bluuebunny/crossref_metadata_2025 huggingface dataset:
CACNA1A Gene Variants in Eight Chinese Patients With a Wide Range of Phenotypes
Authors: Zhang Linxia , Wen Yongxin , Zhang Qingping , Chen Yan , Wang Jiaping , Shi Kaili , Du Lijun , Bao Xinhua
Published: December 2020

Background: The CACNA1A gene encodes the voltage-dependent P/Q-type calcium channel subunit alpha-1A, which is widely expressed throughout the CNS. The biological roles of the P/Q channel are diverse and the phenotypic spectrum caused by CACNA1A mutations is wide. The aim of this study is to demonstrate its phenotypic diversity and analyze the genotype-phenotype correlations in a cohort of Chinese patients.

Methods: Patients with hemiplegic migraine, cerebellar ataxia, developmental delay, or epilepsy without known causes were tested by trios whole-exome sequencing. Patients with pathogenic CACNA1A gene variants were recruited. The clinical information of the patients was collected, and the association between the genotype and the phenotype was investigated.

Results: In total, eight patients (six females and two males) were found to have CACNA1A gene variants. All the variants were de novo including six missense variants and one frameshift variant. Four de novo missense variants were found in five patients located in the S4, S5, or S6 transmembrane segments of Domain II and III (p.R1352Q, p.G701V, p.A713T, p.V1393M). All of them were correlated with severe phenotypes, including three with sporadic hemiplegic migraine type 1 and epilepsy, and two with developmental and epileptic encephalopathy. The other two missense variants, p.Y62C and p.F1814L, located in the cytoplasmic side of the N-terminus and C-terminus, respectively. The variant p.Y62C was associated with severe hemiconvulsion-hemiplegia-epilepsy syndrome, and p.F1814L was associated with relatively mild phenotypes. All the missense variants were speculated as gain-of-function (GOF) mutations. The only frameshift variant, p.Q681Rfs*100, a lose-of-function (LOF) mutation, was found in a patient with episodic ataxia type 2. Meanwhile, all the patients had developmental delay ranging from mild to severe, as well as cerebellar ataxia including one with congenital ataxia, one with episodic ataxia, and six with non-progressive ataxia.

Conclusions:CACNA1A variants could lead to a wide spectrum of neurological disorders including epileptic or non-epileptic paroxysmal events, cerebellar ataxia, and developmental delay. The variants could be both GOF and LOF mutations. There appeared to be some correlations between genotypes and phenotypes.
FGIN-1-27 Inhibits Melanogenesis by Regulating Protein Kinase A/cAMP-Responsive Element-Binding, Protein Kinase C-β, and Mitogen-Activated Protein Kinase Pathways
Authors: Lv Jinpeng , Jiang Songzhou , Yang Ying , Zhang Ximei , Gao Rongyin , Cao Yan , Song Guoqiang
Published: December 2020

FGIN-1-27 is a synthetic mitochondrial diazepam binding inhibitor receptor (MDR) agonist that has demonstrated pro-apoptotic, anti-anxiety, and steroidogenic activity in various studies. Here we report, for the first time, the anti-melanogenic efficacy of FGIN-1-27 in vitro and in vivo. FGIN-1-27 significantly inhibited basal and α-melanocyte-stimulating hormone (α-MSH)-, 1-Oleoyl-2-acetyl-sn-glycerol (OAG)- and Endothelin-1 (ET-1)-induced melanogenesis without cellular toxicity. Mushroom tyrosinase activity assay showed that FGIN-1-27 did not directly inhibit tyrosinase activity, which suggested that FGIN-1-27 was not a direct inhibitor of tyrosinase. Although it was not capable of modulating the catalytic activity of mushroom tyrosinase in vitro, FGIN-1-27 downregulated the expression levels of key proteins that function in melanogenesis. FGIN-1-27 played these functions mainly by suppressing the PKA/CREB, PKC-β, and MAPK pathways. Once inactivated, it decreased the expression of MITF, tyrosinase, TRP-1, TRP-2, and inhibited the tyrosinase activity, finally inhibiting melanogenesis. During in vivo experiments, FGIN-1-27 inhibited the body pigmentation of zebrafish and reduced UVB-induced hyperpigmentation in guinea pig skin, but not a reduction of numbers of melanocytes. Our findings indicated that FGIN-1-27 exhibited no cytotoxicity and inhibited melanogenesis in both in vitro and in vivo models. It may prove quite useful as a safer skin-whitening agent.
In Vitro Effects of St. John’s Wort Extract Against Inflammatory and Oxidative Stress and in the Phagocytic and Migratory Activity of Mouse SIM-A9 Microglia
Authors: Bonaterra Gabriel A. , Mierau Olga , Hofmann Johanna , Schwarzbach Hans , Aziz-Kalbhenn Heba , Kolb Christiane , Kinscherf Ralf
Published: December 2020

Introduction: Herbal medicinal plants as Hypericum perforatum L., known as St. John’s wort (SJW) have been in use for a long time. SJW that is specifically used for the treatment of depressive disorders. Inflammatory cytokines derived from microglia play an important role in the regulation of the synthesis and reuptake of glutamate and influence synaptic function, morphology and neuronal plasticity. The present study was performed to investigate, whether STW3-VI, a special SJW extract has protective effects on mouse SIM-A9 microglia against cytotoxic and proinflammatory effects of ROS, glutamate, NMDA or cortisol. Additionally, we investigated the effects of SJW on migratory and phagocytic properties of microglia.

