bluuebunny/crossref_metadata_2025 huggingface dataset:
The Velocity Field Underneath a Breaking Rogue Wave: Laboratory Experiments Versus Numerical Simulations
Authors: Alberello Alberto , Iafrati Alessandro
Published: April 2019

Wave breaking is the most characteristic feature of the ocean surface. Physical investigations (in the field and at laboratory scale) and numerical simulations have studied the driving mechanisms that lead to wave breaking and its effects on hydrodynamic loads on marine structures. Despite computational advances, accurate numerical simulations of the complex breaking process remain challenging. Validation of numerical codes is routinely performed against experimental observations of the surface elevation. However, it is still uncertain whether simulations can accurately reproduce the velocity field under breaking waves due to the lack of ad-hoc measurements. In the present work, the velocity field recorded with a Particle Image Velocimetry method during experiments conducted in a unidirectional wave tank is directly compared to the results of a corresponding numerical simulation performed with a Navier–Stokes (NS) solver. It is found that simulations underpredict the velocity close to the wave crest compared to measurements. Higher resolutions seem necessary in order to capture the most relevant details of the flow.
Lifetimes of Rogue Wave Events in Direct Numerical Simulations of Deep-Water Irregular Sea Waves
Authors: Kokorina Anna , Slunyaev Alexey
Published: April 2019

The issue of rogue wave lifetimes is addressed in this study, which helps to detail the general picture of this dangerous oceanic phenomenon. The direct numerical simulations of irregular wave ensembles are performed to obtain the complete accurate data on the rogue wave occurrence and evolution. Purely collinear wave systems, moderately crested, and short-crested sea states have been simulated by means of the high-order spectral method for the potential Euler equations. As rogue waves are transient and poorly reflect the physical effects, we join instant abnormally high waves in close locations and close time moments to new objects, rogue events, which helps to retrieve the abnormal occurrences more stably and more consistently from the physical point of view. The rogue event lifetime probability distributions are calculated based on the simulated wave data. They show the distinctive difference between rough sea states with small directional bandwidth on one part, and small-amplitude sea states and short-crested states on the other part. The former support long-living rogue wave patterns (the corresponding probability distributions have heavy tails), though the latter possess exponential probability distributions of rogue event lifetimes and generally produce much shorter rogue wave events.
Tracking Biodistribution of Myeloid-Derived Cells in Murine Models of Breast Cancer
Authors: Li Jun , Mai Junhua , Hinkle Louis , Lin Daniel , Zhang Jingxin , Liu Xiaoling , Ramirez Maricela R. , Zu Youli , Lokesh Ganesh L. , Volk David E. , Shen Haifa
Published: April 2019

A growing tumor is constantly secreting inflammatory chemokines and cytokines that induce release of immature myeloid cells, including myeloid-derived suppressor cells (MDSCs) and macrophages, from the bone marrow. These cells not only promote tumor growth, but also prepare distant organs for tumor metastasis. On the other hand, the myeloid-derived cells also have phagocytic potential, and can serve as vehicles for drug delivery. We have previously identified thioaptamers that bind a subset of MDSCs with high affinity and specificity. In the current study, we applied one of the thioaptamers as a probe to track myeloid cell distribution in the bone, liver, spleen and tumor in multiple murine models of breast cancer including the 4T1 syngeneic model and MDA-MB-231 and SUM159 xenograft models. Information generated from this study will facilitate further understanding of tumor growth and metastasis, and predict biodistribution patterns of cell-mediated drug delivery.
Computational Methods for Detection of Differentially Methylated Regions Using Kernel Distance and Scan Statistics
Authors: Dunbar Faith , Xu Hongyan , Ryu Duchwan , Ghosh Santu , Shi Huidong , George Varghese
Published: April 2019

Motivation: Researchers in genomics are increasingly interested in epigenetic factors such as DNA methylation because they play an important role in regulating gene expression without changes in the sequence of DNA. Abnormal DNA methylation is associated with many human diseases. Results: We propose two different approaches to test for differentially methylated regions (DMRs) associated with complex traits, while accounting for correlations among CpG sites in the DMRs. The first approach is a nonparametric method using a kernel distance statistic and the second one is a likelihood-based method using a binomial spatial scan statistic. The kernel distance method uses the kernel function, while the binomial scan statistic approach uses a mixed-effects model to incorporate correlations among CpG sites. Extensive simulations show that both approaches have excellent control of type I error, and both have reasonable statistical power. The binomial scan statistic approach appears to have higher power, while the kernel distance method is computationally faster. The proposed methods are demonstrated using data from a chronic lymphocytic leukemia (CLL) study.
Molecular Characterization of a Dirofilaria immitis Cysteine Protease Inhibitor (Cystatin) and Its Possible Role in Filarial Immune Evasion
Authors: Dong Xiaowei , Xu Jing , Song Hongyu , Liu Yuchen , Wu Maodi , Zhang Haojie , Jing Bo , Lai Weimin , Gu Xiaobin , Xie Yue , Peng Xuerong , Yang Guangyou
Published: April 2019

