bluuebunny/crossref_metadata_2025 huggingface dataset:
Experimental Pharmacology of Hirunorm: a Novel Synthetic Peptide Thrombin Inhibitor
Authors: Cirillo Rocco , Lippi Annalisa , Subissi Alessandro , Agnelli Giancarlo , Criscuoli Marco
Published: July 2018

Enhanced thrombin activity has been associated with coronary thrombosis and with acute and long-term complications following coronary balloon angioplasty. Blocking thrombin activity with specific inhibitors is proposed as a promising antithrombotic therapy. We describe the anticoagulant and antithrombotic properties of hirunorm, a novel synthetic 26-aminoacid peptide thrombin inhibitor, in comparison with r-hirudin and hirulog-1. Hirunorm was equipotent to hirulog-1 and 1/30 as potent as r-hirudin in blocking a-thrombin amidolytic activity (IC50 = 10 ± 2,15 ± 1 and 0.3 ± 0.1 nM, respectively), but it did not affect trypsin, plasmin and t-PA activities at 10 μM. All the compounds inhibited clot-bound thrombin to clots prepared by thrombin hydrolysis of purified fibrinogen in buffer. Hirunorm and hirulog-1 showed similar species-dependent potency in doubling basal in vitro clotting times of human, rat and rabbit plasma (EC200 varied 70 to 200 nM for TT, 0.7 to 16 μM for aPTT and 0.8 to 17 μM for PT), while r-hirudin was always at least three times more active. When assayed by HPLC or by bioassay of the intact peptide, hirunorm was stable against a-thrombin and plasma hydrolases, but it was catabolized by rat liver and kidney enzymes. Venous thrombosis was produced in anaesthetized rats by vena cava ligation following a procoagulant serum injection. Intravenous and subcutaneous hirunorm inhibited venous thrombosis at doses (≤0.3 mg/kg) two-three times higher than those of r-hirudin. Hirulog-1 was as active as hirunorm only after i. v. infusion. Arterial thrombosis was obtained in the anaesthetized rat by chemical (FeCl2) stimulation of a common carotid and i.v. infused hirunorm (1-3 mg/kg/30 min) inhibited it dose-dependently; r-hirudin was partly active only at 3 mg/kg, but hirulog-1 was inactive at either dose. Full antithrombotic doses of hirunorm did not affect the bleeding time as measured from punctured mesenteric vessels, in anaesthetized rats. In conclusion, hirunorm is a potent peptide thrombin inhibitor endowed with antithrombotic activity in models of venous and arterial thrombosis.
The Interaction of Protein S with the Phospholipid Surface Is Essential for the Activated Protein C-independent Activity of Protein S
Authors: Wijnen Merel van , Stam Jeanette G , van't Veer Cornells , Meijers Joost C M , Reitsma Pieter H , Bertina Rogier M , Bouma Bonno N
Published: July 2018

Protein S is a vitamin-K dependent glycoprotein involved in the regulation of the anticoagulant activity of activated protein C (APC). Recent data showed a direct anticoagulant role of protein S independent of APC, as demonstrated by the inhibition of prothrombinase and tenase activity both in plasma and in purified systems. This anticoagulant effect of protein S can be explained either by a direct interaction of protein S with one of the components of the complexes and/or by the interference with the binding of these components to phospholipid surfaces.

During our investigation we noted that protein S preparations purified in different ways and derived from different sources, expressed discrepant APC cofactor and direct anticoagulant activity. In order to elucidate these differences and to study the mechanism of the APC-inde-pendent activity of protein S, we compared the protein S preparations in phospholipid-binding properties and anticoagulant activity. The dissociation constant for the binding of protein S to immobilized phospholipids ranged from 7 to 74 nM for the different protein S preparations. APC-independent inhibition of both prothrombinase and tenase activity performed on phospholipid vesicles and in plasma showed a strong correlation with the affinity for phospholipids. The APC-independent activity could be abolished by monoclonal antibodies that were either calcium-dependent and/or directed against epitopes in the Gla-region of protein S, suggesting that the protein S-phospholipid interaction is crucial for the APC-independent anticoagulant function of protein S. Protein S preparations with a low APC-independent activity expressed a high APC-cofactor activity suggesting that the affinity of protein S for phospholipids is of less importance in the expression of APC-cofactor activity of protein S.

We conclude that high affinity interactions of protein S with the membrane surface are essential for the direct anticoagulant activity of protein S and we suggest that inhibition of the prothrombinase and the tenase complex by protein S is a consequence of the occupation of the phospholipid surface by protein S molecules.
Haemostatic Parameters and Lifestyle Factors in Elderly Men in Italy and The Netherlands
Authors: Bijnen Fransje C H , Feskens Edith J M , Giampaoli Simona , Menotti Alessandro , Fidanza Flaminio , Hornstra Gerard , Caspersen Carl J , Mosterd Willem L , Kromhout Daan
Published: July 2018

The association between plasma fibrinogen, factor VII, factor X, activated partial thromboplastin time, antithrombin III and the lifestyle factors cigarette smoking, alcohol use, fat intake and physical activity was assessed in 802 men aged 70-90 years in Zutphen (The Netherlands), Montegiorgio and Crevalcore (Italy).