Results: Pre-treatment (48 h) of microglia with STW3-VI (5 or 10 μg/ml)—in contrast to desipramine—inhibited the H2O2-induced TNF-α release by 20–40%. Pre-treatment (48 h) of microglia with STW3-VI (5 or 10 μg/ml) delayed the 3 or 4 mM H2O2-induced intracellular ROS level by 26.9 and 44.4%, respectively. Furthermore, pre-treatment (48 h) of microglia with STW3-VI (5 μg/ml) - in contrast to desipramine - lowered the glutamate-induced cytotoxicity by 13.2%. Besides, pre-treatment (48 h) of microglia with STW3-VI (5 or 10 μg/ml) or desipramine (5 µM) inhibited the NMDA-induced decrease of the viability by 16.5–28.8% or 12%, respectively. Finally, pre-treatment (48 h) of microglia with STW3-VI (5 or 10 μg/ml)—in contrast to desipramine - reduced the cortisol-induced cytotoxicity by 15.5 and 12.9%. Treatment of microglia with STW3-VI (10 or 100 μg/ml) increased the migratory and the phagocytic capacities by 100 and 40%.

Conclusion: Our data provide evidence that STW3-VI—in contrast to desipramine - protects microglia from oxidative stress, NMDA- or glutamate-induced cytotoxicity, and has anti-inflammatory properties that are accompanied by improvement of their migratory and phagocytic capacity. These protective (particularly the anti-inflammatory) properties may be beneficial in the treatment of depressive disorders.
Hydroxychloroquine as Prophylaxis for COVID-19: A Review
Authors: Monti Manuela , Vertogen Bernadette , Masini Carla , Donati Caterina , Lilli Claudia , Zingaretti Chiara , Musuraca Gerardo , De Giorgi Ugo , Cerchione Claudio , Farolfi Alberto , Cortesi Pietro , Viale Pierluigi , Martinelli Giovanni , Nanni Oriana
Published: December 2020

The impact of the COVID-19 pandemic worldwide has led to a desperate search for effective drugs and vaccines. There are still no approved agents for disease prophylaxis. We thus decided to use a drug repositioning strategy to perform a state-of-the-art review of a promising but controversial drug, hydroxychloroquine (HCQ), in an effort to provide an objective, scientific and methodologically correct overview of its potential prophylactic role. The advantage of using known drugs is that their toxicity profile is well known and there are fewer commercial interests (e.g., expired patents), thus allowing the scientific community to be freer of constraints. The main disadvantage is that the economic resources are almost always insufficient to promote large multinational clinical trials. In the present study, we reviewed the literature and available data on the prophylactic use of HCQ. We also took an in-depth look at all the published clinical data on the drug and examined ongoing clinical trials (CTs) from the most important CT repositories to identify a supporting rationale for HCQ prophylactic use. Our search revealed a substantial amount of preclinical data but a lack of clinical data, highlighting the need to further assess the translational impact of in vitro data in a clinical setting. We identified 77 CTs using a multiplicity of HCQ schedules, which clearly indicates that we are still far from reaching a standard of care. The majority of the CTs (92%) are randomized and 53% are being conducted in a phase 3 or 2/3 setting. The comparator is placebo or control in 55 (77%) of the randomized studies. Forty-eight (62%) CTs expect to enroll up to 1,000 subjects and 50 (71%) plan to recruit healthcare workers (HCW). With regard to drug schedules, 45 (58.5%) CTs have planned a loading dose, while 18 (23.4%) have not; the loading dose is 800 mg in 19 trials (42.2%), 400 mg in 19 (42.2%), 600 mg in 4 (8.9%) and 1,200 mg in 1 (2.2%). Forty trials include at least one daily schedule, while 19 have at least one weekly schedule. Forty-one (53.2%) will have a treatment duration of more than 30 days. Awaiting further developments that can only derive from the results of these prospective randomized CTs, the take-home message of our review is that a correct methodological approach is the key to understanding whether prophylactic HCQ can really represent an effective strategy in preventing COVID-19.
Influence of Fluoropolymer Film Modified With Nanoscale Photoluminophor on Growth and Development of Plants
Authors: Ivanyuk Veronika V. , Shkirin Alexey V. , Belosludtsev Konstantin N. , Dubinin Mikhail V. , Kozlov Valeriy A. , Bunkin Nikolay F. , Dorokhov Aleksey S. , Gudkov Sergey V.
Published: December 2020

The red and blue ranges of the optical spectrum are most suitable for plant photosynthesis. Moreover, quanta of red light stimulate photosynthesis more than quanta of blue light. In northern latitudes, the average daily intensity of the red and blue parts of the spectrum is usually not sufficient for many plants. To increase the productivity of greenhouses in northern latitudes, a technology has been developed for fluoropolymer films with photoconversion nanoparticles (quantum dots) that convert UV radiation and violet light into red light. The use of photoconversion fluoropolymer films promotes an increase in the biomass of plants grown in greenhouses at high latitudes. The greatest effect is observed when tomato plants are grown under photoconversion fluoropolymer films. The biomass of tomato berry obtained from one bush grown under films is 20% higher than the biomass of berry of control tomato plants.
A Roadmap for the Molecular Farming of Viral Glycoprotein Vaccines: Engineering Glycosylation and Glycosylation-Directed Folding
Authors: Margolin Emmanuel , Crispin Max , Meyers Ann , Chapman Ros , Rybicki Edward P.
Published: December 2020

