The model of the heart was coupled to lumped-parameter circulatory systems to simulate realistic ventricular loading conditions. Finally, the impact of ventricular assistance was investigated by incorporating a pump with pressure-flow characteristics of an LVAD (HeartMate II™ operating between 8 and 12 k RPM) in parallel to the left ventricle. This allowed us to investigate the mechanical impact of acute left ventricular assistance at multiple operating-speeds on right ventricular mechanics and septal wall motion. Our findings show that left ventricular assistance reduces myofiber stress in the left ventricle and, to a lesser extent, right ventricle free wall, while increasing leftward septal-shift with increased operating-speeds. These effects were achieved with secondary, potentially negative effects on the interventricular septum which showed that support from LVADs, introduces unnatural bending of the septum and with it, increased localized stress regions. Left ventricular assistance unloads the left ventricle significantly and shifts the right ventricular pressure-volume-loop toward larger volumes and higher pressures; a consequence of left-to-right ventricular interactions and a leftward septal shift. The methods and results described in the present study are a meaningful advancement of computational efforts to investigate heart-failure therapies in silico and illustrate the potential of computational models to aid understanding of complex mechanical and hemodynamic effects of new therapies.Predictive computation models offer the potential to uncover the mechanisms of treatments whose actions cannot be easily determined by experimental or imaging techniques. This is particularly relevant for investigating left ventricular mechanical assistance, a therapy for end-stage heart failure, which is increasingly used as more than just a bridge-to-transplant therapy. The high incidence of right ventricular failure following left ventricular assistance reflects an undesired consequence of treatment, which has been hypothesized to be related to the mechanical interdependence between the two ventricles. To investigate the implication of this interdependence specifically in the setting of left ventricular assistance device (LVAD) support, we introduce a patient-specific finite-element model of dilated chronic heart failure. The model geometry and material parameters were calibrated using patient-specific clinical data, producing a mechanical surrogate of the failing  In view of the growing number and dismal prognosis of patients with end-stage heart failure, interest in emerging mechanical therapies such as left ventricular assistance devices (LVADs) has intensified. LVADs are used as a bridge to transplant, bridge to decision, destination therapy and, increasingly, as a bridge to recovery. The latter is fueled by the nearly ubiquitous demonstration that left ventricular (LV) unloading provided by LVADs causes reverse remodeling and, in a small percentage of patients, induces myocardial recovery to the point where devices can be explanted  data sets were obtained as part of the Aliskiren Study in Post-MI Patients to Reduce Remodeling (ASPIRE) trial  were imported and processed in Simpleware ScanIP . Geometrically detailed segmentations of the LV and RV were created relying on a combination of well-established techniques, including region growing, level-set thresholding, and morphological smoothing  was fixed in the longitudinal direction. Furthermore, the nodes on the endocardial annulus were constrained by coupling the average translation and rotation of the nodes to a fixed point in space located at the annulus center. This prevents rigid body rotation while allowing the annulus relative motion to inflate and contract during the cardiac cycle.The passive material response of the cardiac tissue uses an anisotropic hyperelastic formulation proposed by Holzapfel and Ogden  is defined through eight material parameters F is the deformation gradient, J is the determinant of the deformation gradient, J = det(F) and where The expression of these strain invariants can now be defined as:f0 and s0 are orthogonal vectors in the fiber and sheet direction in the reference configuration. Equation (2) is defined through J and a penalty term D, which is a multiple of the bulk modulus (D = 2/K). For deformation that perfectly preserves volume, J = 1.Where This passive material model, Equations (1–2), ensures that the material exhibits the well-documented exponential and anisotropic response to strain TF. We consider an active contractile stress in the fiber direction, resulting in a total Cauchy stress in the fiber direction,  by combining this with to the passive stress state in this direction (σpf):We consider the total Cauchy stress to be an additive contribution of passive and active components. The passive Cauchy stress, ns ∈ , such that the total Cauchy stress in the sheet direction is:Biaxial investigations on actively contracting rabbit myocardium revealed significant stress development in the cross-fiber direction that could not be completely attributed to myofiber dispersion or deformation effects  of 254 ml and an end-systolic volume (ESV) of 224 ml. End-diastolic pressure (EDP) and end-systolic pressure (ESP) were assumed to be 23 and 86 mmHg, respectively. These classify the patient with a severely dilated LV (>200 ml), elevated EDP  and exponential  terms