Infection with canine heartworm (Dirofilaria immitis), spread via mosquito vectors, causes coughing, asthma, pneumonia, and bronchitis in humans and other animals. The disease is especially severe and often fatal in dogs and represents a serious threat to public health worldwide. Cysteine protease inhibitors (CPIs), also known as cystatins, are major immunomodulators of the host immune response during nematode infections. Herein, we cloned and expressed the cystatin Di-CPI from D. immitis. Sequence analysis revealed two specific cystatin-like domains, a Q-x-V-x-G motif, and a SND motif. Phylogenetic analysis indicates that Di-CPI is a member of the second subgroup of nematode type II cystatins. Probing of D. immitis total proteins with anti-rDi-CPI polyclonal antibody revealed a weak signal, and immunofluorescence-based histochemical analysis showed that native Di-CPI is mainly localized in the cuticle of male and female worms and the gut of male worms. Treatment of canine peripheral blood mononuclear cells (PMBCs) with recombinant Di-CPI induced a Th2-type immune response characterized by high expression of the anti-inflammatory factor interleukin-10. Proliferation assays showed that Di-CPI inhibits the proliferation of canine PMBCs by 15%. Together, the results indicate that Di-CPI might be related to cellular hyporesponsiveness in dirofilariasis and may help D. immitis to evade the host immune system.
Dynamics of the Zones of Strong Earthquake Epicenters in the Arctic–Asian Seismic Belt
Authors: Imaeva Lyudmila P. , Imaev Valery S. , Koz’min Boris M.
Published: April 2019

Our comprehensive study of the Russian Arctic region aims to clarify the features and types of seismotectonic deformation of the crust in the Arctic–Asian Seismic Belt, specifically in the zones of strong earthquakes in the Laptev Sea Segment, the Kharaulakh Segment, and the Chersky Seismotectonic Zone. We have analyzed modern tectonic structures and active fault systems, as well as tectonic stress fields reconstructed by tectonophysical analysis of the Late Cenozoic faults and folds. The investigated neotectonic structures are ranked with respect to the regional classification principles. Changes in the crustal stress–strain state in the lithospheric plate boundaries between the Eurasian, North American, and Okhotsk Sea Plates are analyzed, and regularities of such changes are discovered. A set of models has been constructed for the studied segments of plate boundaries with account of the dynamics of the regional geological structures. The models can give a framework for the assessment of potential seismic risks of seismogenerating structures in the Russian Arctic region.
Clinical Evaluation of Sarcoidosis in Community Members with World Trade Center Dust Exposure
Authors: Hena Kerry M. , Murphy Scarlett , Zhang Yian , Shao Yongzhao , Kazeros Angeliki , Reibman Joan
Published: April 2019

Background: Sarcoidosis is a granulomatous disease involving intrathoracic and extrathoracic organs. Genetic and environmental factors, such as exposure to World-Trade Center (WTC) dust after 9/11, may play a role in clinical presentation. Characterization of sarcoidosis in community members with exposure to the WTC dust can provide further insight into the relationship between environmental exposure and sarcoidosis. Methods: Patients with documented sarcoidosis were identified in the WTC Environmental Health Center (EHC), a treatment program for community members. Demographic and clinical data were collected from standardized questionnaires and chart review. Organ involvement was assessed with a standard instrument. Results: Among patients in the WTC EHC, 87 were identified with sarcoidosis after 9/11. Sarcoidosis cases were more likely African-American, local workers, and had more respiratory symptoms, compared with non-sarcoidosis WTC EHC patients. Many (46%) had ≥ Scadding stage 3 on chest imaging, and had reduced lung function measures. Extrathoracic involvement was identified in 33/87 (38%) with a diversity of organs involved. Conclusions: WTC-exposed sarcoidosis in community members is often characterized by severe pulmonary disease and a high rate of diverse extrathoracic involvement. Further analysis is required to characterize the course of disease progression or resolution.
Hygiene During Childbirth: An Observational Study to Understand Infection Risk in Healthcare Facilities in Kogi and Ebonyi States, Nigeria
Authors: Buxton Helen , Flynn Erin , Oluyinka Olutunde , Cumming Oliver , Esteves Mills Joanna , Shiras Tess , Sara Stephen , Dreibelbis Robert
Published: April 2019

Background: Infections acquired during labour and delivery are a significant cause of maternal and child morbidity and mortality. Adherence to hand hygiene protocols is a critical component of infection prevention strategies, but few studies have closely examined the hand hygiene of health care providers with sufficient detail to understand infection risks and prioritize prevention strategies. Methods: This observational study was conducted in six healthcare facilities in Nigeria. In each, five women were observed from the onset of labour through to delivery of the placenta. Hand hygiene infection risk was estimated for all procedures requiring aseptic technique compared against adherence to proper hand hygiene protocol and potential recontamination events. Results: Hands were washed with soap and sterile gloves applied with no observed recontamination before only 3% of all observed procedures requiring aseptic technique. There was no significant difference in hygiene compliance between midwives and doctors nor facilities or states. Adherence to proper hygiene protocol was observed more in morning compared to afternoon and night shifts. Conclusions: This study highlights that hand hygiene remains a barrier to delivering high-quality and safe care in health facilities. Improving hygiene practices during labour and delivery will require strategies that extend beyond infrastructure provision.
Unmet Mental Health Care Needs among Asian Americans 10–11 Years After Exposure to the World Trade Center Attack
Authors: Kung Winnie W. , Wang Xiaoran , Liu Xinhua , Goldmann Emily , Huang Debbie
Published: April 2019