Smoking was positively associated with fibrinogen, also after adjustment for other lifestyle factors, age, use of anticoagulants and aspirin like drugs, body mass index, and history of myocardial infarction. Alcohol use was associated with increased levels of factor X and decreased levels of antithrombin III. Fat intake was positively associated with antithrombin III. Between cohorts, considerable differences were observed in levels of haemostatic parameters and the lifestyle factors. Compared to the mediterranean cohorts the Zutphen cohort showed the highest levels of fibrinogen and factor VII. Differences in lifestyle factors could, however, not explain differences between cohorts in levels of any of the haemostatic parameters, despite the observed associations between lifestyle factors and haemostatic parameters.
Inhibitory Effect of Prostacyclin and Nitroprusside on Type IIB von Willebrand Factor-promoted Platelet Activation
Authors: Francesconi Mariangela , Casonato Alessandra , Pagan Stefano , Donella-Deana Arianna , Pontara Elena , Girolami Antonio , Deana Renzo
Published: July 2018

Von Willebrand disease (vWD) of type IIB is a hereditary haemorragic disorder characterised by an excessive interaction of von Willebrand factor (vWF) with the platelet receptor GPIb which promotes platelet activation and aggregation through a phospholipase A2-mediated release of arachidonic acid.

The present report shows that prostacyclin and nitroprusside, vaso-dilator-compounds that enhance the cAMP and cGMP concentration respectively, cause a drastic inhibition of the type IIB vWF-induced platelet responses including increase of cytosolic Ca2+ concentration, phosphorylation of pleckstrin (47 kDa) and myosin light chain (20 kDa), secretion of ATP and serotonin, and aggregation parallel to a decrease of arachidonic acid release. Type IIB vWF also elicits tyrosine phosphorylation of proteins with apparent molecular mass of 60,74,82 and 130 kDa. Prostacyclin, which induces per se tyrosine-phosphoryla-tion of proteins of about 38 and 45 kDa, inhibits drastically the type IIB vWF-promoted tyrosine-phosphorylation of the 74 kDa protein while inhibits slightly that of 60 kDa band. The protein tyrosine-kinase inhibitor genistein causes a little decrease in the type IIB vWF-induced release of arachidonic acid.

It is concluded that the inhibition exerted by prostacyclin and nitroprusside on type IIB vWF-elicited platelet activation seems to be largely ascribable to prevention of the phospholipase A2 activation with the ensuing decrease of the subsequent protein tyrosine phosphorylation
Pre- and Postoperative State of Coagulation and Fibrinolysis in Plasma of Patients with Benign and Malignant Colorectal Disease - A Preliminary Study
Authors: Iversen L H , Okholm M , Thorlacius-Ussing O
Published: July 2018

This study was carried out in order to compare the coagulation balance in patients with colorectal cancer before and after surgical removal of tumor with an age matched non-malignancy control group. Furthermore, it was studied whether preoperative coagulation state in cancer patients was correlated to the postoperative development of deep venous thrombosis (DVT) diagnosed by venography. Plasma was collected preoperatively in 93 cancer patients and 30 controls, and postoperatively on day one, two, seven, and ninety in 88 cancer patients and 18 controls. Prothrombin fragment 1 + 2 (F1 + 2), thrombin-antithrombin complex (TAT), and total fibrin(ogen) degradation products (TDP) were quantitated in plasma by enzyme linked immunosorbent assays (ELISA). As compared to controls, patients admitted for cancer treatment displayed significantly higher levels of F1 + 2 and TAT. Patients suffering from advanced colorectal cancer had significantly higher levels of TAT and TDP as compared to patients with localized colorectal cancer. Twenty-three percent of cancer patients developed DVT postoperatively. Preoperatively these patients displayed significantly higher TDP levels, and postoperatively higher levels of F1 + 2, TAT, and TDP compared to cancer patients without DVT. The marked activation of blood coagulation and fibrinolysis observed in all patients following major abdominal surgery was even more pronounced in patients not cured for cancer.
Proneness to Formation of Tight and Rigid Fibrin Gel Structures in Men with Myocardial Infarction at a Young Age
Authors: Fatah Kamaran , Silveira Angela , Tornvall Per , Karpe Fredrik , Blombäck Margareta , Hamsten Anders
Published: July 2018

The native fibrin gel structure formed in vitro from plasma samples was examined by liquid permeation of hydrated fibrin gel networks in 38 unselected men who had suffered a myocardial infarction before the age of 45 years and in 88 age-matched population-based control men. Both the fibrin gel porosity (permeability coefficient, Ks) and the calculated fiber mass-length ratio varied considerably within the two groups, but were generally lower in the patients. Ks was 8.3 ± 5.2 cm2 × 109 (mean ± SD) in the patient group and 12.5 ± 5.7cm2 × 109 among controls (p <0.001). The corresponding figures for fiber mass-length ratio were 13.1 ± 7.7 and 16.5 ± 7.5 Dalton/ cm × 10−13, respectively (p <0.01). Around 50% of the patients had Ks values below the 10th percentile of the control group. A strong inverse correlation was seen between plasma plasminogen activator inhibitor-1 (PAI-1) activity and Ks (r = -0.603, p <0.001) or fiber mass-length ratio (r = -0.565, p <0.001) in the patient group. Corresponding weaker associations of PAI-1 with fibrin gel properties were also present in the control group. In addition, inverse relationships of very low density lipoprotein (VLDL) triglyceride concentrations to Ks (r = -0.362, p <0.001) and fiber mass-length ratio (r = -0.283, p <0.01) were found among the controls. Proneness to formation of tight and rigid fibrin gel networks with abnormal architecture in vitro is in vivo associated with myocardial infarction at a young age. Impaired fibrinolytic function secondary to a raised plasma PAI-1 activity level is associated with abnormal fibrin gel structure.
The Effects on Platelet Aggregation and Prostanoid Biosynthesis of two Parenteral Analgesics: Ketorolac Tromethamine and Dipyrone
Authors: geisslinger G , Peskar B A , Pallapies D , Sittl R , Levy M , Brune K
Published: July 2018

