0.01) and QuickDASH score decreased (p = 0.02) significantly after 1 year. Radiological evaluation after 12 months showed a collapse of the scaphometacarpal distance of 45% (p = 0.01). Osteolysis of the distal scaphoid pole and/or proximal metacarpal was seen in 6 out of 10 cases. Because of the osteolysis, the use of the poly-L/D-lactide scaffold was discontinued in our practice.

Conclusion In this limited series, total trapeziectomy with the use of the poly-L/D-lactide scaffold provides significant pain reduction and improvement of overall function. Radiographic evaluation shows significant collapse of the scaphometacarpal distance after 1 year and frequent signs of osteolysis. We do not encourage the use of the poly-L/D-lactide scaffold with total trapeziectomy before long-term clinical and radiological follow-ups of the osteolysis are available.
Hydro-social metabolism: scaling of birth rate with regional water use
Authors: Pande Saket , Pandit Akshay
Published: July 2018

Population growth is often intuitively linked with proportionally higher use of fresh water resources. However, this implies that water use per capita does not change with population growth. We not only find that birth rates of regions are negatively related with its water use per capita (i.e., higher birth rate is associated with lower water use), but also that birth rates scale with the latter with a negative power. We use population and water withdrawal data from 1950 to 2005 at irregular 5-year intervals; with virtual water content, virtual water trade and agricultural production data from 1960 to 2000 for the seven continents to investigate the scaling relationship and interpret it through the lens of metabolism theory. Our analysis reveals that the scaling exponent lies between −1/3 and −1/2. Deviations from the power relationship are observed for Europe and Africa, which are attributed to lower than expected and higher than expected birth rates, respectively. Europe’s deviation from the average scaling relationship may be due to the higher rate of return on human capital in industrialized countries. But why Africa deviates, while other developing and developed regions follow the power relationship more closely, remains a puzzle.
Hermite-Hadamard type inequalities pertaining conformable fractional integrals and their applications
Authors: Iqbal Arshad , Khan Muhammad Adil , Ullah Sana , Chu Yu Ming , Kashuri Artion
Published: July 2018

In this paper, our main aim is to give results for conformable fractional integral version of Hermite-Hadamard inequality and their applications for mid-point formula and means. First, we prove an identity associated with the Hermite-Hadamard inequality for conformable fractional integrals. By using this identity and convexity of different classes of functions and some well-known inequalities, we obtain several results for the inequality. At the end, we also present applications for some error estimations of the mid-point formula and means. The results obtained in this paper generalize the earlier results. Furthermore, the identity can be used to obtain results for Hermite-Hadamard inequality for some other generalized classes of convex functions.
Revised identification of the G-levels in gold doped Si by Laplace deep level transient spectroscopy
Authors: Gwozdz K. , Kolkovsky Vl. , Kolkovsky V. , Weber J.
Published: July 2018

In this study, we re-examine the electronic levels G1-G4 of gold-hydrogen complexes in Si by Laplace deep level transient spectroscopy. In Au doped n- and p-type Si, we analyse the depth profiles of the levels after wet-chemical etching, study their annealing behaviour and detect changes of their emission rates in the electrical field. We give evidence that G1, G3, and G2 are the double acceptor, acceptor, and donor level of the Au complex with one hydrogen atom, whereas G4 belongs to the Au complex with two hydrogen atoms and is probably an acceptor level.
Assessing the vulnerability of organic matter to C mineralisation in pasture and cropping soils of New Zealand
Authors: McNally Sam , Beare Mike , Curtin Denis , Tregurtha Craig , Qiu Weiwen , Kelliher Francis , Baldock Jeff
Published: July 2018

In New Zealand, pastoral soils have substantial organic carbon (OC) stocks, which may be vulnerable to loss from disturbance and environmental perturbations. We assessed OC vulnerability using two approaches. For the first approach, we postulated that the OC deficit of continuously cropped soils relative to nearby pastoral soils would provide a measure of the quantity of potentially vulnerable OC in pastures. As a test, soils were sampled to a depth of 15 cm at 149 sites and the total organic carbon (TOC) and particulate organic carbon (POC) contents were measured. The second approach involved measurement of OC mineralisation in a laboratory assay (98 day aerobic incubation at 25°C). For the pastoral soils, the mean TOC and POC was about twice that of the cropped soils. On average, 89% more OC was mineralised from the pastoral soils compared with the cropped counterparts. However, the quantity of OC mineralised in pasture soils was small relative to the potential for OC loss inferred from the difference in TOC between pastoral and cropped soils. Carbon mineralisation was explained using a two-pool exponential model with rate constants of the ‘fast’ and ‘slow’ pools equating to 0.36 ± 0.155 and 0.007 ± 0.003 day–1 respectively. The larger, slow OC pool correlated strongly with hot water extractable OC whereas the fast pool was related to OC extracted using cold water. Our results suggest that water extraction (using cold and hot water) can provide a rapid estimate of the quantity of mineralisable OC across a wide range of New Zealand soils.
Problematizing ‘wickedness’: a critique of the wicked problems concept, from philosophy to practice
Authors: Turnbull Nick , Hoppe Robert
Published: July 2018