This study investigated the prevalence of unmet mental health care needs (UMHCN) and their associated factors among 2344 Asian Americans directly exposed to the World Trade Center (WTC) attack 10–11 years afterwards. Given the pervasive underutilization of mental health services among Asians, their subjective evaluation of unmet needs could provide more nuanced information on disparities of service. We used the WTC Health Registry data and found that 12% of Asian Americans indicated UMHCN: 69% attributing it to attitudinal barriers, 36% to cost barriers, and 29% to access barriers. Among all the factors significantly related to UMHCN in the logistic model, disruption of health insurance in the past year had the largest odds ratio (OR = 2.37, 95% confidence interval: 1.61–3.48), though similar to functional impairment due to mental disorders. Post-9/11 mental health diagnosis, probable mental disorder and ≥14 poor mental health days in the past month were also associated with greater odds of UMHCN, while greater social support was associated with lower odds. Results suggest that continued outreach efforts to provide mental health education to Asian communities to increase knowledge about mental illness and treatment options, reduce stigmatization of mental illness, and offer free mental health services are crucial to address UMHCN.
An Evaluation of a Low-Intensity Cognitive Behavioral Therapy mHealth-Supported Intervention to Reduce Loneliness in Older People
Authors: Jarvis Mary Ann , Padmanabhanunni Anita , Chipps Jennifer
Published: April 2019

There is a high prevalence of loneliness among older people, especially in residential care settings. Loneliness is often accompanied by maladaptive cognitions which can affect the maintenance and establishment of meaningful social connections. This study implemented and evaluated a low-intensity Cognitive Behavior Therapy (LI-CBT) mHealth-supported intervention which targeted maladaptive cognitions in older people (≥60 years) experiencing loneliness. The three-month intervention using WhatsApp was implemented with older people in four inner-city residential care facilities. The intervention included three components: technology acceptance, psycho-education, and individualized positively worded messages addressing maladaptive cognitions. The intervention was evaluated using a randomized control design. Key outcomes were measured pre-, post-, and one month after the intervention. There were significant changes in social cognition (YSQ-SF T0–T1–T2, X2 = 9.69, p = 0.008) and loneliness levels (total loneliness T0–T1–T2, X2 = 14.62, p = 0.001), and an increase in WhatsApp usage (T0 = 26% vs. T1 = 60%, X2=15.22, p = 0.019). At 1-month follow-up, even with a significant reduction in WhatsApp usage, a significant reduction in loneliness was maintained. LI-CBT delivered via instant messaging may be effective in reducing loneliness experienced by older people.
Social Capital and the Improvement in Functional Ability among Older People in Japan: A Multilevel Survival Analysis Using JAGES Data
Authors: Amemiya Airi , Saito Junko , Saito Masashige , Takagi Daisuke , Haseda Maho , Tani Yukako , Kondo Katsunori , Kondo Naoki
Published: April 2019

We investigated the contextual effects of community social capital on functional ability among older people with functional disability in Japan, and the cross-level interaction effects between community social capital and individual psychosocial characteristics. We used data from the Japan Gerontological Evaluation Study for 1936 men and 2207 women nested within 320 communities and followed for 46 months. We used objective data for functional ability trajectories derived from the national long-term care-insurance system, and a validated measure of health-related community social capital comprising three components: civic participation, social cohesion, and reciprocity. A multilevel survival analysis with a community-level random intercept showed that in communities with high civic participation, women who actively participated in any community group showed greater functional ability improvement than did women who did not participate (pinteraction = 0.05). In communities with high social cohesion, older men who perceived that their communities’ social cohesion was high showed greater functional ability improvement than men who perceived it to be low (pinteraction = 0.02). Community social capital can thus affect functional ability improvements variously, depending on individual psychosocial characteristics and gender. Community interventions aiming to foster social capital should focus on people who are excluded from existing opportunities to participate.
Smartphone Use and Physical Activity among College Students in Health Science-Related Majors in the United States and Thailand
Authors: Penglee Nattika , Christiana Richard W. , Battista Rebecca A. , Rosenberg Ed
Published: April 2019

Smartphone use among college students is prevalent across the world. Recently, research has begun to investigate the relationship between smartphone use and physical activity. This study examined the amount of time spent using a smartphone and the physical activity (PA) levels among college students majoring in health science-related disciplines in the United States (US) and Thailand. Using convenience sampling, college students in the US (n = 242) and Thailand (n = 194) completed an online survey, in Fall 2016, assessing smartphone usage and PA. Data were analyzed using chi-square tests and two-way ANOVA (p < 0.05). US students reported more days per week ( U = 15,150.0 , p = 0.00 , r = 0.33 ) and greater duration of PA ( U = 11,234.0 , p = 0.00 , r = 0.33 ) than Thai students while Thai students used smartphones more per day than US students ( U = 13,137.5 , p = 0.00 , r = 0.40 ). No difference existed for years of smartphone use ( U = 22,207.0 , p = 0.27 ). Greater smartphone use per day inversely related to days per week of engaging in PA among Thai students ( X 2 ( 3 ) = 10.55 , p = 0.01 , ε 2 = 0.06 ), but not among US students ( X 2 ( 3 ) = 2.39 , p = 0.50 ). The high smartphone use among college students, especially in Thailand, may be a barrier to PA as well as a strategy for PA promotion in higher education settings. Research should examine the best techniques for smartphone application development to promote PA in college settings.
The Influence of Land Use Change on Ecosystem Service Value in Shangzhou District
Authors: Yuan Keyue , Li Fei , Yang Haijuan , Wang Yiming
Published: April 2019