The pharmacokinetics and effects on platelet function of dipyrone (1.0 g; 2.5 g; i. v.) and ketorolac tromethamine (30 mg; i.m.) were studied in a three-way crossover study in twelve healthy subjects. The biosynthesis of thromboxane A2 in clotting whole blood ex vivo as well as collagen-induced platelet aggregation were determined before and up to 48 h after administration. Both prostanoid biosynthesis and platelet aggregation were inhibited by ketorolac tromethamine for a significantly longer period of time than by both doses of dipyrone. The changes in platelet functions correlated well with the serum concentrations of ketorolac or 4-methylaminoantipyrine and 4-aminoantipyrine. Using the sigmoidal Emax model the mean serum concentration (SD) of ketorolac, 4-methylaminoantipyrine and 4-aminoantipyrine inhibiting platelet TXB2 generation by 50% (EC50) in vitro was found to be 0.088 ± 0.031, 1.2 ± 0.3 and 10.2 ± 3.4 µg ml-1, respectively. In conclusion the recovery of platelet function after dipyrone administration is faster as compared to ketorolac tromethamine. This is in line with clinical observations and may be an advantage when these drugs are given as postoperative analgesics at the doses tested.
Antithrombotic Effect of Human Recombinant Tissue Factor Pathway Inhibitor on Endotoxin-induced Intravascular Coagulation in Rats: Concerted Effect with Antithrombin
Authors: Kamikubo Yu-ichi , Hamuro Tsutomu , Matsuda Jun-ichi , Shinya Noriko , Miyamoto Seiji , Funatsu Akinobu , Kato Hisao
Published: July 2018

In our current study, we examined the antithrombotic effect of Chinese hamster ovary cell-derived human recombinant tissue factor pathway inhibitor (h-rTFPI) by intravenous injection of h-rTFPI with or without antithrombin into endotoxin-treated rats. An injection of h-rTFPI at a high dose (4 mg/kg of h-rTFPI or three doses of 1 mg/kg) significantly prevented the decrease of fibrinogen and factor VIII and the increase of fibrin/fibrinogen degradation products and glutamic-pyruvic transaminase in rats, while a single injection of 1 mg/kg of h-rTFPI or 250 U/kg of antithrombin did not significantly prevent intravascular coagulation. However, a simultaneous injection of 1 mg/kg of h-rTFPI and 250 U/kg of antithrombin did significantly prevent intravascular coagulation. From the studies on the clearance rate and immunohistochemical staining of injected h-rTFPI into normal rats, we found that most of the intravenously-injected h-rTFPI was localized on the central vein and sinusoids in the liver and catabolized via the proteoglycan-mediated pathway with a half-life of 48 min. These results suggest that h-rTFPI and antithrombin prevented endotoxin-induced intravascular coagulation in concert by binding to the vascular walls of the liver and by inhibiting fibrin formation on Kupffer cells in hepatic sinusoids.
Coagulation and Fibrinolytic Profile of Paediatric Patients Undergoing Cardiopulmonary Bypass
Authors: Chan Anthony K C , Leaker Michael , Burrows Frederick A , Williams William G , Gruenwald Colleen E , Whyte Linda , Adams Margaret , Brooker Lu Ann , Adams Helen , Mitchell Lesley , Andrew Maureen
Published: July 2018

The haemostatic system and the use of heparin during cardiopulmonary bypass (CPB) have been studied extensively in adults but not in children. Results from adult trials cannot be extrapolated to children because of age-dependent physiologic differences in haemostasis. We studied 22 consecutive paediatric patients who underwent CPB at The Hospital for Sick Children, Toronto. Fibrinogen, factors II, V, VII, VIII, IX, XI, XII, prekallikrein, protein C, protein S, antithrombin (AT), heparin cofactor II, α2-macroglobulin, plasminogen, α2-antiplas- min, tissue plasminogen activator (tPA), plasminogen activator inhibitor, thrombin-AT complexes (TAT), D-dimer, heparin (by both anti-factor Xa assay and protamine titration) and activated clotting time (ACT) were assayed perioperatively. The timing of the sampling was: pre heparin, post heparin, after initiation of CPB, during hypothermia, post hypothermia, post protamine reversal and 24 h post CPB. Plasma concentrations of all haemostatic proteins decreased by an average of 56% immediately following the initiation of CPB due to haemodilution. During CPB, the majority of procoagulants, inhibitors and some components of the fibrinolytic system (plasminogen, α2AP) remained stable. However, plasma concentrations of TAT and D-dimers increased during CPB showing that significant activation of the coagulation and fibrinolytic systems occurred. Mechanisms responsible for the activation of haemostasis are likely complex. However, low plasma concentrations of heparin (<2.0 units/ml in 45% of patients) during CPB were likely a major contributing etiology. ACT values showed a poor correlation (r = 0.38) with heparin concentrations likely due to concurrent haemodilution of haemostatic factors, activation of haemostatic system, hypothermia and activation of platelets. In conclusion, CPB in paediatric patients causes global decreases of components of the coagulation and fibrinolytic systems, primarily by haemodilution and secondarily by consumption.
Differential Expression of Low Mr GTP-binding Proteins in Human Megakaryoblastic Leukemia Cell Line, MEG-01, and their Possible Involvement in the Differentiation Process
Authors: Nagata Koh-ichi , Okano Yukio , Nozawa Yoshinori
Published: July 2018

