bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of Motor Primitive-Based Adaptive Control for Lower Limb Exoskeletons
Authors: Nunes Polyana F. , Ostan Icaro , Siqueira Adriano A. G.
Published: January 2021

In order to assist after-stroke individuals to rehabilitate their movements, research centers have developed lower limbs exoskeletons and control strategies for them. Robot-assisted therapy can help not only by providing support, accuracy, and precision while performing exercises, but also by being able to adapt to different patient needs, according to their impairments. As a consequence, different control strategies have been employed and evaluated, although with limited effectiveness. This work presents a bio-inspired controller, based on the concept of motor primitives. The proposed approach was evaluated on a lower limbs exoskeleton, in which the knee joint was driven by a series elastic actuator. First, to extract the motor primitives, the user torques were estimated by means of a generalized momentum-based disturbance observer combined with an extended Kalman filter. These data were provided to the control algorithm, which, at every swing phase, assisted the subject to perform the desired movement, based on the analysis of his previous step. Tests are performed in order to evaluate the controller performance for a subject walking actively, passively, and at a combination of these two conditions. Results suggest that the robot assistance is capable of compensating the motor primitive weight deficiency when the subject exerts less torque than expected. Furthermore, though only the knee joint was actuated, the motor primitive weights with respect to the hip joint were influenced by the robot torque applied at the knee. The robot also generated torque to compensate for eventual asynchronous movements of the subject, and adapted to a change in the gait characteristics within three to four steps.
Hypoxic Transformation of Immune Cell Metabolism Within the Microenvironment of Oral Cancers
Authors: Chaudhary Amrita , Bag Swarnendu , Arora Neeraj , Radhakrishnan Vivek S. , Mishra Deepak , Mukherjee Geetashree
Published: January 2021

Oral squamous cell carcinoma (OSCC) includes tumors of the lips, tongue, gingivobuccal complex, and floor of the mouth. Prognosis for OSCC is highly heterogeneous, with overall 5-year survival of ~50%, but median survival of just 8–10 months for patients with locoregional recurrence or metastatic disease. A key feature of OSCC is microenvironmental oxygen depletion due to rapid growth of constituent tumor cells, which triggers hypoxia-associated signaling events and metabolic adaptations that influence subsequent tumor progression. Better understanding of leukocyte responses to tissue hypoxia and onco-metabolite expression under low-oxygen conditions will therefore be essential to develop more effective methods of diagnosing and treating patients with OSCC. This review assesses recent literature on metabolic reprogramming, redox homeostasis, and associated signaling pathways that mediate crosstalk of OSCC with immune cells in the hypoxic tumor microenvironment. The likely functional consequences of this metabolic interface between oxygen-starved OSCC and infiltrating leukocytes are also discussed. The hypoxic microenvironment of OSCC modifies redox signaling and alters the metabolic profile of tumor-infiltrating immune cells. Improved understanding of heterotypic interactions between host leukocytes, tumor cells, and hypoxia-induced onco-metabolites will inform the development of novel theranostic strategies for OSCC.
Immigrant Naturalisation, Employment and Occupational Status in Western Europe
Authors: Hoxhaj Rezart , Vink Maarten , Breuer Tijana
Published: January 2021

Does citizenship facilitate access to employment and higher status jobs? Existing studies have produced mixed results across mostly single case studies in Europe and North America. To investigate whether this heterogeneity depends on varying institutional and socio-economic conditions, in this paper we analyse the labour market outcomes of immigrants who have naturalised in 13 West European countries. Our empirical analysis draws on data from the 2014 European Labour Force Survey Ad Hoc Module on immigrants. In order to cope with the selective nature of the naturalisation process, we employ a bivariate probit model that accounts for unobserved characteristics of naturalising immigrants. Our main results show a positive relationship across these destination countries between citizenship and the probability of employment, as well as between citizenship and occupational status, but only for immigrant men from developing countries. For women and for migrants from developed countries, we observe no significant differences between citizens and non-citizens. Liberalising the access to citizenship does not diminish the positive returns on employment from naturalisation. For immigrant men from developing countries there is evidence of a trade-off between easier access to citizenship and the returns on occupational status.
Closing a Backdoor to Dual Citizenship: The German Citizenship Law Reform of 2000 and the Abolishment of the “Domestic Clause”
Authors: Falcke Swantje , Vink Maarten
Published: January 2021

The German citizenship law underwent a paradigmatic amendment in 2000. One often overlooked change of this reform was the abolishment of the domestic clause (“Inlandsklausel”) that implied a substantial restriction to de facto dual citizenship acceptance. Combining data from the German Socio-Economic Panel Study (waves 1993–2006) with original data on origin country policies on dual citizenship and citizenship reacquisition, we analyse the impact of the abolishment of the domestic clause on naturalization rates. We apply a difference-in-difference design to investigate the causal impact of this element of the reform which has remained under-studied. We do not find an impact of the abolishment of the domestic clause on naturalization rates, neither among the general migrant population, nor among Turkish migrants who are alleged to be targeted specifically by this reform. These results suggest that a more restrictive approach to dual citizenship did not dissuade migrants from acquiring German citizenship after 2000.
Orthotic Insoles Improve Gait Symmetry and Reduce Immediate Pain in Subjects With Mild Leg Length Discrepancy
Authors: Menez Charlotte , L'Hermette Maxime , Coquart Jeremy
Published: January 2021

Background: Mild leg length discrepancy can lead to musculoskeletal disorders; however, the magnitude starting from which leg length discrepancy alters the biomechanics of gait or benefits from treatment interventions is not clear.

