Optimistic and possible contribution of nanomaterial on biomedical applications: A review.

Nanomaterials have many advantages over bulk materials, including enhanced surface-to-volume proportion as well as magnetic traits. It has been a steady rise in research with using nanomaterials in various biomedical fields in the past few decades. Constructing nanomaterials has emerged as a leading research primary concern in order to discover specialized biomedical applications. Since, their advantageous properties including chemical stability, non-toxicity, bio - compatibility, relatively high magnetization, and strong magnetic vulnerability, nanoparticles of iron oxide had already influenced implementations in different biomedical fields. Nanomaterials can be divided up into four nanomaterials such as metallic nanomaterials, bimetallic or alloy nanomaterials, metal oxide nanomaterials, as well as magnetic nanomaterials. Hence, the purpose of this review is to conduct such in discussion on emerging advancements in nanomaterials for biomedical, with such a special emphasis upon those options of nanomaterials including metallic nanomaterials: Au and Ag, bimetallic nanomaterials: Fe-Co and Fe-Pt, and metal oxides: TiO2 and CeO2. Securing this information gap will result in a better comprehension of the contribution of nanomaterial type and subsequent huge-scale applications in aspects of both their potential and challenges.

Remediation competence of nanoparticles amalgamated biochar (nanobiochar/nanocomposite) on pollutants: A review.

Advanced biochar blended nanoparticles substances, such as nano biochar or nanocomposites, have provided long-term solutions to a wide range of modern-day problems. Biochar blended nano-composites can be created to create better composite materials that combine the benefits of biochar and nanoparticles. Such materials have been typically improved with active functional groups, porous structure, active surface area, catalytic deterioration ability, as well as easy recovery or separation of pollutants. Such biochar-basednanocomposites have good adsorption properties for a variety of pollutants in various form of polluted medium (soil and water contamination). Catalytic nanoparticle encapsulated biochar, can perform concurrently the adsorption (by biochar) as well as catalytic degradation (nanoparticles) functions for pollutants removal from polluted sites. In this review, the advanced and practically feasible techniques involved in the biochar blended nanoparticles-based nanocomposites have been discussed with environmental applications. Furthermore, the mechanisms involved in this composite material in remediation, as well as the advantages and disadvantages of biochar blended nanoparticles-based nanocomposites, were discussed, and future directions for study in this field were suggested.

SARS-CoV-2 Infectivity and Neurological Targets in the Brain.

The gateway for invasion by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into human host cells is via the angiotensin-converting enzyme 2 (ACE2) transmembrane receptor expressed in multiple immune and nonimmune cell types. SARS-CoV-2, that causes coronavirus disease 2019 (COVID-19; CoV-19) has the unusual capacity to attack many different types of human host cells simultaneously via novel clathrin- and caveolae-independent endocytic pathways, becoming injurious to diverse cells, tissues and organ systems and exploiting any immune weakness in the host. The elicitation of this multipronged attack explains in part the severity and extensive variety of signs and symptoms observed in CoV-19 patients. To further our understanding of the mechanism and pathways of SARS-CoV-2 infection and susceptibility of specific cell- and tissue-types and organ systems to SARS-CoV-2 attack in this communication we analyzed ACE2 expression in 85 human tissues including 21 different brain regions, 7 fetal tissues and 8 controls. Besides strong ACE2 expression in respiratory, digestive, renal-excretory and reproductive cells, high ACE2 expression was also found in the amygdala, cerebral cortex and brainstem. The highest ACE2 expression level was found in the pons and medulla oblongata in the human brainstem, containing the medullary respiratory centers of the brain, and may in part explain the susceptibility of many CoV-19 patients to severe respiratory distress.

MicroRNA (miRNA)-Mediated Pathogenetic Signaling in Alzheimer's Disease (AD).