The expression of various low Mr GTP-binding proteins at various states of differentiation of a human megakaryoblastic leukemia cell line, MEG-01, was analyzed using thermocycle amplification of mRNA and immunoblotting. MEG-01 cells were found to express mRNAs of rap1A, rap1B, rap2B, ralA, rhoA, rac1, rac2, CDC42Hs, rab1, rab3B, rab6, ram and ran, but not rab4, and the proteins of Rap 1, Rap2, RhoA, Rac1, Rac2, Rab3B, Rab4, Rab6 and Rab8 were expressed. Differentiation of MEG-01 cells induced by 100 nM 12-O-tetradecanoylphorbol-13-acetate revealed the considerable increases in mRNA expression of rap1B, rab3B, rabA, ram and ran whereas the levels of rap2B, rhoA and rac1 decreased. During the differentiation process, significant changes in protein levels of Rap1, RhoA, Rac1, Rac2, Rab3B, Rab4 and Rab6 were observed among three subcellular (cytosol, Triton X-100-soluble membrane and -insoluble cytoskeleton) fractions. The present investigation may be useful for the study of the megakaryocyte differentiation.
Acute von Willebrand Factor Secretion from the Endothelium In Vivo: Assessment through Plasma Propeptide (vWf:AgII) Levels
Authors: Vischer Ulrich M , Ingerslev Jørgen , Wollheim Claes B , Mestries Jean-Claude , Tsakiris Dimitrios A , Haefeli Walter E , Kruithof Egbert K O
Published: July 2018

Elevated plasma concentrations of von Willebrand factor (vWf) are increasingly recognized as a cardiovascular risk factor, and are used as a marker of endothelial activation. However, the factors which determine the rate of vWf release from the endothelium in vivo have not been defined clearly. In addition, vWf plasma levels may also be influenced by adhesion of vWf to the vascular wall or to platelets, and by its rate of degradation. The propeptide of vWf (also called vWf:AgII) is stored and released in equimolar amounts with vWf. In the present study we attempted to determine whether this propeptide could be a more reliable marker of endothelial secretion than vWf itself. To accomplish this we developed an ELISA based on monoclonal antibodies. The propeptide levels in normal plasma were found to be 0.7 µg/ml, more than 10 times lower than vWf itself. Administration of desmopressin (DDAVP) induced a rapid relative increase in propeptide (from 106 to 879%) and in vWf (from 112 to 272%). However, the increases in vWf and propeptide were equivalent when expressed in molar units. A time course study indicated a half-life of the propeptide of 3 h or less. In a baboon model of disseminated intravascular coagulation (DIC) induced by FXa, vWf increased by less than 100%, whereas the propeptide concentrations increased by up to 450%. In view of the massive thrombin generation (as assessed by fibrinogen depletion), the increases in vWf are small, compared to the strong secretory response to thrombin and fibrin previously observed in vitro. Our results suggest that due to its rapid turnover, the propeptide could provide a sensitive plasma marker of acute endothelial secretion.
C5a Induces Tissue Factor Activity on Endothelial Cells
Authors: Ikeda Kei , Nagasawa Kohei , Horiuchi Takahiko , Tsuru Tomomi , Nishizaka Hiroaki , Niho Yoshiyuki
Published: July 2018

Tissue factor (TF) is an integral membrane glycoprotein that serves as a cofactor for blood coagulation factor VIIa. The induction of TF on the surface of endothelial cells is initiated by various stimuli including lipopolysaccharide, interleukin-1β, and tumor necrosis factor α. We have demonstrated that recombinant human C5a induces TF activity in a dose-dependent fashion in human umbilical vein endothelial cells (HUVEC). Peak activity (4.9-fold increase) was obtained 3-6 h after treatment with 10 μM C5a. TF mRNA as assessed by RT-PCR method was also significantly increased (3.75-fold) after 3 h incubation with C5a, suggesting that C5a induces TF activity on HUVEC, at least in part, by enhancing the level of TF mRNA. The increase in TF activity by C5a was inhibited by methylprednisolone. The induction of TF on endothelial cells by C5a may represent one of many potential interrelationships between the inflammatory and coagulation schemes.
Developing theories of change for social programmes: co-producing evidence-supported quality improvement
Authors: Ghate Deborah
Published: July 2018