were subject to uniform scaling by parameters A and B, a scalar and an exponential multiplier, respectively. These values were found by minimizing the error between the in silico diastolic PV course resulting from loading the LV of the FE model to the analytical Klotz curve . Material parameters were calibrated using ABAQUS as the forward solver, and an in-house PYTHON script containing the sequential least squares programming (SLSQP) optimization algorithm . Calibrated material parameters are presented in Table Once passive parameters were found, the active parameter TThe FE model of the heart was coupled to lumped models of the pulmonary and systemic circulatory systems used in our previous study  between the inflow and outflow cavities to which the pump connects. Experimental datasets specifying flow rates for a range of dP between 0 and 200 mmHg at discrete intervals of 2 mmHg were incorporated in the simulated flow profile for each operating speed. Flow rates were interpolated and extrapolated linearly between discrete values to account for a continuous flow-rate description for any dP the model may encounter. This is illustrated in Figure SD) unless otherwise stated. The differences between results were evaluated using analysis of variance (ANOVA) with differences considered statistically significant with p < 0.05. Time points in the cardiac cycle, such as end diastole, were defined for the untreated case and compared to the time points in simulations with LVAD support. End diastole and end systole, for each ventricle, were identified from the pressure-volume curves as the points immediately preceding isovolumetric contraction and relaxation respectively.For this study, cardiac function was simulated for a patient with chronic heart failure. LVAD therapy was introduced by simulating the effect of a HeartMate II operating at speeds ranging from 8–12 k RPM in 1 k RPM increments. Pressure and volume measurements of the ventricular chambers were recorded and compared to quantify ventricular loading and output performance. Myofiber stress was recorded and quantified for each simulation and compared to analyze the efficacy of treatment in the LV and potential harm of treatment to the septal wall and RV. Stress data in this study are expressed as mean ± standard of deviation , as shown in Table These functional changes are also captured in pressure-volume loops of each chamber, as shown in Figure The interventricular septum is in constant motion throughout the cardiac cycle. We tracked the midpoint motion at the base of the septal wall and quantified the leftward shift of this point in reference to the line through the anterior and posterior LV-septal-RV junctions. This measure of septal shift is positive when the septal wall bulges into the LV cavity, zero when the septal wall forms a straight line and negative when the wall bulges into the RV cavity. A time course of this measure over two heartbeats, shown in Figure SD are presented in Table p < 0.001). The improvements to RV mean myofiber stress were less substantial, with both end diastole and end systole myofiber stress in the RV remaining relatively unaffected.The volumetric-averaged myofiber stress  was calculated at end diastole and end systole, and the mean ± Myofiber stress distributions and overall geometry are presented in Figure A quantitative analysis of myofiber stress distribution in a segmented region of the vulnerable septal wall is presented in Figure We describe a geometrically and physically realistic model of an end-stage failing heart with representative systolic and diastolic myocardial material properties coupled to lumped parameter Windkessel-like models of the pulmonary and systemic circulations. This permitted study of heart mechanics and dynamics under realistic loading conditions i.e., pre-load and afterload of each ventricle. Finally, we simulated the effects of LVAD support by using experimentally recorded pressure-flow characteristics of a commonly used device. The present model represents a significant improvement over our prior modeling efforts  before computational models contribute to clinical decision making. We intend to address this in our future work. Finally, as noted, we studied only one combination of myocardial properties, RV and LV and vascular properties. Every patient is unique and conclusions arrived at are not generalizable.We described results of an FEA model based on the anatomy of an end-stage failing heart coupled to systemic and pulmonary vascular systems and an LVAD. We demonstrated the anatomic and hemodynamic impact of increasing LVAD speed on global pump function, regional stress distributions, and septal position. Realistic results were obtained in terms of ventricular deformation and PV loops. We demonstrated the expected findings that RV systolic and diastolic properties are affected by LVAD-induced unloading of the LV, resulting in RV dysfunction at high LVAD operating speeds. Having established the foundation of this model, we are poised to address the important question of whether and under what conditions, septal shift, and reduced LV pressure generation are important mechanisms of the development of RV failure following LVAD implantation. The specific conditions studied in the present model demonstrate that septal shifting alone is not sufficient to induce RV failure. Simulations spanning a wide range of conditions are required