The Effectiveness of Facial Neuromuscular Retraining on Patients with Facial Nerve Dysfunction: A Mental Health and Quality of Life Analysis.

Background: Facial muscle dysfunction can have drastic psychosocial effects. Objectives: To evaluate the impacts of customized neuromuscular retraining on mental health, quality of life (QoL), facial muscle function, and synkinesis. Methods: Thirty patients with facial nerve dysfunction completed a course of neuromuscular retraining. Patients' mental health, QoL, facial muscle function, and synkinesis were evaluated using Patient Health Questionnaire (PHQ-9), Facial Clinimetric Evaluation (FaCE) scale, electronic, clinician-graded facial function scale (eFACE), and Synkinesis Assessment Questionnaire (SAQ) at the initial and final visits. Scores were compared before and after treatment. Results: Patients (n = 30) included had a mean age of 59.4 ± 13.4 years (range 32.3-82.8) and were mostly female (22/30, 73.3%). The most common etiology was Iatrogenic facial nerve paralysis (11/20, 36.7%). Most patients had postfacial paralysis synkinesis (15/30, 50%), while 10 had complete flaccid paralysis. The median house-Brackmann score was 2 (range 1-6). The mean duration of facial palsy was 39.5 ± 106.9 (range 1-576 months). The duration of follow-up after the initial treatment session was 5.5 months, including 10 sessions. After neuromuscular retraining median PHQ-9 scores improved from 5 (range 0-25) to 3 (range 0-20) (p = 0.002). Mean FaCE PROM scores increased from 47.7 ± 11.5 to 56.5 ± 8.8 (p = 0.001). The mean eFACE score increased from 55.8 ± 15.1 to 71.7 ± 13.6 (p < 0.001). Median SAQ score was lower at the final visit (34.6 ± 13.4) compared to the initial visit (47.7 ± 17.8; p < 0.001). Conclusion: Customized neuromuscular retraining may improve patient-reported mental health, QoL, and facial muscle function and reduce synkinesis in facial nerve dysfunction.

Evaluation of a decellularized bronchial patch transplant in a porcine model.

Biological scaffolds for airway reconstruction are an important clinical need and have been extensively investigated experimentally and clinically, but without uniform success. In this study, we evaluated the use of a decellularized bronchus graft for airway reconstruction. Decellularized left bronchi were procured from decellularized porcine lungs and utilized as grafts for airway patch transplantation. A tracheal window was created and the decellularized bronchus was transplanted into the defect in a porcine model. Animals were euthanized at 7 days, 1 month, and 2 months post-operatively. Histological analysis, immunohistochemistry, scanning electron microscopy, and strength tests were conducted in order to evaluate epithelialization, inflammation, and physical strength of the graft. All pigs recovered from general anesthesia and survived without airway obstruction until the planned euthanasia timepoint. Histological and electron microscopy analyses revealed that the decellularized bronchus graft was well integrated with native tissue and covered by an epithelial layer at 1 month. Immunostaining of the decellularized bronchus graft was positive for CD31 and no difference was observed with immune markers (CD3, CD11b, myeloperoxidase) at two months. Although not significant, tensile strength was decreased after one month, but recovered by two months. Decellularized bronchial grafts show promising results for airway patch reconstruction in a porcine model. Revascularization and re-epithelialization were observed and the immunological reaction was comparable with the autografts. This approach is clinically relevant and could potentially be utilized for future applications for tracheal replacement.

Immunohistochemical Analysis of Lymphocyte Populations in Acute Skin Rejection: The University Health Network Addition to the Banff Classification.

Acute rejection in vascularized composite allotransplantation has been identified using the Banff 2007 working classification. We propose an addition to this classification based on histological and immunological assessment within the skin and subcutaneous tissue. Methods: Biopsies from vascularized composite transplant patients were obtained at scheduled visits and whenever skin changes occurred. Histology and immunohistochemistry were performed on all samples, looking at infiltrating cells. Results: Observations were made specifically related to each component of the skin, including the epidermis, dermis, vessels, and subcutaneous tissue. Our findings led to the establishment of the University Health Network addition of skin rejection. Conclusions: The high rate of rejection where the skin is involved requires novel techniques for early detection. The University Health Network skin rejection addition can serve as an adjunct to the Banff classification.

Perfusion decellularization for vascularized composite allotransplantation.

