bluuebunny/crossref_metadata_2025 huggingface dataset:
Dual RNA-seq provides insight into the biology of the neglected intracellular human pathogenOrientia tsutsugamushi
Authors: Mika-Gospodorz Bozena , Giengkam Suparat , Westermann Alexander J. , Wongsantichon Jantana , Kion-Crosby Willow , Chuenklin Suthida , Wang Loo Chien , Sunyakumthorn Piyanate , Sobota Radoslaw M. , Subbian Selvakumar , Vogel Jörg , Barquist Lars , Salje Jeanne
Published: August 2019

Emerging and neglected diseases pose challenges as their biology is frequently poorly understood, and genetic tools often do not exist to manipulate the responsible pathogen. Organism agnostic sequencing technologies offer a promising approach to understand the molecular processes underlying these diseases. Here we apply dual RNA-seq toOrientia tsutsugamushi(Ot), an obligate intracellular bacterium and the causative agent of the vector-borne human disease scrub typhus. Half the Ot genome is composed of repetitive DNA, and there is minimal collinearity in gene order between strains. Integrating RNA-seq, comparative genomics, proteomics, and machine learning, we investigated the transcriptional architecture of Ot, including operon structure and non-coding RNAs, and found evidence for wide-spread post-transcriptional antisense regulation. We compared the host response to two clinical isolates and identified distinct immune response networks that are up-regulated in response to each strain, leading to predictions of relative virulence which were confirmed in a mouse infection model. Thus, dual RNA-seq can provide insight into the biology and host-pathogen interactions of a poorly characterized and genetically intractable organism such as Ot.
Long-termin vivoimaging ofDrosophilalarvae
Authors: Kakanj Parisa , Eming Sabine A. , Partridge Linda , Leptin Maria
Published: August 2019

TheDrosophilalarva has been used to investigate many processes in cell biology, including morphogenesis, physiology, responses to drugs and new therapeutic compounds. Despite its enormous potential as a model system, it has technical limitations in cases where longer-term live imaging is necessary, because of the lack of efficient methods for immobilising larvae for extended periods. We describe here a simple procedure for anaesthetisation and long-termin vivoimaging of the epidermis and other larval organs including gut, imaginal discs, neurons, fat body, tracheae and haemocytes, and show a procedure for probing cell properties by laser ablation. We include a survey of different anaesthetics, showing that short exposure to diethyl ether is the most effective for long-term immobilisation of larvae. This method does not require specific expertise beyond basicDrosophilagenetics and husbandry, and confocal microscopy. It enables high-resolution studies of many systemic and sub-cellular processes in larvae.
Cell culture dimensionality influences mesenchymal stem cell fate through cadherin-2 and cadherin-11
Authors: Passanha Fiona R , Geuens Thomas , Konig Simon , van Blitterswijk Clemens A , LaPointe Vanessa LS
Published: August 2019

The acquisition of a specific cell fate is one of the core aims of tissue engineering and regenerative medicine. Significant evidence shows that aggregate cultures have a positive influence on fate decisions, presumably through cell-cell interactions, but little is known about the specific mechanisms. To investigate the difference between cells cultured as a monolayer and as aggregates, we started by looking at cadherin expression, an important protein involved in cell adhesion, during the differentiation of bone marrow-derived human mesenchymal stem cells (hMSCs) in aggregate and monolayer cultures. We observed that proliferating hMSCs in monolayer culture express cadherin-2 and undergo a switch to cadherin-11 over time, which was not evident in the aggregate cultures. By knocking down cadherin-2 and cadherin-11, we found that both cadherins were required for adipogenic differentiation in a monolayer as well as aggregate culture. However, during osteogenic differentiation, low levels of cadherin-2 were found to be favorable for cells cultured as a monolayer and as aggregates, whereas cadherin-11 was dispensable for cells cultured as aggregates. Together, these results provide compelling evidence for the important role that cadherins play in regulating the differentiation of hMSCs and how this is affected by the dimensionality of cell culture.
Prevalence and mutational determinants of high tumor mutation burden in breast cancer
Authors: Barroso-Sousa R. , Jain E. , Cohen O. , Kim D. , Buendia-Buendia J. , Winer E. , Lin N. , Tolaney S.M. , Wagle N.
Published: August 2019

Background

High tumor mutation burden (TMB) has been associated with benefit to immunotherapy in multiple tumor types. However, the prevalence of hypermutated breast cancer is not well described. The aim of this study is to evaluate frequency, mutational patterns, and genomic profile of hypermutated breast cancer.

Patients and Methods

We used de-identified data from individuals with primary or metastatic breast cancer from six different publicly available genomic studies. The prevalence of hypermutated breast cancer was determined among 3969 patients’ samples that underwent whole exome sequencing or gene panel sequencing. Samples were classified as having high TMB if they had ≥10 mutations per megabase (mut/Mb). An additional 8 patients were identified from a Dana-Farber Cancer Institute cohort for inclusion in the hypermutated cohort. Among patients with high TMB, the mutational patterns, and genomic profile were determined. A subset of patients was treated with regimens containing PD-1 inhibitors.