Land use change has an impact on the ecosystem service value because it changes the structure and function of ecosystems. This paper analyzed the changes in land use during the period from 2000 to 2015 in Shangzhou district, and used the equivalent value of ecological services per unit area of land ecosystem combining the natural and economic conditions of Shangzhou district. Based on this method, the ecological service value of Shangzhou district was estimated, and the impact of land use change on the ecological service value was analyzed. The results showed that: (1) the main types of land use in Shangzhou district were grassland, woodland and farmland, among which the contribution rate of woodland to the value of local ecosystem services was the highest; (2) the overall trend in the ecosystem service value in Shangzhou district increased between 2000 and 2015, from 10.74 × 108 yuan in 2000 to 20.32 × 108 yuan in 2015, which is the result of the combined effects of regional economic development and changes in the natural environment and land use patterns; and (3) the main reason for the value increase of ecosystem services in Shangzhou district between 2000 and 2015 was that the grain-for-green policy transformed a considerable amount of farmland into woodland, while the main reasons for a decline in value was the expansion of built-up land that occupied other types of land.
Health Literacy: Current Status and Challenges in the Work of Family Doctors in Bosnia and Herzegovina
Authors: Todorovic Nevena , Jovic-Vranes Aleksandra , Djikanovic Bosiljka , Pilipovic-Broceta Natasa , Vasiljevic Nadja , Racic Maja
Published: April 2019

Health literacy (HL) has become an important area of research. The aim of this study was to evaluate the HL of primary healthcare patients in the Republic of Srpska (RS), Bosnia and Herzegovina (B&H) and to identify socioeconomic and health factors associated with HL. This cross-sectional study among 768 patients was conducted in two healthcare centres between March and May 2017, using the Short Test of Functional Health Literacy in Adults (S-TOFHLA). Analysis was done using descriptive and inferential statistics (a chi-squared test and logistic regression). Inadequate and marginal HL was found in 34,6% of respondents. Socioeconomic and self-reported health factors were significantly related to HL. An age of 55 years and over (OR 1.02), living in a rural environment (OR 2.25), being divorced (OR 3.32), being insufficiently physically active (OR 1.29), having poor income (OR 1.96), having more than three chronic diseases (OR 1.94), and poor health (OR 1.59) were significantly corelated with inadequate and marginal HL. The results of our study indicate that a low level of HL is related to the elderly, having a divorce, having a rural residence, poor income, having more than three chronic diseases, poor health, and insufficient physical activity. Further evaluation, monitoring, and activities to improve HL are of great importance for patients’ health outcomes.
Using Patient-Reported Outcomes to Predict Revision Arthroplasty Following Femoral Neck Fracture: Enhancing the Value of Clinical Registries through Data Linkage
Authors: Ekegren Christina L , de Steiger Richard , Edwards Elton R , Page Richard S , Hau Raphael , Liew Susan , Oppy Andrew , Gabbe Belinda J
Published: April 2019

The aim of this study was to determine the association between patient-reported outcome measures (PROMs) six months following femoral neck fracture after a low fall and future arthroplasty, and the factors associated with this. Six-month post-fracture PROMs were collected from the Victorian Orthopaedic Trauma Outcomes Registry (VOTOR) for patients aged >55 years who were admitted for a femoral neck fracture after a low fall between March 2007 and June 2015. These cases were linked with those registered by Australian Orthopaedic Association National Joint Replacement Registry (AOANJRR) up to October 2016. Multivariable analysis was performed using a Cox proportional hazards model to determine factors associated with future arthroplasty, including six-month PROMs. Of the 7077 hip fracture patients registered by VOTOR during the study period, 2325 met the inclusion criteria. Internal fixation being used for the initial hip fracture surgery, being younger and having no pre-injury disability were all independently associated with future revision or conversion to arthroplasty. Out of all PROMs, reporting pain and discomfort six months post-fracture was associated with a 9.5-fold increase in the risk of future arthroplasty (95% CI: 3.81, 23.67). The value of clinical registries can be enhanced via data linkage, in this case by using PROMs to predict arthroplasty following femoral neck fracture.
Prediction of Bioactive Peptides from Chlorella sorokiniana Proteins Using Proteomic Techniques in Combination with Bioinformatics Analyses
Authors: Tejano Lhumen A. , Peralta Jose P. , Yap Encarnacion Emilia S. , Panjaitan Fenny Crista A. , Chang Yu-Wei
Published: April 2019