Vascularized composite allotransplantation is becoming the emerging standard for reconstructive surgery treatment for patients with limb trauma and facial injuries involving soft tissue loss. Due to the complex immunogenicity of composite grafts, patients who undergo vascularized composite allotransplantation are reliant on lifelong immunosuppressive therapy. Decellularization of donor grafts to create an extracellular matrix bio-scaffold provides an immunomodulatory graft that preserves the structural and bioactive function of the extracellular matrix. Retention of extracellular matrix proteins, growth factors, and signaling cascades allow for cell adhesion, migration, proliferation, and tissue regeneration. Perfusion decellularization of detergents through the graft vasculature allows for increased regent access to all tissue layers, and removal of cellular debris through the venous system. Grafts can subsequently be repopulated with appropriate cells through the vasculature to facilitate tissue regeneration. The present work reviews methods of decellularization, process parameters, evaluation of adequate cellular and nuclear removal, successful applications of perfusion decellularization for use in vascularized composite allotransplantation, and current limitations.

Evaluation of Early Markers of Ischemia-reperfusion Injury and Preservation Solutions in a Modified Hindlimb Model of Vascularized Composite Allotransplantation.

BACKGROUND: Ischemia-reperfusion injury plays an important role in vascularized composite allotransplantation (VCA). Currently, there is no ideal preservation solution for VCA. In this study, we investigated the effects of 4 different preservation solutions on different tissues within an allogeneic hindlimb rat model. METHODS: Sprague Dawley rat hindlimbs were flushed and placed at 4°C for 6 h in heparinized saline, histidine-tryptophan-ketoglutarate, University of Wisconsin (UW), and Perfadex and heterotopically transplanted for ease of ambulation. Apoptosis, necrosis, and the extracellular matrix of the tissues within the allograft were analyzed 2 h posttransplantation using immunohistochemistry, terminal deoxynucleotidyl transferase 2'-deoxyuridine 5'-triphosphate nick-end labeling (TUNEL) assay, and enzyme-linked immunoassay. RESULTS: Higher expression of cleaved caspase 3, a significant increase of high-mobility group box 1 and TUNEL-positive apoptotic cells were observed in the muscle and vessels preserved with heparinized saline compared with UW and Perfadex following reperfusion. Higher expression of TUNEL-positive apoptotic cells was observed in the skin at 12 h of ischemia and in the nerve following reperfusion with histidine-tryptophan-ketoglutarate as a preservation solution. CONCLUSIONS: Our data suggest that UW and Perfadex are preferred solutions in VCA. The vessels within the allografts appear to be very susceptible, with laminins and CD31 playing a role in ischemia-reperfusion injury.

Short-Term Preclinical Application of Functional Human Induced Pluripotent Stem Cell-Derived Airway Epithelial Patches.

Airway pathologies including cancer, trauma, and stenosis lack effective treatments, meanwhile airway transplantation and available tissue engineering approaches fail due to epithelial dysfunction. Autologous progenitors do not meet the clinical need for regeneration due to their insufficient expansion and differentiation, for which human induced pluripotent stem cells (hiPSCs) are promising alternatives. Airway epithelial patches are engineered by differentiating hiPSC-derived airway progenitors into physiological proportions of ciliated (73.9 ± 5.5%) and goblet (2.1 ± 1.4%) cells on a silk fibroin-collagen vitrigel membrane (SF-CVM) composite biomaterial for transplantation in porcine tracheal defects ex vivo and in vivo. Evaluation of ex vivo tracheal repair using hiPSC-derived SF-CVM patches demonstrate native-like tracheal epithelial metabolism and maintenance of mucociliary epithelium to day 3. In vivo studies demonstrate SF-CVM integration and maintenance of airway patency, showing 80.8 ± 3.6% graft coverage with an hiPSC-derived pseudostratified epithelium and 70.7 ± 2.3% coverage with viable cells, 3 days postoperatively. The utility of bioengineered, hiPSC-derived epithelial patches for airway repair is demonstrated in a short-term preclinical survival model, providing a significant leap for airway reconstruction approaches.

High resolution melting analysis and detection of Leishmania resistance: the role of multi drug resistance 1 gene.

