Artocarpus hirsutus Lam. is a tropical evergreen recalcitrant tree species. The seed development is marked by the histodifferentiation phase which is highly complicated by the interaction of a variety of proteins involved in embryo expansion and differentiation. For a better understanding of the physiology and biochemistry of this process, protein expression profiling was carried out by SDS-PAGE followed by LC-MS/MS technique. 410 proteins could be identified from the spectral data belonging to different classes viz. cell division, metabolism, stress, transport, folding, hormone metabolism etc. Simultaneous analysis of Plant growth regulators (PGRs) by LC-MS/MS technique revealed that IAA, GA4, trans-Zeatin (tZ), ABA, Jasmonic acid (JA), Epibrasinolide (24epiBl) and salicylic acid (SA) were found to be in elevated levels in the young embryos. However, proteome analysis only showed JA biosynthetic proteins. Results indicate that most of the PGRs involved in histodifferentiation process of recalcitrant embryo development is provided by the maternal tissue rather than de-novo synthesis in young embryo, a possible answer to a much debated question. Comparison of seeds to analyse protein expression during histodifferentiation in Artocarpus hirsutus with Phaseolus vulgaris showed almost similar processes are happening in both the species. However the accumulation of some proteins showed different pattern.

Enzymatic catalysis is one of the fundamental processes that drives the dynamic landscape of post-translational modifications (PTMs), expanding the structural and functional diversity of proteins. Here, we assessed enzyme specificity using a top-down ion mobility spectrometry (IMS) and tandem mass spectrometry (MS/MS) workflow. We successfully applied trapped IMS (TIMS) to investigate site-specific N-ε-acetylation of lysine residues of full-length histone H4 catalyzed by histone lysine acetyltransferase KAT8. We demonstrate that KAT8 exhibits a preference for N-ε-actylation of residue K16, while also installing N-ε-acetyl groups on residues K5 and K8 as the first degree of acetylation. Achieving TIMS resolving power values of up to 300, we fully separated mono-acetylated regioisomers (H4K5ac, H4K8ac, and H4K16ac). Each of these regioisomers produce unique MS/MS fragment ions, enabling estimation of their individual mobility distributions and the exact localization of the N-ε-acetylation sites. This study highlights the potential of top-down TIMS-MS/MS for conducting enzymatic assays at the intact protein level and, more generally, for separation and identification of isomeric proteoforms and precise PTM localization.

Citation mining, citation searching or snowball searches have been recommended as a supplementary search method in the conduct of systematic searches for evidence retrieval as part of systematic review production. However, manual methods are extremely costly and time-consuming, with limited empirical evidence for their utility, and limited guidance on how best to incorporate the method during systematic review production. Encouragingly, the advent of programmatic access to bibliographic databases has enabled exploration of automated citation mining for a potentially scalable and replicable approach. Thus, the study aims to simulate and evaluate the use of exclusively automated citation searching methods for evidence retrieval compared to reference standard boolean logic-based methods, and to explore the factors that influence performance. Methods: A total of 30 systematic reviews will be retrieved from the Cochrane Database of Systematic Reviews, Campbell Systematic Reviews and the Collaboration for Environmental Evidence (CEE). Baseline characteristics will be extracted, including the performance of the reference standard boolean search strategy in terms of recall, precision and F(1-3)-score for each sample review. Seed articles from the background and methods section of each sample review and their baseline characteristics will then be extracted, and automated citation searching will be conducted for different seed article and database combinations (Semantic Scholar, OpenAlex). Each seed article candidate will be ranked according to recall, and the top 10 seed articles will be combined in all possible combinations and evaluated. The end performance of automated citation searching will then be compared against the reference standard Boolean strategy for each sample review. The association of factors related to i) automated citation search parameters, ii) characteristics related to review question, and iii) characteristics related to the initial starting set of seed articles will be evaluated. Empirical guidance surrounding the use of automated citation searching will then be generated.

A document by Simone Fontana. Click on the document to view its contents.