Research question: The aim of the current study was to examine the immediate effects of orthotic insoles on gait symmetry and pain on mild leg length discrepancy according to two groups of the leg length discrepancy (i.e., LLD ≤ 1 cm vs. LLD > 1 cm).

Methods: Forty-six adults with mild leg length discrepancy were retrospectively included and classified into two groups (GLLD≤1cm or GLLD>1cm). All subjects underwent routine 3D gait analysis with and without orthotic insoles. The symmetry index was calculated to assess changes in gait symmetry between the right and left limbs. Pain was rated without (in standing) and with the orthotic insoles (after 30 min of use) on a visual analog scale.

Results: There was a significant improvement in the symmetry index of the pelvis in the frontal plane (p = 0.001) and the ankle in the sagittal plane (p = 0.010) in the stance with the orthotic insoles independent from the group. Pain reduced significantly with the orthotic insoles independently from the group (p < 0.001).

Significance: Orthotic insoles significantly improved gait symmetry in the pelvis in the frontal plane and the ankle in the sagittal plane, as well as pain in all subjects (both LLD ≤ 1 cm and LLD > 1 cm) suggesting that it may be appropriate to treat even mild leg length discrepancy.
Biological Maturity Status in Elite Youth Soccer Players: A Comparison of Pragmatic Diagnostics With Magnetic Resonance Imaging
Authors: Leyhr Daniel , Murr Dennis , Basten Lajos , Eichler Katrin , Hauser Thomas , Lüdin Dennis , Romann Michael , Sardo Giuseppe , Höner Oliver
Published: January 2021

The influence of biological maturity status (BMS) on talent identification and development within elite youth soccer is critically debated. During adolescence, maturity-related performance differences within the same age group may cause greater chances of being selected for early maturing players. Therefore, coaches need to consider players' BMS. While standard methods for assessing BMS in adolescents are expensive and time-consuming imaging techniques (i.e., X-ray and MRI), there also exist more pragmatic procedures. This study aimed to evaluate commonly used methods to assess BMS within a highly selected sample of youth soccer players. A total of N = 63 elite male soccer players (U12 and U14) within the German Soccer Association's talent promotion program completed a test battery assessing BMS outcomes. Utilizing MRI diagnostics, players' skeletal age (SAMRI) was determined by radiologists and served as the reference method. Further commonly used methods included skeletal age measured by an ultrasound device (SAUS), the maturity offset (MOMIR), and the percentage of adult height (PAHKR). The relation of these alternative BMS outcomes to SAMRI was examined using different perspectives: performing bivariate correlation analyses (1), modeling BMS as a latent variable (BMSlat) based on the multiple alternative diagnostics (2), and investigating individual differences in agreement (3). (1) Correlations of SAMRI and the further BMS variables ranked from r = 0.80 to r = 0.84 for the total sample and were lower for U12 (0.56 ≤ r ≤ 0.66), and U14 (0.61 ≤ r ≤ 0.74) (2). The latent structural equation modeling (SEM) (R2 = 51%) revealed a significant influence on BMSlat for MOMIR (β = 0.51, p <0.05). The additional contribution of PAHKR (β = 0.27, p = 0.06) and SAUS (β = −0.03, p = 0.90) was rather small (3). The investigation of individual differences between the reference method and alternative diagnostics indicated a significant bias for MOMIR (p <0.01). The results support the use of economical and time-efficient methods for assessing BMS within elite youth soccer. Bivariate correlation analyses as well as the multivariate latent variable approach highlight the measures' usefulness. However, the observed individual level differences for some of the utilized procedures led to the recommendation for practitioners to use at least two alternative assessment methods in order to receive more reliable information about players' BMS within the talent promotion process.
Perspectives on the Treatment of Lumbar Disc Degeneration: The Value Proposition for a Cell-Based Therapy, Immunomodulatory Properties of Discogenic Cells and the Associated Clinical Evaluation Strategy
Authors: Silverman Lara Ionescu , Heaton Will , Farhang Niloofar , Saxon Lindsey Hart , Dulatova Galina , Rodriguez-Granrose Daniel , Flanagan Flagg , Foley Kevin T.
Published: January 2021

Low back pain (LBP) is a serious medical condition that affects a large percentage of the population worldwide. One cause of LBP is disc degeneration (DD), which is characterized by progressive breakdown of the disc and an inflamed disc environment. Current treatment options for patients with symptomatic DD are limited and are often unsuccessful, so many patients turn to prescription opioids for pain management in a time when opioid usage, addiction, and drug-related deaths are at an all-time high. In this paper, we discuss the etiology of lumbar DD and currently available treatments, as well as the potential for cell therapy to offer a biologic, non-opioid alternative to patients suffering from the condition. Finally, we present an overview of an investigational cell therapy called IDCT (Injectable Discogenic Cell Therapy), which is currently under evaluation in multiple double-blind clinical trials overseen by major regulatory agencies. The active ingredient in IDCT is a novel allogeneic cell population known as Discogenic Cells. These cells, which are derived from intervertebral disc tissue, have been shown to possess both regenerative and immunomodulatory properties. Cell therapies have unique properties that may ultimately lead to decreased pain and improved function, as well as curb the numbers of patients pursuing opioids. Their efficacy is best assessed in rigorous double-blinded and placebo-controlled clinical studies.
Case Report: A Review of the Literature on Spinal Intradural Hemangiopericytoma With Spinal Cord Infiltration and a Case Report
Authors: Chunyang Liu , Huiqin Zhu , Mo Sun , Yubo Wang , Xianfeng Zhang
Published: January 2021

Purpose: Primary spinal intradural hemangiopericytoma (HPC) with spinal cord infiltration is rare. The purposes of this study were to investigate the clinical features of intradural HPC with spinal cord infiltration and to explore the related factors affecting tumor recurrence.