For much of the past two decades, expensive and often imported evidence-based programmes (EBPs) developed by clinician-researchers have been much in vogue in the family and parenting support field, as in many other areas of social provision. With their elaborate infrastructures, voluminous research bases and strict licensing criteria, they have seemed to offer certainty of success over less packaged, less well-evidenced locally developed approaches. Yet recently, evaluation research is showing that success is not assured. EBPs can and regularly do fail, at substantial cost to the public purse. In times of severe resource pressure, a pressing question is, therefore, whether lower cost, home-grown, practitioner-developed programmes—the sort often overlooked by policy-makers —can deliver socially significant and scientifically convincing outcomes at lower cost and at least on a par with their better resourced cousins. This paper shows how the application of techniques increasingly used in implementation science (the science of effective delivery) could help level the playing field. Processes for doing this including co-produced theory of change development and validation are illustrated with reference to the Family Links Ten Week Nurturing Programme (FLNP-10), a popular manualised group-based parenting support programme, designed and disseminated since the 1990s by a UK-based purveyor organisation. The paper draws out general principles for formulating and structuring strong theories of change for practice improvement projects. The work shows that novel application of implementation science-informed techniques can help home-grown programmes to compete scientifically by strengthening their design and delivery, and preparing the ground for better and fairer evaluation.
Cantilever-based microring lasers embedded in a deformable substrate for local strain gauges
Authors: Zhou Taojie , Liu Xiu , Cui Yuzhou , Cheng Yuansen , Fang Xuan , Zhang Weilin , Xiang Boyuan , Zhang Zhaoyu
Published: July 2018

A cantilever-based microring laser structure was proposed for easily integrating III-V active layer into mechanically stretchable substrates. Local strain gauges were demonstrated by embedding cantilever-based microring lasers in a deformable polymer substrate. The characterizations of microscale local strain gauges had been studied from both simulated and experimental results. The lasing wavelength of strain gauges was blue-shift and linear tuned by stretching the flexible substrate. Gauge factor being ∼11.5 nm per stretching unit was obtained for a cantilever-based microring laser with structural parameters R=1.25 μm, W1=450 nm and W2=240 nm. Such microring lasers embedded in a flexible substrate are supposed to function not only as strain gauges for monitoring the micro- or nano-structured deformation, but also tunable light sources for photonic integrated circuits.
Bugs, ants, wasps, moths and other insect species.
Authors: Roques A. , Preda C. , Augustin S. , Auger-Rozenberg M. A.
Published: July 2018

A total of 43 insect species non-native to Europe are so far considered to affect human welfare through their biting, urticating and allergenic properties, or by causing domestic nuisances. They involve several orders. In Hymenoptera, species in two families, Formicidae (ants) and Vespidae (wasps and hornets), are known to cause disturbance and health problems. Several moth species (Lepidoptera) have urticating larvae which may induce painful urticarial and allergic reactions. Bugs in five Hemipteran families have direct impacts on health, such as bed bugs and kissing bugs, which are vectors of pathogens, but most are considered to be household pests, causing nuisances to people when invading houses or aggregating on walls. Several non-native species of cockroaches that develop in synanthropic habitats have body parts, saliva or faeces containing powerful indoor allergens, and they can also facilitate mechanical transmission of pathogens to humans. Some species of Siphonaptera (fleas) and Phthiraptera (lice), which are obligate ectoparasitic insects of warm-blooded animals, are of high importance for human health because they cause itches and skin infection, and transmit major diseases such as bubonic plague and murine typhus.
Invasive freshwater invertebrates and fishes: impacts on human health.
Authors: Souty-Grosset C. , Anastácio P. , Reynolds J. , Tricarico E.
Published: July 2018

Inland waters are subject to more widespread biotic invasions than terrestrial ecosystems. During the last century, 756 aquatic species were introduced in Europe, frequently carrying new parasites for native fauna and humans. The consequences of such invasions are the loss of the invaders' original parasites, the introduction of new parasites, or new intermediate hosts or vectors for existing parasites. Many parasites are water-borne and need aquatic species to complete their transmission cycles. The list of 100 of the World's Worst Invasive Alien Species (Lowe et al., 2000) does not take into account human health problems, so a risk assessment of the consequences of invasive freshwater alien species requires more attention. Here we review the direct and indirect impacts of invasive freshwater alien species on human health. Direct impacts include the injuries or allergies and new contaminants (bacteria, toxins), and their role as intermediate hosts to human parasites. Indirect impacts include the effects of the chemicals needed to control these aliens, changes to ecosystem services making the invaded area less suitable for recreational human use and damage to cultivation/aquaculture affecting human well-being in developing countries. A clear management response is urgently needed to halt their spread and reduce or minimize the risk of human and wildlife disease.
Risks for human health related to invasive alien reptiles and amphibians.
Authors: Pauwels O. S. G. , Pantchev N.
Published: July 2018

More than 100 amphibian and reptile species have established populations outside their natural geographical range, mostly as a consequence of the international pet trade. About 40 zoonoses are associated with reptiles and amphibians. The main zoonotic risks from alien invasive reptiles and amphibians are salmonellosis and probably also vibriosis from a bacteriological point of view, pentastomids, sparganosis and potentially trichinellosis from a parasitical point of view, and West Nile virus. There are also new and emerging pathogens, e.g. atypical Brucella spp., with zoonotic potential. Transmission of pathogens from introduced reptile and amphibian species to humans is limited by the important physiological differences between them and humans, the secretive or shy habits of most introduced species and the rarity of direct contact (with the notable exception of a few exotic species eaten by humans). Locally, alien reptiles include venomous species and large species able to inflict bites of medical concern. In certain areas some species (mainly anuran amphibians) are generating noise pollution affecting human well-being. Given the continued increase of invasive alien population establishments with time, the spread of alien arthropod vectors and aggravating factors such as climate change, it is expected that alien reptiles and amphibians and their associated pathogens will generate more public health concern in the future.
A hybridde novogenome assembly of the honeybee,Apis mellifera, with chromosome-length scaffolds
Authors: Wallberg Andreas , Bunikis Ignas , Pettersson Olga Vinnere , Mosbech Mai-Britt , Childers Anna K. , Evans Jay D. , Mikheyev Alexander S. , Robertson Hugh M. , Robinson Gene E. , Webster Matthew T.
Published: July 2018