Results

The median TMB was 2.63 mut/Mb. Median TMB significantly varied according to tumor subtype (HR-/HER2-> HER2+ > HR+/HER2-,p< 0.05) and sample type (metastatic > primary,p2.2×10−16). Hypermutated tumors were found in 198 patients (5%), with an enrichment in metastatic versus primary tumors (8.4% versus 2.9%, p = 6.5 × 10−14). APOBEC activity (59.2%), followed by mismatch repair deficiency (MMRd; 36.4%), were the most common mutational processes among hypermutated tumors. Three patients with hypermutated breast cancer—including two with a dominant APOBEC activity signature and one with a dominant MMRd signature—treated with pembrolizumab-based therapies derived an objective and durable response to therapy.

Conclusion

Hypermutation occurs in 5% of all breast cancers, with an enrichment in metastatic tumors. Different mutational signatures are present in this population, with APOBEC activity being the most common dominant process. Preliminary data suggest that hypermutated breast cancers are more likely to benefit from PD-1 inhibitors.

Key Message

High tumor mutation burden is found in 5% of all breast cancers and is more common in metastatic tumors. While different mutational signatures are present in hypermutated tumors, APOBEC activity is the most common dominant process. Preliminary data suggest that those tumors are more likely to benefit from PD-1 inhibitors.
Empathic contagious pain and consolation in laboratory rodents: species and sex comparisons
Authors: Du Rui , Luo Wen-Jun , Geng Kai-Wen , Li Chun-Li , Yu Yang , He Ting , Wei Na , Chen Jun
Published: August 2019

Laboratory rodents are gregarious in nature and have a feeling of empathy when witnessing a familiar conspecific in pain. The rodent observers express two levels of empathic responses: observational contagious pain (OCP) and consolation. Here we examined the sex and species difference of OCP and consolation in male and female mice and rats. We observed no species difference in both OCP and consolation, but significant species difference in general social (allo-mouth and/or allo-tail sniffing) and non-social (self-grooming) behaviors. For sex difference, male mouse observers showed more allolicking and allogrooming behaviors toward a familiar conspecific in pain during and longer time increase in pain sensitivity after the PDSI than female mouse observers. However, no sex difference was observed in rats. Our results highlighted an evolutionary view of empathy that social animals including rodents also have the ability to feel, recognize, understand and share the other’s distressing states.
Traction forces control cell-edge dynamics and mediate distance-sensitivity during cell polarization
Authors: Messi Zeno , Bornert Alicia , Raynaud Franck , Verkhovsky Alexander
Published: August 2019

Traction forces are generated by cellular actin-myosin system and transmitted to the environment through adhesions. They are believed to drive cell motion, shape changes, and extracellular matrix remodeling [1–3]. However, most of the traction force analysis has been performed on stationary cells, investigating forces at the level of individual focal adhesions or linking them to static cell parameters such as area and edge curvature [4–10]. It is not well understood how traction forces are related to shape changes and motion, e.g. forces were reported to either increase or drop prior to cell retraction [11–15]. Here, we analyze the dynamics of traction forces during the protrusion-retraction cycle of polarizing fish epidermal keratocytes and find that forces fluctuate in concert with the cycle, increasing during the protrusion phase and reaching maximum at the beginning of retraction. We relate force dynamics to the recently discovered phenomenological rule [16] that governs cell edge behavior during keratocyte polarization: both traction forces and the probability of switch from protrusion to retraction increase with the distance from the cell center. Diminishing traction forces with cell contractility inhibitor leads to decreased edge fluctuations and abnormal polarization, while externally applied force can induce protrusion-retraction switch. These results suggest that forces mediate distance-sensitivity of the edge dynamics and ultimately organize cell-edge behavior leading to spontaneous polarization. Actin flow rate did not exhibit the same distance-dependence as traction stress, arguing against its role in organizing edge dynamics. Finally, using a simple model of actin-myosin network, we show that force-distance relationship may be an emergent feature of such networks.
Spatial variation in introgression along a toad hybrid zone in France
Authors: van Riemsdijk I. , Arntzen J.W. , Bucciarelli G. , McCartney-Melstad E. , Rafajlović M. , Scott P.A. , Toffelmier E. , Shaffer H. B. , Wielstra B.
Published: August 2019

The barrier effect is a restriction of gene flow between diverged populations by barrier genes. Restriction of gene flow and asymmetric introgression over multiple transects indicates species wide (genetic) adaptations, whereas transect-specific barrier loci may indicate local adaptation to gene flow. Asymmetric introgression can be caused by selection, hybrid zone movement, asymmetric reproductive isolation, or a combination of these. We study two widely separated transects (northwest and southeast France) for the 900 km long hybrid zone betweenBufo bufoandB. spinosustoads, using ~1200 markers from restriction-site associated DNA (RAD) sequencing data. Genomic and geographic clines were used to identify outlier markers which show restricted or elevated introgression. Twenty-six barrier markers are shared between transects (the union of 56 and 123 barrier markers identified in each transect), which is more than would be expected by chance. However, the number of barrier markers is twice as high in the southeast transect. In the northwest transect a high amount of (asymmetric) introgression fromB. spinosusintoB. bufois consistent with hybrid zone movement or asymmetric reproductive isolation. In the southeast transect, introgression is symmetric and consistent with a stable hybrid zone. Differences between transects may be related to genetic sub-structure withinB. bufo. A longer period of secondary contact in southeast France appears to result in a relatively stronger barrier effect than in the northwest. TheBufohybrid zone provides an excellent opportunity to separate a general barrier to gene flow from local reductions in gene flow.
β11-12 linker isomerization governs Acid-sensing ion channel desensitization and recovery
Authors: Rook Matthew , Williamson Abby , Lueck John D. , Musgaard Maria , MacLean David M.
Published: August 2019