Methods: We report a case of intramedullary HPC with intramedullary infiltration of the thoracic spine. The relevant literature was searched for with PubMed, and clinical data were extracted from the included studies. Clinical patient data were described and statistically analyzed. Then, Kaplan-Meier (KM) curves were used to describe the relapse-free survival (RFS) of patients in different groups, and the log-rank test was used for evaluation.

Results: A total of 11 cases of spinal intradural HPC with spinal cord infiltration were included (including the case described in this report). Further data analysis showed that sex (P = 0.249), age (P = 0.876), tumor location (P = 0.524), and postoperative radiotherapy (P = 0.12) had no significant influence on RFS. The range of tumor resection (P = 0.004) and the WHO grade (P = 0.014) significantly affect the patient RFS.

Conclusion: RFS was higher in patients with total tumor resection than in patients with subtotal tumor resection. The patients with lower WHO grade have better RFS. Total tumor resection is the primary objective of surgical treatment of spinal intradural HPC with spinal infiltration. Long-term postoperative follow-up is considered necessary.
The Evolving E-cigarette: Comparative Chemical Analyses of E-cigarette Vapor and Cigarette Smoke
Authors: Cunningham Anthony , McAdam Kevin , Thissen Jesse , Digard Helena
Published: January 2021

Background: E-cigarette designs, materials, and ingredients are continually evolving, with cotton wicks and diverse coil materials emerging as the popular components of atomisers. Another recent development is the use of nicotine salts in e-liquids to replicate the form of nicotine found in cigarette smoke, which may help cigarette smokers to transition to e-cigarettes. However, scientific understanding of the impact of such innovations on e-cigarette aerosol chemistry is limited.

Methods: To address these knowledge gaps, we have conducted a comparative study analyzing relevant toxicant emissions from five e-cigarettes varying in wick, atomiser coil, and benzoic acid content and two tobacco cigarettes, quantifying 97 aerosol constituents and 84 smoke compounds, respectively. Our focus was the potential for benzoic acid in e-liquids and cotton wicks to form aerosol toxicants through thermal degradation reactions, and the potential for nickel–iron alloy coils to catalyze degradation of aerosol formers. In addition, we analyzed e-cigarette emissions for 19 flavor compounds, thermal decomposition products, and e-liquid contaminants that the FDA has recently proposed adding to the established list of Harmful and Potentially Harmful Constituents (HPHCs) in tobacco products.

Results: Analyses for benzene and phenol showed no evidence of the thermal decomposition of benzoic acid in the e-cigarettes tested. Measurements of cotton decomposition products, such as carbonyls, hydrocarbons, aromatics, and PAHs, further indicated that cotton wicks can be used without thermal degradation in suitable e-cigarette designs. No evidence was found for enhanced thermal decomposition of propylene glycol or glycerol by the nickel–iron coil. Sixteen of the 19 FDA-proposed compounds were not detected in the e-cigarettes. Comparing toxicant emissions from e-cigarettes and tobacco cigarettes showed that levels of the nine WHO TobReg priority cigarette smoke toxicants were more than 99% lower in the aerosols from each of five e-cigarettes as compared with the commercial and reference cigarettes.

Conclusions: Despite continuing evolution in design, components and ingredients, e-cigarettes continue to offer significantly lower toxicant exposure alternatives to cigarette smoking.
Lactobacillus salivarius, a Potential Probiotic to Improve the Health of LPS-Challenged Piglet Intestine by Alleviating Inflammation as Well as Oxidative Stress in a Dose-Dependent Manner During Weaning Transition
Authors: Sun Zeyang , Li Haihua , Li Yupeng , Qiao Jiayun
Published: January 2021