Background

The ability to generate long sequencing reads and access long-range linkage information is revolutionizing the quality and completeness of genome assemblies. Here we use a hybrid approach that combines data from four genome sequencing and mapping technologies to generate a new genome assembly of the honeybeeApis mellifera. We first generated contigs based on PacBio sequencing libraries, which were then merged with linked-read 10x Chromium data followed by scaffolding using a BioNano optical genome map and a Hi-C chromatin interaction map, complemented by a genetic linkage map.

Results

Each of the assembly steps reduced the number of gaps and incorporated a substantial amount of additional sequence into scaffolds. The new assembly (Amel_HAv3) is significantly more contiguous and complete than the previous one (Amel_4.5), based mainly on Sanger sequencing reads. N50 of contigs is 120-fold higher (5.381 Mbp compared to 0.053 Mbp) and we anchor >98% of the sequence to chromosomes. All of the 16 chromosomes are represented as single scaffolds with an average of three sequence gaps per chromosome. The improvements are largely due to the inclusion of repetitive sequence that was unplaced in previous assemblies. In particular, our assembly is highly contiguous across centromeres and telomeres and includes hundreds ofAvaIandAluIrepeats associated with these features.

Conclusions

The improved assembly will be of utility for refining gene models, studying genome function, mapping functional genetic variation, identification of structural variants, and comparative genomics.
Cicada endosymbionts contain tRNAs that are processed despite having genomes that lack tRNA processing activities
Authors: Van Leuven James T. , Mao Meng , Bennett Gordon M. , McCutcheon John P.
Published: July 2018

Gene loss and genome reduction are defining characteristics of nutritional endosymbiotic bacteria. In extreme cases, even essential genes related to core cellular processes such as replication, transcription, and translation are lost from endosymbiont genomes. Computational predictions on the genomes of the two bacterial symbionts of the cicadaDiceroprocta semicincta, “CandidatusHodgkinia cicadicola” (Alphaproteobacteria) and “Ca. Sulcia muelleri” (Betaproteobacteria), find only 26 and 16 tRNA, and 15 and 10 aminoacyl tRNA synthetase genes, respectively. Furthermore, the original “Ca.Hodgkinia” genome annotation is missing several essential genes involved in tRNA processing, such as RNase P and CCA tRNA nucleotidyltransferase, as well as several RNA editing enzymes required for tRNA maturation. How “Ca. Sulcia” and “Ca. Hodgkinia” preform basic translation-related processes without these genes remains unknown. Here, by sequencing eukaryotic mRNA and total small RNA, we show that the limited tRNA set predicted by computational annotation of “Ca. Sulcia” and “Ca. Hodgkinia” is likely correct. Furthermore, we show that despite the absence of genes encoding tRNA processing activities in the symbiont genomes, symbiont tRNAs have correctly processed 5’ and 3’ ends, and seem to undergo nucleotide modification. Surprisingly, we find that most “Ca. Hodgkinia”and “Ca. Sulcia” tRNAs exist as tRNA halves. Finally, and in contrast with other related insects, we show that cicadas have experienced little horizontal gene transfer that might complement the activities missing from the endosymbiont genomes. We conclude that “Ca. Sulcia” and “Ca. Hodgkinia” tRNAs likely function in bacterial translation, but require host-encoded enzymes to do so.
Identification of pyroptosis inhibitors that target a reactive cysteine in gasdermin D
Authors: Hu Jun Jacob , Liu Xing , Zhao Jingxia , Xia Shiyu , Ruan Jianbin , Luo Xuemei , Kim Justin , Lieberman Judy , Wu Hao
Published: July 2018

Inflammasomes are multi-protein signalling scaffolds that assemble in response to invasive pathogens and sterile danger signals to activate inflammatory caspases (1/4/5/11), which trigger inflammatory death (pyroptosis) and processing and release of pro-inflammatory cytokines1,2. Inflammasome activation contributes to many human diseases, including inflammatory bowel disease, gout, type II diabetes, cardiovascular disease, Alzheimer’s disease, and sepsis, the often fatal response to systemic infection3–6. The recent identification of the pore-forming protein gasdermin D (GSDMD) as the final pyroptosis executioner downstream of inflammasome activation presents an attractive drug target for these diseases7–11. Here we show that disulfiram, a drug used to treat alcohol addiction12, and Bay 11-7082, a previously identified NF-κB inhibitor13, potently inhibit GSDMD pore formation in liposomes and inflammasome-mediated pyroptosis and IL-1β secretion in human and mouse cells. Moreover, disulfiram, administered at a clinically well-tolerated dose, inhibits LPS-induced septic death and IL-1β secretion in mice. Both compounds covalently modify a conserved Cys (Cys191 in human and Cys192 in mouse GSDMD) that is critical for pore formation8,14. Inflammatory caspases employ Cys active sites, and many previously identified inhibitors of inflammatory mediators, including those against NLRP3 and NF-κB, covalently modify reactive cysteine residues15. Since NLRP3 and noncanonical inflammasome activation are amplified by cellular oxidative stress16–22, these redox-sensitive reactive cysteine residues may regulate inflammation endogenously, and compounds that covalently modify reactive cysteines may inhibit inflammation by acting at multiple steps. Indeed, both disulfiram and Bay 11-7082 also directly inhibit inflammatory caspases and pleiotropically suppress multiple processes in inflammation triggered by both canonical and noncanonical inflammasomes, including priming, puncta formation and caspase activation. Hence, cysteine-reactive compounds, despite their lack of specificity, may be attractive agents for reducing inflammation.
Transient Hsp90 suppression promotes a heritable change in protein translation
Authors: Tsvetkov Peter , Brune Zarina , Eisen Timothy J. , Heinrich Sven , Newby Greg A. , Hallacli Erinc , Kayatekin Can , Pincus David , Lindquist Susan
Published: July 2018

