bluuebunny/crossref_metadata_2025 huggingface dataset:
Urethral‐fixation technique improves early urinary continence recovery in patients who undergo retropubic radical prostatectomy
Authors: Ficarra Vincenzo , Crestani Alessandro , Rossanese Marta , Palumbo Vito , Calandriello Mattia , Novara Giacomo , Praturlon Silvio , Giannarini Gianluca
Published: April 2016

Objectives

To describe step‐by‐step an original urethrovesical anastomosis technique (urethral fixation) in patients undergoing retropubic radical prostatectomy (RRP), to compare their early urinary continence recovery with those in a control group receiving a standard anastomosis technique and to identify the predictors of early urinary continence recovery.

Patients and methods

We compared 70 patients who underwent RRP with the urethral‐fixation technique with a contemporary control group of 51 patients who received RRP with a standard urethrovesical anastomosis. In the urethral‐fixation group, the urethrovesical anastomosis was made using eight single sutures. Specifically, to avoid retraction and/or deviations, we fixed the urethral stump laterally to the medial portion of levator ani muscle. Also, to maintain the normal position in the context of the pelvic floor, we fixed the urethral sphincter deeper to the medial dorsal raphe using a 3‐0 polydioxanone suture at the 6 o'clock position before completing the incision of the urethral wall. Urinary continence recovery was evaluated at 1, 4, 8 and 12 weeks after catheter removal. Patients self‐reporting no urine leak were considered continent. Uni‐ and multivariable analyses were used to identify predictors of urinary incontinence at the different follow‐up time‐points.

Results

The evaluated groups had comparable preoperative variables. At 1 week after catheter removal, 32 (45.7%) patients in the urethral‐fixation group and 10 (19.6%) in the control group were continent (P = 0.01). At 4 weeks after catheter removal, 46 (65.7%) patients in the urethral‐fixation group and 16 (31.4%) in the control group were continent (P = 0.001). At 8 weeks after catheter removal, 59 (84.3%) patients in the urethral‐fixation group and 21 (41.2%) in the control group were continent (P < 0.001). Finally, at 12 weeks after catheter removal, 63 (90%) patients in the urethral‐fixation group and 32 (62.7%) in the control group were continent (P = 0.001). The urethral‐fixation technique was an independent predictor of urinary continence recovery at 1 week [odds ratio (OR) 4.305; P = 0.002); 4 weeks (OR 4.784; P < 0.001); 8 weeks (OR 7.678; P < 0.001) and 12 weeks (OR 5.152; P = 0.001) after catheter removal.

Conclusions

The urethral‐fixation technique significantly improves early urinary continence recovery in comparison with the standard technique. Moreover, our study confirmed that this surgical technique is an independent predictor of urinary continence recovery at 1, 4, 8 and 12 weeks after catheter removal.
Recent advances in immuno‐oncology and its application to urological cancers
Authors: Mataraza Jennifer M. , Gotwals Philip
Published: April 2016

Recent advances in immuno‐oncology have the potential to transform the practice of medical oncology. Antibodies directed against negative regulators of T‐cell function (checkpoint inhibitors), engineered cell therapies and innate immune stimulators, such as oncolytic viruses, are effective in a wide range of cancers. Immune‘based therapies have had a clinically meaningful impact on the treatment of advanced melanoma, and the lessons regarding use of single agents and combinations in melanoma may be applicable to the treatment of urological cancers. Checkpoint inhibitors, cytokine therapy and therapeutic vaccines are already showing promise in urothelial bladder cancer, renal cell carcinoma and prostate cancer. Critical areas of future immuno‐oncology research include the prospective identification of patients who will respond to current immune‐based cancer therapies and the identification of new therapeutic agents that promote immune priming in tumours, and increase the rate of durable clinical responses.
Multiyear dual nitrate isotope signatures suggest that N‐saturated subtropical forested catchments can act as robust N sinks
Authors: Yu Longfei , Zhu Jing , Mulder Jan , Dörsch Peter
Published: April 2016

In forests of the humid subtropics of China, chronically elevated nitrogen (N) deposition, predominantly as ammonium (NH4+), causes significant nitrate (NO3−) leaching from well‐drained acid forest soils on hill slopes (HS), whereas significant retention of NO3− occurs in near‐stream environments (groundwater discharge zones, GDZ). To aid our understanding of N transformations on the catchment level, we studied spatial and temporal variabilities of concentration and natural abundance (δ15N and δ18O) of nitrate (NO3−) in soil pore water along a hydrological continuum in the N‐saturated Tieshanping (TSP) catchment, southwest China. Our data show that effective removal of atmogenic NH4+ and production of NO3− in soils on HS were associated with a significant decrease in δ15N‐NO3−, suggesting efficient nitrification despite low soil pH. The concentration of NO3− declined sharply along the hydrological flow path in the GDZ. This decline was associated with a significant increase in both δ15N and δ18O of residual NO3−, providing evidence that the GDZ acts as an N sink due to denitrification. The observed apparent 15N enrichment factor (ε) of NO3− of about −5‰ in the GDZ is similar to values previously reported for efficient denitrification in riparian and groundwater systems. Episode studies in the summers of 2009, 2010 and 2013 revealed that the spatial pattern of δ15N and δ18O‐NO3− in soil water was remarkably similar from year to year. The importance of denitrification as a major N sink was also seen at the catchment scale, as largest δ15N‐NO3− values in stream water were observed at lowest discharge, confirming the importance of the relatively small GDZ for N removal under base flow conditions. This study, explicitly recognizing hydrologically connected landscape elements, reveals an overlooked but robust N sink in N‐saturated, subtropical forests with important implications for regional N budgets.
Mineral chemistry of a Cenozoic igneous complex, the Urumieh–Dokhtar magmatic belt, Iran: Petrological implications for the plutonic rocks
Authors: Honarmand Maryam , Omran Nematollah Rashidnejad , Neubauer Franz , Nabatian Ghasem , Emami Mohammad Hashem , Bernroider Manfred , Ahmadian Jamshid , Ebrahimi Mohammad , Liu Xiaoming
Published: April 2016