The concept of ‘wicked problems’ is a major current in the fields of policy analysis and planning. However, the basis of the concept has been insufficiently examined. This re-examination of its conceptual basis explains the origins of the limitations and flaws in the wicked problems concept. This paper analyses and rejects the notion of ‘wicked problems’ on philosophical and practical grounds. We argue instead that the policy sciences already had better conceptualizations of public problems before Rittel and Webber’s flawed formulation. We return to this literature, and build upon it by reframing ‘wickedness’ in terms of higher and lower levels of problematicity in problem structuring efforts. In doing so, we offer an alternative, novel combination of the philosophy of questioning and the policy work approach to policy practice. ‘Wickedness’ is re-conceptualized as problematicity, conceived as the distance between those who question or inquire into a policy problem. This is primarily a political distance, articulated in terms of ideas, interests, institutions and practices. High problematicity arises only when wide political distances are explicitly maintained, such that partial answers cannot be reached. Practitioners deal with problematicity by a dual practical strategy of balancing closing-down and opening-up sub-questions to the problem in order to structure them such that they become amenable to action through partial answers. This simultaneously incorporates a politics of negotiating political distance via partisan adjustment and serial strategic analysis. The argument constitutes a theoretically and practically superior alternative to the ‘wicked problems’ perspective.
The 21st Century Cures Act and electronic health records one year later: will patients see the benefits?
Authors: Lye Carolyn T , Forman Howard P , Daniel Jodi G , Krumholz Harlan M
Published: July 2018

While federal regulation provides patients the right to access their electronic health records and promotes increased use of health information technology, patient access to electronic health records remains limited. The 21st Century Cures Act, signed into law over a year ago, has important provisions that could significantly improve access and availability of health data. Specifically, the provisions call for partnerships among health information exchange networks, educational and research initiatives, and health information technology certification requirements that encourage interoperability. The article reviews the potential benefits and concerns regarding implementation of these provisions, particularly the difficulty of aligning incentives and requirements for data sharing and the question of whether currently proposed rules and guidance will support the goal of improved patient access and health information exchange. Researchers, clinicians, and patients have the power to advocate for improved patient access and interoperability as policy development and implementation of the 21st Century Cures Act continues.
In Vitro Activity of Ceftolozane/Tazobactam vs Nonfermenting, Gram-Negative Cystic Fibrosis Isolates
Authors: Forrester Jeanne B , Steed Lisa L , Santevecchi Barbara A , Flume Patrick , Palmer-Long Gloria E , Bosso John A
Published: July 2018

Ceftolozane/tazobactam (C/T) was tested and compared against 93 nonfermenting, Gram-negative clinical isolates from cystic fibrosis specimens. Based on current breakpoints for intra-abdominal and urinary tract infections (which may not be appropriate for pulmonary infections), C/T was found to be the most active agent against P. aeruginosa (95.7% susceptible), followed by piperacillin/tazobactam (89.4% susceptible). For other Gram-negative pathogens included, C/T had varying activity.
Coronal Imbalance After Three-Column Osteotomy in Thoracolumbar Congenital Kyphoscoliosis
Authors: Xu Liang , Chen Xi , Qiao Jun , Chen Zhonghui , Shi Benlong , Li Song , Du Changzhi , Zhou Qingshuang , Zhu Zezhang , Qiu Yong , Sun Xu
Published: July 2018

Study Design.

Retrospective radiographical clinical study.

Objective.

To investigate the incidence and risk factors of coronal imbalance (CI) after three-column osteotomy (3-CO) in patients with thoracolumbar congenital kyphoscoliosis (CKS).

Summary of Background Data.