Acid-sensing ion channels (ASICs) are neuronal sodium-selective channels activated by reductions in extracellular pH. Structures of the three presumptive functional states, high-pH resting, low-pH desensitized, and toxin-stabilized open, have all been solved for chicken ASIC1. These structures, along with prior functional data, suggest that the isomerization or flipping of the β11-12 linker in the extracellular, ligand-binding domain is an integral component of the desensitization process. To test this, we combined fast perfusion electrophysiology, molecular dynamics simulations and state-dependent non-canonical amino acid cross-linking. We find that both desensitization and recovery can be accelerated by orders of magnitude by mutating resides in this linker or the surrounding region. Furthermore, desensitization can be suppressed by trapping the linker in the resting state, indicating that isomerization of the β11-12 linker is not merely a consequence of, but a necessity for the desensitization process in ASICs.
Comparison of Leptospira species isolated from environmental water and soil in Japan
Authors: Masuzawa Toshiyuki , Uno Rurika , Matsuhashi Nana , Yamaguchi Masaya , Xu Jun , Nakamura Shuichi
Published: August 2019

Leptospira was isolated from environmental water in central Japan using selective medium comprising five antibiotics, namely sulfamethoxazole, trimethoprim, amphotericin B, fosfomycin, and 5‐fluorouracil. Of 100 water samples 57 (57%) were culture‐positive and 50 pure cultures were isolated. Of the 50 cultures isolated from water 48 were classified into a saprophytic clade on the basis of 16S ribosomal RNA gene sequences. However, it was previously reported that isolates from soil in Japan belonged to pathogenic, intermediate, and saprophytic clades, the current findings suggest less diversity of Leptospira species in environmental water than that in soil in Japan.
Impact value of laparoscopic ultrasound over triphasic computed tomography in the management of hepatic focal lesions
Authors: Seleem Mohammed I. , Salaam Mohammed A.
Published: August 2019

Aim

Intraoperative ultrasound has an established role in hepatic resections for patients who undergo surgery for potential liver resection. The aim of the present study was to evaluate the impact of laparoscopic intraoperative ultrasonography (LUS) findings compared to pre‐operative triphasic computed tomography (TCT) findings in the management of hepatic focal lesions.

Patient and methods

Eighty‐seven patients with 192 hepatic focal lesions underwent preoperative TCT and intraoperative LUS evaluation. Data in each case were collected, including number, site and size of hepatic focal lesions.

Results

There were 112 lesions detected on preoperative TCT, and 80 additional new lesions (42 per cent) were detected in 50 patients, as confirmed by LUS. Twenty‐three lesions (12 per cent) in 20 patients were treated by resection or ablation, and correct staging of the tumours was possible in 61 patients (70 per cent).

Conclusion

The use of LUS can be considered a cornerstone for any contemplated hepatic surgery, as it corrects the staging of these lesions. In addition, the present study proved that LUS is more sensitive than TCT in detecting liver tumours, especially smaller lesions and those on the inferior and diaphragmatic surfaces of the liver.
A pace and shape perspective on fertility
Authors: Baudisch Annette , Stott Iain
Published: August 2019

Ageing is ubiquitous to all organisms, but ageing does not always mean senescence. Counter to most evolutionary theories of ageing, the patterns of mortality and reproduction may remain unchanged or improve with age, as well as deteriorate. Describing this diversity presents a challenge to eco‐evolutionary demography. The pace–shape framework of mortality tackled this challenge to qualify and quantify orthogonal components of ageing patterns in mortality. Here, we extend this framework to fertility.

Analogous to the logic of the mortality framework, we define a perspective, a framework and novel methods for the pace and shape of fertility. These distinguish between orthogonal components of time‐scale (pace) and distribution (shape) of reproduction over adult life span.

Our pace and shape framework mirrors that of mortality, through a shift of perspective from the mother giving birth, to the offspring being born. Our new measures overcome many problems associated with measuring natural fertility trajectories, have both a clear biological and mathematical interpretation, can be intuitively visualized and satisfy and extend important conditions of the pace–shape paradigm.

A comprehensive framework of fertility pace–shape facilitates ecological and evolutionary research addressing interactions and trade‐offs between components of birth and death patterns, across the whole tree of life. The burgeoning emergence of large comparative demographic data sources across wide environmental, geographical, temporal and phylogenetic ranges, combined with pace–shape measures, opens the door to comparative analyses of ageing which were never possible before.
Telomere length and aging‐related outcomes in humans: A Mendelian randomization study in 261,000 older participants
Authors: Kuo Chia‐Ling , Pilling Luke C. , Kuchel George A. , Ferrucci Luigi , Melzer David
Published: August 2019