BACKGROUND: Pentavalent antimonial compounds are currently used to treat leishmaniasis and resistance to these drugs is a serious problem. Multidrug resistance protein is an efflux pump of the cell membrane that expels foreign compounds. This study designed to evaluate the mutations in the multi-drug resistance 1 (MDR1) gene, in biopsy specimens of Leishmania tropica, with high resolution melting (HRM) method. In this experimental study, genomic DNA was extracted from 130 patients with skin leishmaniasis. Then, nucleotide changes were investigated throughout the gene using HRM and sequencing methods. The samples categorized in 5 groups by differences in the melting temperature (Tm). RESULT: The nucleotide changes analysis showed that 61% of the samples of different groups that were unresponsive to drug had mutations in the MDR1 gene, which were also confirmed by the sequencing method. These mutations can be one of the factors responsible for non-responsiveness to the treatment. CONCLUSION: According to the findings, it seems that mutation in MDR1 gene could be responsible for drug resistance to pentavalent antimonial compounds. Furthermore, HRM method can be used to diagnose drug resistance in leishmaniasis. It is also recommended that further studies be done regarding the importance of drug resistance in the leishmania affected patients.

Corrigendum to "Enchondroma protuberans of the hand: A case report" [Radiology Case Reports 15 (2020) 943-946].

[This corrects the article DOI: 10.1016/j.radcr.2020.04.026.].

A Concise Review and Required Precautions for COVID-19 Outbreak in Diagnostic and Interventional Radiology.

A global outbreak of a novel coronavirus (COVID-19) pneumonia began in December 2019 in Wuhan, China. The World Health Organization (WHO) announced a pandemic on March 11, 2020. The rapid rise in the case numbers and mortality led to the saturation of hospitals in many countries. COVID-19 patients usually present with fever, fatigue, dry cough, and dyspnea. Given the shortage of diagnostic kits in many countries and very high sensitivity of computed tomography (CT) for diagnosis of COVID-19 in clinically suspicious patients, the chest CT has been implemented among the primary initial methods of diagnosis before the confirmatory laboratory tests. This puts radiologists and radiology staff on the front line of this alarming pandemic. This report summarizes chest CT findings of COVID-19 patients to facilitate diagnosis and reviews a list of necessary precautions and safety measures for diagnostic and interventional radiology personnel. These precautionary plans are extremely important to avoid contamination of the health-care providers, as well as cross-contamination between patients.

Differentiating glioblastoma multiforme from cerebral lymphoma: application of advanced texture analysis of quantitative apparent diffusion coefficients.

PURPOSE: The purpose of this study was to differentiate glioblastoma multiforme from primary central nervous system lymphoma using the customised first and second-order histogram features derived from apparent diffusion coefficients.Methods and materials: A total of 82 patients (57 with glioblastoma multiforme and 25 with primary central nervous system lymphoma) were included in this study. The axial T1 post-contrast and fluid-attenuated inversion recovery magnetic resonance images were used to delineate regions of interest for the tumour and peritumoral oedema. The regions of interest were then co-registered with the apparent diffusion coefficient maps, and the first and second-order histogram features were extracted and compared between glioblastoma multiforme and primary central nervous system lymphoma groups. Receiver operating characteristic curve analysis was performed to calculate a cut-off value and its sensitivity and specificity to differentiate glioblastoma multiforme from primary central nervous system lymphoma. RESULTS: Based on the tumour regions of interest, apparent diffusion coefficient mean, maximum, median, uniformity and entropy were higher in the glioblastoma multiforme group than the primary central nervous system lymphoma group (P ≤ 0.001). The most sensitive first and second-order histogram feature to differentiate glioblastoma multiforme from primary central nervous system lymphoma was the maximum of 2.026 or less (95% confidence interval (CI) 75.1-99.9%), and the most specific first and second-order histogram feature was smoothness of 1.28 or greater (84.0% CI 70.9-92.8%). Based on the oedema regions of interest, most of the first and second-order histogram features were higher in the glioblastoma multiforme group compared to the primary central nervous system lymphoma group (P ≤ 0.015). The most sensitive first and second-order histogram feature to differentiate glioblastoma multiforme from primary central nervous system lymphoma was the 25th percentile of 0.675 or less (100% CI 83.2-100%) and the most specific first and second-order histogram feature was the median of 1.28 or less (85.9% CI 66.3-95.8%). CONCLUSIONS: Texture analysis using first and second-order histogram features derived from apparent diffusion coefficient maps may be helpful in differentiating glioblastoma multiforme from primary central nervous system lymphoma.