Four different types of specimens were prepared from the center and the periphery of two large IN706 forged discs of commercial scale, and low cycle fatigue (LCF) tests were conducted at 650 oC. The IN706 specimens with lower LCF lives showed relatively large fraction of cleavage-like fracture along linearly aligned η (Ni 3Ti) precipitates in the area of crack propagation. The amount of η precipitates for those specimens with greater LCF lives was negligible, and the fracture mode of crack propagation was dominantly intergranular. Crack initiation was mainly by persistent slip band (PSB) cracking at the surface, and no notable difference was found for each specimen. The correlation between tensile properties, grain size and LCF lives of IN706 at 650 oC appeared to be not as significant as expected.

Background: Mycoplasma pneumoniae pneumonia (MPP) is common among children, but the impact of atopy on MPP severity in children is unknown. This study investigated whether atopic vs. nonatopic children had greater MPP severity. Methods: Of 539 children (ages 3-14 years) diagnosed with MPP between January 2018 and December 2021 in the First Affiliated Hospital of Anhui Medical University and enrolled in this study, 195 were atopic and 344 were nonatopic. Of them, 204 had refractory MPP, and 335 had general MPP. Data on demographic and clinical characteristics, laboratory findings, clinical treatments, lung function, and fibrobronchoscopy results were analyzed. Results: Significantly more boys with MPP were atopic than nonatopic ( P<0.05). More atopic (than nonatopic) children presented with prolonged fever and hospitalization, severe extra-pulmonary complications, steroid treatment, wheezing, and increased IgE levels (all P<0.05). In atopic (vs. nonatopic) children with MPP, the incidence of mucosal inflammation with lymphoid follicular hyperplasia and segmental pneumonia was significantly increased and required bronchoscopy-assisted and steroid therapy. Compared with nonatopic children, more atopic children developed refractory MPP ( P<0.05). Prolonged fever and hospitalization, severe extra-pulmonary complications, lymphocyte count, procalcitonin and lactate dehydrogenase levels, and percentages of atopy were all significantly higher ( P<0.05) among children with refractory MPP vs. general MPP. Spearman correlation analysis showed strong associations between atopy and male sex, length of hospital stay, fever duration, IgE level, wheezing, segmental pneumonia, refractory MPP, and treatment with hormones or bronchoscopy ( P<0.05). Conclusions: Atopy may be a risk factor for and was positively correlated with the severity of MPP in children.

Introduction: Elective flexible bronchoscopy (FB) is now widely available and standard practice for a variety of indications in children with respiratory conditions. However, there is limited evidence regarding the utility of elective FB in children. This systematic review aimed to determine the utility of FB on its impact in clinical decision making and patient-reported outcome measures (PROMs). Methods: We searched Pubmed, Cochrane central register of controlled trials, Embase, World Health Organisation Clinical Trials Registry Platform and Cochrane database of systematic reviews from inception to 20 th April 2023. We included systematic reviews (SRs) and randomised controlled trials (RCTs) that used parallel group design (comparing use of elective FB versus no FB, or a wait-list approach [early FB versus usual wait FB]) in children aged ≤ 18 years. Our protocol was prospectively registered and used Cochrane methodology for systemic reviews of interventions. Results: Our search identified 859 articles; 102 duplicates were removed, and 753 articles were excluded by title and abstract. Four full text articles were reviewed and subsequently excluded, as none met the inclusion criteria outlined in our PICO framework. Conclusions: Given the paucity of data, there is need for high-quality evidence to support the routine use of elective FB in children with respiratory conditions. We recommend clinicians adhere to current international guidelines until such evidence emerges. Registration: PROSPERO CRD42021291305.

Objective: Home pulse oximetry is often prescribed to children with chronic disease upon hospital discharge. Children monitored at home may generate >20 alarms every 8 hours, contributing to premature discontinuation of monitoring. We aimed to improve the home oximetry ordering process using clinical decision support (CDS), supporting more liberal oxygen saturation (SpO 2) alarm limits. Methods: Within a large single-center improvement project to increase informativeness of alarms in the hospital and in patients’ homes, we compared home care oximetry orders of discharged children pre-post CDS implementation. Order parameters included low SpO 2 limit, specification of intensity of use, an intervention plan, pulse oximetry probe prescription, and order completeness. We extracted order details 6 months pre-CDS and 6 months post-CDS with a one-month washout period. The CDS intervention used a letter template to include all required home oximeter order elements and provide more liberal age-specific default alarm limits. Results: There were 100 orders in the pre-CDS epoch (7/1/2021-12/31/2021) and 112 orders in the post-CDS epoch (2/1/2022-7/31/2022). The median low SpO 2 alarm limit post-CDS implementation (87%, IQR 87%-90%) was significantly lower than pre-CDS (90%, IQR 90%-90%, p=<0.001). In the post-CDS epoch significantly more orders included an intervention plan (80.4% versus 31%, p<0.001), prescribed pulse oximeter probes (85.7% versus 52.0%, p<0.001), and were complete (68.8% versus 13.0%, p<0.001). Conclusions: CDS implementation resulted in a significant decrease in median low SpO 2 limit and a significant increase in home oximetry order completeness. These changes may decrease home oximetry alarm burden and improve caregiver experiences with home oximetry.