Inherited genetic variation influencing leukocyte telomere length provides a natural experiment for testing associations with health outcomes, more robust to confounding and reverse causation than observational studies. We tested associations between genetically determined telomere length and aging‐related health outcomes in a large European ancestry older cohort. Data were from n = 379,758 UK Biobank participants aged 40–70, followed up for mean of 7.5 years (n = 261,837 participants aged 60 and older by end of follow‐up). Thirteen variants strongly associated with longer telomere length in peripheral white blood cells were analyzed using Mendelian randomization methods with Egger plots to assess pleiotropy. Variants in TERC, TERT, NAF1, OBFC1, and RTEL1 were included, and estimates were per 250 base pairs increase in telomere length, approximately equivalent to the average change over a decade in the general white population. We highlighted associations with false discovery rate‐adjusted p‐values smaller than .05. Genetically determined longer telomere length was associated with lowered risk of coronary heart disease (CHD; OR = 0.95, 95% CI: 0.92–0.98) but raised risk of cancer (OR = 1.11, 95% CI: 1.06–1.16). Little evidence for associations were found with parental lifespan, centenarian status of parents, cognitive function, grip strength, sarcopenia, or falls. The results for those aged 60 and older were similar in younger or all participants. Genetically determined telomere length was associated with increased risk of cancer and reduced risk of CHD but little change in other age‐related health outcomes. Telomere lengthening may offer little gain in later‐life health status and face increasing cancer risks.
Increase in nonnative understorey vegetation cover after nonnative conifer removal and passive restoration
Authors: Sample Martha , Aslan Clare E. , Policelli Nahuel , Sanford Robert L. , Nielsen Erik , Nuñez Martín A.
Published: August 2019

Nonnative conifers are widespread in the southern hemisphere, where their use as plantation species has led to adverse ecosystem impacts sometimes intensified by invasion. Mechanical removal is a common strategy used to reduce or eliminate the negative impacts of nonnative conifers, and encourage native regeneration. However, a variety of factors may preclude active ecological restoration following removal. As a result, passive restoration – unassisted natural vegetation regeneration – is common following conifer removal. We asked, ‘what is the response of understorey cover to removal of nonnative conifer stands followed by passive restoration?' We sampled understorey cover in three site types: two‐ to ten‐year‐old clearcuts, native forest and current plantations. We then grouped understorey species by origin (native/nonnative) and growth form, and compared proportion and per cent cover of these groups as well as of bare ground and litter between the three site types. For clearcuts, we also analysed the effect of time since clearcut on the studied variables. We found that clearcuts had a significantly higher average proportion of nonnative understorey vegetation cover than native forest sites, where nonnative vegetation was nearly absent. The understorey of clearcut sites also averaged more overall vegetation cover and more nonnative vegetation cover (in particular nonnative shrubs and herbaceous species) than either plantation or native forest sites. Notably, 99% of nonnative shrub cover in clearcuts was the invasive nonnative species Scotch broom (Cytisus scoparius). After ten years of passive recovery since clearcutting, the proportion of understorey vegetation cover that is native has not increased and remains far below the proportion observed in native forest sites. Reduced natural regeneration capacity of the native ecosystem, presence of invasive species in the surrounding landscape and land‐use legacies from plantation forestry may inhibit native vegetation recovery and benefit opportunistic invasives, limiting the effectiveness of passive restoration in this context.

Abstract in Spanish is available with online material.
Nitrated food proteins induce a regulatory immune response associated with allergy prevention after oral exposure in a Balb/c mouse food allergy model
Authors: Samadi Nazanin , Klems Martina , Heiden Denise , Bauer Renate , Kitzmüller Claudia , Weidmann Eleonore , Ret Davide , Ondracek Anna S. , Duschl Albert , Horejs‐Hoeck Jutta , Untersmayr Eva
Published: August 2019

Background

Food allergy is associated with a high personal health and economic burden. For immunomodulation toward tolerance, food compounds could be chemically modified, for example, by posttranslational protein nitration, which also occurs via diet‐derived nitrating agents in the gastrointestinal tract.

Objective

We sought to analyze the effect of pretreatment with nitrated food proteins on the immune response in a mouse food allergy model and on human monocyte‐derived dendritic cells (moDCs) and PBMCs.

Methods

The model allergen ovalbumin (OVA) was nitrated in different nitration degrees, and the secondary structures of proteins were determined by circular dichroism (CD). Allergy‐preventive treatment with OVA, nitrated OVA (nOVA), and maximally nitrated OVA (nOVAmax) were performed before mice were immunized with or without gastric acid‐suppression medication. Antibody levels, regulatory T‐cell (Treg) numbers, and cytokine levels were evaluated. Human moDCs or PBMCs were incubated with proteins and evaluated for expression of surface markers, cytokine production, and proliferation of Th2 as well as Tregs.

Results

In contrast to OVA and nOVA, the conformation of nOVAmax was substantially changed. nOVAmax pretreated mice had decreased IgE as well as IgG1 and IgG2a levels and Treg numbers were significantly elevated, while cytokine levels remained at baseline level. nOVAmax induced a regulatory DC phenotype evidenced by a decrease of the activation marker CD86 and an increase in IL‐10 production and was associated with a higher proliferation of memory Tregs.

Conclusion

Oral pretreatment with highly nitrated proteins induces a tolerogenic immune response in the food allergy model and in human immune cells.
Effects of poloxamer‐based thermo‐sensitive sol–gel agent on urethral stricture after transurethral resection of the prostate for benign prostatic hyperplasia: a multicentre, single‐blinded, randomised controlled trial
Authors: Chung Jae Hoon , Kim Kyu Shik , Choi Jae Duck , Kim Tae Hyo , Lee Ki Soo , Oh Cheol Young , Noh Joon Hwa , Kim Jun Seok , Kim Won Tae , Lee Seung Hwan , Kim Jae Heon , Kim Tae Nam , Huh Wan , Lee Seung Wook
Published: August 2019

Objective

To evaluate the effectiveness of poloxamer‐based thermo‐sensitive sol–gel instillation, after transurethral resection of the prostate (TURP), for preventing urethral stricture.