The Niyasar plutonic complex, one of the Cenozoic magmatic assemblages in the Urumieh‐Dokhtar magmatic belt, was the subject of detailed petrographic and mineralogical investigations. The Niyasar magmatic complex is composed of Eocene to Oligocene mafic rocks and Miocene granitoids. Eleven samples, representing the major rock units in the Niyasar magmatic complex and contact aureole were chosen for mineral chemical studies and for estimation of the pressure, temperature, and oxygen fugacity conditions of mineral crystallization during emplacement of various magmatic bodies. The analyzed samples are composed of varying proportions of quartz, plagioclase, K‐feldspar, hornblende, biotite, titanite, magnetite, apatite, zircon, garnet, and clinopyroxene. Application of the Al‐in‐hornblende barometer indicates pressures of around 0.2 to 0.4 kbar for the Eocene–Oligocene mafic bodies and around 0.5 to 1.7 kbar for the Miocene granitoids. Hornblende‐plagioclase thermometry yields relatively low temperatures (661–780 °C), which probably reflect late stage re‐equilibration of these minerals. The assemblage titanite–magnetite–quartz as well as hornblende composition were used to constrain the oxygen fugacity and H2O content during the crystallization of the parent magmas in the Miocene plutons. The results show that the Miocene granitoids crystallized from magmas with relatively high oxygen fugacity and high H2O content (~5 wt% H2O). The Miocene granitoids show similar range of oxygen fugacity, H2O contents and mineral chemical compositions, which indicate a common source for their magmas. Although the crystallization pressures of the Miocene plutons discriminate various categories of plutonic bodies emplaced at depths of about 5.7–6.5 km (Marfioun pluton), about 4.2 km (Ghalhar pluton) and 1.9–2.3 km (Poudalg pluton), they were later uplifted to the same level by vertical displacement of faults. The emplacement depths of the Niyasar plutons suggest that the central part of the Urumieh‐Dokhtar magmatic belt has experienced an uplift rate of ca. 0.25–0.4 mm/yr from the Miocene onwards.
Effect of Different Firing Temperatures on Structural Changes in Porcelain
Authors: Tripathi Arvind , Bagchi Soumyojeet , Singh Juhi , Gaurav Vivek , Pal Singh Negi Mahendra
Published: April 2016

Purpose

To study the structural changes occurring in the dental porcelain mass fired at various firing temperatures using scanning electron microscopy (SEM) and X‐ray diffraction (XRD) spectroscopy. Also, additional tests, namely compressive strength, abrasion resistance analysis, and the amount of oxides released, were conducted at different firing temperatures.

Materials and Methods

Six groups (40 specimens in each group) of porcelain mass were prepared. The dimensions and weight of all the specimens were kept constant. The specimens were then heat treated at different firing temperatures (660°C, 760°C, 860°C, 900°C, 960°C, 990°C). Half of the specimens of each group were subjected to a compressive strength test on a universal testing machine and then finely ground using an electrochemical grinder to prepare for XRD analysis. The other half of the specimens was weighed to analyze the amount of oxides released after each firing cycle. Following this, the specimens underwent an abrasion resistance test on a Nanovea Tribometer. The unaltered surface was scanned using SEM. The data (numerical and graphical) for all the tests were recorded and analyzed using one‐way ANOVA and post hoc Tukey test.

Results

The specimens fired at 900°C exhibited superior compressive strength and abrasion resistance. The quantity of oxides released by the specimens fired at 900°C was the least compared to specimens heat treated at the other firing temperatures. XRD analysis proved that the oxide released by the porcelain mass was calcium aluminum chromium oxide. Also, the fewer peaks obtained in the XRD graphs of specimens fired at 900°C signified lesser porosities in the porcelain specimens. SEM analysis depicted a homogeneous mass of porcelain at 900°C.

Conclusion

All the above findings validate the objective of studying the physical and internal structural changes of dental porcelain when subjected to an increasing firing temperature gradient. The specimens fired at 900°C exhibited superior strength and abrasion resistance. SEM analysis depicted a homogeneous mass of dental porcelain, implying that firing was complete at 900°C.
Conversion of human fibroblasts into functional cardiomyocytes by small molecules
Authors: Cao Nan , Huang Yu , Zheng Jiashun , Spencer C. Ian , Zhang Yu , Fu Ji-Dong , Nie Baoming , Xie Min , Zhang Mingliang , Wang Haixia , Ma Tianhua , Xu Tao , Shi Guilai , Srivastava Deepak , Ding Sheng
Published: April 2016

Making cardiac cells from fibroblasts

Reprogramming noncardiac cells into functional cardiomyocytes without any genetic manipulation could open up new avenues for cardiac regenerative therapies. Cao
et al.
identified a combination of nine small molecules that could epigenetically activate human fibroblasts, efficiently reprogramming them into chemically induced cardiomyocytes (ciCMs). The ciCMs contracted uniformly and resembled human cardiomyocytes. This method may be adapted for reprogramming multiple cell types and have important implications in regenerative medicine.