to fully address this important question.in vivo. Such an approach has the potential for predicting hemodynamics, such as the degree of unloading achievable, and the risk of developing right heart failure following LVAD implantation. Advances in patient-specific modeling in other fields of cardiology are already having an impact on clinical practice and it is anticipated that new applications will emerge, especially in the field of heart failure. The methods and results described in the present study have potential to meaningfully advance such efforts.Beyond this, the methods are generalizable in the sense that patient-specific geometries and vascular properties can be incorporated into the model, with the ultimate purpose of gaining insights into LVAD effects Patient cardiac MR data was obtained as part of the Aliskiren Study in Post-MI Patients to Reduce Remodeling (ASPIRE) trial. The study protocol was written by members of the executive committee of the trial and was approved by the ethics committees at each participating site. Individual patients provided written informed consent in accordance with the Declaration of Helsinki and anonymized data were sent to a core laboratory for analysis.KS, TF, DB, and JG were involved in the conception and design of study. KS created the computational models. Acquisition of various data critical to model creation was performed by SS and DB. The analysis and interpretation of modeling results was performed by all authors. KS wrote the first draft of the manuscript. DB and JG wrote sections of the manuscript. All authors contributed to manuscript revision, read and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Nucleic acids, proteins, and polysaccharides are the most important classes of biopolymers. The inherent properties of biomacromolecules are contrary to those of well-defined small molecules consequently raising a number of specific challenges which become particularly apparent if biomacromolecules are treated as objects in quantitative analysis. At the same time, their specific functional ability of molecular recognition and self-organization  enables us to make biomacromolecules serving as molecular tools in biochemistry and molecular biology, or as precisely controllable dimensional platforms for nanometrological applications. Given the complexity of biomacromolecules, quantitative analysis is not limited to the measurement of their concentration but also involves the determination of numerous descriptors related to structure, interaction, activity, and function. Among the biomacromolecules, glycans set examples that quantitative characterization is not necessarily directed to the measurement of amount-of-substance concentration but instead involves the determination of relative proportions (molar ratios) of structural features for comparison with theoretical models. This article addresses current activities to combine optical techniques such as Raman spectroscopy with isotope dilution approaches to realize reference measurement procedures for the quantification of protein biomarkers as an alternative to mass spectrometry-based techniques. Furthermore, it is explored how established ID-MS protocols are being modified to make them applicable for quantifying virus proteins to measure the HIV viral load in blood samples. As an example from the class of carbohydrates, the challenges in accurate determination of substitution patterns are outlined and discussed. Finally, it is presented that biomacromolecules can also serve as tools in quantitative measurements of dimensions with an example of DNA origami to generate defined dimensional standards to be used for calibration in super-resolution fluorescence microscopy. Biomacromolecules are naturally occurring macromolecular compounds that make up the essential building blocks of nearly all types of biological systems. Nucleic acids, proteins, and polysaccharides are the most important classes of biopolymers which are formed through condensation of their monomeric units: nucleotides, amino acids, and monosaccharides. The huge number of monomers linked together during polycondensation and the countless combinations that exist for sequencing them lay the foundation for an enormous diversity and structural complexity. Thus, biomacromolecules often exhibit a high degree of functionality and the ability for branching and/or subsequent chemical modification that facilitates the formation of linear and non-linear 2D/3D biopolymer architectures .The inherent properties of biomacromolecules are therefore contrary to those of well-defined small molecules consequently raising several specific challenges which become particularly apparent if biomacromolecules are treated as objects in quantitative analysis. At the same time, this specific functional ability of molecular recognition and self-organization  enables us to make biomacromolecules serving as molecular tools in biochemistry and molecular biology, or as precisely controllable dimensional platforms for nanometrological applications. Beyond that, the ability for molecular interactions and self-organization accompanied by a change of physical properties makes biopolymers a valuable basis for designed biomaterials, e.g., in medicine or for use in controlled drug release. Since the desired properties are also related to the dimensional structure of domains, the components of such functional biomaterials must