Intestinal health is a critical issue for piglets during their weaning transition period. Previous reports have emphasized the promise of distinct probiotics in improving the enteric health. Here in this research, a newly isolated Lactobacillus salivarius strain was pretreated to Lipopolysaccharide (LPS)-challenged piglets and its association with integrity of the intestinal barrier coupled with effective dosage were expected to be signified. In the present study, 72 piglets (Landrace × Yorkshiere × Duroc) were randomly allotted to four groups, each group with six replicates. The subjects in the control group were provided with basal diet while those in other tested groups with extra 0.05, 0.1, and 0.2% L. salivarius, respectively. Fourteen days later, LPS was intraperitoneally injected and sodium pentobarbital was then delivered to euthanize those LPS-challenged piglets. An increase of average daily gain and body weight along with an apparent decline of diarrhea rate were observed in L. salivarius-treated groups. Both 0.1 and 0.2% L. salivarius supplement in total diet had the capability to markedly elevate levels of CAT, GSH-Px, SOD, anti-inflammatory cytokine from the serum as well as tight junction proteins (Claudin-1, Occludin, and ZO-1) extracted from intestine in LPS-challenged piglets. These changes were accompanied by the obvious downregulation of D-lactic acid, DAO, MDA and pro-inflammatory mediators in the serum, including IL-1β, IL-6, IFN-γ, and TNF-α. Meanwhile, the expression levels of TLR2 and TLR4 in spleen and mesenteric lymph nodes were significantly lower whereas the oxidation-related gene, ho-1 was up-regulated with L. salivarius administration. Our findings suggested that relatively high dose L. salivarius (0.1–0.2%) could regulate the progression of inflammatory response and oxidative stress when individuals were exposed to LPS, thus probably offering valuable assistance in restoring barrier function and improving overall performance.
Evaluation of the Control Options of Bovine Tuberculosis in Ethiopia Using a Multi-Criteria Decision Analysis
Authors: Gutema Fanta D. , Agga Getahun E. , Makita Kohei , Smith Rebecca L. , Mourits Monique , Tufa Takele B. , Leta Samson , Beyene Tariku J. , Asefa Zerihun , Urge Beksissa , Ameni Gobena
Published: January 2021

Bovine tuberculosis (BTB) is a zoonotic bacterial infection caused by Mycobacterium bovis and is characterized by the development of granulomatous lesions in the lymph nodes, lungs and other tissues. It poses serious public health impacts and food security challenges to the agricultural sector in terms of dairy and meat productions. In Ethiopia, BTB has been considered as a priority disease because of its high prevalence in urban and peri-urban dairy farms. However, there has not been any national control program in the country. Thus, in order to initiate BTB control program in the country, information on control options is needed to tailor the best option for the Ethiopian situation. The objective of this study was to identify, evaluate and rank various BTB control options in Ethiopia using a multi-criteria decision analysis based on preference ranking organization method for enrichment evaluations (PROMETHEE) approach while accounting for the stakeholders' preferences. Control options were evaluated under two scenarios: with (scenario 1) and without (scenario 2) bacillus Calmette–Guérin (BCG) vaccination. Nine potential control options were identified that include combinations of three control options (1) test and slaughter with or without government support, (2) test and segregation, and (3) BCG vaccination. Under scenario 1, BCG vaccination, BCG vaccination and test and slaughter with partial compensation by government, and BCG vaccination and test and slaughter with full compensation by government were the top three ranked control options. Under scenario 2, test and slaughter with full compensation by government was the preferred control option, followed by test and segregation supported by test and slaughter with full government compensation, and test and slaughter with half compensation by government. Irrespective of the variability in the weighting by the stakeholders, the sensitivity analysis showed the robustness of the ranking method. In conclusion, the study demonstrated that BCG vaccination, and test and slaughter with full compensation by government were the two most preferred control options under scenarios 1 and 2, respectively. National level discussions were strongly recommended for further concretization and implementation of these control measures.
Psychological and Physiological Stress in Hens With Bone Damage
Authors: Rokavec Neža , Zupan Šemrov Manja
Published: January 2021

Abnormalities in bone development in humans and non-humans can lead to impaired physical and psychological health; however, evidence is lacking regarding the role of individual psychosocial factors in the development of poor bone conditions. Addressing this lack of knowledge, we used low-productive laying hens (n = 93) and assessed behavioral responses to an open-field test [at 17, 18, 29, 33 weeks of age (wa)], an aerial predator test (at 39 wa), and a social reinstatement test (at 42 wa). Bone condition was assessed using a palpation technique on five occasions (at 16, 29, 33, 45, 58 wa), with half of the hens experiencing damage (deviations, fractures, or both) at 29 wa and all hens by 58 wa. Corticosterone (CORT) concentration in feathers (at 16, 33, 58 wa) and body weight (at 23, 47, 58 wa) were also investigated. We hypothesized that lighter birds (at 23 wa) with higher CORT (at 16 wa) and open field-induced fear collected before the onset of lay (at 17 and 18 wa) are associated with a worse bone condition when in lay. We also hypothesized that those birds with more damage at the peak of laying (at 29 wa) would be lighter at 47 and 58 wa and more fearful by showing higher open field-induced (at 29 and 33 wa) and predator-induced fear responses, however, acting less socially toward conspecifics. These hens were also expected to have higher CORT (at 33 and 58 wa). Our results show no association between open-field fear level and fear behavior, CORT concentration, or body weight on the one hand (all measured before starting to lay) and bone damage at 29 wa on the other. When in lay, bone damage was associated with more pecking and less crossing zones when faced with an open-field situation at 29 wa and improved sociality at 42 wa. This study provides the first evidence of a relationship of bone health with fear, sociality, and stress response. When in poor bone condition, our hens had enhanced psychological stress measured by fear behavior reactivity but not physiological stress measured as feather CORT concentration.
Effect of Different Levels of Energy Diet Restriction on Energy Balance, Leptin and CL Development, Vascularization, and Function in South American Camelids
Authors: Norambuena Cecilia , Hernandez Francisca , Alfaro Jorge , Silva Gonzalo , Topp Shirley , Ratto Marcelo
Published: January 2021