Alzheimer's disease (AD) is an expanding health and socioeconomic concern in industrialized societies, and the leading cause of intellectual impairment in our aging population. The cause of AD remains unknown, and there are currently no effective treatments to stop or reverse the progression of this uniquely human and age-related neurological disorder. Elucidation of the AD mechanism and factors that contribute to the initiation, progression, and spreading of this chronic and fatal neurodegeneration will ultimately result in improved and effective diagnostics and therapeutic strategies.microRNAs (miRNAs) comprise a relatively recently discovered category of 20-24 nucleotide non-coding RNAs that function post-transcriptionally in shaping the transcriptome of the cell, and in doing so, contribute to the molecular-genetics and phenotype of human CNS health and disease. To date about 2550 unique mature human miRNAs have been characterized, however only highly selected miRNA populations appear to be enriched in different anatomical compartments within the CNS.This general commentary for the 'Special Issue: 40th Year of Neurochemical Research' will bring into perspective (i) some very recent findings on the extraordinary biophysics and signaling properties of CNS miRNA in AD and aging human brain; (ii) how specific intrinsic biophysical attributes of miRNAs may play defining roles in the establishment, proliferation and spreading of the AD phenotype; and (iii) how miRNAs can serve as prospective therapeutic targets and biomarkers potentially useful in the clinical management of this terminal neurological disease whose incidence in our rapidly aging population is reaching epidemic proportions.

Energy behaviour for DNA translocation through graphene nanopores.

Nanoparticles have considerable promise for many applications in electronics, energy storage, bioscience and biotechnologies. Here we use applied mathematical modelling to exploit the basic principles of mechanics and the 6-12 Lennard-Jones potential function together with the continuum approach, which assumes that a discrete atomic structure can be replaced by an average constant atomic surface density of atoms that is assumed to be smeared over each molecule. We identify a circular hole in a graphene sheet as a nanopore and we consider the molecular interaction energy for both single-strand and double-strand DNA molecules assumed to move through the circular hole in a graphene sheet to determine the radius b of the hole that gives the minimum energy. By minimizing the interaction energy, we observe that the single-strand DNA and double-strand DNA molecules penetrate through a graphene nanopore when the pore radii b> 7.8Å and b> 12.7Å, respectively. Our results can be adopted to offer new applications in the atomic surface processes and electronic sensing.

Modeling interactions between C60 antiviral compounds and HIV protease.

Fullerenes have generated a great deal of interest in recent years, due to their properties and potential applications in many fields, including medicine. In this paper, we study an antiviral fullerene compound which may be used to treat the human immunodeficiency virus (HIV). We formulate a mathematical model which can describe the interaction energy between the C[Formula: see text] antiviral compounds and the HIV. In particular, this paper predicts the energy and force arising from the interaction between HIV active region and the antiviral molecule which is attached to the external surface of a fullerene C[Formula: see text]. These interactions are calculated based on the structure of the antiviral molecules. Our results show that the binding of fullerene C[Formula: see text] to the antiviral molecules increases the efficiency of the compound to prohibit the activity of HIV.

Fatty acid transport protein 4 (FATP4) prevents light-induced degeneration of cone and rod photoreceptors by inhibiting RPE65 isomerase.

Although rhodopsin is essential for sensing light for vision, it also mediates light-induced apoptosis of photoreceptors in mouse. RPE65, which catalyzes isomerization of all-trans retinyl fatty acid esters to 11-cis-retinol (11cROL) in the visual cycle, controls the rhodopsin regeneration rate and photoreceptor susceptibility to light-induced degeneration. Mutations in RPE65 have been linked to blindness in affected children. Despite such importance, the mechanism that regulates RPE65 function remains unclear. Through unbiased expression screening of a bovine retinal pigment epithelium (RPE) cDNA library, we have identified elongation of very long-chain fatty acids-like 1 (ELOVL1) and fatty acid transport protein 4 (FATP4), which each have very long-chain fatty acid acyl-CoA synthetase (VLCFA-ACS) activity, as negative regulators of RPE65. We found that the VLCFA derivative lignoceroyl (C24:0)-CoA inhibited synthesis of 11cROL, whereas palmitoyl (C16:0)-CoA promoted synthesis of 11cROL. We further found that competition of FATP4 with RPE65 for the substrate of RPE65 was also involved in the mechanisms by which FATP4 inhibits synthesis of 11cROL. FATP4 was predominantly expressed in RPE, and the FATP4-deficient RPE showed significantly higher isomerase activity. Consistent with these results, the regeneration rate of 11-cis-retinaldehyde and the recovery rate for rod light sensitivity were faster in FATP4-deficient mice than wild-type mice. Moreover, FATP4-deficient mice displayed increased accumulation of the cytotoxic all-trans retinaldehyde and hypersusceptibility to light-induced photoreceptor degeneration. Our findings demonstrate that ELOVL1, FATP4, and their products comprise the regulatory elements of RPE65 and play important roles in protecting photoreceptors from degeneration induced by light damage.