We believe that the data in this letter clearly demonstrate that even with CFTR2 expansion to 719 variants, striving to achieve equity of early diagnosis of CF via screening requires states to perform a sweat test in all infants with a high IRT level and one identified CFTR variant. This recommended policy can be debated but sweat testing overload should not be the argued as the barrier and CF specialists need to recognize that CFTR2 may never include all of the very rare, “private” pathogenic variants nor will next generation sequencing cover the structural variants such as deletions and duplications.

As we know life expectancy in CP can improve by both preventive measurements as initiation of early therapy concerning respiratory morbidity. The prevalence of PA in this prospective study of children with CP is low, gram-negative bacteria were most commonly found. Therefore, it is recommended to repeat and expand this study since the prevalence of respiratory tract infections is again increasing in the post-covid era.

Children with home invasive mechanical ventilation face numerous difficulties, including endocrine problems that can arise as a consequence of their condition. Endocrine problems seen in children treated with HIMV may develop due to the underlying disease, drugs used, or prolonged mechanical ventilation. This manuscript will discuss the most common endocrine problems encountered in children with home invasive mechanical ventilation, including problems in glucose metabolism, thyroid dysfunction, bone metabolism, adrenal dysfunctions, growth and puberty. Close monitoring, multidisciplinary care, and regular assessments are essential to optimize the endocrin system functions of children requiring HMV. By understanding these complications, it can develop effective management strategies to optimize the health and well-being of these vulnerable individuals.

Molnupiravir (also known as EIDD-2801/MK-4482), which is used as an antiviral drug has its mechanism of action by incorporating into the viral genome increasing errors, mismatching, and misdirecting the viral polymerase, leading to the accumulation of deleterious errors and halting viral RNA replication of SARS-CoV-2 and other RNA viruses. Our meta-analysis aims to evaluate virology profile, and adverse effects associated with the use of molnupiravir on a large patient population. Following PRISMA guidelines performed a thorough literature search of electronic and medical databases (MEDLINE and Cochrane CENTRAL) from their establishment to January 2023 without any limitations on time, language, or sample size. The random effects model was utilized to calculate the weighted mean difference (WMD) and its associated 95% confidence intervals (CIs) to pool continuous outcomes of interest. Using a random effects model, odds ratio, and accompanying 95% confidence intervals (CIs). Molnupiravir 800 mg at day 5 is significant in creating viral RNA error rate (WMD: 4.91; 95% CI; [1.19, 8.63] p=0.01; I 2=0%). (P-value for subgroup differences = 0.05). A significant outcome was reported with 400mg molnupiravir (WMD: 2.27; 95% CI; 2.27 [0.50, 4.65] p=0.02; I 2=0%). Significant outcome for mean change in SARS-COV-2 RNA viral load from baseline in nasopharyngeal sample at 800 mg molnupiravir on day 3 (WMD: -0.22; 95% CI; [-0.35,-0.08] p=0.002; I 2=0%), day 5 (WMD: -0.32; 95% CI; [-0.53,-0.11] p=0.003; I 2=24%) and overall pooled analysis (WMD: -0.17; 95% CI; [-0.29, 0.33] p=0.003; I 2=32%). Similarly for 400 mg at DAY 5 and overall analysis comparing the molnupiravir group to the placebo group, a significant reduction in viral RNA load was seen from baseline. (WMD: -0.46; 95% CI; [-0.77,-0.15] p=0.004; I 2=0%), (WMD: -0.28; 95% CI; [-0.49,-0.07] p=0.009; I 2=0%). Molnupiravir 400mg significantly reduced the incidence of death as compared to the placebo group. (RR: 0.17; 95% CI; [0.07, 0.43] p=0.0002; I 2=0%). In our meta-analysis, we conclude that molnupiravir is effective in treating SARS-COV-2 patients with respect to eliminating the virus from the host through their mechanism of action. Thereby, widely used and appropriate to treat SARS-COV-2.