Immunization with recombinant glycoprotein-based vaccines is a promising approach to induce protective immunity against viruses. However, the complex biosynthetic maturation requirements of these glycoproteins typically necessitate their production in mammalian cells to support their folding and post-translational modification. Despite these clear advantages, the incumbent costs and infrastructure requirements with this approach can be prohibitive in developing countries, and the production scales and timelines may prove limiting when applying these production systems to the control of pandemic viral outbreaks. Plant molecular farming of viral glycoproteins has been suggested as a cheap and rapidly scalable alternative production system, with the potential to perform post-translational modifications that are comparable to mammalian cells. Consequently, plant-produced glycoprotein vaccines for seasonal and pandemic influenza have shown promise in clinical trials, and vaccine candidates against the newly emergent severe acute respiratory syndrome coronavirus-2 have entered into late stage preclinical and clinical testing. However, many other viral glycoproteins accumulate poorly in plants, and are not appropriately processed along the secretory pathway due to differences in the host cellular machinery. Furthermore, plant-derived glycoproteins often contain glycoforms that are antigenically distinct from those present on the native virus, and may also be under-glycosylated in some instances. Recent advances in the field have increased the complexity and yields of biologics that can be produced in plants, and have now enabled the expression of many viral glycoproteins which could not previously be produced in plant systems. In contrast to the empirical optimization that predominated during the early years of molecular farming, the next generation of plant-made products are being produced by developing rational, tailor-made approaches to support their production. This has involved the elimination of plant-specific glycoforms and the introduction into plants of elements of the biosynthetic machinery from different expression hosts. These approaches have resulted in the production of mammalian N-linked glycans and the formation of O-glycan moietiesin planta. More recently, plant molecular engineering approaches have also been applied to improve the glycan occupancy of proteins which are not appropriately glycosylated, and to support the folding and processing of viral glycoproteins where the cellular machinery differs from the usual expression host of the protein. Here we highlight recent achievements and remaining challenges in glycoengineering and the engineering of glycosylation-directed folding pathways in plants, and discuss how these can be applied to produce recombinant viral glycoproteins vaccines.
Crisis Without Borders: What Does International Law Say About Border Closure in the Context of Covid-19?
Authors: Chetail Vincent
Published: December 2020

This paper is assessing the legality of border closures decided by a vast number of countries with the view of limiting the spread of Covid-19. Although this issue has raised diverging interpretations in relation to International Health Regulations and regional free movement agreements, international human rights law provides a clear-cut answer: the rule of law stops neither at the border nor in times of emergency. Against this normative framework, border control can and must be carried out with the twofold purpose of protecting public health and individual rights, whereas border closure is unable to do so because it is by essence a collective and automatic denial of admission without any other form of process. This paper argues that blanket entry bans on the ground of public health are illegal under international human rights law. They cannot be reconciled with the most basic rights of migrants and refugees, including the principle of non-refoulement and access to asylum procedures, the prohibition of collective expulsion, the best interests of the child and the principle of non-discrimination. The paper concludes on the ways to better integrate at the borders public health and human rights imperatives in due respect with the rule of law. In both law and practice, public health and migrant's rights are not mutually exclusive. They can reinforce each other within a comprehensive human rights based approach to health and migration policies.
Pedestrians’ Understanding of a Fully Autonomous Vehicle’s Intent to Stop: A Learning Effect Over Time
Authors: Hochman Michal , Parmet Yisrael , Oron-Gilad Tal
Published: December 2020

This study explored pedestrians’ understanding of Fully Autonomous Vehicles (FAVs) intention to stop and what influences pedestrians’ decision to cross the road over time, i.e., learnability. Twenty participants saw fixed simulated urban road crossing scenes with a single FAV on the road as if they were pedestrians intending to cross. Scenes differed from one another in the FAV’s, distance from the crossing place, its physical size, and external Human-Machine Interfaces (e-HMI) message by background color (red/green), message type (status/advice), and presentation modality (text/symbol). Eye-tracking data and decision measurements were collected. Results revealed that pedestrians tend to look at the e-HMI before making their decision. However, they did not necessarily decide according to the e-HMIs’ color or message type. Moreover, when they complied with the e-HMI proposition, they tended to hesitate before making the decision. Overall, a learning effect over time was observed in all conditions regardless of e- HMI features and crossing context. Findings suggest that pedestrians’ decision making depends on a combination of the e-HMI implementation and the car distance. Moreover, since the learning curve exists in all conditions and has the same proportion, it is critical to design an interaction that would encourage higher probability of compatible decisions from the first phase. However, to extend all these findings, it is necessary to further examine dynamic situations.
Using Smartphones When Eating Increases Caloric Intake in Young People: An Overview of the Literature
Authors: La Marra Marco , Caviglia Giorgio , Perrella Raffaella
Published: December 2020

Recent literature highlights that the use of smartphones during meals increases the number of calories ingested in young people. Although the distraction interferes with physiological signals of hunger and satiety, a social facilitation effect has also been suggested. Cognition is a pivotal component in regulating food intake, and activities requiring high perceptual demands should be discouraged during meals.
Centrality of Pregnancy and Prenatal Attachment in Pregnant Nulliparous After Recent Elective or Therapeutic Abortion
Authors: Smorti Martina , Ponti Lucia , Bonassi Lucia , Cattaneo Elena , Ionio Chiara
Published: December 2020

Background

There are two types of voluntary interruption of pregnancy: elective and therapeutic abortion. These forms are different for many reasons, and it is reasonable to assume that they can have negative consequences that can last until a subsequent gestation. However, no study has analyzed the psychological experience of gestation after a previous abortion, distinguishing the two forms of voluntary interruption of pregnancy.