be thoroughly characterized with respect to topological dimensions.Given the complexity of biomacromolecules, quantitative analysis is not limited to the measurement of their concentration but also involves the determination of numerous descriptors related to structure, interaction, activity, and function. However, even the determination of the amount-of-substance concentration of a biomacromolecule requires strategies different from those commonly used for the corresponding monomeric constituents.Several proteins, for instance, can serve as specific early warning indicators of critical body malfunctions related to common widespread diseases like cancer, cardiovascular diseases, or neurodegenerative diseases. The concentration of these protein biomarkers in body fluids like blood serum is, therefore, frequently measured in medical diagnostics. The techniques most widely used in laboratory medicine rely on immunochemical coupling reactions between antibodies and the target proteins realized by means of commercial test kits. SI traceability is realized via primary reference methods . AlthougThe dimensions of biological macromolecules typically are within nanometer length scales. Measurement methods used in nanometrology such as SFM, TEM, and SEM are therefore frequently applied for measuring physical, chemical, and dimensional properties of biomacromolecules. On the other hand, nanometer molecular dimensions in connection with their inherent ability of molecular recognition and self-assembly predestine them not only to be objects of analysis but also becoming versatile measuring tools. By the DNA origami technique, custom-shaped regular symmetric structures can be generated . In combIn case of the multifunctional and less uniform polysaccharides and their derivatives, we have to consider various types of dispersity: of molecular weight and if applicable of various sugar constituents in the macromolecular chains, of branching and of location of substituents which can only be quantitatively described with probabilities, as patterns and profiles. Among the biomacromolecules, the glycans set examples that quantitative characterization is not necessarily directed to the measurement of amount-of-substance concentration but instead involves the determination of relative proportions (molar ratios) of structural features for comparison with theoretical models, or the average dimension of domains , 7.This paper points out how Raman spectroscopy is currently being combined with isotope dilution approaches for realizing optical techniques as a possible alternative to ID-MS. Furthermore, it addresses current activities aiming at modifying ID-MS protocols to make them applicable for quantifying virus proteins to measure the viral load of HIV in blood samples. As an example from the class of carbohydrates, the challenges in the accurate determination of substitution patterns are outlined and discussed. Finally, it is shown that and how biomacromolecules can also serve as tools in quantitative measurements of dimensions is presented on the example of DNA origami to generate defined dimensional test samples that can be used for calibration in super-resolution fluorescence microscopy.Over the last two decades, complex biomolecules such as proteins increasingly gained in importance due to their ability to act as biomarker for supporting early diagnosis and monitoring of widespread diseases. The most commonly used standard techniques for their quantification in human body fluids are immunochemical-based test kits. Nevertheless, the high diversity and complexity of protein markers can lead to varying results depending on the assay design. To assure both SI traceability and comparability of the results of routine laboratory tests, reference measurement procedures are urgently needed.13C or 15N (or both) serves as an internal standard (spike) which is added to the sample in a known amount. Thus, the molecular mass is the distinctive feature between analyte and spike resulting in signal ratios if mass-sensitive techniques are used for detection. Neither MS nor Raman methods are limited to small molecules as has recently been demonstrated with the example of measuring the total hemoglobin (Hb) concentration in blood, one of the most frequently measured diagnostic parameter in laboratory medicine [While most established reference methods in clinical chemistry are based on mass spectrometry , recent medicine . This laRaman spectroscopy provides direct access to structural information of large and diverse molecules such as proteins. Based on molecular vibrations, a mass discrimination between analyte and spike is obtained from a shift of vibrational frequencies while the set of specific vibrations additionally provides a means for unambiguous identification of a compound including possible structural modifications of otherwise similar compounds.To achieve highest sensitivity, Raman spectroscopy is combined with nanotechnologies resulting in plasmon resonant signal enhancement, known as surface-enhanced Raman scattering (SERS). Coupling of the analyte to suitable metallic nanostructures or nanoparticles does not only improve the detection limits down to the sub μg/g level, but also opens the opportunity for integrating these methods into lab-on-a-chip devices . However15N2-DTNB -infected patients, monitoring of the viral load during therapy monitoring is implemented by the