Chlorella is one of the most nutritionally important microalgae with high protein content and can be a good source of potential bioactive peptides. In the current study, isolated proteins from Chlorella sorokiniana were subjected to in silico analysis to predict potential peptides with biological activities. Molecular characteristics of proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and proteomics techniques. A total of eight proteins were identified by proteomics techniques from 10 protein bands of the SDS-PAGE. The predictive result by BIOPEP’s profile of bioactive peptides tools suggested that proteins of C. sorokiniana have the highest number of dipeptidyl peptidase-IV (DPP IV) inhibitors, with high occurrence of other bioactive peptides such as angiotensin-I converting enzyme (ACE) inhibitor, glucose uptake stimulant, antioxidant, regulating, anti-amnestic and antithrombotic peptides. In silico analysis of enzymatic hydrolysis revealed that pepsin (pH > 2), bromelain and papain were proteases that can release relatively larger quantity of bioactive peptides. In addition, combinations of different enzymes in hydrolysis were observed to dispense higher numbers of bioactive peptides from proteins compared to using individual proteases. Results suggest the potential of protein isolated from C. sorokiniana could be a source of high value products with pharmaceutical and nutraceutical application potential.
Effects of Naphthazarin (DHNQ) Combined with Lawsone (NQ-2-OH) or 1,4-Naphthoquinone (NQ) on the Auxin-Induced Growth of Zea mays L. Coleoptile Segments
Authors: Rudnicka Małgorzata , Ludynia Michał , Karcz Waldemar
Published: April 2019

Naphthoquinones, plants secondary metabolites are known for their antibacterial, antifungal, anti-inflammatory, anti-cancer and anti-parasitic properties. The biological activity of naphthoquinones is connected with their ability to generate reactive oxygen species and to modify biological molecules at their nucleophilic sites. In our research, the effect of naphthazarin (DHNQ) combined with 2-hydroxy-1,4-naphthoquinone (NQ-2-OH) or 1,4-naphthoquinone (1,4-NQ) on the elongation growth, pH changes of the incubation medium, oxidative stress and redox activity of maize coleoptile cells were investigated. This paper describes experiments performed with maize (Zea mays L.) coleoptile segments, which is a classical model system to study plant cell elongation growth. The data presented clearly demonstrate that lawsone and 1,4-naphthoquinone combined with naphthazarin, at low concentrations (1 and 10 nM), reduced the endogenous and IAA-induced (Indole-3-Acetic Acid) elongation growth of maize coleoptile segments. Those changes in growth correlated with the proton concentration in the incubation medium, which suggests that the changes in the growth of maize coleoptile segments observed in the presence of naphthoquinones are mediated through the activity of PM H+-ATPase. The presence of naphthoquinones induced oxidative stress in the maize coleoptile tissue by producing hydrogen peroxide and causing changes in the redox activity. Moreover, the incubation of maize segments with both naphthoquinones combined with naphthazarin resulted in lipid peroxidation and membrane damage. The regulation of PM H+-ATPase activity, especially its inhibition, may result from two major types of reaction: first, a direct interaction between an enzyme and naphthoquinone, which leads to the covalent modification of the protein thiols and the generation of thioethers, which have been found to alter the activity of the PM H+-ATPases; second, naphthoquinones induce reactive oxygen species (ROS) production, which inhibits PM H+-ATPases by increasing cytosolic Ca2+. This harmful effect was stronger when naphthazarin and 1,4-naphthoquinone were added together. Taking these results into account, it can be suggested that by combining naphthoquinones in small quantities, an alternative to synthetic pesticides could be developed.
Complementary Transcriptomic and Proteomic Analysis Reveals a Complex Network Regulating Pollen Abortion in GMS (msc-1) Pepper (Capsicum annuum L.)
Authors: Cheng Qing , Li Ting , Ai Yixin , Lu Qiaohua , Wang Yihao , Sun Liang , Shen Huolin
Published: April 2019

Pepper (Capsicum annuum L.) is a globally important horticultural crop. Use of the genic male-sterile (GMS) line enables efficient commercial hybrid pepper seed production. However, the mechanisms of pepper GMS functioning remain unclear. In this study, we used proteomic and transcriptomic analysis to identify proteins and genes related to genic male sterility. A total of 764 differentially expressed proteins (DEPs) and 1069 differentially expressed genes (DEGs) were identified in the proteomic and transcriptomic level respectively, and 52 genes (hereafter “cor-DEGs-DEPs” genes) were detected at both levels. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis identified 13 DEPs and 14 DEGs involved in tapetum and pollen development. Among the 13 DEPs identified, eight were involved in pollen exine formation, and they were all up-regulated in the fertile line 16C1369B. For the 14 DEGs identified, ABORTED MICROSPORES (AMS) and DEFECTIVE IN TAPETAL DEVELOPMENT AND FUNCTION1 (TDF1) were involved in tapetum development, and both are possibly regulated by Msc-1. All of these genes were detected and confirmed by qRT-PCR. The presence of these genes suggests their possible role in tapetum and pollen exine formation in GMS pepper. Most key genes and transcription factors involved in these processes were down-regulated in the sterile line 16C1369A. This study provides a better understanding of GMS (msc-1) molecular functioning in pepper.
Cryopreservation Differentially Alters the Proteome of Epididymal and Ejaculated Pig Spermatozoa
Authors: Perez-Patiño Cristina , Barranco Isabel , Li Junwei , Padilla Lorena , Martinez Emilio A. , Rodriguez-Martinez Heriberto , Roca Jordi , Parrilla Inmaculada
Published: April 2019

Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate to the loss of sperm function during cryopreservation remains unsolved. The present study aimed to clarify this issue evaluating differential changes in the proteome of fresh and frozen-thawed pig spermatozoa retrieved from the cauda epididymis and the ejaculate of the same boars, with clear differences in cryotolerance. Spermatozoa were collected from 10 healthy, sexually mature, and fertile boars, and cryopreserved using a standard 0.5 mL-straw protocol. Total and progressive motility, viability, and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT spermatozoa were analyzed using a LC-ESI-MS/MS-based Sequential Window Acquisition of All Theoretical Spectra approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein–protein networks highlighted a set of proteins quantitatively altered in ejaculated spermatozoa, directly involved in mitochondrial functionality which would explain why ejaculated spermatozoa deteriorate during cryopreservation.
Hydrodynamic Behavior of the Intrinsically Disordered Potyvirus Protein VPg, of the Translation Initiation Factor eIF4E and of their Binary Complex
Authors: Walter Jocelyne , Barra Amandine , Doublet Bénédicte , Céré Nicolas , Charon Justine , Michon Thierry
Published: April 2019

Protein intrinsic disorder is involved in many biological processes and good experimental models are valuable to investigate its functions. The potyvirus genome-linked protein, VPg, displays many features of an intrinsically disordered protein. The virus cycle requires the formation of a complex between VPg and eIF4E, one of the host translation initiation factors. An in-depth characterization of the hydrodynamic properties of VPg, eIF4E, and of their binary complex VPg-eIF4E was carried out. Two complementary experimental approaches, size-exclusion chromatography and fluorescence anisotropy, which is more resolving and revealed especially suitable when protein concentration is the limiting factor, allowed to estimate monomers compaction upon complex formation. VPg possesses a high degree of hydration which is in agreement with its classification as a partially folded protein in between a molten and pre-molten globule. The natively disordered first 46 amino acids of eIF4E contribute to modulate the protein hydrodynamic properties. The addition of an N-ter His tag decreased the conformational entropy of this intrinsically disordered region. A comparative study between the two tagged and untagged proteins revealed the His tag contribution to proteins hydrodynamic behavior.
Bone Marrow Endothelial Cells Influence Function and Phenotype of Hematopoietic Stem and Progenitor Cells after Mixed Neutron/Gamma Radiation
Authors: Cary Lynnette , Noutai Daniel , Salber Rudolph , Fadiyimu Opeyemi , Gross Arthur , Almeida-Porada Graca , Kidane Yared , Whitnall Mark
Published: April 2019

The bone marrow (BM) microenvironment plays a crucial role in the maintenance and regeneration of hematopoietic stem (HSC) and progenitor cells (HSPC). In particular, the vascular niche is responsible for regulating HSC maintenance, differentiation, and migration of cells in and out of the BM. Damage to this niche upon exposure to ionizing radiation, whether accidental or as a result of therapy, can contribute to delays in HSC recovery and/or function. The ability of BM derived-endothelial cells (BMEC) to alter and/or protect HSPC after exposure to ionizing radiation was investigated. Our data show that exposure of BMEC to ionizing radiation resulted in alterations in Akt signaling, increased expression of PARP-1, IL6, and MCP-1, and decreased expression of MMP1 and MMP9. In addition, global analysis of gene expression of HSC and BMEC in response to mixed neutron/gamma field (MF) radiation identified 60 genes whose expression was altered after radiation in both cell types, suggesting that a subset of genes is commonly affected by this type of radiation. Focused gene analysis by RT-PCR revealed two categories of BMEC alterations: (a) a subset of genes whose expression was altered in response to radiation, with no additional effect observed during coculture with HSPC, and (b) a subset of genes upregulated in response to radiation, and altered when cocultured with HSPC. Coculture of BMEC with CD34+ HSPC induced HSPC proliferation, and improved BM function after MF radiation. Nonirradiated HSPC exhibited reduced CD34 expression over time, but when irradiated, they maintained higher CD34 expression. Nonirradiated HSPC cocultured with nonirradiated BMEC expressed lower levels of CD34 expression compared to nonirradiated alone. These data characterize the role of each cell type in response to MF radiation and demonstrate the interdependence of each cell’s response to ionizing radiation. The identified genes modulated by radiation and coculture provide guidance for future experiments to test hypotheses concerning specific factors mediating the beneficial effects of BMEC on HSPC. This information will prove useful in the search for medical countermeasures to radiation-induced hematopoietic injury.
Altered Glutamate Receptor Ionotropic Delta Subunit 2 Expression in Stau2-Deficient Cerebellar Purkinje Cells in the Adult Brain
Authors: Pernice Helena F. , Schieweck Rico , Jafari Mehrnoosh , Straub Tobias , Bilban Martin , Kiebler Michael A. , Popper Bastian
Published: April 2019

Staufen2 (Stau2) is an RNA-binding protein that is involved in dendritic spine morphogenesis and function. Several studies have recently investigated the role of Stau2 in the regulation of its neuronal target mRNAs, with particular focus on the hippocampus. Here, we provide evidence for Stau2 expression and function in cerebellar Purkinje cells. We show that Stau2 downregulation (Stau2GT) led to an increase of glutamate receptor ionotropic delta subunit 2 (GluD2) in Purkinje cells when animals performed physical activity by voluntary wheel running compared with the age-matched wildtype (WT) mice (C57Bl/6J). Furthermore, Stau2GT mice showed lower performance in motor coordination assays but enhanced motor learning abilities than did WT mice, concomitantly with an increase in dendritic GluD2 expression. Together, our results suggest the novel role of Stau2 in Purkinje cell synaptogenesis in the mouse cerebellum.
Activation of PPARs Modulates Signalling Pathways and Expression of Regulatory Genes in Osteoclasts Derived from Human CD14+ Monocytes
Authors: Kasonga Abe , Kruger Marlena C. , Coetzee Magdalena
Published: April 2019