Objective

This study aims to explore the level of prenatal attachment and centrality of pregnancy in nulliparous low-risk pregnant women with a recently (<3 years) previous elective or therapeutic abortion.

Methods

A total of 34 nulliparous pregnant women with a history of abortion (23 elective and 11 therapeutic abortion), aged from 27 to 48 years (mean = 37.17), were recruited in the maternity ward of a public hospital of the metropolitan area of Tuscany and Lombardy (Italy) during the third trimester of gestation. The participants filled out a battery of questionnaires aimed at assessing prenatal attachment and centrality of pregnancy.

Results

Analyses of variance showed that women with a history of elective abortion reported a higher centrality of pregnancy than women with a past therapeutic abortion. On the contrary, women with a past therapeutic abortion reported higher prenatal attachment.

Conclusion

Elective and therapeutic abortions are different experiences that impact the way women experience a subsequent pregnancy. Future research should further investigate the psychological experience of gestation after abortion.
Psychological Status Among Anesthesiologists and Operating Room Nurses During the Outbreak Period of COVID-19 in Wuhan, China
Authors: Li Xin-yi , Wang Jin , Zhang Rui-xian , Chen Luhua , He Colin K. , Wang Cheng-yao , Ke Jian-juan , Wang Yan-lin , Zhang Zong-ze , Song Xue-min
Published: December 2020

Background: Coronavirus Disease 2019 (COVID-19) caused by a novel strain of coronavirus (SARS-CoV-2) posed a major threat to public health. Anesthesiologists and operating room (OR) nurses are at high risk of occupational exposure to SARS-CoV-2 and developing COVID-19. We conducted a single-center survey to investigate the psychological status and perceived social support among operation room (OR) medical staffs during the outbreak of Coronavirus Disease 2019 (COVID-19).

Methods: A total of 197 OR medical staffs were enrolled in the survey. The authors performed a cohort study during the period of Wuhan lockdown and then conducted a longitudinal follow-up after lifting of lockdown. The Patient Health Questionaire-9 (PHQ-9) was used to assess for depression and Generalized Anxiety Disorder-7 (GAD-7) for anxiety. The Multidimensional Scale of Perceived Social Support (MSPSS) was used to assess perceived social support. We compared the psychological status of OR medical staffs before and after lifting of Wuhan lockdown.

Results: During the period of city lockdown, 177 (89.8%) had close contact with confirmed COVID-19 cases. The prevalence of depression and anxiety in OR medical staffs was 41.6 and 43.1% under Wuhan lockdown, while 13.2 and 15.7% after lifting of lockdown (P = 0.002, P = 0.004). Logistic regression analysis showed that being female, living in suburb areas, shortage of protective equipment and close contact with COVID-19 patients were associated with a higher risk of depression and anxiety. Perceived social support was negatively correlated with depression and anxiety severity in the OR medical staffs (P < 0.05).

Conclusions: OR medical staffs exhibited high incidence of anxiety and depression faced with the high risk of exposure to COVID-19 patients. More social support and social recognition for anesthesiologists and OR nurses might potentially help them relieve their psychological pressure.
Psychological, Social and Health-Related Challenges in Spanish Older Adults During the Lockdown of the COVID-19 First Wave
Authors: Rodríguez-González Raquel , Facal David , Martínez-Santos Alba-Elena , Gandoy-Crego Manuel
Published: December 2020

Background: The global pandemic of COVID-19 has required a population lockdown. Spain has one of the oldest/most aging populations in the world and was one of the most affected countries. We aim to describe the psychological and social implications as well as health-related behaviors as a result of the lockdown in community-dwelling older adults.

Materials and Methods: Observational cross-sectional study. A total of 528 participants of over 60 years of age were recruited using snowball sampling technique during the lockdown of the COVID-19 first wave using an anonymous self-administered questionnaire composed of descriptive questions and validated scales for resilience (Brief Resilient Coping Scale) and emotions (Scale of Positive And Negative Experience).

Results: Most participants (76.9%) live with other people and have an open space at home (64%). Only 33.7% continued doing activities to promote healthy aging, 65.7% did less physical activity and 25.6% increased their intellectual activity. Most of them (83%) used electronic communication with family and friends to a greater extent, and left the house to run basic errands. Greater scores on resilience showed significant negative correlations with age and negative feelings, and positive correlations with the size of the social network and positive feelings. Lacking an open space at home was associated with more negative feelings.