Science
, this issue p.
1216
NAD
+
repletion improves mitochondrial and stem cell function and enhances life span in mice
Authors: Zhang Hongbo , Ryu Dongryeol , Wu Yibo , Gariani Karim , Wang Xu , Luan Peiling , D’Amico Davide , Ropelle Eduardo R. , Lutolf Matthias P. , Aebersold Ruedi , Schoonjans Kristina , Menzies Keir J. , Auwerx Johan
Published: April 2016

A dietary supplement protects aging muscle

The oxidized form of cellular nicotinamide adenine dinucleotide (NAD
+
) is critical for mitochondrial function, and its supplementation can lead to increased longevity. Zhang
et al.
found that feeding the NAD
+
precursor nicotinamide riboside (NR) to aging mice protected them from muscle degeneration (see the Perspective by Guarente). NR treatment enhanced muscle function and also protected mice from the loss of muscle stem cells. The treatment was similarly protective of neural and melanocyte stem cells, which may have contributed to the extended life span of the NR-treated animals.

Science
, this issue p.
1436
; see also p.
1396
Draft Genome Sequence of
Microdochium bolleyi
, a Dark Septate Fungal Endophyte of Beach Grass
Authors: David Aaron S. , Haridas Sajeet , LaButti Kurt , Lim Joanne , Lipzen Anna , Wang Mei , Barry Kerrie , Grigoriev Igor V. , Spatafora Joseph W. , May Georgiana
Published: April 2016

Here, we present the genome sequence of the dark septate fungal endophyte
Microdochium bolleyi
(
Ascomycota, Sordariomycetes, Xylariales
). The assembled genome size was 38.84 Mbp and consisted of 173 scaffolds and 13,177 predicted genes.
Complete Genome Sequence of Middle East Respiratory Syndrome Coronavirus Isolated from a Dromedary Camel in Egypt
Authors: Kandeil Ahmed , Shehata Mahmoud M. , El Shesheny Rabeh , Gomaa Mokhtar R. , Ali Mohamed A. , Kayali Ghazi
Published: April 2016

We generated the near-full genome sequence of Middle East respiratory syndrome coronavirus (MERS-CoV) from a collected nasal sample of dromedary camel in Egypt. The newly characterized Egyptian strain has high similarity to the previously characterized Egyptian virus and both of viruses fell into a cluster distinct from other MERS-CoVs.
C. N. Yang on Teaching and Research in Physics
Authors: Shi Yu , Waxman David
Published: April 2016

This document is based on five conversations between Prof. C. N. Yang and others in Beijing in 1986. In the conversations, Yang gave his views on the state and development of physics at that time, and the relationship between physics and philosophy. The conversations also contain Yang’s reminiscences on the creation of Yang–Mills theory and his advice to young people, especially those in China.
Light Higgs bosons in NMSSM at the LHC
Authors: Guchait Monoranjan , Kumar Jacky
Published: April 2016

The next-to-minimal supersymmetric Standard Model (NMSSM) with an extended Higgs sector offers at least one Higgs boson as the Standard Model (SM)-like Higgs with a mass around 125 GeV. In this work, we revisit the mass spectrum and couplings of non-SM-like Higgs bosons taking into consideration most relevant constraints and identify the relevant parameter space. The discovery potential of these non-SM-like Higgs bosons, apart from their masses, is guided by their couplings with gauge bosons and fermions which are very much parameter space sensitive. We evaluate the rates of productions of these non-SM-like Higgs bosons at the LHC for a variety of decay channels in the allowed region of the parameter space. Although [Formula: see text], [Formula: see text] modes appear to be the most promising decay channels, but for a substantial region of parameter space the two-photon decay mode has a remarkably large rate. In this study we emphasize that this diphoton mode can be exploited to find the non-SM-like Higgs bosons of the NMSSM and can also be a potential avenue to distinguish the NMSSM from the MSSM. In addition, we discuss briefly the various detectable signals of these non-SM Higgs bosons at the LHC.
Stochastic resonance in two-frequency signal systems
Authors: Song Hui , Huang Shengming , Liu Xuemei
Published: April 2016

Signal-to-noise ratio (SNR) in two-frequency signal systems is investigated in the presence of the correlated multiplicative and white additive noises. From the Wiener–Khinchin theorem, the exact expression of the SNR is obtained. It is found that stochastic resonance (SR) appears when two frequencies are almost the same, while SR gradually disappears when the frequency difference increases. We also found that there exists an optimized value of the frequency at which SNR takes its maximal value.
One-pot room temperature synthesizing Cu- and Mn-doped ZnSe nanocrystals by a rapid photochemical method
Authors: Bahador A. R. , Molaei M. , Karimipour M.
Published: April 2016