Insight into the significance of blood flow inside stenosis coronary jointed with bypass vein: The case of anemic, normal, and hypertensive individuals.

BACKGROUND AND OBJECTIVE: Recently, transplant of bypass vein is employed to charge blood fluid inside vein after stenosis position. Because, bypass vein can compensate consequences of stenosis in reducing blood flow within vessels. Therefore, analyses of shear stress for anemia, normal, and hypertensive individuals can prepare a valuable understanding in mechanical geometry which is applicable to design of transplantation especially for critical condition of shear stress. In this work, the transplantation of vessels is simulated in Fluent software, and user-defined function is used to indicate the blood properties as a non-Newtonian fluid based on Carreau fluid model. METHODS: Generally, shear stress profiles are studied for three cases of anemia, normal, and hypertensive individuals. Also, stenosis with the severity of 30% is simulated before the junction of host and grafted vessels. Finally, the results of shear stress on the walls (WSS) are reported with respect to three divided sections. Section one related to the distance from the stenosis position to the joining position of veins. Section two is the complete distance of the transplantation of veins, which blood flows are mixed. Section three is related to the distance after transplantation of host and grafted vessels. RESULTS: It was reported that flow separation causes the velocity of blood flow increases, which enhances shear stress. Moreover, increasing velocity in a hypertensive individual can exacerbate the shear stress. Maximum shear stress is as much as 105 Pa, 125 Pa, and 220 Pa in order of anemia, normal, and hypertensive individuals with abbreviations of LHD, NHD, and HHD, orderly. CONCLUSIONS: A comparison of maximum shear stress values in the Heel and Toe section showed that transplantation of veins can be a critical position of failure, which is introduced potentially as a sensible position for medical treatment and related surgeries.

Numerical study for blood rheology inside an artery: The effects of stenosis and radius on the flow behavior.

BACKGROUND AND OBJECTIVE: In this work, using Sisko model, blood flow is simulated inside an artery which have cone shape of stenosis with different angles of φ = 0.25, φ = 0.5, φ = 0.75, φ = 1 and φ = 1.25 degree, respectively. METHODS: In the first step, an artery radius of 0.002 m is fixed to study the effects of cone shape of arterial stenosis on the flow behavior. Then, stenosis angle of φ = 0.5 degree is fixed to study the effects of different Artery radii of 0.002 m, 0.0025 m, 0.003m, and 0.0035 m orderly on the flow behavior. For simulation the blood flow, Sisko model is used. Afterward, stenosis angle of φ = 0.5 degrees with a radius of 0.002m is fixed for investigating the influences of different behavior of blood fluid by manipulation of constant parameters of the Sisko model. RESULTS: It is reported that with increasing arterial stenosis angle, maximum blood flow velocity is sharply increased in central region of artery from 0.12 m/s to 0.16 m/s, 0.25 m/s, 0.36 m/s and 0.56 m/s in order of increasing stenosis angles from φ = 0.25 to φ = 0.5, φ = 0.75, φ = 1 and φ = 1.25 degree, respectively. Also, maximum shear stress of artery wall are as much as 64 Pa, 42 Pa, 24 Pa, 18 Pa and 16Pa respectively in order of stenosis angles of φ = 0.25, φ = 0.5, φ = 0.75, φ = 1 and φ = 1.25 degree. On the other side, the effect of increasing artery radius is against the influences of stenosis angle, and contradiction of these parameters is affected by the stress tension and viscosity of blood. CONCLUSIONS: Variations of blood behavior from non-Newtonian to Newtonian shows that shear stress in blood stream in the stenosis artery with non-Newtonian blood is higher than that of Newtonian blood due to differences in their viscous behaviors and reactions in exposure of stenosis and artery wall effects.

Blood flow analysis inside different arteries using non-Newtonian Sisko model for application in biomedical engineering.