Patients and Methods

In all, 198 patients underwent TURP for benign prostatic hyperplasia. Recruited patients were randomly divided into two groups: groups A and B. Patients in Group A (100 patients, experimental group) received poloxamer‐based thermo‐sensitive sol–gel instillation and patients in the Group B (98 patients, control group) received lubricant instillation after TURP. Each patient was evaluated at 4 (V1), 12 (V2), and 24 weeks (V3) after TURP. The effectiveness of poloxamer‐based thermo‐sensitive sol–gel instillation was evaluated based on the International Prostate Symptom Score (IPSS), IPSS‐Quality of Life (QoL), Overactive bladder questionnaire (OAB‐q), maximum urinary flow rate (Qmax), post‐void residual urine volume (PVR), and cystoscopy.

Results

Amongst the initial 198 participants, 80 patients in Group A and 83 in Group B completed the study. There were no significant differences in IPSS‐QoL and OAB‐q between the groups. However, Qmax was significantly different between groups A and B, at a mean (SD) of 18.92 (9.98) vs 15.58 (9.24) mL/s (P = 0.028) at 24 weeks after TURP. On cystoscopic examination, urethral stricture after TURP was found in two of the 80 patients in Group A and 10 of 83 in Group B (P = 0.023).

Conclusions

Poloxamer‐based thermo‐sensitive sol–gel instillation after TURP lowered the incidence of urethral stricture.
Considerations for improved performance of competition association assays analysed with the Motulsky–Mahan's “kinetics of competitive binding” model
Authors: Georgi Victoria , Dubrovskiy Alexey , Steigele Stephan , Fernández‐Montalván Amaury E.
Published: August 2019

Background and Purpose

Target engagement dynamics can influence drugs' pharmacological effects. Kinetic parameters for drug:target interactions are often quantified by evaluating competition association experiments—measuring simultaneous protein binding of labelled tracers and unlabelled test compounds over time—with Motulsky–Mahan's “kinetics of competitive binding” model. Despite recent technical improvements, the current assay formats impose practical limitations to this approach. This study aims at the characterisation, understanding and prevention of these experimental constraints, and associated analytical challenges.

Experimental Approach

Monte Carlo simulations were used to run virtual kinetic and equilibrium tracer binding and competition experiments in both normal and perturbed assay conditions. Data were fitted to standard equations derived from the mass action law (including Motulsky–Mahan's) and to extended versions aiming to cope with frequently observed deviations of the canonical traces. Results were compared to assess the precision and accuracy of these models and identify experimental factors influencing their performance.

Key Results

Key factors influencing the precision and accuracy of the Motulsky–Mahan model are the interplay between compound dissociation rates, measurement time and interval frequency, tracer concentration and binding kinetics and the relative abundance of equilibrium complexes in vehicle controls. Experimental results produced recommendations for better design of tracer characterisation experiments and new strategies to deal with systematic signal decay.

Conclusions and Implications

Our data advances our comprehension of the Motulsky–Mahan kinetics of competitive binding models and provides experimental design recommendations, data analysis tools, and general guidelines for its practical application to in vitro pharmacology and drug screening.
The multifunctional peptide DN‐9 produced peripherally acting antinociception in inflammatory and neuropathic pain via μ‐ and κ‐opioid receptors
Authors: Xu Biao , Zhang Mengna , Shi Xuerui , Zhang Run , Chen Dan , Chen Yong , Wang Zilong , Qiu Yu , Zhang Ting , Xu Kangtai , Zhang Xiaoyu , Liedtke Wolfgang , Wang Rui , Fang Quan
Published: August 2019

Background and Purpose

Considerable effort has recently been directed at developing multifunctional opioid drugs to minimize the unwanted side effects of opioid analgesics. We have developed a novel multifunctional opioid agonist, DN‐9. Here, we studied the analgesic profiles and related side effects of peripheral DN‐9 in various pain models.

Experimental Approach

Antinociceptive effects of DN‐9 were assessed in nociceptive, inflammatory, and neuropathic pain. Whole‐cell patch‐clamp and calcium imaging assays were used to evaluate the inhibitory effects of DN‐9 to calcium current and high‐K+‐induced intracellular calcium ([Ca2+]i) on dorsal root ganglion (DRG) neurons respectively. Side effects of DN‐9 were evaluated in antinociceptive tolerance, abuse, gastrointestinal transit, and rotarod tests.

Key Results

DN‐9, given subcutaneously, dose‐dependently produced antinociception via peripheral opioid receptors in different pain models without sex difference. In addition, DN‐9 exhibited more potent ability than morphine to inhibit calcium current and high‐K+‐induced [Ca2+]i in DRG neurons. Repeated treatment with DN‐9 produced equivalent antinociception for 8 days in multiple pain models, and DN‐9 also maintained potent analgesia in morphine‐tolerant mice. Furthermore, chronic DN‐9 administration had no apparent effect on the microglial activation of spinal cord. After subcutaneous injection, DN‐9 exhibited less abuse potential than morphine, as was gastroparesis and effects on motor coordination.