Receptor interacting protein kinase-mediated necrosis contributes to cone and rod photoreceptor degeneration in the retina lacking interphotoreceptor retinoid-binding protein.

Interphotoreceptor retinoid-binding protein (IRBP) secreted by photoreceptors plays a pivotal role in photoreceptor survival with an unknown mechanism. A mutation in the human IRBP has been linked to retinitis pigmentosa, a progressive retinal degenerative disease. Mice lacking IRBP display severe early and progressive photoreceptor degeneration. However, the signaling pathway(s) leading to photoreceptor death in IRBP-deficient mice remains poorly understood. Here, we show that amounts of tumor necrosis factor-α (TNF-α) in the interphotoreceptor matrix and retinas of Irbp(-/-) mice were increased more than 10-fold and fivefold, respectively, compared with those in wild-type mice. Moreover, TNF-α receptor 1, an important membrane death receptor that mediates both programmed apoptosis and necrosis, was also significantly increased in Irbp(-/-) retina, and was colocalized with peanut agglutinin to the Irbp(-/-) cone outer segments. Although these death signaling proteins were increased, the caspase-dependent and independent apoptotic pathways were mildly activated in the Irbp(-/-) retinas, suggesting that other cell death mechanism(s) also contributes to the extensive photoreceptor degeneration in Irbp(-/-) retina. We found that receptor interacting protein 1 and 3 (RIP1 and RIP3) kinases, the intracellular key mediators of TNF-induced cellular necrosis, were elevated at least threefold in the Irbp(-/-) retinas. Moreover, pharmacological inhibition of RIP1 kinase significantly prevented cone and rod photoreceptor degeneration in Irbp(-/-) mice. These results reveal that RIP kinase-mediated necrosis strongly contributes to cone and rod degeneration in Irbp(-/-) mice, implicating the TNF-RIP pathway as a potential therapeutic target to prevent or delay photoreceptor degeneration in patients with retinitis pigmentosa caused by IRBP mutation.

Offset configurations for single- and double-strand DNA inside single-walled carbon nanotubes.

Nanotechnology is a rapidly expanding research area, and it is believed that the unique properties of molecules at the nano-scale will prove to be of substantial benefit to mankind, especially so in medicine and electronics. Here we use applied mathematical modelling exploiting the basic principles of mechanics and the 6-12 Lennard-Jones potential function together with the continuum approximation, which assumes that intermolecular interactions can be approximated by average atomic surface densities. We consider the equilibrium offset positions for both single-strand and double-strand DNA molecules inside a single-walled carbon nanotube, and we predict offset positions with reference to the cross-section of the carbon nanotube. For the double-strand DNA, the potential energy is determined for the general case for any helical phase angle ϕ, but we also consider a special case when ϕ = π, which leads to a substantial simplification in the analytical expression for the energy. As might be expected, our results confirm that the global minimum energy positions for a single-strand DNA molecule and a double-strand DNA molecule will lie off axis and they become closer to the tube wall as the radius of the tube increases.

High performance hydrogen storage from Be-BTB metal-organic framework at room temperature.

The metal-organic framework beryllium benzene tribenzoate (Be-BTB) has recently been reported to have one of the highest gravimetric hydrogen uptakes at room temperature. Storage at room temperature is one of the key requirements for the practical viability of hydrogen-powered vehicles. Be-BTB has an exceptional 298 K storage capacity of 2.3 wt % hydrogen. This result is surprising given that the low adsorption enthalpy of 5.5 kJ mol(-1). In this work, a combination of atomistic simulation and continuum modeling reveals that the beryllium rings contribute strongly to the hydrogen interaction with the framework. These simulations are extended with a thermodynamic energy optimization (TEO) model to compare the performance of Be-BTB to a compressed H2 tank and benchmark materials MOF-5 and MOF-177 in a MOF-based fuel cell. Our investigation shows that none of the MOF-filled tanks satisfy the United States Department of Energy (DOE) storage targets within the required operating temperatures and pressures. However, the Be-BTB tank delivers the most energy per volume and mass compared to the other material-based storage tanks. The pore size and the framework mass are shown to be contributing factors responsible for the superior room temperature hydrogen adsorption of Be-BTB.

Intracerebral propagation of Alzheimer's disease: strengthening evidence of a herpes simplex virus etiology.