The incidence and risk factors of postoperative CI have been reported in adolescent idiopathic and degenerative lumbar scoliosis. However, limited data exists for patients with CKS after 3-CO.

Methods.

We reviewed a consecutive series of patients with CKS who underwent posterior-only 3-CO. Coronal curve patterns were classified according to absolute C7 translation values into: Type A, C7 translation is less than 30 mm; Type B, C7 translation more than or equal to 30 mm and C7 plumb line (C7PL) shifted to the concave side of the main curve; and Type C, C7 translation more than or equal to 30 mm and C7PL shifted to the convex side. CI was defined as C7 translation on either side more than or equal to 30 mm. According to C7 translation at the latest follow-up, patients was subdivided into an imbalanced group and a balanced group.

Results.

One-hundred-thirty patients (mean age, 17.7 ± 5.2 yr) were recruited. The mean follow-up was 41.3 ± 18.5 months. Twenty-six patients (20%) were identified as having CI at the latest follow-up. Compared with the balanced group, the imbalanced group had a larger proportion of preoperative Type C pattern, higher main curve correction, and greater lowest instrumented vertebra (LIV) tilt before and after surgery. Multiple logistic regression showed that risk factors for CI were preoperative type C pattern, preoperative LIV tilt more than or equal to 23.5° and immediate postoperative LIV tilt more than or equal to 12.3°.

Conclusion.

The incidence of CI in patients with CKS after 3-CO was 20%. A preoperative type C pattern, preoperative LIV tilt more than or equal to 23.5°, and immediate postoperative LIV tilt more than or equal to 12.3° were found to be associated with CI at the latest follow-up.

Level of Evidence: 3
Dose Escalation in Stereotactic Body Radiation Therapy for Pancreatic Cancer
Authors: Zaorsky Nicholas G. , Lehrer Eric J. , Handorf Elizabeth , Meyer Joshua E.
Published: July 2018

Objective:

To determine whether increasing biologically effective dose (BED) with stereotactic body radiation therapy (SBRT) is associated with improved local control (LC) or toxicities in patients with locally advanced pancreatic cancer.

Methods:

A PICOS/PRISMA/MOOSE selection protocol was used to identify 15 studies across 12 institutions in 5 countries where patients received definitive SBRT for nonmetastatic disease. Biologically equivalent doses were calculated with an α/β of 10 (ie, BED10) for LC and acute toxicity and 3 (ie, BED3) for late toxicity. Fixed and random effects models were used to characterize LC and grade 3/4 toxicities by BED.

Results:

There were 508 patients included with a median follow-up time of 9.1 months. The median dose was 30 Gy, and the most common regimen was 30 Gy/5 fractions. There was no significant difference in LC rates at 1 year between the BED10<70 Gy versus ≥70 Gy groups, with an estimate of 0.60 (95% confidence interval [CI], 0.36-0.81) versus 0.83 (95% CI, 0.63-0.97), respectively. There was no significant difference in acute toxicity rates between the BED10<70 Gy versus ≥70 Gy groups, with an estimate of 0.02 (95% CI, 0.00-0.08) versus 0.05 (95% CI, 0.00-0.22), respectively. Given the dose distribution across studies, 3 intervals were used to characterize BED3. There were no significant differences in late toxicity among those receiving BED3<100, 100 to 200, or >200 Gy.

Conclusions:

SBRT for pancreatic cancer results in LC rates of 60% to 83% and clinically significant toxicity of <7%. Increasing BED10 beyond 70 Gy was not associated with increased rates of 1-year LC or acute toxicity. Increasing BED3 beyond 100 Gy was not associated with increased rates of late toxicity.
Programmed genome editing of the omega-1 ribonuclease 1 of the blood fluke,Schistosoma mansoni
Authors: Ittiprasert Wannaporn , Mann Victoria H. , Karinshak Shannon E. , Coghlan Avril , Rinaldi Gabriel , Sankaranarayanan Geetha , Chaidee Apisit , Tanno Toshihiko , Kumkhaek Chutima , Prangtaworn Pannathee , Mentink-Kane Margaret , Cochran Christina J. , Driguez Patrick , Holroyd Nancy , Tracey Alan , Rodpai Rutchanee , Everts Bart , Hokke Cornelis H. , Hoffmann Karl F. , Berriman Matthew , Brindley Paul J.
Published: July 2018