Conclusions and Implications

DN‐9 produces potent analgesia with minimal side effects, which strengthen the candidacy of peripherally acting opioids with multifunctional agonistic properties to enter human studies to alleviate the current highly problematic misuse of classic opioids on a large scale.
Structural evolution and the partitioning of deformation during basin growth and inversion: A case study from the Mizen Basin Celtic Sea, offshore Ireland
Authors: Rodríguez‐Salgado Pablo , Childs Conrad , Shannon Patrick M. , Walsh John J.
Published: August 2019

The Celtic Sea basins lie on the continental shelf between Ireland and northwest France and consist of a series of ENE–WSW trending elongate basins that extend from St George’s Channel Basin in the east to the Fastnet Basin in the west. The basins, which contain Triassic to Neogene stratigraphic sequences, evolved through a complex geological history that includes multiple Mesozoic rift stages and later Cenozoic inversion. The Mizen Basin represents the NW termination of the Celtic Sea basins and consists of two NE–SW‐trending half‐grabens developed as a result of the reactivation of Palaeozoic (Caledonian, Lower Carboniferous and Variscan) faults. The faults bounding the Mizen Basin were active as normal faults from Early Triassic to Late Cretaceous times. Most of the fault displacement took place during Berriasian to Hauterivian (Early Cretaceous) times, with a NW–SE direction of extension. A later phase of Aptian to Cenomanian (Early to Late Cretaceous) N–S‐oriented extension gave rise to E–W‐striking minor normal faults and reactivation of the pre‐existing basin bounding faults that propagated upwards as left‐stepping arrays of segmented normal faults. In common with most of the Celtic Sea basins, the Mizen Basin experienced a period of major erosion, attributed to tectonic uplift, during the Paleocene. Approximately N–S Alpine regional compression‐causing basin inversion is dated as Middle Eocene to Miocene by a well‐preserved syn‐inversion stratigraphy. Reverse reactivation of the basin bounding faults was broadly synchronous with the formation of a set of near‐orthogonal NW–SE dextral strike‐slip faults so that compression was partitioned onto two fault sets, the geometrical configuration of which is partly inherited from Palaeozoic basement structure. The segmented character of the fault forming the southern boundary of the Mizen Basin was preserved during Alpine inversion so that Cenozoic reverse displacement distribution on syn‐inversion horizons mirrors the earlier extensional displacements. Segmentation of normal faults therefore controls the geometry and location of inversion structures, including inversion anticlines and the back rotation of earlier relay ramps.
Serum lipidomics for exploring biomarkers of bortezomib therapy in patients with multiple myeloma
Authors: Maekawa Keiko , Ri Masaki , Nakajima Miki , Sekine Akihiro , Ueda Ryuzo , Tohkin Masahiro , Miyata Naoki , Saito Yoshiro , Iida Shinsuke
Published: August 2019

Although the proteasome inhibitor bortezomib (BTZ) shows excellent efficacy in multiple myeloma (MM), a fraction of patients has a suboptimal or no response to this agent. In addition, BTZ‐induced peripheral neuropathy (BiPN), a frequent side‐effect of this therapy, limits its use in some patients. This study aimed to explore serum lipid biomarker candidates to predict the response to BTZ and the severity of BiPN. Fifty‐nine serum samples were collected from patients with MM prior to receiving BTZ plus low‐dose dexamethasone therapy. Serum levels of phospholipids, sphingolipids, neutral lipids, and polyunsaturated fatty acids and their oxidation products were measured by a comprehensive lipidomic study. Overall, 385 lipid metabolites were identified in patients’ sera; lower levels of several glycerophospholipids, sphingolipids, and cholesteryl esters were associated with a poor treatment response. Metabolites related to platelet‐activating factor biosynthesis and cholesterol metabolism appeared particularly relevant. Furthermore, several lysophosphatidylcholines, phosphatidylcholines, ceramides, neutral lipids, and oxidative fatty acids were significantly increased or decreased in patients with BiPN grades ranging from G0 to G3. Among these compounds, mediators reportedly inducing myelin breakdown and stimulating inflammatory responses were prominent. Although further study is necessary to validate these biomarker candidates, our results contribute to the development of predictive biomarkers for response to BTZ treatment, or ensuing severe BiPN, in patients with MM.
Triterpenes from Poria cocos are revealed as potential retinoid X receptor selective agonists based on cell and in silico evidence
Authors: Xu Hui , Wang Yuchen , Zhao Junnan , Jurutka Peter W. , Huang Dechun , Liu Liangyun , Zhang Lange , Wang Suilou , Chen Yadong , Cheng Shujie
Published: August 2019

Poria cocos is an edible and medicinal fungus that is widely used in Traditional Chinese Medicines as well as in modern applications. Retinoid X receptor (RXR) occupies a central place in nuclear receptor signaling, and a pharmacological RXR‐dependent pathway is involved in myeloid cell function. Here, structural information for 82 triterpenes from P. cocos and 17 known RXR agonists was collected in a compound library and retrieved for a molecular docking study. Three triterpenes, 16α‐hydroxytrametenolic acid (HTA), pachymic acid (PA), and polyporenic acid C (PPAC), were identified as novel RXR‐specific agonists based on luciferase reporter assays and in silico evidence. Treatment with HTA, PA, and PPAC significantly induced differentiation of the human promyelocytic leukemia cell line HL‐60 with EC50 values of 21.0 ± 0.52, 6.7 ± 0.37, and 9.4 ± 0.65 μM, respectively. These effects were partly blocked by the RXR antagonist UVI3003, suggesting that an RXR‐dependent pathway may play an important role in their anti‐acute promyelocytic leukemia (APL) effects. Taken together, triterpenes from P. cocos are revealed as naturally occurring RXR selective agonists with the potential for anti‐cancer activity. These results suggest a novel approach to the treatment or prevention of APL.
Discovery of small molecule agonists targeting neuropeptide Y4 receptor using homology modeling and virtual screening
Authors: Kang Ning , Wang Xiao‐Lei , Zhao Yaxue
Published: August 2019