quantification of viral RNA by quantitative real-time PCR assays (qRT-PCR). In these assays, the fluorescent intensities measured at the threshold of the amplification curves are compared with those of a series of external standards with known copy numbers. The latest technological developments in quantitative molecular approaches are digital PCR (dPCR) methods, which enable absolute quantification of infectious agents without external standards. The more accurate quantification of DNA molecules by dPCR is based on the partitioning of DNA at limiting dilution resulting in zero or one molecules per reaction. The capabilities of dPCR for improved reproducible and traceable measurements have been demonstrated in several applications , 15. NevConsequently, the traceable quantification of viral proteins might be an appropriate alternative approach to overcome present shortcomings in viral molecular PCR tools, and isotope dilution mass spectrometry (ID-MS) would be a further powerful tool in this regard Fig. D. With IFurthermore, the development of independent approaches to virus quantification provides a more detailed knowledge about the correlations between viral protein and RNA amounts and possible discrepancies between these values and measurement failures. In this manner, exact measurements of viral RNA and protein amounts combined with calculations of HIV particle sizes might be used for the development and characterization of certified HIV reference materials which up to now are not available. In addition, the developed and validated ID-MS method can be used as reference for calibration of conventional qPCR assays.In contrast to proteins and DNA with their identical copies of defined macromolecules, polysaccharides—the most abundant class of biopolymers—usually do not possess a distinct size and sequence. Rather, they show various types of dispersity with respect to molecular weight, structure, and chemistry, including branching, sequence, and average composition of constituents. Furthermore, their polyfunctionality invites to chemical modification to obtain new materials, e.g., for biomedical and construction applications. To elucidate structure-property relationships, the structure has to be described in form of quantitative patterns and profiles with the highest resolution possible. As an example, water-solubility and thermoreversible gelation of methyl cellulose (MC) do not only depend on the average degree of substitution (DS) with methyl groups, but also on their location on the cellulose-constituting monomer, glucose, and on their distribution along and over the macromolecules . ConsequN-acetyl groups in chitosan. Beside specific depolymerization, appropriate derivatization and separation techniques, mass spectrometry (MS) is the method of choice for quantitative characterization of these types of complex samples. Due to the sensitivity of soft ionization techniques and mass analyzers to the chemistry and mass of analytes, mixtures of large heterogeneity with respect to DP and substitution pattern profiles cannot be analyzed quantitatively without further modifications  of analytes caused by different surface activities is overcome by this way of “isotopic labeling” [m/z. Discrimination due to discrepancies of complexation affinities to sodium are solved by introduction of tags carrying a permanent charge such as Girard’s T or aminobenzoic acid  maltooligosaccharides, the absolute ion intensities increased up to the surface saturation point of droplets generated by electrospray from which absolute intensities remained constant with certain discrepancy. Furthermore, it is possible to improve the quantitative MS analysis by employing an appropriate internal standard addition method, thus neutralizing the effect of out of control measurement conditions [In case of partially abeling” . Such prabeling” . Surfacecid Fig. B. Couplicid Fig. C, but alf ESI-MS . With innditions .Figure While we have just shown how biomacromolecules can serve as objects, we will now focus on their use as a tool in nanometrology. We therefore discuss DNA origami because they are currently one of the most prominent nanoscopic examples.DNA is a useful tool in quantitative and qualitative analysis. Due to its simple structure compared to proteins and enzymes, it can be controlled quite well. The most important advantage of DNA is the strong and specific hybridization of two single strands (Watson-Crick base pairing). Depending on the length of the DNA strand and the selection of bases, the strength of the bond between two single-stranded DNA (ssDNA) pieces can be adjusted. The size and properties of double-stranded DNA are well known and can be used for the controlled self-assembly of nanodevices (DNA origami) with very high yields of correctly folded shapes after purification . The syn6–1010) can be obtained.The DNA origami technique was introduced 2006 by Paul Rothemund  and a skAnother key aspect of the DNA origami besides the fast and easy assembly of billions of identical structures in one experiment is the easy way of adding dyes, nanoparticles, proteins and other small molecules of interest. Thus, a dye linked to DNA can be inserted into the DNA origami while folding  microscopy distances smaller than the diffraction limit of optical light can be resolved. The most common super-resolution microscopy techniques are structured illumination microscopy (SIM), stimulated