In this work, a one-pot, rapid, green and room temperature photochemical synthesis of transition metal (TM; Cu, Mn)-doped ZnSe nanocrystals (NCs) was reported. NCs were successfully characterized using Fourier transform-infrared (FT-IR), photoluminescence (PL) and UV-Visible (UV-Vis) spectroscopy, transmission electron microscopy (TEM), X-ray diffractometry (XRD) and energy dispersive X-ray spectra (EDX). FT-IR spectra confirmed the capping of ZnSe by thioglycolic acid (TGA) molecules. XRD and TEM analysis demonstrated zinc blend phase NCs with an average size of around 3 nm. Band gap of ZnSe NCs was about 3.6 eV which it was decreased by increasing the illumination time. PL spectra of ZnSe NCs showed a broad emission with two peaks located at 380 nm and 490 nm related to excitonic and trap states emission, respectively. For ZnSe:Cu NCs, excitonic emission disappeared completely and PL intensity of trap states emission increased with the increase in the Cu[Formula: see text] ion concentration so that for precursor ratio of Cu:Zn 1%, optimal value of PL intensity was obtained. For ZnSe:Mn NCs, the excitonic emission decreased gradually with the increase in the impurity concentration whereas trap state emission increased. Moreover, a peak about 590 nm was appeared from 4T1-6A1 transition of the Mn[Formula: see text] impurity, demonstrating the Mn incorporation inside the ZnSe NCs structure.
A Stabilizing Optimal ℓ-Exclusion Algorithm
Authors: Karaata Mehmet Hakan , Hadid Rachid
Published: April 2016

In this paper, we present a simple permission-based fair stabilizing solution to the [Formula: see text]-exclusion problem in tree networks. The [Formula: see text]-exclusion problem is a generalization of the mutual exclusion problem where [Formula: see text] processes, instead of 1, are allowed to use a shared resource (enter the critical section) simultaneously. The proposed algorithm is optimal in terms of waiting times of processes to enter critical sections, i.e., between two entries of a process to its critical section, no other process can enter its critical section more than once after stabilization. Since our algorithm is stabilizing, it does not require initialization and withstands transient faults. The stabilization time of the algorithm is [Formula: see text] rounds and the waiting time is [Formula: see text], where h and n are the height and the size of the tree, respectively. In addition, this algorithm satisfies all the requirements of the [Formula: see text]-exclusion problem: [Formula: see text] and liveness.
Compressive Sensing of Multichannel Electrocardiogram Signals in Wireless Telehealth System
Authors: Hua Jing , Zhang Hua , Liu Jizhong , Zhou Junlong
Published: April 2016

Due to the capacity of compressing and recovering signal with low energy consumption, compressive sensing (CS) has drawn considerable attention in wireless telemonitoring of electrocardiogram (ECG) signals. However, most existing CS methods are designed for reconstructing single channel signal, and hence difficult to reconstruct multichannel ECG signals. In this paper, a spatio-temporal sparse model-based algorithm is proposed for the reconstruction of multichannel ECG signals by not only exploiting the temporal correlation in each individual channel signal, but also the spatial correlation among signals from different channels. In addition, a dictionary learning (DL) approach is developed to enhance the performance of the proposed reconstruction algorithm by using the sparsity of ECG signals in some transformed domain. The approach determines a dictionary by learning local dictionaries for each channel and merging them to form a global dictionary. Extensive simulations were performed to validate the proposed algorithms. Simulation results show that the proposed reconstruction algorithm has a better performance in recovering multichannel ECG signals as compared to the benchmarking methods. Moreover, the reconstruction performance of the algorithm can be further improved by using a dictionary matrix, which is obtained from the proposed DL algorithm.
Damage Assessment of Two-Way RC Slab Subjected to Blast Load Using Mode Approximation Approach
Authors: Huang Xin , Bao Huirong , Hao Yifei , Hao Hong
Published: April 2016

Significant research efforts have been invested on studying the response and damage of structures subjected to blast loads for better life and property protections. The single-degree-of-freedom (SDOF) approach has been widely adopted to simplify the structural response analysis for engineering design purpose. However, such an approach under certain circumstances oversimplifies the structural behavior and might not give reliable predictions of structural responses to blast loads. On the other hand, although detailed high fidelity finite element (FE) approach is able to give relatively accurate predictions of structural response, it is unfortunately not straightforward for application and very time-consuming, which impedes its application among engineers. Therefore, a method that can assure not only reliability but also efficiency is highly needed for design practice. In the present study, mode approximation method with Pressure–Impulse (P-I) diagrams is applied to analyze response and damage of RC slab due to blast load. Slab under analysis is assumed rigid-plastic and simply supported. Shear failure, bending failure and combined failure modes are considered based on different failure modes. Critical equations for structural shear and bending failures are derived respectively with appropriate failure criteria. P–I diagrams are then developed for quick damage assessments. The analytical results are verified by comparing with high fidelity numerical simulations. The reliability and efficiency of using this approach for design and analyzing RC slab response under blast loads are demonstrated.
The Haustorium, a Specialized Invasive Organ in Parasitic Plants
Authors: Yoshida Satoko , Cui Songkui , Ichihashi Yasunori , Shirasu Ken
Published: April 2016

Parasitic plants thrive by infecting other plants. Flowering plants evolved parasitism independently at least 12 times, in all cases developing a unique multicellular organ called the haustorium that forms upon detection of haustorium-inducing factors derived from the host plant. This organ penetrates into the host stem or root and connects to its vasculature, allowing exchange of materials such as water, nutrients, proteins, nucleotides, pathogens, and retrotransposons between the host and the parasite. In this review, we focus on the formation and function of the haustorium in parasitic plants, with a specific emphasis on recent advances in molecular studies of root parasites in the Orobanchaceae and stem parasites in the Convolvulaceae.
Mapping Transcriptional Networks in Plants: Data-Driven Discovery of Novel Biological Mechanisms
Authors: Gaudinier Allison , Brady Siobhan M.
Published: April 2016