Discussion: Older people are a vulnerable group severely affected by this pandemic crisis at multiple levels, requiring specific interventions to minimize the effects of changes in lifestyle that may be harmful. Detecting needs is essential to improve care and support from community health and social services, both nowadays and in future similar situations.
Community Environments That Promote Intergenerational Interactions vs. Walking Among Older Adults
Authors: Zhong Sinan , Lee Chanam , Lee Hanwool
Published: December 2020

Background: Intergenerational interactions and walking are two of the most beneficial forms of activities for older adults. As older adults spend most of their time at or near home, the characteristics of the proximate residential environments are particularly important for supporting those activities. This study aims to (1) explore places used for various social interactions older adults engage in, (2) examine specific neighborhood environmental features associated with intergenerational interactions, and (3) compare similarities and differences in environmental correlates of intergenerational interactions vs. walking.

Methods: This cross-sectional study analyzed self-reported survey data from 455 community-dwelling adults aged 65+ from Austin, Texas, as well as Geographic Information System (GIS) measures capturing the neighborhood environment around each participant's home. Descriptive statistics were used for Aim 1. Multivariable binary logistic models were used for Aims 2 and 3, to identify environmental variables predicting the odds of participating in intergenerational interactions (with children 1+ times/week, and with children, teenagers, or adults 1+ times/week) in one's neighborhood, as well as walking 1+ times/week for transportation or recreation purposes.

Results: Participants had a mean age of 73 years, and were primarily female (72.1%) and non-Hispanic white (72.8%). Older adults interacted frequently with adults (79.2%, 1+ times/week) and other older adults (66.9%) in their neighborhood, while less frequently with children (28.0%) and teenagers (21.9%). Recreational walking (73.3%, 1+ times/week) was more popular for older adults than transportation walking (43.8%). Multivariable analyses showed that neighborhood perceptions, transportation infrastructure, land uses, land covers, population densities, development activities, and composite scores were significant predictors of intergenerational activities. Both similarities and differences were found in terms of the neighborhood environmental factors associated with intergenerational interactions vs. walking although differences were more evident in the domains of land covers, development activities, and composite scores.

Conclusions: Given the significant health benefits, promoting intergenerational interactions and walking among older adults should be a national/global responsibility. Further work is needed to improve our understanding of the specific social and physical environmental facilitators as well as barriers to creating intergenerational communities that can support healthy living of all generations.
Coffee Microbiota and Its Potential Use in Sustainable Crop Management. A Review
Authors: Duong Benoit , Marraccini Pierre , Maeght Jean-Luc , Vaast Philippe , Lebrun Michel , Duponnois Robin
Published: December 2020

Intensive coffee production is accompanied by several environmental issues, including soil degradation, biodiversity loss, and pollution due to the wide use of agrochemical inputs and wastes generated by processing. In addition, climate change is expected to decrease the suitability of cultivated areas while potentially increasing the distribution and impact of pests and diseases. In this context, the coffee microbiota has been increasingly studied over the past decades in order to improve the sustainability of the coffee production. Therefore, coffee associated microorganisms have been isolated and characterized in order to highlight their useful characteristics and study their potential use as sustainable alternatives to agrochemical inputs. Indeed, several microorganisms (including bacteria and fungi) are able to display plant growth-promoting capacities and/or biocontrol abilities toward coffee pests and diseases. Despite that numerous studies emphasized the potential of coffee-associated microorganisms under controlled environments, the present review highlights the lack of confirmation of such beneficial effects under field conditions. Nowadays, next-generation sequencing technologies allow to study coffee associated microorganisms with a metabarcoding/metagenomic approach. This strategy, which does not require cultivating microorganisms, now provides a deeper insight in the coffee-associated microbial communities and their implication not only in the coffee plant fitness but also in the quality of the final product. The present review aims at (i) providing an extensive description of coffee microbiota diversity both at the farming and processing levels, (ii) identifying the “coffee core microbiota,” (iii) making an overview of microbiota ability to promote coffee plant growth and to control its pests and diseases, and (iv) highlighting the microbiota potential to improve coffee quality and waste management sustainability.
Birth of a Live Cria After Transfer of a Vitrified-Warmed Alpaca (Vicugna pacos) Preimplantation Embryo
Authors: Lutz Jennifer C. , Johnson Susan L. , Duprey Kimberly J. , Taylor Paul J. , Vivanco-Mackie Henry William , Ponce-Salazar Daniel , Miguel-Gonzales Marlene , Youngs Curtis R.
Published: December 2020