This review explores interpersonal emotion regulation and its implications for work performance. It provides a comprehensive review of the literature, encompassing the definition and conceptualization of interpersonal emotion regulation, models and theories explaining its relationship with work performance, key constructs, and mechanisms involved, factors influencing its effectiveness at individual, organizational, and social levels, as well as methodological approaches used in studying this phenomenon. The study highlights the positive effects of interpersonal emotion regulation, including enhanced emotional well-being, improved job satisfaction, increased task performance and productivity, and enhanced teamwork and collaboration. It also discusses the negative effects, such as emotional labor and burnout, suppression of authentic emotions, and strain on interpersonal relationships and team dynamics. Practical implications for organizations and managers are presented, emphasizing strategies for promoting positive interpersonal emotion regulation and the importance of employee well-being and job satisfaction. The study concludes by suggesting future research directions, including developing context-specific models, exploring mediating and moderating factors, and evaluating intervention programs.

The main proteases (M pro) are highly conserved cysteine-rich proteins that can be covalently modified by numerous natural and synthetic compounds. Herein, we constructed an integrative approach to efficiently discover covalent inhibitors of M pro from complex herbal matrices. This work begins with biological screening of sixty clinically used antiviral herbal medicines, among which Lonicera japonica (LJ) demonstrated the strongest anti-M pro effect (IC 50 = 37.82 μg/mL). Mass spectrometry-based chemical analysis and chemoproteomic profiling revealed that LJ extract contains at least 50 constituents, of which 22 exhibited the capability to covalently modify M pro. We subsequently verified the anti-M pro effects of these covalent binders. Gallic acid and quercetin were found to potently inhibit SARS-CoV-2 M pro in dose- and time- dependent manners, with the IC 50 values below 10 µM. The inactivation kinetics, binding affinity and binding mode of gallic acid and quercetin were further characterized by fluorescence resonance energy transfer, surface plasmon resonance, and covalent docking simulations. Overall, this study established a practical approach for efficiently discovering the covalent inhibitors of M pro from herbal medicines by integrating target-based high-throughput screening and mass spectrometry-based assays, which would greatly facilitate the discovery of key anti-viral constituents from medicinal plants.

Considering that individuals psychological adjustment is negatively affected after traumatic events such as earthquakes, it is important to examine the effect of the need for information about negative news on individuals mental well-being. The aim of this study is to examine the effect of psychological adjustment on mental well-being and to investigate the mediating role of doomscrolling and emotion dysregulation. A total of 407 Turkish adults (264 women, 143 men) participated in the study. The average age of the participants is about 35 years old. Participants completed scales for psychological adjustment, doomscrolling, emotion dysregulation, and mental well-being. Analysis was conducted using structural equation modeling (SEM). The mediating roles of doomscrolling and emotion dysregulation in the relationship between psychological adjustment and mental well-being were examined. Doomscrolling and emotion dysregulation fully mediated psychological adjustment and mental well-being. The results, psychological adjustment, mental well-being, doomscrolling and emotion dysregulation were discussed in line with the information in the literature.

According to the IPCC, the observed and projected changes in temperature and precipitation patterns are changing. There is a need to know the probable states of vegetation in the future to ensure meeting the goals of sustainable development of productive activities promoted by the United Nations. Our model species, Rubus geoides Sm, is distributed in many places in the southern Patagonia region. Since the first European settlements the distribution of this species has become more restricted, because of overgrazing and the change of land use by mining, forestry, livestock, expansion of cities, and the interest in functional and nutraceutical food that makes people harvest fruits directly from natural populations. We use a multidisciplinary approach by the union of scientific knowledge from morphology, genetics, and niche modeling to establish the dynamics of the species in the past and under future climate change scenarios. This effort can help determine the state of vegetation resources and give them the tools for decision-makers for conservation and management efforts and their food potential under climate change scenarios. Our results showed a significant difference in blooming among populations revealed by gene flow from specific populations. Also, we observed moderate genetic differentiation among populations, mixed ancestry in some populations, private alleles in specific sites, and species expansion demonstrated by the genetic analysis and the potential distribution model. This paper discusses the evolution of this species, considering the dispersal after the last glacial maximum (LGM) and which challenges we have in front of the change in land use and the future of this species in this changing climate.