CRISPR/Cas9 based genome editing has yet been reported in parasitic or indeed any species of the phylum Platyhelminthes. We tested this approach by targeting omega-1 (ω1) ofSchistosoma mansonias a proof of principle. This secreted ribonuclease is crucial for Th2 priming and granuloma formation, providing informative immuno-pathological readouts for programmed genome editing. Schistosome eggs were either exposed to Cas9 complexed with a synthetic guide RNA (sgRNA) complementary to exon 6 of ω1 by electroporation or transduced with pseudotyped lentivirus encoding Cas9 and the sgRNA. Some eggs were also transduced with a single stranded oligodeoxynucleotide donor transgene that encoded six stop codons, flanked by 50 nt-long 5’-and 3’-microhomology arms matching the predicted Cas9-catalyzed double stranded break (DSB) within ω1. CRISPResso analysis of amplicons spanning the DSB revealed ∼4.5% of the reads were mutated by insertions, deletions and/or substitutions, with an efficiency for homology directed repair of 0.19% insertion of the donor transgene. Transcripts encoding ω1 were reduced >80% and lysates of ω1-edited eggs displayed diminished ribonuclease activity indicative that programmed editing mutated the ω1 gene. Whereas lysates of wild type eggs polarized Th2 cytokine responses including IL-4 and IL-5 in human macrophage/T cell co-cultures, diminished levels of the cytokines followed the exposure to lysates of ω1-mutated schistosome eggs. Following injection of schistosome eggs into the tail vein of mice, the volume of pulmonary granulomas surrounding ω1-mutated eggs was 18-fold smaller than wild type eggs. Programmed genome editing was active in schistosomes, Cas9-catalyzed chromosomal breakage was repaired by homology directed repair and/or non-homologous end joining, and mutation of ω1 impeded the capacity of schistosome eggs both to drive Th2 polarization and to provoke formation of pulmonary circumoval granulomas. Knock-out of ω1 and the impaired immunological phenotype showcase the novel application of programmed gene editing in and functional genomics for schistosomes.
Comparative genomic analysis revealed rapid differentiation in the pathogenicity-related gene repertoires betweenPyricularia oryzaeandPyricularia pennisetiisolated from aPennisetumgrass
Authors: Zheng Huakun , Zhong Zhenhui , Shi Mingyue , Zhang Limei , Lin Lianyu , Hong Yonghe , Fang Tian , Zhu Yangyan , Guo Jiayuan , Zhang Limin , Fang Jie , Lin Hui , Norvienyeku Justice , Chen Xiaofeng , Lu Guodong , Hu Hongli , Wang Zonghua
Published: July 2018

Backgrounds

Pyriculariais a multispecies complex that could infect and cause severe blast disease on diverse hosts, including rice, wheat and many other grasses. Although the genome size of this fungal complex is small [~40 Mbp forPyricularia oryzae(syn.Magnaporthe oryzae), and ~45 Mbp forP. grisea], the genome plasticity allows the fungus to jump and adapt to new hosts. Therefore, deciphering the genome basis of individual species could facilitate the evolutionary and genetic study of this fungus. However, except for theP. oryzaesubgroup, many other species isolated from diverse hosts, such as thePennisetumgrasses, remain largely uncovered genetically.

Results

Here, we report the genome sequence of a pyriform-shaped fungal strainP. pennisetiP1609 isolated from aPennisetumgrass (JUJUNCAO) using PacBio SMRT sequencing technology. We performed a phylogenomic analysis of 28 Magnaporthales species and 5 non-Magnaporthales species and addressed P1609 into aPyriculariasubclade that is distant fromP. oryzae. Comparative genomic analysis revealed that the pathogenicity-related gene repertoires were fairly different between P1609 and theP. oryzaestrain 70-15, including the cloned avirulence genes, other putative secreted proteins, as well as some other predictedPathogen-Host Interaction(PHI) genes. Genomic sequence comparison also identified many genomic rearrangements.