emission depletion (STED) and localization-based microscopy . The advantage of fluorescence-based SR microscopy techniques over conventional techniques  is the possibility to measure biological structures without their destruction, even in living cells.Until the concept of DNA origami nanorulers was presented, the resolution limit was testified using, e.g., actin filaments or short dsDNA. The resolution is then determined as the smallest distance resolvable between two actin filaments or two dyes on the dsDNA strand. The drawbacks of actin filaments  can be overcome by the use of dsDNA. Therefore, the DNA origami became an important tool in super-resolution microscopy. With DNA origami nanorulers, a distance between dyes in the range of 6 to 360 nm is achievable and the quantification of the resolution of a microscopy system under ambient conditions is possible.x is proportional to the inverse square root of the number of emitted photons per localization: There are various techniques available for SR microscopy. In the following, we will focus on dSTORM to explain the application of DNA origami nanorulers. In the first step, the DNA origami is immobilized on a glass surface . By the use of wide-field fluorescence illumination, hundreds of molecules can be observed simultaneously Fig. E. This rA significant example for the use of DNA origami nanorulers is described by Balzarotti et al. . They teAll in all, DNA origami nanorulers provide the possibility to measure multiple identical distances at the same time. In addition, the DNA origamis can be easily modified to test for different distances as well as different wavelengths.The methods and applications discussed in this perspective are developing strategies for the analysis of biomacromolecules at the nanoscale. Quantification of biomacromolecules that serve as diagnostic markers is a challenging task in medical diagnostics. In this context, the availability of reference measurement procedures is of major importance since only those methods are suitable to adequately provide the necessary quality assurance in clinical laboratory medicine. Alternatives to ID-MS-based techniques such as ID-SERS are suitable to complement the portfolio of existing metrological tools. This would in any case be justified, since the huge number, diversity and complexity of potential biomarkers will most probably not be covered by a single universal analytical approach. Further development of existing MS methods towards the determination of viral loads would also help to improve reliability and comparability of routine test procedures for disease monitoring. Both the patients’ well-being and the healthcare system would likewise benefit from the availability of such a metrological basis in clinical chemistry.In case of complex mixtures of polysaccharide derivatives, mass spectrometry is a promising technique for determination of structural features, e.g., substituent pattern or nanoscale block lengths of blockwise-structured polysaccharide derivatives, provided proper isotopic modification is employed to suppress the chemical and instrumental discriminations. Developing a robust, yet highly accurate method for structural analysis of such complex derivatives provides better understanding of structure-property relationships, and forges the way to preparation of tailor-made materials.Self-assembled DNA-origami nanorulers are promising nanostructures that serve as a tool for measurements at the nanoscale. They are commonly used in super-resolution microscopy of biological samples to test the resolution under physiological conditions. Due to the huge number of variations that can be applied to control the design of a DNA-origami nanoruler, any possible distance between arbitrary chosen dyes can be realized. A further step in the nanoruler development is to use them in biological samples as a comparison tool for the estimation of nanometer distances.


We review how the hyperspectral dark field analysis gives us quantitative insights into the manner that different nanoscale materials interact with their environment and how this relationship is directly expressed in an optical readout. We engage classification tools to identify dominant spectral signatures within a scene or to qualitatively characterize nanoparticles individually or in populations based on their composition and morphology. Moreover, we follow up the morphological evolution of nanoparticles over time and in different biological environments to better understand and establish a link between the observed nanoparticles’ changes and cellular behaviors. Dark field microscopy anchored with hyperspectral imagery (HSI) is a novel optical approach with great potential in bio-related disciplines because it allows the identification and quantitative determination of specific components in the biological milieu. The optical approaches are universally employed methods as they offer a direct and reliable observation, however, they are limited by the resolution of the optical system . Bright HSI is a powerful multitool expanding into the realm of biological and medical sciences, which holds a proven record of success in astronomy, geosciences, agriculture, surveillance, resource management, and environmental monitoring, among other applications. HSI yields a hyperdata cube with continuous spectral and spatial information in one measurement, engaging non-contact and remote