Phenotypic characterization of the Rwandan stinging nettle (Urtica massaica Mildbr.) with emphasis on leaf morphological differences. Authors : Nduwamungu, J.1, Munyandamutsa, P.1, Senyanzobe, J.M.V.1, Ruhimbana, C.1, Ugirabe, M.A1, Mahoro, J1., Dusingize, M.C1., Kabarungi, M.1, Irimaso, E.1, Maniraho, E1., Nsabimana, P.1, Mugunga, C.1, and Mugemangango, C.2 1. College of Agriculture, Animal Sciences and Veterinary Medicine 2. College of Sciences and Technology. Corresponding author: nzobe2020@gmail.com Abstract Patterns of intraspecific variation based on environmental conditions in which populations live may reflect adaptive responses to their habitat. The Rwandan stinging nettle plant grows in most parts of Rwanda both in the wild and domestication forms. While the plant can easily be identified through its leaves and life form, it has been observed that the leaf morphology slightly varied from one region to another. This study aimed to investigate morphological variations, particularly in leaf morphology of the Rwandan stinging nettle growing in the lowland, midland, and highland. Specimens of the stinging nettle were taken from different sites located in the three altitudinal zones. Plant heights and leaf lengths varied from one site to another and the statistical analysis revealed that the average plant heights, as well as leaf lengths of mature stinging nettle samples from highland, midland, and lowland, were significantly different. The results showed that there were morphological differences, particularly in leaves among the three altitudinal zones. The most prominent difference was in the main vein of the stinging nettle. Changes in leaf morphology can be linked to differences in environment and nutrient availability between the three habitats which could have enabled the species to evolve differently. However, the genetic basis of these phenotypic changes needs to be examined in future research to establish their heritability for future populations of the stinging nettle plant in Rwanda. Key words: Morphometrics, stinging nettle, traits, habitat, Rwanda.

Background and Purpose. The sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a) depression substantially contributes to diastolic dysfunction in heart failure (HF), suggesting that SERCA2a stimulation may be a “causal” HF therapy. Istaroxime is a drug endowed with both a SERCA2a stimulatory activity and a Na+/K+ pump inhibitory activity for the acute HF treatment. Its main metabolite PST3093 shows a more favorable therapeutic profile as compared to the parent drug, but it is still unsuitable for chronic usage. Novel PST3093 derivatives have been recently developed for oral (chronic) HF treatment; compound 8 was selected among them and here characterized. Experimental Approach. Effects of compound 8 were evaluated in a context of SERCA2a depression, by using streptozotocin-treated rats, a well-known model of diastolic dysfunction. The impact of SERCA2a stimulation by compound 8 was assessed both at the cellular level ad in vivo, following i.v. infusion (acute effects) or oral administration (chronic effects). Key Results. As expected from SERCA2a stimulation, compound 8 induced SR Ca2+ compartmentalization in STZ myocytes. In-vivo echocardiographic analysis during i.v. infusion and after repeated oral administration of compound 8, detected a significant improvement of diastolic function. Moreover, compound 8 did not affect electrical activity of healthy guinea-pig myocytes, in line with the absence of off-target effects. Finally, compound 8 was well tolerated in mice with no evidence of acute toxicity. Conclusion and Implications. The pharmacological evaluation of compound 8 indicates that it may be a safe and selective drug for a “causal” treatment of chronic HF by restoring SERCA2a activity.