The heat shock protein 90 (Hsp90) chaperone functions as a protein-folding buffer and plays a unique role promoting the evolution of new heritable traits. To investigate the role of Hsp90 in modulating protein synthesis, we screened more than 1200 proteins involved in mRNA regulation for physical interactions with Hsp90 in human cells. Among the top hits was CPEB2, which strongly binds Hsp90 via its prion domain, reminiscent of the prion-like regulation of translation of Aplysia CPEB. In a yeast model of CPEB prion-dependent translation regulation, transient inhibition of Hsp90 amplified CPEB2 prion activity and resulted in persistent translation of the CPEB reporter. Remarkably, inhibition of Hsp90 was sufficient to induce a heritable change in protein translation that persisted for 30 generations, even in the absence of exogenous CPEB. Although we identified a variety of perturbations that enhanced translation of the reporter, only Hsp90 inhibition led to persistent activation. Thus, transient loss of Hsp90 function leads to the non-genetic inheritance of a novel translational state. We propose that, in addition to sculpting the conformational landscape of the proteome, Hsp90 promotes phenotypic variation by modulating protein synthesis.
Osmia bicornis is rarely an adequate regulatory surrogate species. Comparing its acute sensitivity towards multiple insecticides with regulatory Apis mellifera endpoints
Authors: Uhl Philipp , Awanbor Osarobo , Schulz Robert S. , Brühl Carsten A.
Published: July 2018

Bee species provide essential ecosystem services and maintain floral biodiversity. However, there is an ongoing decline of wild and domesticated bee species. Since agricultural pesticide use is a key driver of this process, there is a need for a protective risk assessment. To achieve a more protective registration process, two wild bee species, Osmia bicornis and Bombus terrestris, were proposed by the European Food Safety Authority as additional test surrogates. We investigated the acute toxicity (median lethal dose, LD50) of multiple commercial insecticide formulations towards the red mason bee (O. bicornis) and compared these values to honey bee (Apis mellifera) regulatory endpoints. In two thirds of all cases O. bicornis was less sensitive than the honey bee. By applying an assessment factor of 10 on the honey bee endpoint a protective level was achieved for 87% (13 out 15) of all evaluated products. Our results show that O. bicornis as a non-sensitive species is rarely an adequate additional surrogate species for lower tier risk assessment. Given the currently limited database, the honey bee seems sufficiently protective in acute scenarios as long as a reasonable assessment factor is applied. However, additional surrogate species such as O. bicornis and B. terrestris are still relevant for ecologically meaningful higher tier studies.
MicroRNA miR-7 and miR-17-92 in POMC neurons are associated with sex-specific regulation of diet-induced obesity
Authors: Gao Yanxia , Li Jiaheng , Zhang Zhen , Zhang Ruihan , Pollock Andrew , Sun Tao
Published: July 2018

Proopiomelanocortin (POMC) neurons in the arcuate nucleus (ARC) in mammalian hypothalamus play important roles in regulating appetite, energy expenditure, and glucose and fat metabolisms. Diet-induced obesity often show sex-specific difference. But the underlying mechanisms remain unclear. Here we show that microRNA (miRNA) miR-7 and miR-17-92 are expressed in the mouse ARC, and mostly in POMC neurons. Knockdown of miR-7 and knockout of miR-17-92 specifically in POMC neurons aggravate diet-induced obesity only in females and males, respectively. Moreover, gene expression profile analysis identifies sex-differential genes in male and female ARCs in wildtype adult mice. Interestingly, these genes that normally show low-expression in the female and male ARCs display upregulated expression in female miR-7 knockdown and male miR-17-92 knockout mice, respectively. Our results demonstrate an important role of miRNAs in regulating sex-specific diet-induced obesity, likely through modulating expression of target genes that show sex-differential expression in the ARC of hypothalamus.
Structured environments fundamentally alter dynamics and stability of ecological communities
Authors: Vallespir Nick , Ursell Lowery & Tristan
Published: July 2018