sensing. Moreover, this method is sensitive to subtle spectral changes ensuring a thorough discrimination of chemical or biological entities, or tracking agents' changes over time . The comHEDFM has a powerful resolving capability and is designed to provide in situ optical observation and spectral characterization of a wide range of nanomaterials as they interact with both biological and material-based matrixes, a key aspect for many bioanalytical applications. The unique interaction between light and matter generates characteristic profiles of scattered light, serving as spectral signatures for each target. Dark field illumination based on an annular cardioid condenser and highly collimated light at oblique angles on the sample, generates images with enhanced contrast and signal-to-noise ratio up to ten times (×10) higher than the conventional dark field optics ,7. The iHEDFM allows real-time analysis and can be successfully implemented in various biomedical areas, for instance, bionanomaterials synthesis, nanodrug delivery, and nanotoxicology ,11. The The hyperspectral data cube acquisition provides three-dimensional data with two spatial  and one spectral (z) dimensions. The hyperspectral image thereby is considered the dark field image along with the spectral information associated with each pixel of the image. Different strategies may be employed for the data cube acquisition, depending on the aimed resolution of the method. In general, the commonly utilized methods in HSI from higher to lower resolution are: point mapping or “whiskbroom”, line scanning or “pushbroom”, plane scanning or “staredown”, and the non-scanning or “snapshot” method . The imaNormally, a hypercube may contain hundreds of thousands of pixels with the size of a few nanometers each. A raw hypercube includes unwanted information, nonresponsive pixels, and spikes because of cosmic radiation that need to be carefully removed. Next, thresholding techniques are employed to visualize the image background at a wavelength that ensures enough contrast between the target and the background. The region-of-interest (ROI) tool converts the selected pixels into a spectral library or endmember collection file for the subsequent mapping analysis. The peak location classifier detects the pixels in a specific wavelength range allowing quantitative examination. For example, the size characterization of spherical gold nanoparticles (AuNPs) is possible across a wide optical window. The image pixels are analyzed in a desired wavelength range and are further classified upon their spectral responses. Colors can be associated to each range for the ease of identification of spectrally different nanoparticle subpopulations or spectral changes in individual nanoparticles. The Spectral Angle Mapper (SAM) is an automated mapping method acting as a threshold of spectral angle. The algorithm determines the spectral similarity and provides information about the location and analogy of endmember pixels in an input image. The spectral angle between the endmember and the image pixels defines the degree of similarity between known and unknown pixels. The SAM classifier is a powerful exploratory tool enabling the investigation of a new scene. In this review, we introduce the reader to how HEDFM can track interfacial changes at the level of individual and collective nanoparticles. Factors affecting the interface dispersant (NP)-dispersion medium are, for example, the composition of the medium, NPs’ stabilization procedure, NPs’ surface chemistry. This technique provides a tool to correlate changes in the physicochemical properties of colloidal nanoparticles caused by environmental  and non-environmental factors, such as the time, upon the identification of the corresponding spectral signatures. High speed and reliable screening, localization, and chemical identification of various components is the key in nanomedicine and biotechnology. Morphology and composition define the scattering properties of each compound which are then associated to a characteristic spectral signature. The surface of engineered nanoparticles synthesized using bottom-up protocols is normally modified  to provide colloidal stability in water-based media and to further functionalize the NP’s surface. The surface chemistry governs the interaction of the NPs with the environment , therefore its nature defines the NPs’ optical response. In aqueous conditions, many protective coatings present sensitive functional groups for the electrostatic stabilization of the particles that are highly affected by pH. We engaged a detailed HEDFM analysis to examine these differences using gold-copper sulfide nanoparticles (AuCuS NPs)  8]. Our. Our8]. All optical processes, including the scattering of light, are heavily influenced by the properties of the medium across the pathway of light. Biological environments are complex aqueous media rich in enzymes, proteins, sugars, peptides, electrolytes, among other molecules affecting the properties of the bulk in terms of ionic strength, viscosity, and refractive index. We wanted to study the impact of the composition of the dispersion medium on the NPs, so we carried out a spectral analysis of gold nanostars (AuNSs) in ethanol (EtOH) (a standard medium used for the storage of NPs) and phosphate-buffered saline (PBS) (a standard medium used in bio-related applications). The generated mean spectrum in ethanol