In plants, systems biology approaches have led to the generation of a variety of large data sets. Many of these data are created to elucidate gene expression profiles and their corresponding transcriptional regulatory mechanisms across a range of tissue types, organs, and environmental conditions. In an effort to map the complexity of this transcriptional regulatory control, several types of experimental assays have been used to map transcriptional regulatory networks. In this review, we discuss how these methods can be best used to identify novel biological mechanisms by focusing on the appropriate biological context. Translating network biology back to gene function in the plant, however, remains a challenge. We emphasize the need for validation and insight into the underlying biological processes to successfully exploit systems approaches in an effort to determine the emergent properties revealed by network analyses.
Autophagy induced by exogenous bile acids is therapeutic in a model of α-1-AT deficiency liver disease
Authors: Tang Youcai , Fickert Peter , Trauner Michael , Marcus Nancy , Blomenkamp Keith , Teckman Jeffrey
Published: April 2016

The bile acid nor-ursodeoxycholic acid (norUDCA) has many biological actions, including antiapoptotic effects. Homozygous PIZZ α-1-antitrypsin (A1AT)-deficient humans are known to be at risk for liver disease, cirrhosis, and liver cancer as a result of the accumulation of the toxic, A1AT mutant Z protein within hepatocytes. This accumulation triggers cell death in the hepatocytes with the largest mutant Z-protein burdens, followed by compensatory proliferation. Proteolysis pathways within the hepatocyte, including autophagy, act to reduce the intracellular burden of A1AT Z protein. We hypothesized that norUDCA would reduce liver cell death and injury in A1AT deficiency. We treated groups of PiZ transgenic mice and wild-type mice with norUDCA or vehicle, orally, and examined the effects on the liver. The PiZ mouse is the best model of A1AT liver injury and recapitulates many features of the human liver disease. Mice treated with norUDCA demonstrated reduced hepatocellular death by compensatory hepatocellular proliferation as determined by bromodeoxyuridine incorporation (3.8% control, 0.88% treated, P < 0.04). Ki-67 staining as a marker for hepatocellular senescence and death was also reduced ( P < 0.02). Reduced apoptotic signaling was associated with norUDCA, including reduced cleavage of caspases-3, -7, and -8 (all P < 0.05). We determined that norUDCA was associated with a >70% reduction in intrahepatic mutant Z protein ( P < 0.01). A 32% increase in hepatic autophagy associated with norUDCA was the likely mechanism. norUDCA administration is associated with increased autophagy, reduced A1AT protein accumulation, and reduced liver injury in a model of A1AT deficiency.
Context-dependent effects of SOCS3 in angiotensin II-induced vascular dysfunction and hypertension in mice: mechanisms and role of bone marrow-derived cells
Authors: Li Ying , Kinzenbaw Dale A. , Modrick Mary L. , Pewe Lecia L. , Faraci Frank M.
Published: April 2016

Carotid artery disease is a major contributor to stroke and cognitive deficits. Angiotensin II (Ang II) promotes vascular dysfunction and disease through mechanisms that include the IL-6/STAT3 pathway. Here, we investigated the importance of suppressor of cytokine signaling 3 (SOCS3) in models of Ang II-induced vascular dysfunction. We examined direct effects of Ang II on carotid arteries from SOCS3-deficient (SOCS3+/−) mice and wild-type (WT) littermates using organ culture and then tested endothelial function with acetylcholine (ACh). A low concentration of Ang II (1 nmol/l) did not affect ACh-induced vasodilation in WT but reduced that of SOCS3+/−mice by ∼50% ( P < 0.05). In relation to mechanisms, effects of Ang II in SOCS3+/−mice were prevented by inhibitors of STAT3, IL-6, NF-κB, or superoxide. Systemic Ang II (1.4 mg/kg per day for 14 days) also reduced vasodilation to ACh in WT. Surprisingly, SOCS3 deficiency prevented most of the endothelial dysfunction. To examine potential underlying mechanisms, we performed bone marrow transplantation. WT mice reconstituted with SOCS3+/−bone marrow were protected from Ang II-induced endothelial dysfunction, whereas reconstitution of SOCS3+/−mice with WT bone marrow exacerbated Ang II-induced effects. The SOCS3 genotype of bone marrow-derived cells did not influence direct effects of Ang II on vascular function. These data provide new mechanistic insight into the influence of SOCS3 on the vasculature, including divergent effects depending on the source of Ang II. Bone marrow-derived cells deficient in SOCS3 protect against systemic Ang II-induced vascular dysfunction.
Surfactant dysfunction during overexpression of TGF-β1 precedes profibrotic lung remodeling in vivo
Authors: Lopez-Rodriguez Elena , Boden Caroline , Echaide Mercedes , Perez-Gil Jesus , Kolb Martin , Gauldie Jack , Maus Ulrich A. , Ochs Matthias , Knudsen Lars
Published: April 2016