The objective was to determine the effect of energy diet restriction on energy balance, systemic leptin and corpus luteum (CL) vascularization, development, and function in South American camelids. In experiment 1, adult llamas were randomly assigned to receive a diet of 70% of their maintenance energy requirements (MER) (Restricted group, n = 7) or fed ad libitum (Control group, n = 7) during 28 days. Body live weight (BLW) and body condition score (BCS) were recorded, blood samples were collected every 2 weeks to measure plasma leptin concentrations, and energy metabolites were quantified. In experiment 2, adult alpacas were randomly assigned to receive a diet of 40% MER for 21 days (Restricted group, n = 7) or fed ad libitum (Control group, n = 7). Then, ovulation was induced with gonadorelin acetate (day = 0), and trans-rectal ultrasonography (7.5 MHz) was performed using B and Doppler mode to record the diameter of the pre-ovulatory follicle, ovulation, CL diameter, and vascularization from Days 0 to 13. Blood samples were collected every 48 h from Days 1 to 13 to quantify plasma leptin and progesterone concentrations. In experiment 1, energy diet restriction of 70% MER did not affect plasma leptin concentration and metabolic parameters of the Restricted group. In experiment 2, the Restricted group had a lower BCS (p < 0.001), a smaller diameter of the CL on Days 5 and 7 (p < 0.05), and a smaller maximum diameter of the CL (10.2 ± 0.6 mm) than the Control group (12.1 ± 0.6 mm; p = 0.04). Low energy restriction of 70% MER for 28 days did not affect the energy balance of llamas (Experiment 1). Moderate energy restriction of 40% MER for 21 days negatively affected energy balance (BCS), and CL development but not its vascularization, leptin, and progesterone concentrations. These species must be submitted to longer periods or a higher level of energy restriction to impair ovarian function.
The Effect of Intense Exercise on Equine Serum Proteoglycan-4/Lubricin
Authors: Matheson Austyn , Regmi Suresh C. , Jay Gregory D. , Schmidt Tannin A. , Scott W. Michael
Published: January 2021

Objective:Local biological and biomechanical-stimuli modulate proteoglycan-4 secretion within synovial joints. For the horse, changes to proteoglycan-4 concentration and function are notable in acute joint injury and osteoarthritis. Proteoglycan-4 (also known as Lubricin) is present in the blood, however the effect of exercise on equine serum levels is unknown. The overall objective of this study was, therefore, to investigate the effect of intense exercise on serum proteoglycan-4 in thoroughbred horses.

Methods:Samples of blood were taken from thoroughbreds (n= 12) during a chuckwagon racing event (Alberta, Canada). The chuckwagon race is a sprint racing event where teams of horses pull a combined 1,325 lbs (601 kg) of wagon and driver around a 5/8th mile (1 km) of dirt track, racing at full gallop to the finish. Blood samples were collected 30-min before the race start, and several timepoints post-race: 5-min, 90-min, 3-h, 12-h, and 23-h. Proteoglycan-4 concentrations in serum were quantified by enzyme-linked-immunosorbent-assay using recombinant-human proteoglycan-4 standards and anti-proteoglycan-4 mAb 9G3. The molecular weight of immunoreactive proteoglycan-4 in serum was assessed by western blot.

Results:Proteoglyan-4 in serum demonstrated the expected high MW immunoreactivity to mAb 9G3, consistent with that of full length PRG4. Serum proteoglycan-4 decreased five-minutes post-race from baseline concentration (0.815 ± 0.175 to 0.466 ± 0.090 μg/mL, μ ± SEM,p< 0.01).

Conclusions:The concentration of serum proteoglycan-4 in horses decreased significantly five min post-exercise. A potential explanation for this finding could be increased proteoglycan-4 clearance from the circulation. Further investigations could extend to complete the detailed characterization of proteoglycan-4 structure and its potential function within the blood as it relates to joint health and exercise.
High-Zinc Supplementation of Weaned Piglets Affects Frequencies of Virulence and Bacteriocin Associated Genes Among Intestinal Escherichia coli Populations
Authors: Johanns Vanessa C. , Epping Lennard , Semmler Torsten , Ghazisaeedi Fereshteh , Lübke-Becker Antina , Pfeifer Yvonne , Eichhorn Inga , Merle Roswitha , Bethe Astrid , Walther Birgit , Wieler Lothar H.
Published: January 2021