The alpaca (Vicugna pacos) is an important species for the production of fiber and food. Genetic improvement programs for alpacas have been hindered, however, by the lack of field-practical techniques for artificial insemination and embryo transfer. In particular, successful techniques for the cryopreservation of alpaca preimplantation embryos have not been reported previously. The objective of this study was to develop a field-practical and efficacious technique for cryopreservation of alpaca preimplantation embryos using a modification of a vitrification protocol originally devised for horses and adapted for dromedary camels. Four naturally cycling non-superovulated Huacaya females serving as embryo donors were mated to males of proven fertility. Donors received 30 μg of gonadorelin at the time of breeding, and embryos were non-surgically recovered 7 days after mating. Recovered embryos (n = 4) were placed individually through a series of three vitrification solutions at 20°C (VS1: 1.4 M glycerol; VS2: 1.4 M glycerol + 3.6 M ethylene glycol; VS3: 3.4 M glycerol + 4.6 M ethylene glycol) before loading into an open-pulled straw (OPS) and plunging directly into liquid nitrogen for storage. At warming, each individual embryo was sequentially placed through warming solutions (WS1: 0.5 M galactose at 37°C; WS2: 0.25 M galactose at 20°C), and warmed embryos were incubated at 37°C in 5% CO2 in humidified air for 20–22 h in 1 ml Syngro® holding medium supplemented with 10% (v/v) alpaca serum to perform an initial in vitro assessment of post-warming viability. Embryos whose diameter increased during culture (n = 2) were transferred individually into synchronous recipients, whereas embryos that did not grow (n = 2) were transferred together into a single recipient to perform an in vivo assessment of post-warming viability. Initial pregnancy detection was performed ultrasonographically 29 days post-transfer when fetal heartbeat could be detected, and one of three recipients was pregnant (25% embryo survival rate). On November 13, 2019, the one pregnant recipient delivered what is believed to be the world's first cria produced from a vitrified-warmed alpaca embryo.
Phosphoproteomics to Characterize Host Response During H3N2 Canine Influenza Virus Infection of Dog Lung
Authors: Liu Yongbo , Fu Cheng , Ye Shaotang , Liang Yingxin , Qi Zhonghe , Yao Congwen , Wang Zhen , Wang Ji , Cai Siqi , Tang Shiyu , Chen Ying , Li Shoujun
Published: December 2020

Avian-origin H3N2 canine influenza viruses (CIVs) cause severe contagious respiratory disease in dogs, and quickly adapt to new environments. To further understand the mechanism of virus infection and host-virus interactions, we characterized the complete phosphoproteome of dogs infected with H3N2 CIV. Nine-week-old Beagle dogs were inoculated intranasally with 106 EID50 of A/canine/Guangdong/04/2014 (H3N2) virus. Lung sections were harvested at 5 days post-inoculation (dpi) and processed for global and quantitative analysis of differentially expressed phosphoproteins. A total of 1,235 differentially expressed phosphorylated proteins were identified in the dog lung after H3N2 CIV infection, and 3,016 modification sites were identified among all differentially expressed proteins. We then performed an enrichment analysis of functional annotations using Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene ontology (GO) database analyses to predict the functions of the identified differential phosphoproteins. Our data indicate that H3N2 CIV infection causes dramatic changes in the host protein phosphorylation of dog lungs. To our knowledge, this is the first study to assess the effect of H3N2 CIV infection on the phosphoproteome of beagles. These data provide novel insights into H3N2-CIV-triggered regulatory phosphorylation circuits and signaling networks and may improve our understanding of the mechanisms underlying CIV pathogenesis in dogs.
Sex Differences in Group Composition and Habitat Use of Iberian Free-Range Pigs
Authors: Dalmau Antoni , Martínez-Macipe Míriam , Manteca Xavier , Mainau Eva
Published: December 2020

The aim of the present work was to study group size, group composition and habitat use of Iberian pigs along the year when reared outdoor. This consists of a regimen in which animals are reared free range from 2 months of age until at least 14 months of age. In a first stage, animals are supplemented with concentrates, and in a second, called montanera, pigs eat just natural resources in areas with no more than two pigs per hectare. In these systems, males are castrated to avoid boar taint and females spayed to avoid the attraction and mounting by wild boars. The study was carried out in five different farms allocated in the south-west of Spain during 2 consecutive years, from March 2012 to February 2014, under the montanera regimen, and with a total of 995 animals observed (498 males and 497 females). The data were analyzed with SAS by means of general models and proc mixed. Mean group size along the year was of 17 ± 12.9 individuals, but this was significantly lower (P < 0.05) during the montanera (12 ± 0.8) and at midday (13 ± 0.8). Groups were bigger (P < 0.05) when they were more than 50 m from a tree (23 ± 1.8), or <10 m from the shelter (25 ± 1.5), the feeding area (31 ± 3.1) and the water-bath area (25 ± 1.5). Nine percent of the groups were solitary animals, being higher (P = 0.0286) during the montanera (11%) than the rest of the year (8%) and being formed in 68% by males. Males were less involved in mixed groups than were females (75% vs. 91%), especially in spring, where the largest (P < 0.0001) male groups were found. Female groups were less frequent and smaller (P < 0.0001) than were male and mixed groups. In conclusion, although males were castrated at a very young age, they showed a different behavior than females, forming in bachelor groups during the spring and being less involved in mixed groups and with more solitary animals. During the montanera, when animals were feeding on acorns and other natural resources, groups were smaller and closer to the trees, solitary males reaching a maximum percent.
The Expression of Proteins Related to Serotonin Pathway in Pulmonary Arteries of Dogs Affected With Pulmonary Hypertension Secondary to Degenerative Mitral Valve Disease
Authors: Sakarin Siriwan , Surachetpong Sirilak Disatian , Rungsipipat Anudep
Published: December 2020

Background: Pulmonary hypertension (PH) can cause medial thickening, a hallmark of pulmonary arterial remodeling. The serotonin (5HT) pathway has been suggested as a factor associated with PH by inducing pulmonary arterial smooth muscle cells (SMCs) proliferation, a major cause of medial thickening. This study aims to demonstrate the expression of molecules in the 5HT pathway in the pulmonary artery of dogs affected with PH secondary to degenerative mitral valve disease (DMVD) compared to DMVD and healthy control dogs.