Transforming growth factor-β1 (TGF-β1) is involved in regulation of cellular proliferation, differentiation, and fibrogenesis, inducing myofibroblast migration and increasing extracellular matrix synthesis. Here, TGF-β1 effects on pulmonary structure and function were analyzed. Adenovirus-mediated gene transfer of TGF-β1 in mice lungs was performed and evaluated by design-based stereology, invasive pulmonary function testing, and detailed analyses of the surfactant system 1 and 2 wk after gene transfer. After 1 wk decreased static compliance was linked with a dramatic alveolar derecruitment without edema formation or increase in the volume of septal wall tissue or collagen fibrils. Abnormally high surface tension correlated with downregulation of surfactant proteins B and C. TTF-1 expression was reduced, and, using PLA (proximity ligand assay) technology, we found Smad3 and TTF-1 forming complexes in vivo, which are normally translocated into the nucleus of the alveolar epithelial type II cells (AE2C) but in the presence of TGF-β1 remain in the cytoplasm. AE2C show altered morphology, resulting in loss of total apical surface area per lung and polarity. These changes of AE2C were progressive 2 wk after gene transfer and correlated with lung compliance. Although static lung compliance remained low, the volume of septal wall tissue and collagen fibrils increased 2 wk after gene transfer. In this animal model, the primary effect of TGF-β1 signaling in the lung is downregulation of surfactant proteins, high surface tension, alveolar derecruitment, and mechanical stress, which precede fibrotic tissue remodeling and progressive loss of AE2C polarity. Initial TTF-1 dysfunction is potentially linked to downregulation of surfactant proteins.
Effects of mannose-binding lectin on pulmonary gene expression and innate immune inflammatory response to ozone
Authors: Ciencewicki Jonathan M. , Verhein Kirsten C. , Gerrish Kevin , McCaw Zachary R. , Li Jianying , Bushel Pierre R. , Kleeberger Steven R.
Published: April 2016

Ozone is a common, potent oxidant pollutant in industrialized nations. Ozone exposure causes airway hyperreactivity, lung hyperpermeability, inflammation, and cell damage in humans and laboratory animals, and exposure to ozone has been associated with exacerbation of asthma, altered lung function, and mortality. The mechanisms of ozone-induced lung injury and differential susceptibility are not fully understood. Ozone-induced lung inflammation is mediated, in part, by the innate immune system. We hypothesized that mannose-binding lectin (MBL), an innate immunity serum protein, contributes to the proinflammatory events caused by ozone-mediated activation of the innate immune system. Wild-type ( Mbl+/+) and MBL-deficient ( Mbl−/−) mice were exposed to ozone (0.3 ppm) for up to 72 h, and bronchoalveolar lavage fluid was examined for inflammatory markers. Mean numbers of eosinophils and neutrophils and levels of the neutrophil attractants C-X-C motif chemokines 2 [ Cxcl2 (major intrinsic protein 2)] and 5 [ Cxcl5 (limb expression, LIX)] in the bronchoalveolar lavage fluid were significantly lower in Mbl−/− than Mbl+/+ mice exposed to ozone. Using genome-wide mRNA microarray analyses, we identified significant differences in transcript response profiles and networks at baseline [e.g., nuclear factor erythroid-related factor 2 (NRF2)-mediated oxidative stress response] and after exposure (e.g., humoral immune response) between Mbl+/+ and Mbl−/− mice. The microarray data were further analyzed to discover several informative differential response patterns and subsequent gene sets, including the antimicrobial response and the inflammatory response. We also used the lists of gene transcripts to search the LINCS L1000CDS2 data sets to identify agents that are predicted to perturb ozone-induced changes in gene transcripts and inflammation. These novel findings demonstrate that targeted deletion of Mbl caused differential levels of inflammation-related gene sets at baseline and after exposure to ozone and significantly reduced pulmonary inflammation, thus indicating an important innate immunomodulatory role of the gene in this model.
Identifying immune mechanisms mediating the hypertension during preeclampsia
Authors: LaMarca Babbette , Cornelius Denise C. , Harmon Ashlyn C. , Amaral Lorena M. , Cunningham Mark W. , Faulkner Jessica L. , Wallace Kedra
Published: April 2016

Preeclampsia (PE) is a pregnancy-associated disorder that affects 5–8% of pregnancies and is a major cause of maternal, fetal, and neonatal morbidity and mortality. Hallmark characteristics of PE are new onset hypertension after 20 wk gestation with or without proteinuria, chronic immune activation, fetal growth restriction, and maternal endothelial dysfunction. However, the pathophysiological mechanisms that lead to the development of PE are poorly understood. Recent data from studies of both clinical and animal models demonstrate an imbalance in the subpopulations of CD4+ T cells and a role for these cells as mediators of inflammation and hypertension during pregnancy. Specifically, it has been proposed that the imbalance between two CD4+ T cell subtypes, regulatory T cells (Tregs) and T-helper 17 cells (Th17s), is involved in the pathophysiology of PE. Studies from our laboratory highlighting how this imbalance contributes to vasoactive factors, endothelial dysfunction, and hypertension during pregnancy will be discussed in this review. Therefore, the purpose of this review is to highlight hypertensive mechanisms stimulated by inflammatory factors in response to placental ischemia, thereby elucidating a role
Accommodation to hyperpolarization of human axons assessed in the frequency domain
Authors: Howells James , Bostock Hugh , Burke David
Published: April 2016