Conclusion

Taken together, our results suggested that the genomic sequence of theP. pennisetiP1609 could be a useful resource for the genetic study of thePennisetum-infectingPyriculariaspecies.
Multifunctional CRISPR/Cas9 with engineered immunosilenced human T cell epitopes
Authors: Ferdosi Shayesteh R. , Ewaisha Radwa , Moghadam Farzaneh , Krishna Sri , Park Jin G. , Ebrahimkhani Mo R. , Kiani Samira , Anderson Karen S.
Published: July 2018

The application of Cas9 for genetic and epigenetic therapies in humans raises concerns over immunogenicity of this foreign protein. We report pre-existing human CD8+ T cell immunity to Streptococcus pyogenes Cas9 in the majority of healthy individuals screened. In a proof-of-principle study, we demonstrate that Cas9 protein can be modified to eliminate immunodominant epitopes through targeted mutation while preserving its function and specificity.
Beyond members of the Flaviviridae family, sofosbuvir also inhibits chikungunya virus replication
Authors: Ferreira André C. , Reis Patrícia A. , de Freitas Caroline S. , Sacramento Carolina Q. , Bôas Hoelz Lucas Villas , Bastos Mônica M. , Mattos Mayara , Loiola Erick Correia , Trindade Pablo , Vieira Yasmine Rangel , Barbosa-Lima Giselle , de Castro Faria Neto Hugo C. , Boechat Nubia , Rehen Stevens K. , Brüning Karin , Bozza Fernando A. , Bozza Patrícia T. , Souza Thiago Moreno L.
Published: July 2018

Chikungunya virus (CHIKV) causes a febrile disease associated with chronic arthralgia, which may progress to neurological impairment. Chikungunya fever (CF) is a consolidated public health problem, in tropical and subtropical regions of the world, where control of CHIKV vector, mosquitos of theAedesgenus, failed. Since there is no vaccine or specific treatment against CHIKV, infected patients receive only palliative care to alleviate pain and arthralgia. Thus, drug repurposing is necessary to identify antivirals against CHIKV. Recently, the structure and activity of CHIKV RNA polymerase was partially resolved, revealing similar aspects with the enzyme counterparner on other positive sense RNA viruses, such as members of the Flaviviridae family. We then evaluated if sofosbuvir, clinically approved against hepatitis C virus RNA polymerase, which also aims to dengue, Zika and yellow fever viruses replication, would inhibit CHIKV replication. Indeed, sofosbuvir was 5-times more selective in inhibiting CHIKV production in human hepatoma cells than ribavirin, a pan-antiviral drug. Although CHIKV replication in human induced pluripotent stem cell (iPS)-derived astrocytes was less sensitive to sofosbuvir’s, compared to hepatoma cells – this drug still impaired virus production and cell death in a MOI-dependent manner. Sofosbuvir also exhibited antiviral activityin vivo, by preventing CHIKV-induced paw oedeme in adult mice, at 20 mg/kg/day, and mortality on neonate mice model, at 40 and 80 mg/kg/day. Our data demonstrates that a prototypic alphavirus, CHIKV, is also susceptible to sofosbuvir. Since this is a clinically approved drug, it could pave the way to become a therapeutic option against CF.
PRSS56 is required for the developmental positioning of ocular angle structures
Authors: Labelle-Dumais Cassandre , Tolman Nicholas G , Paylakhi Seyyedhassan , John Simon WM , Nair K Saidas
Published: July 2018

Angle-closure glaucoma (ACG) is a severe form of glaucoma affecting up to 16 million people worldwide. In ACG, physical blockage of the ocular drainage tissue by the peripheral iris impedes the drainage of aqueous humor resulting in elevated intraocular pressure (IOP) and subsequent optic nerve damage. Despite the high prevalence of ACG, the precise mechanism(s) underlying pathogenesis are only partially understood. We have previously demonstrated that a mutation in the gene encoding the serine protease PRSS56 causes an ACG phenotype in mice. Notably,Prss56mutant mice exhibit a reduced ocular axial length and a lens occupying a larger ocular volume compared to WT mice, recapitulating characteristic features of human AGC. Our findings utilizing mouse genetic models demonstrate that loss of PRSS56 function results in altered configuration of ocular angle structures characterized by a posterior shift in the positioning of the ocular drainage tissue relative to the ciliary body and iris during development, leading to a physical blockage of drainage structure (angle closure) and high IOP. Utilizing a previously employed genetic strategy of rescuing mutantPrss56mediated reduction in ocular size by inactivation of EGR1 (Egr1;Prss56double mutants) we determined the influence of ocular size on developmental positioning of the ocular angle tissues. Our findings suggest that abnormal positioning of the drainage structure as a result of loss of PRSS56 function is uncoupled from its effect on ocular axial length reduction. Furthermore, we demonstrate that the IOP elevation observed inPrss56mutant mice is genetic context-dependent and identify a dominant modifier locus on Chromosome 2 of the C3H/HeJ genome conferring susceptibility to high IOP. Overall, our findings reveal a novel role for PRSS56 in the proper configuration of the iridocorneal angle and provide new insight into the developmental pathways implicated in glaucoma pathogenesis.
VPAC1 couples with TRPV4 channel to promote calcium-dependent gastric cancer progression
Authors: Tang Bo , Wu Jilin , Zhu Michael X. , Sun Xuemei , Liu Jingjing , Xie Rui , Dong Tobias Xiao , Xiao Yufeng , Carethers John M. , Yang Shiming , Dong Hui
Published: July 2018