The dynamics and stability of ecological communities are intimately linked with the specific interactions – like cooperation or predation – between constituent species. In microbial communities, like those found in soils or the mammalian gut, physical anisotropies produced by fluid flow and chemical gradients impact community structure and ecological dynamics, even in structurally isotropic environments. Though natural communities existing in physically unstructured environments is rare, the role of environmental structure in determining community dynamics and stability remains poorly studied. To address this gap, we used modified Lotka-Volterra simulations of competitive microbial communities to characterize the effects of surface structure on community dynamics. We find that environmental structure has profound effects on communities, in a manner dependent on the specific pattern of interactions between community members. For two mutually competing species, eventual extinction of one competitor is effectively guaranteed in isotropic environments. However, addition of environmental structure enables long-term coexistence of both species via local ‘pinning’ of competition interfaces, even when one species has a significant competitive advantage. In contrast, while three species competing in an intransitive loop (as in a game of rock-paper-scissors) coexist stably in isotropic environments, structural anisotropy disrupts the spatial patterns on which coexistence depends, causing chaotic population fluctuations and subsequent extinction cascades. These results indicate that the stability of microbial communities strongly depends on the structural environment in which they reside. Therefore, a more complete ecological understanding, including effective manipulation and interventions in natural communities of interest, must account for the physical structure of the environment.

SIGNIFICANCE

Many microbial communities of ecological and medical importance reside in complex and heterogeneous environments, such as soils or intestinal tracts. While many studies consider the effects of flow or chemical gradients in structuring these communities, how the physical structure of the environment shapes community dynamics and outcomes remains poorly understood. Using simulations of competitive microbial communities, we show that stability and dynamics qualitatively shift in environments with complex surface structures compared to open isotropic environments. Therefore, in addition to biochemical interactions between species, our work suggests that the physical structure of the environment is an equally important determinant of dynamics and stability in microbial communities, in a manner dependent on the specific patterns of interactions within that community.
Inversion and computational maturation of drug response using human stem cell derived cardiomyocytes in microphysiological systems
Authors: Tveito Aslak , Jæger Karoline Horgmo , Huebsch Nathaniel , Charrez Berenice , Edwards Andrew G. , Wall Samuel , Healy Kevin E.
Published: July 2018

While cardiomyocytes differentiated from human induced pluripotent stems cells (hiPSCs) hold great promise for drug screening, the electrophys-iological properties of these cells can be variable and immature, producing results that are significantly different from their human adult counterparts. Here, we describe a computational framework to address this limitation, and show how in silico methods, applied to measurements on immature cardiomyocytes, can be used to both identify drug action and to predict its effect in mature cells. Our synthetic and experimental results indicate that optically obtained waveforms of voltage and calcium from microphysiological systems can be inverted into information on drug ion channel blockage, and then, through assuming functional invariance of proteins during maturation, this data can be used to predict drug induced changes in mature ventricular cells. Together, this pipeline of measurements and computational analysis could significantly improve the ability of hiPSC derived cardiomycocytes to predict dangerous drug side effects.
Can we obtain in vivo transmural mean hoop stress of the aortic wall without knowing patient-specific material properties and residual deformations?
Authors: Liu Minliang , Liang Liang , Liu Haofei , Zhang Ming , Martin Caitlin , Sun Wei
Published: July 2018

It is well known that residual deformations/stresses alter the mechanical behavior of arteries, e.g. the pressure-diameter curves. In an effort to enable personalized analysis of the aortic wall stress, approaches have been developed to incorporate experimentally-derived residual deformations into in vivo loaded geometries in finite element simulations using thick-walled models. Solid elements are typically used to account for “bending-like” residual deformations. Yet, the difficulty in obtaining patient-specific residual deformations and material properties has become one of the biggest challenges of these thick-walled models. In thin-walled models, fortunately, static determinacy offers an appealing prospect that allows for the calculation of the thin-walled membrane stress without patient-specific material properties. The membrane stress can be computed using forward analysis by enforcing an extremely stiff material property as penalty treatment, which is referred to as the forward penalty approach. However, thin-walled membrane elements, which have zero bending stiffness, are incompatible with the residual deformations, and therefore, it is often stated as a limitation of thin-walled models. In this paper, by comparing the predicted stresses from thin-walled models and thick-walled models, we demonstrate that the transmural mean hoop stress is the same for the two models and can be readily obtained from in vivo clinical images without knowing the patient-specific material properties and residual deformations. Computation of patient-specific mean hoop stress can be greatly simplified by using membrane model and the forward penalty approach, which may be clinically valuable.
2-(5-Methyl-1-phenyl-1H-1,2,3-triazol-4-yl)-5-phenyl-1,3,4-oxadiazole
Authors: El-Hiti Gamal A. , Abdel-Wahab Bakr F. , Mostafa Mohamed S. , Khidre Rizk E. , Hegazy Amany S. , Kariuki Benson M.
Published: July 2018

The asymmetric unit of the title compound, C17H13N5O, comprises four independent molecules (A–D). The respective interplanar angles between the phenyl/oxadiazole/methyltriazole/phenyl rings for the four independent molecules are A 8.8 (2), 13.0 (2), 22.5 (2)°; B 6.3 (2), 8.9 (2), 29.0 (1)°; C 4.0 (2), 10.0 (2), 24.5 (2)°; D 3.5 (2), 10.1 (2), 27.2 (2)°. In the crystal, molecules form two separate stacks parallel to the b-axis direction: one consists of A and D molecules, and the other of B and C molecules. Aromatic π–π stacking is observed within each stack, with the shortest centroid–centroid separation being 3.552 (2) Å.