Human axons in vivo were subjected to subthreshold currents with a threshold impedance amplitude profile to allow the use of frequency domain techniques to determine the propensity for resonant behavior and to clarify the relative contributions of different ion channels to their low-frequency responsiveness. Twenty-four studies were performed on the motor and sensory axons of the median nerve in six subjects. The response to oscillatory currents was tested between direct current (DC) and 16 Hz. A resonant peak at ∼2–2.5 Hz was found in the response of hyperpolarized axons, but there was only a small broad response in axons at resting membrane potential (RMP). A mathematical model of axonal excitability developed using DC pulses provided a good fit to the frequency response for human axons and indicated that the hyperpolarization-activated current Ihand the slow potassium current IKsare principally responsible for the resonance. However, the results indicate that if axons are hyperpolarized by more than −60% of resting threshold, the only conductances that are appreciably active are Ihand the leak conductance, i.e., that the activity of these conductances can be studied in vivo virtually in isolation at hyperpolarized membrane potentials. Given that the leak conductance dampens resonance, it is suggested that the −60% hyperpolarization used here is optimal for Ih. As expected, differences between the frequency responses of motor and sensory axons were present and best explained by reduced slow potassium conductance GKs, up-modulation of Ih, and increased persistent Na+current INaP(due to depolarization of RMP) in sensory axons.
Saccadic adaptation to a systematically varying disturbance
Authors: Cassanello Carlos R. , Ohl Sven , Rolfs Martin
Published: April 2016

Saccadic adaptation maintains the correct mapping between eye movements and their targets, yet the dynamics of saccadic gain changes in the presence of systematically varying disturbances has not been extensively studied. Here we assessed changes in the gain of saccade amplitudes induced by continuous and periodic postsaccadic visual feedback. Observers made saccades following a sequence of target steps either along the horizontal meridian (Two-way adaptation) or with unconstrained saccade directions (Global adaptation). An intrasaccadic step—following a sinusoidal variation as a function of the trial number (with 3 different frequencies tested in separate blocks)—consistently displaced the target along its vector. The oculomotor system responded to the resulting feedback error by modifying saccade amplitudes in a periodic fashion with similar frequency of variation but lagging the disturbance by a few tens of trials. This periodic response was superimposed on a drift toward stronger hypometria with similar asymptotes and decay rates across stimulus conditions. The magnitude of the periodic response decreased with increasing frequency and was smaller and more delayed for Global than Two-way adaptation. These results suggest that—in addition to the well-characterized return-to-baseline response observed in protocols using constant visual feedback—the oculomotor system attempts to minimize the feedback error by integrating its variation across trials. This process resembles a convolution with an internal response function, whose structure would be determined by coefficients of the learning model. Our protocol reveals this fast learning process in single short experimental sessions, qualifying it for the study of sensorimotor learning in health and disease.
Assessment of the expression and role of the α1-nAChR subunit in efferent cholinergic function during the development of the mammalian cochlea
Authors: Roux Isabelle , Wu (武靜靜) Jingjing Sherry , McIntosh J. Michael , Glowatzki Elisabeth
Published: April 2016

Hair cell (HC) activity in the mammalian cochlea is modulated by cholinergic efferent inputs from the brainstem. These inhibitory inputs are mediated by calcium-permeable nicotinic acetylcholine receptors (nAChRs) containing α9- and α10-subunits and by subsequent activation of calcium-dependent potassium channels. Intriguingly, mRNAs of α1- and γ-nAChRs, subunits of the “muscle-type” nAChR have also been found in developing HCs (Cai T, Jen HI, Kang H, Klisch TJ, Zoghbi HY, Groves AK. J Neurosci 35: 5870–5883, 2015; Scheffer D, Sage C, Plazas PV, Huang M, Wedemeyer C, Zhang DS, Chen ZY, Elgoyhen AB, Corey DP, Pingault V. J Neurochem 103: 2651–2664, 2007; Sinkkonen ST, Chai R, Jan TA, Hartman BH, Laske RD, Gahlen F, Sinkkonen W, Cheng AG, Oshima K, Heller S. Sci Rep 1: 26, 2011) prompting proposals that another type of nAChR is present and may be critical during early synaptic development. Mouse genetics, histochemistry, pharmacology, and whole cell recording approaches were combined to test the role of α1-nAChR subunit in HC efferent synapse formation and cholinergic function. The onset of α1-mRNA expression in mouse HCs was found to coincide with the onset of the ACh response and efferent synaptic function. However, in mouse inner hair cells (IHCs) no response to the muscle-type nAChR agonists (±)-anatoxin A, (±)-epibatidine, (−)-nicotine, or 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP) was detected, arguing against the presence of an independent functional α1-containing muscle-type nAChR in IHCs. In α1-deficient mice, no obvious change of IHC efferent innervation was detected at embryonic day 18, contrary to the hyperinnervation observed at the neuromuscular junction. Additionally, ACh response and efferent synaptic activity were detectable in α1-deficient IHCs, suggesting that α1is not necessary for assembly and membrane targeting of nAChRs or for efferent synapse formation in IHCs.
High-resolution mapping of a novel rat blood pressure locus on chromosome 9 to a region containing the Spp2 gene and colocalization of a QTL for bone mass
Authors: Nie Ying , Kumarasamy Sivarajan , Waghulde Harshal , Cheng Xi , Mell Blair , Czernik Piotr J. , Lecka-Czernik Beata , Joe Bina
Published: April 2016