Although VPAC1 and its ligand vasoactive intestinal peptide (VIP) are important in gastrointestinal physiology, their involvements in progression of gastrointestinal tumor have not been explored. Here, we found that higher expression of VIP/VPAC1 was observed in gastric cancer compared to the adjacent normal tissues. The increased expression of VIP/VPAC1 in gastric cancer correlated positively with invasion, tumor stage, lymph node, distant metastases, and poor survival. Moreover, high expression of VIP and VPAC1, advanced tumor stage and distant metastasis were independent prognostic factors. VPAC1 activation by VIP markedly induced TRPV4-mediated Ca2+ entry, and eventually promoted gastric cancer progression in a Ca2+ signaling-dependent manner. Inhibition of VPAC1 and its signaling pathway could block the progressive responses. VPAC1/TRPV4/Ca2+ signaling in turn enhanced the expression and secretion of VIP in gastric cancer cells, enforcing a positive feedback regulation mechanism. Taken together, our study demonstrate that VPAC1 is significantly overexpressed in gastric cancer and VPAC1/TRPV4/Ca2+ signaling axis could enforce a positive feedback regulation in gastric cancer progression. VIP/VPAC1 may serve as potential prognostic markers and therapeutic targets for gastric cancer.
Dynamics of multiple interacting excitatory and inhibitory populations with delays
Authors: Kim Christopher M. , Egert Ulrich , Kumar Arvind
Published: July 2018

A network consisting of excitatory and inhibitory (EI) neurons is a canonical model for understanding cortical network activity. In this study, we extend the EI network model and investigate how its dynamical landscape can be enriched when it interacts with another excitatory (E) population with transmission delays. Through analysis and simulations of a rate model and a spiking network model, we study the transition from stationary to oscillatory states by analyzing the Hopf bifurcation structure in terms of two network parameters: 1) transmission delay between the EI subnetwork and the E population and 2) inhibitory couplings that induce oscillatory activity in the EI subnetwork. We find that the critical coupling strength can strongly modulate as a function of transmission delay, and consequently the stationary state is interwoven intricately with oscillatory states generating different frequency modes. This leads to the emergence of an isolated stationary state surrounded by multiple oscillatory states and cross-frequency coupling develops at the bifurcation points. We identify the possible network motifs that induce oscillations and examine how multiple oscillatory states come together to enrich the dynamical landscape.
Genetic mapping with Allele Dosage Information in TetraploidUrochloa decumbens(Stapf) R.D. Webster Reveals Insights into Spittlebug (Notozulia entrerianaBerg) Resistance
Authors: Ulbricht Ferreira Rebecca Caroline , de Castro Lara Letícia Aparecida , Chiari Lucimara , Lima Barrios Sanzio Carvalho , do Valle Cacilda Borges , Valério José Raul , Vilela Torres Fabrícia Zimermann , Franco Garcia Antonio Augusto , de Souza Anete Pereira
Published: July 2018

Urochloa decumbens(Stapf) R.D. Webster is one of the most important African forage grasses in Brazilian beef production. Currently available genetic-genomic resources for this species are restricted mainly due to polyploidy and apomixis. Therefore, crucial genomic-molecular studies such as the construction of genetic maps and the mapping of quantitative trait loci (QTLs) are very challenging and consequently affect the advancement of molecular breeding. The objectives of this work were to (i) construct an integratedU. decumbensgenetic map for a full-sibling progeny using GBS-based markers with allele dosage information, (ii) detect QTLs for spittlebug (Notozulia entreriana) resistance, and (iii) seek putative candidate genes involved in resistance/defense against pathogens. We used theSetaria viridisgenome as reference to align GBS reads and selected 4,240 high-quality SNPs markers with allele dosage information. Of these markers, 1,000 were distributed throughout nine homologous groups with a cumulative map length of 1,335.09 cM and an average marker density of 1.33 cM. We detected QTLs for resistance to spittlebug, an important pasture insect pest, that explained between 4.66% and 6.24% of the phenotypic variation. These QTLs are in regions containing putative candidate genes related to resistance/defense against pathogens. Because this is the first genetic map with SNP autotetraploid dosage data and QTL detection inU. decumbens, it will be useful for future evolutionary studies, genome assembly, and other QTL analyses inUrochloaspp. Moreover, the results might facilitate the isolation of spittlebug-related candidate genes and help clarify the mechanism of spittlebug resistance. These approaches will improve selection efficiency and accuracy inU. decumbensmolecular breeding and shorten the breeding cycle.
Tracking disease resistance deployment in potato breeding by enrichment sequencing
Authors: Armstrong Miles R , Vossen Jack , Lim Tze Yin , Hutten Ronald C B , Xu Jianfei , Strachan Shona M , Harrower Brian , Champouret Nicolas , Gilroy Eleanor M , Hein Ingo
Published: July 2018