To prevent economic losses due to post-weaning diarrhea (PWD) in industrial pig production, zinc (Zn) feed additives have been widely used, especially since awareness has risen that the regular application of antibiotics promotes buildup of antimicrobial resistance in both commensal and pathogenic bacteria. In a previous study on 179 Escherichia coli collected from piglets sacrificed at the end of a Zn feeding trial, including isolates obtained from animals of a high-zinc fed group (HZG) and a corresponding control group (CG), we found that the isolate collection exhibited three different levels of tolerance toward zinc, i.e., the minimal inhibitory concentration (MIC) detected was 128, followed by 256 and 512 μg/ml ZnCl2. We further provided evidence that enhanced zinc tolerance in porcine intestinal E. coli populations is clearly linked to excessive zinc feeding. Here we provide insights about the genomic make-up and phylogenetic background of these 179 E. coli genomes. Bayesian analysis of the population structure (BAPS) revealed a lack of association between the actual zinc tolerance level and a particular phylogenetic E. coli cluster or even branch for both, isolates belonging to the HZG and CG. In addition, detection rates for genes and operons associated with virulence (VAG) and bacteriocins (BAG) were lower in isolates originating from the HZG (41 vs. 65% and 22 vs. 35%, p < 0.001 and p = 0.002, resp.). Strikingly, E. coli harboring genes defining distinct pathotypes associated with intestinal disease, i.e., enterotoxigenic, enteropathogenic, and Shiga toxin-producing E. coli (ETEC, EPEC, and STEC) constituted 1% of the isolates belonging to the HZG but 14% of those from the CG. Notably, these pathotypes were positively associated with enhanced zinc tolerance (512 μg/ml ZnCl2 MIC, p < 0.001). Taken together, zinc excess seems to influence carriage rates of VAGs and BAGs in porcine intestinal E. coli populations, and high-zinc feeding is negatively correlated with enteral pathotype occurrences, which might explain earlier observations concerning the relative increase of Enterobacterales considering the overall intestinal microbiota of piglets during zinc feeding trials while PWD rates have decreased.
Solid-State 2H NMR Study for Deuterated Phenylene Dynamics in a Crystalline Gyroscope-Like Molecule
Authors: Setaka Wataru , Yamaguchi Kentaro , Kira Mitsuo
Published: January 2021

Molecular rotors have earned substantial popularity in recent times, owing to the unique dependence of its crystalline properties on the rotational dynamics of the rotor. We have recently reported the synthesis and crystal structure of a phenylene-bridged macrocage as a gyroscope-like molecule in the crystalline state. The dynamics of the phenylene moiety was probed by solid-state 13C CP/MAS proton dipolar dephasing NMR spectroscopy. Herein, solid-state 2H NMR studies were performed to study the dynamics of the gyroscope-like molecule with a deuterated rotor in the crystalline state. A spectrum with a narrow line shape was obtained at 300 K. The facile exchange among three stationary states, which was observed by X-ray crystallography, was clearly confirmed. Additionally, a crystal-to-crystal phase transition that switches the motion of the rotor was observed in the DSC analysis of the powdered sample.
Use of Nanoparticle Enhanced Phase Change Material for Cooling of Surface Acoustic Wave Sensor
Authors: Abdollahzadeh Jamalabadi Mohammad Yaghoub
Published: January 2021

The use of resonators, filters, interdigital transducers (IDT) and stable sources in electronic industry is widespread today. One of the most used filters are the surface acoustic wave (SAW) type, which is mostly based on Rayleigh waves propagation on the surface. On the other hand, the use of Phase change materials (PCMs) is considered as a heat sink method in the field of thermal cooling of electronic circuits. Recent development in heat transfer is obtained by nanoparticle-enhanced PCM (NEPCM), which is a result of combining nanoparticles with PCMs. Increase of thermal conductivity of NEPCM in comparison with common PCM enhances the heat transfer rate. The aim of the current study is thermal management of SAW for the application of high frequency heating by phase change material. Melting of NEPCMs inside a rectangular cavity next to the SAW cell is used for the cooling purpose. Free convection heat transfer of a NEPCMs in an square cavity is modeled throughout the mass and momentum. Energy governing equations are solved by using the finite element method. Electrohydrodynamic (EHD) forces exist in natural convection heat transfer within the fluid part of the enclosure. The results also show that the NEPCM causes heat transfer improvement up to 10%.
New Advances in Marine Engineering Geology
Authors: Liu Xiaolei , Yang Qing , Wang Yin , Jeng Dong-Sheng , Sturm Hendrik
Published: January 2021

The ocean is the cradle of life and is rich in natural resources [...]
Polymorphisms in Pepsinogen C and miRNA Genes Associate with High Serum Pepsinogen II in Gastric Cancer Patients
Authors: De Re Valli , Zorzi Mariangela , Caggiari Laura , Repetto Ombretta , Brisotto Giulia , Magris Raffaela , Zanussi Stefania , Steffan Agostino , Cannizzaro Renato
Published: January 2021

Background: Pepsinogen (PG) II (PGII) is a serological marker used to estimate the risk of gastric cancer but how PGII expression is regulated is largely unknown. It has been suggested that PGII expression, from the PGC (Progastricsin) gene, is regulated by microRNAs (miRNA), but how PGII levels vary with Helicobacter pylori (H. pylori) infection and miRNAs genotype remains unclear. Methods: Serum levels of PGI and PGII were determined in 80 patients with gastric cancer and persons at risk for gastric cancer (74 first-degree relatives of patients, 62 patients with autoimmune chronic atrophic gastritis, and 2 patients with dysplasia), with and without H. pylori infection. As control from the general population, 52 blood donors were added to the analyses. Associations between PGII levels and genetic variants in PGC and miRNA genes in these groups were explored based on H. pylori seropositivity and the risk for gastric cancer. The two-dimensional difference in gel electrophoresis (2D-DIGE) and the NanoString analysis of messenger RNA (mRNAs) from gastric cancer tissue were used to determine the pathways associated with increased PGII levels. Results: PGII levels were significantly higher in patients with gastric cancer, and in those with H. pylori infection, than in other patients or controls. A PGI/PGII ratio ≤ 3 was found better than PGI < 25 ng/mL to identify patients with gastric cancer (15.0% vs. 8.8%). For two genetic variants, namely rs8111742 in miR-Let-7e and rs121224 in miR-365b, there were significant differences in PGII levels between genotype groups among patients with gastric cancer (p = 0.02 and p = 0.01, respectively), but not among other study subjects. Moreover, a strict relation between rs9471643 C-allele with H. pylori infection and gastric cancer was underlined. Fold change in gene expression of mRNA isolated from gastric cancer tissue correlated well with polymorphism, H. pylori infection, increased PGII level, and pathway for bacteria cell entry into the host. Conclusions: Serum PGII levels depend in part on an interaction between H. pylori and host miRNA genotypes, which may interfere with the cut-off of PGI/PGII ratio used to identify persons at risk of gastric cancer. Results reported new findings regarding the relation among H. pylori, PGII-related host polymorphism, and genes involved in this interaction in the gastric cancer setting.
Utilizing Amino Acid Composition and Entropy of Potential Open Reading Frames to Identify Protein-Coding Genes
Authors: McNair Katelyn , Ecale Zhou Carol L. , Souza Brian , Malfatti Stephanie , Edwards Robert A.
Published: January 2021