Osteoclasts are the sole bone resorbing cell in the body and their over activity is key in the development of osteoporosis. Osteoclastogenesis is mediated by receptor activator of nuclear factor κB ligand (RANKL) signalling pathways. Unsaturated fatty acids (UFA) are known to inhibit osteoclastogenesis by targeting RANKL signalling. However, the mechanisms of action remain unclear. Peroxisome proliferator activated receptors (PPARs) are a family of nuclear receptors, with three known isoforms (PPAR-α, PPAR-β/δ and PPAR-γ), that are known to bind UFAs and are expressed in osteoclasts. In this study, we aimed to determine how different families of UFAs activate PPARs and how PPAR activation influences osteoclast signalling. Human CD14+ monocytes were seeded into cluster plates with RANKL and macrophage colony stimulating factor (M-CSF) in the presence of PPAR agonists or different types of UFAs. All the PPAR agonists were shown to upregulate the activity of their respective receptors. Polyunsaturated fatty acids increased PPAR-α to a greater extent than monounsaturated fatty acids (MUFAs), which favoured PPAR-β/δ activation. All PPAR agonists inhibited osteoclastogenesis. The activation of RANKL signalling pathways and expression of key osteoclast genes were downregulated by PPAR agonists. This study reveals that PPAR activation can inhibit osteoclastogenesis through modulation of RANKL signalling.
Low-Luminance Blue Light-Enhanced Phototoxicity in A2E-Laden RPE Cell Cultures and Rats
Authors: Lin Cheng-Hui , Wu Man-Ru , Huang Wei-Jan , Chow Diana Shu-Lian , Hsiao George , Cheng Yu-Wen
Published: April 2019

N-retinylidene-N-retinylethanolamine (A2E) and other bisretinoids are components of lipofuscin and accumulate in retinal pigment epithelial (RPE) cells—these adducts are recognized in the pathogenesis of retinal degeneration. Further, blue light-emitting diode (LED) light (BLL)-induced retinal toxicity plays an important role in retinal degeneration. Here, we demonstrate that low-luminance BLL enhances phototoxicity in A2E-laden RPE cells and rats. RPE cells were subjected to synthetic A2E, and the effects of BLL on activation of apoptotic biomarkers were examined by measuring the levels of cleaved caspase-3. BLL modulates the protein expression of zonula-occludens 1 (ZO-1) and paracellular permeability in A2E-laden RPE cells. Early inflammatory and angiogenic genes were also screened after short-term BLL exposure. In this study, we developed a rat model for A2E treatment with or without BLL exposure for 21 days. BLL exposure caused fundus damage, decreased total retinal thickness, and caused neuron transduction injury in the retina, which were consistent with the in vitro data. We suggest that the synergistic effects of BLL and A2E accumulation in the retina increase the risk of retinal degeneration. These outcomes help elucidate the associations between BLL/A2E and angiogenic/apoptotic mechanisms, as well as furthering therapeutic strategies.
L-Arginine Inhibited Inflammatory Response and Oxidative Stress Induced by Lipopolysaccharide via Arginase-1 Signaling in IPEC-J2 Cells
Authors: Qiu Yueqin , Yang Xuefen , Wang Li , Gao Kaiguo , Jiang Zongyong
Published: April 2019

This study aimed to explore the effect of L-arginine on lipopolysaccharide (LPS)-induced inflammatory response and oxidative stress in IPEC-2 cells. We found that the expression of toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88), cluster of differentiation 14 (CD14), nuclear factor-kappaBp65 (NF-κBp65), chemokine-8 (IL-8), tumor necrosis factor (TNF-α) and chemokine-6 (IL-6) mRNA were significantly increased by LPS. Exposure to LPS induced oxidative stress as reactive oxygen species (ROS) and malonaldehyde (MDA) production were increased while glutathione peroxidase (GSH-Px) were decreased in LPS-treated cells compared to those in the control. LPS administration also effectively induced cell growth inhibition through induction of G0/G1 cell cycle arrest. However, compared with the LPS group, cells co-treatment with L-arginine effectively increased cell viability and promoted the cell cycle into the S phase; L-arginine exhibited an anti-inflammatory effect in alleviating inflammation induced by LPS by reducing the abundance of TLR4, MyD88, CD14, NF-κBp65, and IL-8 transcripts. Cells treated with LPS+L-arginine significantly enhanced the content of GSH-Px, while they decreased the production of ROS and MDA compared with the LPS group. Furthermore, L-arginine increased the activity of arginase-1 (Arg-1), while Arg-1 inhibitor abolished the protection of arginine against LPS-induced inflammation and oxidative stress. Taken together, these results suggested that L-arginine exerted its anti-inflammatory and antioxidant effects to protect IPEC-J2 cells from inflammatory response and oxidative stress challenged by LPS at least partly via the Arg-1 signaling pathway.
Studies on the Interaction between Poly-Phosphane Gold(I) Complexes and Dihydrofolate Reductase: An Interplay with Nicotinamide Adenine Dinucleotide Cofactor
Authors: Pucciarelli Stefania , Vincenzetti Silvia , Ricciutelli Massimo , Simon Oumarou Camille , Ramadori Anna Teresa , Luciani Lorenzo , Galassi Rossana
Published: April 2019