BACKGROUND AND OBJECTIVE: In the present research, simulation of blood flow is carried out inside the artery with different radiuses of 0.002 m, 0.0025 m, 0.003 m, and 0.0035 m. METHODS: To simulate the blood as non-Newtonian fluid using of Sisko model, different constant heat fluxes are applied on the boundary walls of the artery. Then, the results of velocity, temperature, and Nusselt number are reported versus axial and radial directions. RESULTS: Results show that blood temperature is enhanced with increasing axial distance. Also, maximum temperatures are seen at maximum axial and radial distances from references of entry and central regions of artery. Furthermore, increasing the radius of the artery can increase blood temperature due to a reduction in blood velocity inside the vessel. Consequently, blood particles can spend more time to receive thermal energy, which leads to emerging higher blood temperature. This phenomenon can be important in the oxygenation process inside the human body. It is observed that effect of increasing the radius of the artery can enhance blood temperature as much as 0.001 K. Also, applying constant heat fluxes in order 4 W/m2 to 5 W/m2 and 6 W/m2 on the artery wall brings axial Nusselt values of 0.365-0.4575 and 0.55, respectively. As a result of axial and radial Nusselt numbers, it is reported that because radial Nusslet is unchanged in the central region of the artery, temperature shall be constant in a radius less of 0.0019 m. Therefore, the influences of heat fluxes are ignorable in the central region of the vein. Also, maximum temperatures are reported as much as 310.5 K, 311.1 K, and 311.5 K in order of applying thermal boundary flux of q'' = 400 W/m2, q'' = 800 W/m2 and q'' = 1000 W/m2 respectively. Therefore, applying heat fluxes in the range of investigated can raise the blood temperature as much as 1.5 °C, which is equal to 38.5 °C. Thus, there is no doubt that such a high temperature is dangerous for human health. CONCLUSIONS: As conclusion, the results of this research are important hints for medical diagnostics of oxygenation, hematocrit, polycythemia, and blood disorders.

Simulation of blood flow in arteries with aneurysm: Lattice Boltzmann Approach (LBM).

BACKGROUND AND OBJECTIVE: In most countries, the higher death rates are due to cardiovascular disease and stroke. These problems often derive from irregular blood flow and the circulatory system disorder. METHODS: In this paper, the blood flow is simulated in a created aneurysm in the artery upon using Lattice Boltzmann Method (LBM). Blood is selected as a non-Newtonian fluid which was simulated with power-law model. The lattice Boltzmann results for non-Newtonian fluid flow with power-law model and the curved boundary are compared and validated with previous studies which show a good agreement. In this study, simulations are carried out for two types of aneurysms. For the first aneurysm, three power-law exponents of 0.6, 0.8 and 1.0 at Reynolds number of 100 for three different cases are investigated. RESULTS: The results show that the wall shear stress increases with increasing the power-law exponent. In addition, in the main duct of artery where the velocity is larger, shear stress is lower due to the smaller velocity gradient. For the second Aneurysm, the simulations are done for three Reynolds numbers of 100, 150 and 200, and three Womersley numbers of 4, 12 and 20. The blood flow is pulsating at the inlet such as the real pulsating wave in the blood. Results show that with increasing the Womersley number, the velocity profiles in the middle of the aneurysm are closer at a constant Reynolds number. CONCLUSIONS: With increasing the Reynolds number, the range of vortices and values of velocity and tension grow in the aneurysm.

The effect of the second excitation frequency mode under different conditions on the fluid streaming and microparticles acoustophoresis with the aim of separating biological cells.

BACKGROUND AND OBJECTIVE: In this study, the effect of the second excitation frequency mode under different conditions on the fluid streaming and its microparticles displacement is investigated. METHODS: For this purpose, some variable parameters such as the particle diameter, microchannel aspect ratio, and applied frequency modes have been selected to study. The resulted acoustic streaming was scrutinized to understand the physics of the problem under different geometrical and input conditions. Finally, the effect of the increasing the microparticle size and aspect ratio of the microchannel, simultaneously, has been evaluated. RESULTS: The results demonstrated that increasing the microparticle size accelerates the displacement of the microparticles. On the other hand, changing the aspect ratio affects the formation of the microparticle distribution and it also changes the velocity of the microparticles due to the gradient of the second-order pressure. CONCLUSIONS: The obtained results have wide applications in the military, medical, petrochemical, and other related studies.

Molecular dynamics simulation of Doxorubicin loading with N-isopropyl acrylamide carbon nanotube in a drug delivery system.