Following the molecular characterisation of functional disease resistance genes in recent years, methods to track and verify the integrity of multiple genes in varieties are needed for crop improvement through resistance stacking. Diagnostic resistance gene enrichment sequencing (dRenSeq) enables the high-confidence identification and complete sequence validation of known functional resistance genes in crops. As demonstrated for tetraploid potato varieties, the methodology is more robust and cost-effective in monitoring resistances than whole-genome sequencing and can be used to appraise (trans)gene integrity efficiently. All currently known NB-LRRs effective against viruses, nematodes and the late blight pathogen Phytophthora infestans can be tracked with dRenSeq in potato and hitherto unknown polymorphisms have been identified. The methodology provides a means to improve the speed and efficiency of future disease resistance breeding in crops by directing parental and progeny selection towards effective combinations of resistance genes.
Generating Yeast One-Hybrid DNA-Bait Strains
Authors: Reece-Hoyes John S. , Walhout Albertha J.M.
Published: July 2018

Generating DNA-bait strains for gateway-compatible yeast one-hybrid (Y1H) screens involves three steps. The first is to generate an Entry clone containing the DNA-bait of interest. Gateway cloning is used to clone larger baits, such as promoters, into pDONR-P4-P1R. (An alternative set of steps is also presented in this protocol that describes the creation of Entry clones by annealing primers and performing conventional ligation into pMW#5—a strategy best suited for smaller DNA-baits up to 100 bp.) The second is to transfer this DNA-bait from the Entry clone to the two Y1H reporter Destination vectors, pMW#2 (HIS3) and pMW#3 (LacZ). A two-step process is used because Entry clones generate a versatile resource that can be used for transfer of DNA-baits into a variety of vectors, for instance, upstream of the green fluorescent protein-encoding ORF to study spatiotemporal expression patterns. The final step is to integrate the HIS3 and LacZ reporter constructs into the genome of the Y1H yeast strain, YM4271. The entire process takes 24–32 d, plus sequence confirmation if necessary.
Identifying Interactors from an Activation Domain Prey Library
Authors: Reece-Hoyes John S. , Walhout Albertha J.M.
Published: July 2018

In yeast hybrid assays, the process of identifying preys that interact with the bait of interest involves several steps. First, in this protocol, the bait yeast strain is transformed with a library of activation domain (AD)-prey clones and plated on selective media containing 3-aminotriazole (3AT). This selects transformants containing an AD-prey clone that induces HIS3 reporter expression. Second, these “HIS-positive” colonies are analyzed for LacZ induction (and, optionally, URA3 induction in yeast two-hybrid (Y2H) assays). Third, yeast PCR is used on these “double-positive” colonies to amplify the insert from the AD-prey plasmid. Fourth, some of this PCR product is used to perform a gap-repair retest to confirm the interaction in fresh bait-strain yeast, and the remainder is used for DNA sequencing to determine prey identity for those that successfully retest. Finally, interactions are carefully examined to filter out likely false-positive interactions. This protocol takes 20–43 d plus sequence confirmation to complete.
Preparing Protein Solutions for Immunoblotting
Authors: Litovchick Larisa
Published: July 2018