Materials and Methods: The study included lung samples from the carcasses of 19 older small-breed dogs (Control n = 5, DMVD n = 7, DMVD+PH n = 7). Lung tissue sections were performed Hematoxylin and Eosin staining for measuring the percentage of medial thickness and immunohistochemistry for evaluating the expression of proteins in the 5HT pathway including serotonin transporter (SERT), serotonin 2A receptor (5HT2A), tryptophan hydroxylase 1 (TPH1), extracellular regulated kinase 1/2 (ERK1/2), and phosphorylated ERK1/2 (pERK1/2).

Results: Medial thickening of the pulmonary arteries was found in the DMVD and DMVD+PH groups compared to the control. The medial thickening of the DMVD+PH group was increased significantly compared to that in the DMVD group. Intracytoplasmic expression of proteins related to the 5HT pathway was mainly presented in the medial layer of the pulmonary arteries. The control group showed a low expression of proteins related to the 5HT pathway. An intensive expression of SERT, 5HT2A, TPH1, and ERK1/2 protein was seen in the DMVD and DMVD+PH groups. Interestingly, pERK1/2 was strongly represented only in the DMVD+PH group.

Conclusions: Overexpression of proteins related to the 5HT pathway including SERT, 5HT2A, TPH1, ERK1/2, and pERK1/2 was associated with medial remodeling in dogs affected with secondary to DMVD.
Improvement of Comfort in Suspension Seats with a Pneumatic Negative Stiffness System
Authors: Palomares Eduardo , Morales Angel L. , Nieto Antonio J. , Chicharro Jose M. , Pintado Publio
Published: December 2020

This paper presents a Negative Stiffness System (NSS) for vibration isolation and comfort improvement of vehicle seats, which enhances the performance of optimized traditional passive seat suspensions. The NSS is based on a set of two Pneumatic Linear Actuators (PLAs) added to a seat supported by a pneumatic spring. One end of each PLA is joined to the seat while the other end is joined to the vehicle frame. In static conditions, the PLAs remain horizontal, whereas in dynamic conditions, their vertical forces work against the pneumatic spring, reducing the overall dynamic stiffness and improving passenger comfort. The paper presents a stability analysis of the highly nonlinear dynamic system, as well as the numerical determination of the optimum PLA pressure for a given passenger mass that maximises comfort without instabilities. Finally, the performance of the proposed NSS is compared to that of a traditionally optimized passive seat suspension via simulations of an eight-degree-of-freedom vehicle model traversing several road profiles and speed bumps. Comfort improvements between 10% and 35% are found in all tests considered.
Manufacturing Technology on a Mechatronics Line Assisted by Autonomous Robotic Systems, Robotic Manipulators and Visual Servoing Systems
Authors: Filipescu Adrian , Mincă Eugenia , Filipescu Adriana , Coandă Henri-George
Published: December 2020

This paper proposes the implementation of an assisting technology to a processing/reprocessing mechatronics line (P/RML), comprising the following: two autonomous robotic systems (ARSs), two robotic manipulators (RMs) and three visual servoing systems (VSSs). The P/RML has four line-shaped workstations assisted by two ARSs—wheeled mobile robots (WMRs): one of them equipped with an RM, used for manipulation, and the other one used for transport. Two types of VSSs—eye to hand and eye in hand—are used as actuators for precise positioning of RMs to catch and release the work-piece. The work-piece visits stations successively as it is moved along the line for processing. If the processed piece does not pass the quality test, it is taken from the last stations of the P/RML and it is transported to the first station where it will be considered for reprocessing. The P/RML, assisted by ARSs, RMs and VSSs, was modelled with the synchronized hybrid Petri nets (SHPN). To control the ARSs, we propose the use of trajectory-tracking and sliding-mode control (TTSMC). The precise positioning that allows the picking up and releasing of the work-piece was performed using two types of VSSs. In the case of the first one, termed eye to hand VSS, the cameras have a fixed position, located at the last and the first workstations of the P/RML. For the second one, named eye in hand VSS, the camera is located at the end effector of the RM.
Biochar Reduces the Stability of Soil Aggregates during Intensive Leaching Experiment
Authors: Teutscherova Nikola , Lojka Bohdan , Benito Marta , Masaguer Alberto , Vázquez Eduardo
Published: December 2020

The interplay of different mechanisms shaping the biochar impact on soil structure remains relatively unexplored. We investigated the impact of biochar application to two contrasting soils on the stability of soil aggregates under an intensive intermittent leaching regime. A greenhouse experiment was established using PVC columns filled with 500 g soil from an Acrisol or Calcisol amended with three biochar applications (0, 1 and 2% w/w). The columns were watered weekly (100 mL) during two leaching cycles (each lasting 10 weeks). The amount of leached base cations, the stability of 1–2 mm aggregates fraction and soil chemical properties were determined. Biochar enhanced the leaching of the studied cations, but the content of base cations and effective cation exchange capacity remained higher in the biochar-amended Acrisol when compared to control soil. In both soils, biochar reduced the amount of water-stable aggregates, which seemed to be attributed to the increase of K in the exchange complex in the Acrisol while no significant correlation was detected between aggregation in Calcisol and other variables. The negative impact of biochar on soil aggregation is likely linked to higher sensitivity of biochar-amended soils to aggregate disruption under changing moisture conditions caused by frequent and intensive leaching events. These results highlight the gaps in our understanding of biochar impact on soil aggregation, which have implications for soil erodibility or restoration of degraded lands under changing climate.
Nutrient Extraction in Pansy Fertigated with Pure, Diluted, Depurated and Phytodepurated Leachates from Municipal Solid Waste
Authors: Plaza Blanca María , Maggini Rita , Borghesi Eva , Pardossi Alberto , Lao María Teresa , Jiménez-Becker Silvia
Published: December 2020