BACKGROUND: A faulty human protein, abnormally phosphorylated tau, was recently publicized to spread "like a virus" from neuron to neuron in Alzheimer's patients' brains. For several decades, we have been amassing arguments showing that herpes simplex virus type 1 (HSV-1), not p-tau, propagates this interneuronal, transsynaptic pathologic cascade. METHODS: We reiterate convincing data from our own (and other) laboratories, reviewing the first anatomic foothold neurofibrillary tangles gain in brainstem and/or entorhinal cortex; the chronic immunosurveillance cellularity of the trigeminal ganglia wherein HSV-1 awakens from latency to reactivate; the inabilities of p-tau protein's physical properties to promote it to jump synapses; the amino acid homology between human p-tau and VP22, a key target for phosphorylation by HSV serine/threonine-protein kinase UL13; and the exosomic secretion of HSV-1-infected cells' L-particles, attesting to the cell-to-cell passage of microRNAs of herpesviruses. RESULTS: The now-maturing construct that reactivated HSV-1 best accounts for the intracerebral propagation of AD changes in the human brain should at last seem highly attractive. This hypothesis might even explain statins' apparent mechanism in some studies for lowering AD incidence. CONCLUSION: Provided that funding agencies will quickly ignite a new realm of investigation, the rejuvenated enthusiasm for testing this optimistic construct holds incalculable potential for rapid, efficacious clinical application, through already available and relatively safe antiviral therapeutics.

HSV-1 latent rabbits shed viral DNA into their saliva.

BACKGROUND: Rabbits latent with HSV-1 strain McKrae spontaneously shed infectious virus and viral DNA into their tears and develop recurrent herpetic-specific corneal lesions. The rabbit eye model has been used for many years to assess acute ocular infections and pathogenesis, antiviral efficacy, as well as latency, reactivation, and recurrent eye diseases. This study used real-time PCR to quantify HSV-1 DNA in the saliva and tears of rabbits latent with HSV-1 McKrae. METHODS: New Zealand white rabbits used were latent with HSV-1 strain McKrae and had no ocular or oral pathology. Scarified corneas were topically inoculated with HSV-1. Eye swabs and saliva were taken from post inoculation (PI) days 28 through 49 (22 consecutive days). Saliva samples were taken four times each day from each rabbit and the DNA extracted was pooled for each rabbit for each day; one swab was taken daily from each eye and DNA extracted. Real-time PCR was done on the purified DNA samples for quantification of HSV-1 DNA copy numbers. Data are presented as copy numbers for each individual sample, plus all the copy numbers designated as positive, for comparison between left eye (OS), right eye (OD), and saliva. RESULTS: The saliva and tears were taken from 9 rabbits and from 18 eyes and all tested positive at least once. Saliva was positive for HSV-1 DNA at 43.4% (86/198) and tears were positive at 28.0% (111/396). The saliva positives had 48 episodes and the tears had 75 episodes. The mean copy numbers ± the SEM for HSV-1 DNA in saliva were 3773 ± 2019 and 2294 ± 869 for tears (no statistical difference). CONCLUSION: Rabbits latent with strain McKrae shed HSV-1 DNA into their saliva and tears. HSV-1 DNA shedding into the saliva was similar to humans. This is the first evidence that documents HSV-1 DNA in the saliva of latent rabbits.

Rabbit and mouse models of HSV-1 latency, reactivation, and recurrent eye diseases.

The exact mechanisms of HSV-1 establishment, maintenance, latency, reactivation, and also the courses of recurrent ocular infections remain a mystery. Comprehensive understanding of the HSV-1 disease process could lead to prevention of HSV-1 acute infection, reactivation, and more effective treatments of recurrent ocular disease. Animal models have been used for over sixty years to investigate our concepts and hypotheses of HSV-1 diseases. In this paper we present descriptions and examples of rabbit and mouse eye models of HSV-1 latency, reactivation, and recurrent diseases. We summarize studies in animal models of spontaneous and induced HSV-1 reactivation and recurrent disease. Numerous stimuli that induce reactivation in mice and rabbits are described, as well as factors that inhibit viral reactivation from latency. The key features, advantages, and disadvantages of the mouse and rabbit models in relation to the study of ocular HSV-1 are discussed. This paper is pertinent but not intended to be all inclusive. We will give examples of key papers that have reported novel discoveries related to the review topics.

Geometric influence on Ruderman-Kittel-Kasuya-Yosida interactions in zigzag carbon nanotubes.