A class of gold(I) phosphane complexes have been identified as inhibitors of dihydrofolate reductase (DHFR) from E. coli, an enzyme that catalyzes the reduction of dihydrofolate (DHF) to tetrahydrofolate (THF), using NADPH as a coenzyme. In this work, to comprehend the nature of the interaction at the basis of these inhibitory effects, the binding properties of bis- and tris-phosphane gold(I) chloride compounds in regards to DHFR have been studied by emission spectroscopy and spectrophotometric assays. The lack of cysteine and seleno-cysteine residues in the enzyme active site, the most favorable sites of attack of Au(I) moieties, makes this work noteworthy. The interaction with the gold compounds results into the quenching of the DHFR tryptophan’s emissions and in an enhancement of their intrinsic emission intensities. Moreover, a modulating action of NADPH is highlighted by means of an increase of the gold compound affinity toward the enzyme; in fact, the dissociation constants calculated for the interactions between DHFR and each gold compound in the presence of saturating NADPH were lower than the ones observed for the apo-enzyme. The fluorimetric data afforded to Kd values ranged from 2.22 ± 0.25 µM for (PPh3)2AuCl in the presence of NADPH to 21.4 ± 3.85 µM for 4L3AuTf in the absence of NADPH. By elucidating the energetic aspects of the binding events, we have attempted to dissect the role played by the gold phosphane/protein interactions in the inhibitory activity, resulting in an exothermic enthalpy change and a positive entropic contribution (ΔH° = −5.04 ± 0.08 kcal/mol and ΔS° = 7.34 ± 0.005 cal/mol·K).
The ERK MAPK Pathway Is Essential for Skeletal Development and Homeostasis
Authors: Kim Jung-Min , Yang Yeon-Suk , Park Kwang Hwan , Oh Hwanhee , Greenblatt Matthew B. , Shim Jae-Hyuck
Published: April 2019

Mitogen-activated protein kinases (MAPKs) are a family of protein kinases that function as key signal transducers of a wide spectrum of extracellular stimuli, including growth factors and pro-inflammatory cytokines. Dysregulation of the extracellular signal-regulated kinase (ERK) MAPK pathway is associated with human skeletal abnormalities including Noonan syndrome, neurofibromatosis type 1, and cardiofaciocutaneous syndrome. Here, we demonstrate that ERK activation in osteoprogenitors is required for bone formation during skeletal development and homeostasis. Deletion of Mek1 and Mek2, kinases upstream of ERK MAPK, in osteoprogenitors (Mek1OsxMek2−/−), resulted in severe osteopenia and cleidocranial dysplasia (CCD), similar to that seen in humans and mice with impaired RUNX2 function. Additionally, tamoxifen-induced deletion of Mek1 and Mek2 in osteoprogenitors in adult mice (Mek1Osx-ERTMek2−/−) significantly reduced bone mass. Mechanistically, this corresponded to decreased activation of osteoblast master regulators, including RUNX2, ATF4, and β-catenin. Finally, we identified potential regulators of osteoblast differentiation in the ERK MAPK pathway using unbiased phospho-mass spectrometry. These observations demonstrate essential roles of ERK activation in osteogenesis and bone formation.
Prevention of Deficit in Neuropsychiatric Disorders through Monitoring of Arsenic and Its Derivatives as Well as Through Bioinformatics and Cheminformatics
Authors: Avram Speranta , Udrea Ana Maria , Negrea Adina , Ciopec Mihaela , Duteanu Narcis , Postolache Carmen , Duda-Seiman Corina , Duda-Seiman Daniel , Shaposhnikov Sergey
Published: April 2019

Neuropsychiatric disorders are induced by various risk factors, including direct exposure to environmental chemicals. Arsenic exposure induces neurodegeneration and severe psychiatric disorders, but the molecular mechanisms by which brain damage is induced are not yet elucidated. Our aim is to better understand the molecular mechanisms of arsenic toxicity in the brain and to elucidate possible ways to prevent arsenic neurotoxicity, by reviewing significant experimental, bioinformatics, and cheminformatics studies. Brain damage induced by arsenic exposure is discussed taking in account: the correlation between neuropsychiatric disorders and the presence of arsenic and its derivatives in the brain; possible molecular mechanisms by which arsenic induces disturbances of cognitive and behavioral human functions; and arsenic influence during psychiatric treatments. Additionally, we present bioinformatics and cheminformatics tools used for studying brain toxicity of arsenic and its derivatives, new nanoparticles used as arsenic delivery systems into the human body, and experimental ways to prevent arsenic contamination by its removal from water. The main aim of the present paper is to correlate bioinformatics, cheminformatics, and experimental information on the molecular mechanism of cerebral damage induced by exposure to arsenic, and to elucidate more efficient methods used to reduce its toxicity in real groundwater.