The disposal of municipal solid waste (MSW) in landfills might result in serious environmental pollution. The goal of the present assay was to assess if MSW wastewater (pure or treated) can be used as a potential source for fertilizers, studying its effects on plant biometric parameters as well as on anion concentrations and anion partitioning of the bedding plant Viola × wittrockiana Gams. (pansy), to detect possible harmful impacts on plants. The treatments tested were: W9.0 (pure wastewater, EC 9.0 dS m−1), W4.5 (diluted wastewater, EC 4.5 dS m−1), DW4.5 (depurated wastewater, EC 4.5 dS m−1), PW4.5 (phytodepurated wastewater, EC 4.5 dS m−1), and T (tap water, the control, EC 1.5 dS m−1). The treatment with pure wastewater negatively affected plant fresh weight, water content (on a fresh weight basis), leaf area, plant height and width, and the number of leaves and flowers due to an accumulation of SO42− in plant tissues. Phytodepurated wastewater improved plant fresh weight, water content, plant height and width, the number of leaves and flowers, and reduced the leaf area because this treatment decreased SO42− concentrations in wastewater by half. Pansy plants also showed high levels of nutrients (N, P, and Mg) when compared with fertigation with pure and diluted landfill leachate. These results show that phytodepurated wastewater from MSW can be used for the fertigation of pansy plants.
Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation
Authors: Pittalà Maria Gaetana Giovanna , Reina Simona , Cubisino Salvatore Antonio Maria , Cucina Annamaria , Formicola Beatrice , Cunsolo Vincenzo , Foti Salvatore , Saletti Rosaria , Messina Angela
Published: December 2020

Mitochondria from affected tissues of amyotrophic lateral sclerosis (ALS) patients show morphological and biochemical abnormalities. Mitochondrial dysfunction causes oxidative damage and the accumulation of ROS, and represents one of the major triggers of selective death of motor neurons in ALS. We aimed to assess whether oxidative stress in ALS induces post-translational modifications (PTMs) in VDAC1, the main protein of the outer mitochondrial membrane and known to interact with SOD1 mutants related to ALS. In this work, specific PTMs of the VDAC1 protein purified by hydroxyapatite from mitochondria of a NSC34 cell line expressing human SOD1G93A, a suitable ALS motor neuron model, were analyzed by tryptic and chymotryptic proteolysis and UHPLC/High-Resolution ESI-MS/MS. We found selective deamidations of asparagine and glutamine of VDAC1 in ALS-related NSC34-SOD1G93A cells but not in NSC34-SOD1WT or NSC34 cells. In addition, we identified differences in the over-oxidation of methionine and cysteines between VDAC1 purified from ALS model or non-ALS NSC34 cells. The specific range of PTMs identified exclusively in VDAC1 from NSC34-SOD1G93A cells but not from NSC34 control lines, suggests the appearance of important changes to the structure of the VDAC1 channel and therefore to the bioenergetics metabolism of ALS motor neurons. Data are available via ProteomeXchange with identifier .
Discovery of Spilanthol Endoperoxide as a Redox Natural Compound Active against Mammalian Prx3 and Chlamydia trachomatis Infection
Authors: Dushime Rosine , Zhu Yunhuang , Wu Hanzhi , Saez Daniel , Shukla Kirtikar , Brown-Harding Heather , Biavatti Maique W. , Nelson Kimberly J. , Poole Leslie B. , Lowther William T. , Jones Paul B. , Furdui Cristina M. , Tsang Allen W.
Published: December 2020

Chlamydia trachomatis (Ct) is a bacterial intracellular pathogen responsible for a plethora of diseases ranging from blindness to pelvic inflammatory diseases and cervical cancer. Although this disease is effectively treated with antibiotics, concerns for development of resistance prompt the need for new low-cost treatments. Here we report the activity of spilanthol (SPL), a natural compound with demonstrated anti-inflammatory properties, against Ct infections. Using chemical probes selective for imaging mitochondrial protein sulfenylation and complementary assays, we identify an increase in mitochondrial oxidative state by SPL as the underlying mechanism leading to disruption of host cell F-actin cytoskeletal organization and inhibition of chlamydial infection. The peroxidation product of SPL (SPL endoperoxide, SPLE), envisioned to be the active compound in the cellular milieu, was chemically synthesized and showed more potent anti-chlamydial activity. Comparison of SPL and SPLE reactivity with mammalian peroxiredoxins, demonstrated preferred reactivity of SPLE with Prx3, and virtual lack of SPL reaction with any of the reduced Prx isoforms investigated. Cumulatively, these findings support the function of SPL as a pro-drug, which is converted to SPLE in the cellular milieu leading to inhibition of Prx3, increased mitochondrial oxidation and disruption of F-actin network, and inhibition of Ct infection.