We derive an analytic description of the spin susceptibility in finite length zigzag carbon nanotubes (CNT) with chirality (n, 0). The spin susceptibility is proportional to the Ruderman-Kittel-Kasuya-Yosida (RKKY) interactions which describes indirect carrier mediated exchange coupling between localized magnetic moments. We show that the strongest RKKY interactions are along the edges of the nanotube and in the thermodynamic limit at half filling with spin symmetry the shape of the susceptibility curve about the edge of the CNT can be determined solely by the lattice geometry represented by the parameter n and a parameter L which describes the nanotube length. We also show that the introduction of Zeeman splitting or doping may have no effect on the spin susceptibility, provided n is small. A detailed knowledge of magnetic interactions, such as RKKY interactions, in CNT is of vital importance to the development of nanotechnology applications.

microRNA, the Innate-Immune System and SARS-CoV-2.

The single-stranded viral RNA (ssvRNA) known as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes COVID-19 can be effectively inactivated by a number of natural ribonucleic acid-based host cell defenses. One of the most important of these defenses includes the actions of a class of small non-coding RNAs (sncRNAs) known as microRNAs (miRNAs). Via base-pair complementarity miRNAs are capable of specifically targeting ssvRNA sequences such as SARS-CoV-2 promoting its inactivation and neutralization. RNA-sequencing and bioinformatics analysis indicate that multiple naturally-occurring human miRNAs have extensive complementarity to the SARS-CoV-2 ssvRNA genome. Since miRNA abundance, speciation, and complexity vary significantly amongst human individuals, this may in part explain the variability in the innate-immune and pathophysiological response of different individuals to SARS-CoV-2 and overall susceptibility to ssvRNA-mediated viral infection.

Ferric Ion Diffusion for MOF-Polymer Composite with Internal Boundary Sinks.

Simple and economical ferric ion detection is necessary in many industries. An europium-based metal organic framework has selective sensing properties for solutions containing ferric ions and shows promise as a key component in a new sensor. We study an idealised sensor that consists of metal organic framework (MOF) crystals placed on a polymer surface. A two-dimensional diffusion model is used to predict the movement of ferric ions through the solution and polymer, and the ferric ion association to a MOF crystal at the boundary between the different media. A simplified one-dimensional model identifies the choice of appropriate values for the dimensionless parameters required to optimise the time for a MOF crystal to reach steady state. The model predicts that a large non-dimensional diffusion coefficient and an effective association with a small effective flux will reduce the time to steady-state. The effective dissociation is the most significant parameter to aid the estimation of the ferric ion concentration. This paper provides some theoretical insight for material scientists to optimise the design of a new ferric ion sensor.

Lytic HSV-1 infection induces the multifunctional transcription factor Early Growth Response-1 (EGR-1) in rabbit corneal cells.

BACKGROUND: Herpes simplex virus type-1 (HSV-1) infections can cause a number of diseases ranging from simple cold sores to dangerous keratitis and lethal encephalitis. The interaction between virus and host cells, critical for viral replication, is being extensively investigated by many laboratories. In this study, we tested the hypothesis that HSV-1 lytic infection triggers the expression of important multi-functional transcription factor Egr1. The mechanisms of induction are mediated, at least in part, by signaling pathways such as NFκB and CREB. METHODS: SIRC, VERO, and 293HEK cell lines were infected with HSV-1, and the Egr-1 transcript and protein were detected by RT-PCR and Western blot, respectively. The localization and expression profile of Egr-1 were investigated further by immunofluorescence microscopy analyses. The recruitment of transcription factors to the Egr-1 promoter during infection was studied by chromatin immunoprecipitation (ChIP). Various inhibitors and dominant-negative mutant were used to assess the mechanisms of Egr-1 induction and their effects were addressed by immunofluorescence microscopy. RESULTS: Western blot analyses showed that Egr-1 was absent in uninfected cells; however, the protein was detected 24-72 hours post treatment, and the response was directly proportional to the titer of the virus used for infection. Using recombinant HSV-1 expressing EGFP, Egr-1 was detected only in the infected cells. ChIP assays demonstrated that NFкB and cAMP response element binding protein (CREB) were recruited to the Egr-1 promoter upon infection. Additional studies showed that inhibitors of NFкB and dominant-negative CREB repressed the Egr-1 induction by HSV-1 infection. CONCLUSION: Collectively, these results demonstrate that Egr-1 is expressed rapidly upon HSV-1 infection and that this novel induction could be due to the NFкB/CREB-mediated transactivation. Egr-1 induction might play a key role in the viral gene expression, replication, inflammation, and the disease progression.