Most protein solutions can be mixed with sample buffer and analyzed directly by immunoblotting. Any protein extraction buffer or solvent that is compatible with gel electrophoresis can be used for preparing samples for immunoblotting. Examples of the protein solutions suitable for immunoblotting include purified proteins, detergent lysates of cells, samples prepared by subcellular fractionation, and protein eluates from chromatography columns. The protein of interest must be sufficiently abundant in the sample to allow for efficient detection, but the total amount of protein in the sample should not exceed the loading capacity of the gel. For this reason, protein antigens of very low abundance must be partially purified from the protein mixtures containing high concentrations of the contaminating proteins. In this protocol, solutions containing the protein antigens of interest are mixed with SDS-PAGE sample buffer and boiled to prepare samples for protein gel electrophoresis.
Larval Thymectomy of Xenopus laevis
Authors: Mashoof Sara , Breaux Breanna , Criscitiello Michael F.
Published: July 2018

In jawed vertebrates from sharks to mammals, the thymus is the primary (or central) lymphoid tissue where T cells develop and mature. The particular stromal cell types, cytokine environment, and tissue organization in the thymus are essential for V(D)J recombination, positive selection for major histocompatibility complex recognition, and negative selection against self-peptide recognition of most αβ T cells. The thymectomy operation on Xenopus tadpole larva described here creates a T-cell–deficient model suitable for many immunology studies.
Foreign sequential entry mode choice
Authors: Xu Gangxiang , Guo Bin , Li Wen , Wang Xiaoting
Published: July 2018

Purpose

The purpose of this paper is to use the theoretical perspective of structural inertia as a unique lens to study foreign sequential entry mode choices of multinational firms.

Design/methodology/approach

It adopts quantitative analysis of a sample of 121 Chinese publicly listed firms with 564 foreign entry incidents in the 2001-2012 period to test the hypotheses.

Findings

The empirical results show that multinational firms have a tendency to adopt the same mode in the subsequent entry as the number of prior entry mode choice of a given type (joint venture (JV) in this study) increases. The results support the theoretical prediction that organizations repeat their past activities due to structural inertia. Moreover, such an inertia effect in foreign sequential entry mode choices becomes stronger for older multinational firms, larger multinational firms and state-owned multinational firms.

Research limitations/implications

Consistent with existing research, this study focuses on the entry mode choice between JV and wholly owned subsidiaries. However, it is better to examine the relationship identified in the study for different types of entry mode choices to assess result generalizability.

Practical implications

It reminds managers of multinational firms that they should be cautious to the influence of structural inertia that can be a barrier to strategic flexibility when they make entry mode choices.

Originality/value

The main contribution of this study resides in introducing structural inertia perspective to help understand the determinants of foreign sequential entry mode choices of multinational firms.
Clients, clinics and social justice
Authors: Collinson Robert , Diver Alice , McAvoy Sharon
Published: July 2018

Purpose

The purpose of this paper is to present a case study of an innovative, three-module pathway designed by the Department of Law and Criminology at Edge Hill University (England) in 2014. In addition to supporting the work of its campus pro-bono law clinic, the first-two modules aim to enhance and evidence the legal skills of EHU’s undergraduate LLB students, to embed a deeper awareness of the (legal) ethics needed for sustainable legal practice (within PRME), and to highlight the increasing need for socially responsible advocates, able to defend the rights of marginalised, vulnerable clients.

Design/methodology/approach

The critical analysis of the content and scope of an innovative, work-based learning LLB module pathway, which furthers the aim of the UN Global Compact and the PRME, and ties them firmly to socio-legal issues and advocacy involving recent jurisprudence.

Findings

The case law used within the modules, and the practical work of the students in the campus law clinic, are relevant to social justice issues and to the promotion of PRME values—they promote awareness of human rights principles, highlight the importance of access to legal services and provide students with knowledge of legal ethics. Enhanced employability skills flow from this.

Research limitations/implications

This is a narrow case study but still provides a useful analysis of an innovative, PRME relevant module pathway. The model mirrors international trends in clinical legal education and also offers a template for other law schools keen to promote the concept of ethical, just legal practice.

Practical implications

The paper posits that enhanced employability can flow from real world tasks such as advocacy for marginalised or disadvantaged groups and presents an exemplar for other law schools wishing to embed ethics/clinical law practice into their curriculum.

Social implications

The paper highlights how the campus law clinic serves the public in a deprived region—it raises awareness of human rights and of social justice issues. It has the potential to feed into litigation on social welfare issues (housing, social security, child welfare, etc.).

Originality/value

The discussion of the human rights case law that is used in the Year 2 “bridging module” (which prepares students for working in the law clinic in their final year) is particularly relevant and is analysed in detail, highlighting how this module pathway is aimed at promoting PRME and UN Global Compact principles.