One of the main steps in gene-finding in prokaryotes is determining which open reading frames encode for a protein, and which occur by chance alone. There are many different methods to differentiate the two; the most prevalent approach is using shared homology with a database of known genes. This method presents many pitfalls, most notably the catch that you only find genes that you have seen before. The four most popular prokaryotic gene-prediction programs (GeneMark, Glimmer, Prodigal, Phanotate) all use a protein-coding training model to predict protein-coding genes, with the latter three allowing for the training model to be created ab initio from the input genome. Different methods are available for creating the training model, and to increase the accuracy of such tools, we present here GOODORFS, a method for identifying protein-coding genes within a set of all possible open reading frames (ORFS). Our workflow begins with taking the amino acid frequencies of each ORF, calculating an entropy density profile (EDP), using KMeans to cluster the EDPs, and then selecting the cluster with the lowest variation as the coding ORFs. To test the efficacy of our method, we ran GOODORFS on 14,179 annotated phage genomes, and compared our results to the initial training-set creation step of four other similar methods (Glimmer, MED2, PHANOTATE, Prodigal). We found that GOODORFS was the most accurate (0.94) and had the best F1-score (0.85), while Glimmer had the highest precision (0.92) and PHANOTATE had the highest recall (0.96).
Effects of Spartina alterniflora Invasion on Soil Microbial Community Structure and Ecological Functions
Authors: Cao Minmin , Cui Lina , Sun Huimin , Zhang Xiaomian , Zheng Xiang , Jiang Jiang
Published: January 2021

It has been reported that the invasion of Spartina alterniflora changed the soil microbial community in the mangrove ecosystem in China, especially the bacterial community, although the response of soil fungal communities and soil microbial ecological functions to the invasion of Spartina alterniflora remains unclear. In this study, we selected three different communities (i.e., Spartina alterniflora community (SC), Spartina alterniflora–mangrove mixed community (TC), and mangrove community (MC)) in the Zhangjiangkou Mangrove Nature Reserve in China. High-throughput sequencing technology was used to analyze the impact of Spartina alterniflora invasion on mangrove soil microbial communities. Our results indicate that the invasion of Spartina alterniflora does not cause significant changes in microbial diversity, but it can alter the community structure of soil bacteria. The results of the LEfSe (LDA Effect Size) analysis show that the relative abundance of some bacterial taxa is not significantly different between the MC and SC communities, but different changes have occurred during the invasion process (i.e., TC community). Different from the results of the bacterial community, the invasion of Spartina alterniflora only cause a significant increase in few fungal taxa during the invasion process, and these taxa are at some lower levels (such as family, genus, and species) and classified into the phylum Ascomycota. Although the invasion of Spartina alterniflora changes the taxa with certain ecological functions, it may not change the potential ecological functions of soil microorganisms (i.e., the potential metabolic pathways of bacteria, nutritional patterns, and fungal associations). In general, the invasion of Spartina alterniflora changes the community structure of soil microorganisms, but it may not affect the potential ecological functions of soil microorganisms.
The Epichloë festucae Antifungal Protein Efe-AfpA Is also a Possible Effector Protein Required for the Interaction of the Fungus with Its Host Grass Festuca rubra subsp. rubra
Authors: Wang Ruying , Luo Simin , Clarke Bruce B. , Belanger Faith C.
Published: January 2021

Strong creeping red fescue (Festuca rubra subsp. rubra) is a commercially important low-maintenance turfgrass and is often naturally infected with the fungal endophyte Epichloëfestucae. Epichloë spp. are endophytes of several cool-season grass species, often conferring insect resistance to the grass hosts due to the production of toxic alkaloids. In addition to insect resistance, a unique feature of the strong creeping red fescue/E. festucae symbiosis is the endophyte-mediated disease resistance to the fungal pathogen Clarireedia jacksonii, the causal agent of dollar spot disease. Such disease resistance is not a general feature of other grass/ Epichloë interactions. E. festucae isolates infecting red fescue have an antifungal protein gene Efe-afpA, whereas most other Epichloë spp. do not have a similar gene. The uniqueness of this gene suggests it may, therefore, be a component of the unique disease resistance seen in endophyte-infected red fescue. Here, we report the generation of CRISPR-Cas9 Efe-afpA gene knockouts with the goal of determining if absence of the protein in endophyte-infected Festuca rubra leads to disease susceptibility. However, it was not possible to infect plants with the knockout isolates, although infection was possible with the wild type E. festucae and with complemented isolates. This raises the interesting possibility that, in addition to having antifungal activity, the protein is required for the symbiotic interaction. The antifungal protein is a small secreted protein with high expression in planta relative to its expression in culture, all characteristics consistent with effector proteins. If Efe-AfpA is an effector protein it must be specific to certain interactions, since most Epichloë spp. do not have such a gene in their genomes.
Bacteria Broadly-Resistant to Last Resort Antibiotics Detected in Commercial Chicken Farms
Authors: Jochum Jared M. , Redweik Graham A. J. , Ott Logan C. , Mellata Melha
Published: January 2021