Hydrogen storage inside graphene-oxide frameworks.

In this paper, we use applied mathematical modelling to investigate the storage of hydrogen molecules inside graphene-oxide frameworks, which comprise two parallel graphenes rigidly separated by perpendicular ligands. Hydrogen uptake is calculated for graphene-oxide frameworks using the continuous approximation and an equation of state for both the bulk and adsorption gas phases. We first validate our approach by obtaining results for two parallel graphene sheets. This result agrees well with an existing theoretical result, namely 1.85 wt% from our calculations, and 2 wt% arising from an ab initio and grand canonical Monte Carlo calculation. This provides confidence to the determination of the hydrogen uptake for the four graphene-oxide frameworks, GOF-120, GOF-66, GOF-28 and GOF-6, and we obtain 1.68, 2, 6.33 and 0 wt%, respectively. The high value obtained for GOF-28 may be partly explained by the fact that the benzenediboronic acid pillars between graphene sheets not only provide mechanical support and porous spaces for the molecular structure but also provide the higher binding energy to enhance the hydrogen storage inside graphene-oxide frameworks. For the other three structures, this binding energy is not as large in comparison to that of GOF-28 and this effect diminishes as the ligand density decreases. In the absence of conflicting data, the present work indicates GOF-28 as a likely contender for practical hydrogen storage.

Design parameters for carbon nanobottles to absorb and store methane.

We investigate the internal mechanics for methane storage in a nanobottle, which is assumed to comprise a metallofullerene located inside a carbon nanobottle, which is constructed from a half-fullerene as the base, and two nanotubes which are joined by a nanocone. The interaction potential energy for the metallofullerene is obtained from the 6-12 Lennard-Jones potential and the continuum approximation, which assumes that a discrete atomic structure can be replaced by an average atomic surface density. This potential energy shows that the metallofullerene has two minimum energy positions, which are located close to the neck of the bottle and at the base of the nanobottle, and therefore it may be used as a bottle-stopper to open or to close the nanobottle. At the neck of the bottle, the encapsulated metallofullerene closes the nanobottle, and by applying an external electrical force, the metallofullerene can overcome the energy barrier of the nanotube, and pass from the neck of the nanobottle to the base so that the nanobottle is open. For methane storage, the metallofullerene serves the dual purposes of opening and closing the nanobottle, as well as an attractor for the methane gas. The analytical formulation gives rise to a rapid computational capacity, and enables the direct determination of the optimal dimensions necessary to ensure the correct working function of the nanobottle, and specific ranges for the critical parameters are formulated.

Reference Phantom Method for Ultrasonic Imaging of Thin Dynamic Constructs.

Quantitative ultrasound has a great potential for the non-destructive evaluation of tissue engineered constructs, where the local attenuation and the integrated backscatter coefficient (IBC) can be used for monitoring the development of biological processes. The local determination of both parameters can be achieved using the reference phantom method (RPM). However, its accuracy can be affected when evaluating constructs of evolving sound speed, attenuation and thickness, for example, when evaluating biodegradable hydrogels developing neocartilage. To assess the feasibility of using the RPM under such dynamic conditions while employing a 50-MHz transducer, we conducted a series of experiments on 3-mm-thick acellular hydrogels laden with microspheres. The ultrasonic evaluation procedure used was validated by detecting and compensating for large attenuation variations occurring in the construct, up to 20-fold with respect to the reference phantom, with estimations errors below 1%. We found that sound speed mismatch does not affect the local attenuation estimation, but causes a strong diffraction effect by reducing the backscatter intensity. Such intensity reduction was compensated by determining the IBC percentage change (IBCΔ) as function of sound speed mismatch with respect to the reference phantom (ΔSS), with the equation IBCΔ = (0.63 ± 0.07) ΔSS + (8.54 ± 0.76) 10-3 ΔSS2. The investigated ΔSS interval was up to 120 m/s and using two different concentrations of microspheres, with estimation errors below 7% relative to the construct's actual IBC. Finally, we found that the spectral difference method is sufficient to measure within a few millimetres in depth mismatch, and when combining with sound speed mismatch, we found negligible additional effects. These results pave the way for the use of a generic reference phantom for the evaluation of thin dynamic constructs, thus simplifying the need for using different phantoms depending on the construct's properties.