Neuropeptide Y4 receptor has the most significant effect on body weight and fat mass in its physiological functions, and the activation of Y4 receptor has explicit role on losing weight. The Y4 receptor has been successfully applied in the development of anti‐obesity agent, thus representing a potential therapeutic target for obesity treatment. Here, we reported the first discovery of small molecule agonists targeting Y4 receptor: three Y4 receptor models with active and inactive conformations were built, each model was submitted following structure‐based virtual screening, and finally six hits were identified as Y4 receptor agonists. These results confirm the reliability of the constructed Y4 receptor models and the proposed computational strategy for investigating novel Y4 receptor agonists. These new small molecule Y4 receptor agonists will contribute to the further development of Y4 agonists as potential therapeutics and functional probes.
Prospective randomized controlled clinical study comparing two types of two‐piece dental implants supporting fixed reconstructions—Results at 5 years of loading
Authors: Ioannidis Alexis , Heierle Linda , Hämmerle Christoph H. F. , Hüsler Jürg , Jung Ronald E. , Thoma Daniel S.
Published: August 2019

Objectives

Compare clinical outcomes of two types of dental implants with non‐matching implant–abutment junctions loaded with fixed implant‐borne reconstructions at 5 years of loading.

Materials and methods

In 64 patients, one of two implant systems (S1, S2) was randomly used to support fixed dental prostheses (FDP). At loading (TL), after 1 (T1), 3 (T3) and 5 years (T5), the implant and reconstruction survival, biological and technical complications, radiographic marginal bone levels, clinical outcomes were recorded. The implants of the groups S1 and S2 contained of two‐piece titanium implants with a non‐matching implant–abutment junction. Data were analyzed on the patient level (1 implant/patient) using the non‐parametric Wilcoxon–Mann–Whitney test.

Results

Ninety‐seven implants were placed and loaded with fixed reconstructions in 64 patients. At T5, 29 S1 and 28 S2 implants were available for the patient‐level analysis. Two implants in group S1 had to be removed in the same patient due to severe peri‐implantitis, resulting in a survival rate of 96.1% on the implant level and 96.6% on the patient level. No implant in group S2 was lost. The technical complication rate on patient‐level amounted of 24.2% (S1) and of 6.5% (S2) (p > .05). Biological complications on patient‐level were observed in 15.2% (S1) and 16.1% (S2) (p > .05). From TL to T5, the medians of the mean marginal bone level changes wer a gain of 0.15 mm in group S1 and a loss of 0.14 mm in group S2 (p > .05).

Conclusions and clinical implications

Both implant systems revealed high survival rates and minimal changes of the marginal bone levels during 5 years. Few biological complications occurred in both groups. S1 revealed a high rate of technical complications. Therefore, both implant systems can be recommended for fixed reconstructions.
The up‐to‐11‐year survival and success of implants and abutment teeth under solely implant‐supported and combined tooth–implant‐supported double crown‐retained removable dentures
Authors: Fobbe Hannah , Rammelsberg Peter , Lorenzo Bermejo Justo , Kappel Stefanie
Published: August 2019

Objective

This retrospective analysis was designed to compare the survival and success of dental implants in solely implant‐supported, double crown‐retained removable dentures (DCRDs) and combined tooth–implant‐supported DCRDs, due to a lack of data on the latter.

Material and methods

From 2002, patients were consecutively admitted for treatment at the Department of Prosthodontics, University of Heidelberg. Surgical and prosthodontic treatment and scheduled and unscheduled visits were recorded on standardized documentation forms. Kaplan–Meier analysis was used to estimate success (survival without major complications). Cox regression was applied to assess risk factors.

Results

From this clinical study, 139 DCRDs on 412 implants and 239 teeth met the inclusion criteria. Fifty‐three dentures were implant‐supported (213 implants). Eighty‐six were supported by teeth and implants. The DCRDs were placed in 126 patients. After an observation period of up to 11.2 years (median: 4.2 years), total implant survival was 99.5% for tooth–implant‐supported DCRDs and 93.4% for implant‐supported ones. The estimated cumulative success was 97.2% (standard error/SE ± 1.2%) and 85.9% (SE ± 2.4%) at five years, respectively. Complications included implant or tooth loss, peri‐implantitis, apical periodontitis and tooth fracture. In terms of abutments, results from multiple Cox regression revealed lower failure rates in tooth–implant‐supported dentures (p = .04). No other risk factors were identified.

Conclusions

Within the limitations of this study, survival and success are high for both treatment options. Combining teeth and implants in one DCRD might have a positive effect on the prognosis of the implant and the survival of remaining teeth.
Daratumumab added to standard of care in patients with newly diagnosed multiple myeloma: A network meta‐analysis
Authors: Xu Wenjun , Li DianFang , Sun Yanhua , Ran Xuehong , Wang Baohong , Wu Wei , Sheng Zhixin , Liu Liping
Published: August 2019

Purpose

To investigate the activity and safety of daratumumab added to standard of care and evaluate the relative efficacy of DRd vs DVMP and other regimens on survival endpoints for untreated myeloma, we undertook this meta‐analysis.

Methods

We searched published reports that described the activity and safety of daratumumab added to standard of care for untreated myeloma.