Resistance to last resort antibiotics in bacteria is an emerging threat to human and animal health. It is important to identify the source of these antimicrobial resistant (AMR) bacteria that are resistant to clinically important antibiotics and evaluate their potential transfer among bacteria. The objectives of this study were to (i) detect bacteria resistant to colistin, carbapenems, and β-lactams in commercial poultry farms, (ii) characterize phylogenetic and virulence markers of E. coli isolates to potentiate virulence risk, and (iii) assess potential transfer of AMR from these isolates via conjugation. Ceca contents from laying hens from conventional cage (CC) and cage-free (CF) farms at three maturity stages were randomly sampled and screened for extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae, carbapenem-resistant Acinetobacter (CRA), and colistin resistant Escherichia coli (CRE) using CHROMagar™ selective media. We found a wide-spread abundance of CRE in both CC and CF hens across all three maturity stages. Extraintestinal pathogenic Escherichia coli phylogenetic groups B2 and D, as well as plasmidic virulence markers iss and iutA, were widely associated with AMR E. coli isolates. ESBL-producing Enterobacteriaceae were uniquely detected in the early lay period of both CC and CF, while multidrug resistant (MDR) Acinetobacter were found in peak and late lay periods of both CC and CF. CRA was detected in CF hens only. blaCMY was detected in ESBL-producing E. coli in CC and CF and MDR Acinetobacter spp. in CC. Finally, the blaCMY was shown to be transferrable via an IncK/B plasmid in CC. The presence of MDR to the last-resort antibiotics that are transferable between bacteria in food-producing animals is alarming and warrants studies to develop strategies for their mitigation in the environment.
The Use of MALDI-TOF Mass Spectrometry to Analyze Commensal Oral Yeasts in Nursing Home Residents
Authors: Lu Jang-Jih , Lo Hsiu-Jung , Lee Chih-Hua , Chen Mei-Jun , Lin Chih-Chao , Chen Yin-Zhi , Tsai Ming-Horng , Wang Shao-Hung
Published: January 2021

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a rapid and accurate method to identify microorganisms in clinical laboratories. This study isolates yeast-like microorganisms in the oral washes that are collected from non-bedridden nursing home residents, using CHROMagar Candida plates, and identifies them using Bruker MALDI-TOF MS. The ribosomal DNA sequences of the isolates are then examined. Three hundred and twenty yeast isolates are isolated from the oral washes. Candida species form the majority (78.1%), followed by Trichosporon/Cutaneotrichosporon species (8.8%). Bruker MALDI-TOF MS gives a high-level confidence, with a log(score) value of ≥1.8, and identifies 96.9% of the isolates. There are six inconclusive results (1.9%), and those sequences are verified as rare clinical species, including Candida ethanolica, Cutaneotrichosporon jirovecii, Exophiala dermatitidis, and Fereydounia khargensis. Almost all of the isolates have a regular color on the CHROMagar Candida plates. If the colonies are grouped by color on the plates, a specific dominant yeast species is present in each color group, except for purple or orange isolates. In conclusion, MALDI-TOF MS is verified as a fast, accurate and practical method to analyze oral yeasts in elderly subjects.
Evolutionary Genetics of Mycobacterium Tuberculosis and HIV-1: “The Tortoise and the Hare”
Authors: Santos-Pereira Ana , Magalhães Carlos , Araújo Pedro M. M. , Osório Nuno S.
Published: January 2021

The already enormous burden caused by Mycobacterium tuberculosis and Human Immunodeficiency Virus type 1 (HIV-1) alone is aggravated by co-infection. Despite obvious differences in the rate of evolution comparing these two human pathogens, genetic diversity plays an important role in the success of both. The extreme evolutionary dynamics of HIV-1 is in the basis of a robust capacity to evade immune responses, to generate drug-resistance and to diversify the population-level reservoir of M group viral subtypes. Compared to HIV-1 and other retroviruses, M. tuberculosis generates minute levels of genetic diversity within the host. However, emerging whole-genome sequencing data show that the M. tuberculosis complex contains at least nine human-adapted phylogenetic lineages. This level of genetic diversity results in differences in M. tuberculosis interactions with the host immune system, virulence and drug resistance propensity. In co-infected individuals, HIV-1 and M. tuberculosis are likely to co-colonize host cells. However, the evolutionary impact of the interaction between the host, the slowly evolving M. tuberculosis bacteria and the HIV-1 viral “mutant cloud” is poorly understood. These evolutionary dynamics, at the cellular niche of monocytes/macrophages, are also discussed and proposed as a relevant future research topic in the context of single-cell sequencing.