Gut microbes play an essential role in the nutritional metabolism of insects. In particular, they are critical to host health when challenged with toxic substances, pathogens, or parasites. Therefore, gut microbiota is a potential resource for developing microbial products for pest control and management. Insect gut microbiota vary across the life cycle, and successful application of microbes for pest control requires precision in specific insect developmental stages. However, the dynamic processes of gut microbiota are poorly understood, and the complexity of ecological factors affecting gut microbiota and the nature of the interactions have been little studied in natural populations. In this study, we combined quantitative characterisation of microbial community diversity, taxonomic analysis of microbial communities, and functional prediction to explore gut microbiota dynamics of Locusta migratoria throughout its life cycle. Proteobacteria and Firmicutes were the dominant phyla throughout the life stages of Locusta migratoria; Enterobacter, Pseudocitrobacter, and Enterococcus were consistently present throughout all life stages. Weissella, which is associated with grasshopper aggregation, was significantly higher in L1 and L2 than in the other life stages. In addition, we analysed the functions of the dominant bacterial flora of Locusta migratoria to screen for new control grasshopper gut bacteria. Identifying and interpreting the various ecological sources that shape individual gut bacteria is essential for understanding the role of gut microbiota in animal health. Moreover, the results of this study provide potential targets for future control of harmful grasshoppers and lays the theoretical foundation for pest control.

A document by Eliza Khwairakpam. Click on the document to view its contents.

Distributed algorithms involving quantization have received a lot of interest recently as the quantized communication appears naturally in real applications. For such algorithms, it is non-trivial to select appropriate stepsizes for high performance, due to presence of the noise effect induced from quantization. In this paper, we establish new convergence results for the quantized decentralized gradient descent and we propose a novel strategy of choosing the stepsizes for the high performance of the algorithm. Precisely, under the strongly convexity assumption on the aggregate cost function and the smoothness assumption on each local cost function, we prove the algorithm converges exponentially fast to a small neighborhood of the optimizer whose radius depends on the stepsizes. Then, based on our convergence result, we suggest an effective stepsize selection algorithm which repeats diminishing the stepsizes after a number of specific iterations by a certain rule. Both the convergence results and the effectiveness of the suggested stepsize selection are also verified by the numerical experiments.

Hollow fiber-based membrane filtration has emerged as the dominant technology for cell retention in perfusion processes yet significant challenges in alleviating filter fouling remain unsolved. In this work, the benefits of co-current filtrate flow applied to a tangential flow filtration (TFF) module to reduce or even completely remove Starling recirculation caused by the axial pressure drop within the module was studied by pressure characterization experiments and perfusion cell culture runs. Additionally, a novel concept to achieve alternating Starling flow within unidirectional TFF was investigated. Pressure profiles demonstrated that precise flow control can be achieved with both lab-scale and manufacturing scale filters. TFF systems with co-current flow showed up to 40% higher product sieving compared to standard TFF. The decoupling of transmembrane pressure from crossflow velocity and filter characteristics in co-current TFF alleviates common challenges for hollow-fiber based systems such as limited crossflow rates and relatively short filter module lengths, both of which are currently used to avoid extensive pressure drop along the filtration module. Therefore, co-current filtrate flow in unidirectional TFF systems represents an interesting and scalable alternative to standard TFF or alternating TFF operation with additional possibilities to control Starling recirculation flow.

The research investigates the usage of fins fastened to a solar panel’s rear surface for passive cooling purposes. In the research, solar modules with air conditioning were compared to modules with connected fins acting as a heat sink to cool them. The former served as the benchmark for comparison. The research explores two approaches for cooling modules with fins: ventilation air and still air. The research looked at the heat transfer by photovoltaic panels with and without fins to examine the effects of the environment and solar radiation on the performance of solar cells. The inquiry was carried out ostensibly. According to the findings, the use of fins for cooling reduced cell temperature and improved electrical and thermal efficiency. The estimated performance values of PVT solar cells obtained by using the COMSOL software and the experimental tests carried out during the daytime were reasonably in accord. At midday, the cell temperature decreased by a maximum of 2.8 C o, while electrical and thermal efficiency increased by a maximum of 18.2% and 55.13%, respectively. Good agreement between experimental and numerical results.

In this paper, an adaptive fuzzy event-triggered control (ETC) problem of switched nonlinear systems (SNSs) under actuator failures is studied. Fuzzy logic systems (FLSs) are introduced to approximate the unknown nonlinear terms of the system. To reduce the wastage of communication resources (WCRs), an adaptive fuzzy event-triggered mechanism (ETM) with a state-dependent switching law is designed. In order to ensure the stability of whole systems, the convex combination technology is adopted. The dynamic surface control method is employed to deal with the problem of ‘curse of dimensionality’. Additionally, a single Lyapunov function is constructed to analyze the stability of the system. It is proved that all signals of the closed-loop system (CLS) are bounded. Finally, the effectiveness of the method is verified by simulation results.