Through linkage analysis of the Dahl salt-sensitive (S) rat and the spontaneously hypertensive rat (SHR), a blood pressure (BP) quantitative trait locus (QTL) was previously located on rat chromosome 9. Subsequent substitution mapping studies of this QTL revealed multiple BP QTLs within the originally identified logarithm of odds plot by linkage analysis. The focus of this study was on a 14.39 Mb region, the distal portion of which remained unmapped in our previous studies. High-resolution substitution mapping for a BP QTL in the setting of a high-salt diet indicated that an SHR-derived congenic segment of 787.9 kb containing the gene secreted phosphoprotein-2 ( Spp2) lowered BP and urinary protein excretion. A nonsynonymous G/T polymorphism in the Spp2 gene was detected between the S and S.SHR congenic rats. A survey of 45 strains showed that the T allele was rare, being detected only in some substrains of SHR and WKY. Protein modeling prediction through SWISSPROT indicated that the predicted protein product of this variant was significantly altered. Importantly, in addition to improved cardiovascular and renal function, high salt-fed congenic animals carrying the SHR T variant of Spp2 had significantly lower bone mass and altered bone microarchitecture. Total bone volume and volume of trabecular bone, cortical thickness, and degree of mineralization of cortical bone were all significantly reduced in congenic rats. Our study points to opposing effects of a congenic segment containing the prioritized candidate gene Spp2 on BP and bone mass.
Exome-Scale Discovery of Hotspot Mutation Regions in Human Cancer Using 3D Protein Structure
Authors: Tokheim Collin , Bhattacharya Rohit , Niknafs Noushin , Gygax Derek M. , Kim Rick , Ryan Michael , Masica David L. , Karchin Rachel
Published: April 2016

The impact of somatic missense mutation on cancer etiology and progression is often difficult to interpret. One common approach for assessing the contribution of missense mutations in carcinogenesis is to identify genes mutated with statistically nonrandom frequencies. Even given the large number of sequenced cancer samples currently available, this approach remains underpowered to detect drivers, particularly in less studied cancer types. Alternative statistical and bioinformatic approaches are needed. One approach to increase power is to focus on localized regions of increased missense mutation density or hotspot regions, rather than a whole gene or protein domain. Detecting missense mutation hotspot regions in three-dimensional (3D) protein structure may also be beneficial because linear sequence alone does not fully describe the biologically relevant organization of codons. Here, we present a novel and statistically rigorous algorithm for detecting missense mutation hotspot regions in 3D protein structures. We analyzed approximately 3 × 105 mutations from The Cancer Genome Atlas (TCGA) and identified 216 tumor-type–specific hotspot regions. In addition to experimentally determined protein structures, we considered high-quality structural models, which increase genomic coverage from approximately 5,000 to more than 15,000 genes. We provide new evidence that 3D mutation analysis has unique advantages. It enables discovery of hotspot regions in many more genes than previously shown and increases sensitivity to hotspot regions in tumor suppressor genes (TSG). Although hotspot regions have long been known to exist in both TSGs and oncogenes, we provide the first report that they have different characteristic properties in the two types of driver genes. We show how cancer researchers can use our results to link 3D protein structure and the biologic functions of missense mutations in cancer, and to generate testable hypotheses about driver mechanisms. Our results are included in a new interactive website for visualizing protein structures with TCGA mutations and associated hotspot regions. Users can submit new sequence data, facilitating the visualization of mutations in a biologically relevant context. Cancer Res; 76(13); 3719–31. ©2016 AACR.
Transforming Growth Factor-β Limits Secretion of Lumican by Activated Stellate Cells within Primary Pancreatic Adenocarcinoma Tumors
Authors: Kang Ya'an , Roife David , Lee Yeonju , Lv Hailong , Suzuki Rei , Ling Jianhua , Rios Perez Mayrim V. , Li Xinqun , Dai BingBing , Pratt Michael , Truty Mark J. , Chatterjee Deyali , Wang Huamin , Thomas Ryan M. , Wang Yu , Koay Eugene J. , Chiao Paul J. , Katz Matthew H. , Fleming Jason B.
Published: April 2016

Purpose: Pancreatic ductal adenocarcinoma (PDAC) is lethal cancer whose primary tumor is characterized by dense composition of cancer cells, stromal cells, and extracellular matrix (ECM) composed largely of collagen. Within the PDAC tumor microenvironment, activated pancreatic stellate cells (PSC) are the dominant stromal cell type and responsible for collagen deposition. Lumican is a secreted proteoglycan that regulates collagen fibril assembly. We have previously identified that the presence of lumican in the ECM surrounding PDAC cells is associated with improved patient outcome after multimodal therapy and surgical removal of localized PDAC.

Experimental Design: Lumican expression in PDAC from 27 patients was determined by IHC and quantitatively analyzed for colocalization with PSCs. In vitro studies examined the molecular mechanisms of lumican transcription and secretion from PSCs (HPSCs and HPaSteC), and cell adhesion and migration assays examined the effect of lumican on PSCs in a collagen-rich environment.

Results: Here we identify PSCs as a significant source of extracellular lumican production through quantitative IHC analysis. We demonstrate that the cytokine, TGF-β, negatively regulates lumican gene transcription within HPSCs through its canonical signaling pathway and binding of SMAD4 to novel SBEs identified within the promoter region. In addition, we found that the ability of HPSCs to produce and secrete extracellular lumican significantly enhances HPSCs adhesion and mobility on collagen.

Conclusions: Our results demonstrate that activated pancreatic stellate cells within PDAC secrete lumican under the negative control of TGF-β; once secreted, the extracellular lumican enhances stellate cell adhesion and mobility in a collagen-rich environment. Clin Cancer Res; 22(19); 4934–46. ©2016 AACR.