BACKGROUND AND OBJECTIVE: Doxorubicin is one of the common drugs used for cancer therapy. Molecular dynamics were applied to investigate the loading of Doxorubicin with thermosensitive N-isopropyl acrylamide Carbon nanotube carrier. METHODS: The results showed that the smaller polymer chain length has more decrease of gyration radius. A decrease of gyration radius resulted in more concentrated aggregation with stronger bonds. Therefore, the shorter the polymer chain lengths, the more stable polymer interaction and better Doxorubicin delivery. Smaller polymers also form more hydrogen bonds with the drug leading to stronger and more stable carriers. RESULTS: A lower amount of wall shear stress was found near the inner wall of the artery, distal to the plaque region (stenosis), and in both percentages of stenosis the maximum wall shear stress will accrue in the middle of the stenosis; however it is much more in the higher rate of stenosis. CONCLUSIONS: The results indicated that N-isopropyl acrylamide - Carbon nanotube is suitable for the delivery of Doxorubicin, and five mer N-isopropyl acrylamide is the optimum carrier for Doxorubicin loading.

pH-sensitive loading/releasing of doxorubicin using single-walled carbon nanotube and multi-walled carbon nanotube: A molecular dynamics study.

BACKGROUND AND OBJECTIVE: Doxorubicin is one of the drugs used to treat cancer, and many studies have been conducted to control its release. In this study, carbon nanotubes have been proposed as a doxorubicin carrier, and the effect of carboxyl functional group on the controlled release of doxorubicin has been studied. METHODS: This study has been done by molecular dynamics simulation and was based on changing the pH as a mechanism controller. RESULTS: This work is intended to test the efficacy of this drug carrier for the release of doxorubicin. A comparison was also made between single-walled and double-walled carbon nanotubes to answer the question of which one can be a better carrier for doxorubicin. The study of DOXORUBICIN adsorption and release showed that the DOXORUBICIN adsorption on single-walled carbon nanotube and multi-walled carbon nanotube in neutral pH was stronger than it was in acidic pH, which could be due to the electrostatic interactions between the carboxyl group of nanotubes and DOXORUBICIN. Based on this and according to the investigation of hydrogen bonds, diffusion coefficients, and other results it was clear that the drug release in acidic pH was appropriate for body conditions. Since cancer tissues pH is acidic, this shows the suitability of carbon nanotube in drug delivery and DOXORUBICIN release in cancer tissues. In addition, it was shown that the blood pH (pH = 7) is suitable for DOXORUBICIN loading on the carbon nanotube and carbon nanotube-DOXORUBICIN linkage remained stable at this pH; accordingly, the carbon nanotube could deliver DOXORUBICIN in blood quite well and release it in cancerous tissues. This suggests the carbon nanotubes as a promising drug carrier in the cancer therapy which can be also investigated in experiments. CONCLUSION: It was revealed that the bonds between multi-walled carbon nanotube and DOXORUBICIN was stronger and this complex had a slower release in the cancer tissues compared to the single-walled carbon nanotube; this can be regarded as an advantage over the single-walled carbon nanotube in the DOXORUBICIN delivery and release.

Calculation of the thermal conductivity of human serum albumin (HSA) with equilibrium/non-equilibrium molecular dynamics approaches.

BACKGROUND AND OBJECTIVE: Human serum albumin (HSA) controls the flow of numerous chemical structures and molecules in the cardiovascular system. So, thermal conductivity of this atomic compound is important in medicinal applications. METHODS: In this work, the thermal conductivity of HSA is calculated with equilibrium/non-equilibrium molecular dynamic approaches. In these methods each HSA molecule is exactly represented by C, N, O and S atoms and their implemented dreiding potential. Finally by using Green-Kubo and Fourier's law the thermal conductivity of HSA/H2O mixture is calculated. RESULTS: Our calculated rates for thermal conductivity via equilibrium/non-equilibrium molecular dynamics methods are 0.496 W/m K and 0.448 W/m K, respectively. The calculated thermal conductivity for this structure was very close to the thermal conductivity calculated for water molecules which were reported by other research groups. Furthermore our simulated structures show that thermal conductivity of HAS/H2O mixtures has inverse relation with HAS molecules numbers and temperature of simulated atomic structures. CONCLUSIONS: Comparing thermal conductivity from equilibrium/non-equilibrium molecular dynamics methods for HAS/H2O shows that EMD and NEMD results are reliable and EMD calculated results are higher than NEMD results.