Results

Six daratumumab trials were identified, covering 5106 subjects. Daratumumab containing combinations for untreated myeloma attained an impressive complete response or better (≥CR) rate of 24%, very good partial response or better (≥VGPR) rate of 67%, overall response rate (ORR) of 92%. Daratumumab added to standard of care significantly improved progression free survival (PFS): the HR for PFS was 0.52 [0.44, 0.61], P < .001. The HR for overall survival (OS) was 0.73 [0.52, 1.04], P = .09. In the network meta‐analysis for patients ineligible for autologous stem‐cell transplantation (ASCT), DRd regimen produced significant PFS advantage vs other first‐line treatments (VMP HR:0.39 P < .001, Rd HR:0.55 P < .001, MPT HR:0.38 P < .001, and MP HR:0.22 P < .001); DVMP regimen also produced significant PFS advantage vs VMP (HR:0.50 P < .001), MPT (HR:0.49 P < .001), and MP (HR:0.28 P < .001). Among these first‐line regimens (DRd, DVMP, VMP, Rd, MPT, and MP), DRd regimen had the highest probability to be the best intervention, with 83.4% and 91.0% probability to reach the longest PFS and OS, respectively. Toxicity consisted primarily of myelosuppression. And, the vital non‐hematologic adverse events (AEs) were peripheral sensory neuropathy (41% of all grades) and upper respiratory tract infection (39% of all grades).

Conclusions

Daratumumab added to standard of care could produce clinical benefits in newly diagnosed patients with multiple myeloma. DRd and DVMP could be good combination options for those patients ineligible for ASCT.
Carbon storage in a rainfed Mediterranean vertisol: Effects of tillage and crop rotation in a long‐term experiment
Authors: López‐Bellido Luis , López‐Bellido Rafael , Fernández‐García Purificación , Muñoz‐Romero Verónica , Lopez‐Bellido Francisco Javier
Published: August 2019

The storage of carbon (C) in cultivated soils can be increased with the adoption of different practices. The objective of this study was to determine soil organic carbon (SOC) storage in the 0‐ to 90‐cm depth profile, in four different soil layers (0–15, 15–30, 30–60 and 60–90 cm) in a long‐term (29 years) experiment established in 1986 on a rainfed Mediterranean Vertisol in southern Spain. The treatments studied were: conventional tillage (CT) versus no‐tillage (NT); five 2‐year crop rotations (wheat–chickpea, wheat–sunflower, wheat–bare fallow, wheat–faba bean, and continuous wheat; and nitrogen (N) fertilizer applied to wheat at four rates (0, 50, 100 and 150 kg N ha−1). The SOC accumulation was higher in the 30–60‐cm layer (9.2 Mg ha−1) due to the size of the characteristic cracks of Vertisols under semiarid conditions. Over the 29‐year study period, the SOC in the 0–90‐cm layer increased by 23.6 Mg ha−1 due to the change in residue management. The NT treatment exhibited a higher mean annual rate of organic C accumulation compared with the CT treatment (1.0 and 0.66 Mg ha−1 year−1, respectively) due to the retention of the mulched residue. Additionally, crop rotation influenced the rate of organic C accumulation, with wheat‐faba bean, wheat‐sunflower and continuous wheat exhibiting the highest levels of C storage in comparison to the other treatments. In rainfed Mediterranean agriculture, the selection of no‐tillage along with a rotation with legumes is key to improving soil fertility and increasing C reserves and the rate of C accumulation by soil.

Highlights

The SOC increase over the 29‐year study period was 23.6 Mg ha−1 in the 0–90‐cm soil profile.

The characteristic cracks of Vertisol increase SOC stocks in deeper soil layers.

No tillage accumulated 66.2% more SOC than conventional tillage in the 0–90‐cm profile.

The wheat–sunflower rotation had the highest annual rate of C sequestration (1 Mg ha−1 year−1) and wheat–chickpea the lowest (0.6 Mg ha−1 year−1) over 29 years.
Haplotype analysis encompassing HTT gene in Chinese patients with Huntington's disease
Authors: Li X.‐Y. , Li H.‐L. , Dong Y. , Gao B. , Cheng H.‐R. , Ni W. , Gan S.‐R. , Liu Z.‐J. , Burgunder J.‐M. , Wu Z.‐Y.
Published: August 2019

Background and purpose

Huntington's disease (HD) is a dominantly inherited neurodegenerative disorder with varied prevalence in different populations, which may be associated with specific haplotypes. This study aimed to explore the haplotypes encompassing the HTT gene in the Chinese population.

Methods

A total of 406 individuals with HD and 59 normal relatives from 253 families with HD were enrolled. A total of 29 tag single nucleotide polymorphisms (tSNPs) were selected and genotyped for the haplotype analysis.

Results

In stage one, we used 18 tSNPs to replicate the distribution of three major haplogroups (A, B, C). We found that risk‐associated haplogroup variants A1 and A2, enriched on Caucasian HD chromosomes, were totally absent from both Chinese HD and control chromosomes, and the distributions of haplogroups between HD and control chromosomes were similar. Therefore, in stage two, we used 29 tSNPs (including the18 tSNPs) to define new haplogroups (I, II, III) and found that haplogroup I accounted for 61.4% on HD chromosomes and 34.4% on control chromosomes, indicating that haplogroup I was enriched on Chinese HD chromosomes.

Conclusions

This is the first haplotype analysis encompassing HTT in the Chinese population. The results contribute to explaining the low prevalence of HD in China and provide a better understanding of genetic diversity in the HTT region.