showed a band located around 631 nm, while, in PBS, the band maximum slightly redshifted to 645 nm . This coNext, we exposed plasmonic nanoparticles with different physicochemical properties compared to the nanostars (around 80 nm) to ethanol and cell growing medium (GM), a more complex medium with high ionic strength like the PBS and higher viscosity due to the presence of macromolecules. To this end, we used polymeric hollow nanocapsules whose cavity was covered with discrete gold nanoparticles (PM–AuNPs) . t = 0 month . The highest concentration of nanostructures was subsequently employed in the photothermal ablation assays, demonstrating their potential for cancer treatment. Jenkins and co-workers used HEDFM to examine the colloidal stability of nanoparticles in blood [t = 5 min and a data cube for t = 18 h of incubation. The generated spectral libraries concluded that the AuNPs presented higher colloidal stability over time. In the case of AgNPs, the researchers evidenced the presence of agglomerated particles in the blood, pointing out their potential toxicity for in vivo studies upon intravenous administration.The ability to internalize hybrid plasmonic–superparamagnetic nanostructures by human breast cancer cells was reported in the work of Sotiriou et al . A spectin blood . The stuThe investigation and analysis of nanomaterials in tissue sections using HEDFM require special attention. The steps required for the preparation of the samples are prone to change the scattering properties of the nanomaterial. For example, Sen et al. described a shift in the scattering spectra of gold-nanorods-labeled leucocytes (GNRLL) after exposing them to mounting media which presented a high refractive index . They alWen et al. were able to spectrally differentiate the presence of three different nanosystems inside brain histological sections ,23. TheyNanomedicine is an emerging discipline capable to offer new perspectives to current medical limitations. Novel imaging technologies are necessary tools to improve the fundamental knowledge in bio-nano-interactions promoting advances at the diagnostic and therapeutic levels. In this context, the HEDFM presents straightforward and rapid analysis of nanomaterials in their colloidal dispersion state. The latter is of great importance to better understand the biointeractions of nanomaterials and study their fate in the native aqueous environment of any biological system. However, the huge amount of the generated data forming the hypercube require copious processing and careful analysis to extract appropriate conclusive information. Many research reports use HEDFM to present spectral differences or similarities, lacking both a linkage with the physicochemical background and a profound interpretation of the findings. Nanomaterials with well-defined optical properties are, by far, better candidates for reproducible and representative analyses minimizing the variations derived from particle polydispersity. As a final remark, we anticipate that the HEDFM technology will be part of the growing toolbox for the bionanotechnological development in modern health sciences.This review highlights the great potential of HEDFM technology to shed light on the dynamic interplay between nanomaterials and biological systems. Nanocomposites, at the single particle level or collectively, can be tracked by exploiting the dark field optics and characterized by their spectral signatures. Furthermore, with this technique, a non-invasive and label-free bioanalysis is possible, spanning from live cell and tissue studies to in vivo imaging of small organisms. We engaged HEDFM analysis to elucidate the relationship between nanomaterials and their environment through the monitoring of the optical response. We considered intrinsic factors of the nanomaterials, such as morphology and composition, as well as extrinsic stimuli related to their environment and to time. Examples of nanomaterial interactions in cells, blood, and tissues were presented for their biomedical relevance, and to show the applicability of the methodology in complex biological matrixes. To facilitate a comprehensive understanding of the methodology, we introduced the reader to the hyperspectral data acquisition and analysis.


To identify the fetal stem cell (FSC) response to maternal renal injury with emphasis on renal integrity improvement and Y chromosome detection in damaged maternal kidney.Eight non-green fluorescent protein (GFP) transgenic Sprague-Dawley rats were mated with GFP-positive transgenic male rats. Renal damage was induced on the right kidney at gestational day 11. The same procedure was performed in eight non-pregnant rats as control group. Three months after delivery, right ne- phrectomy was performed in order to evaluate the injured kidney. The fresh perfused kidneys were stained with anti-GFP antibody. Polymerase chain reaction (PCR) assay was also performed for the Y chromosome detection. Cell culture was performed to detect the GFP-positive cells. Technetium-99m-DMSA renal scan and single-photon emission computed tomography (SPECT) were performed after renal damage induction and 3 months later to evaluate the improvement of renal integrity.The presence of FSCs was confirmed by immune histochemical staining as well as immunofluorescent imaging of the damaged part. Gradient PCR of female rat purified DNA demonstrated the presence of Y-chromosome in the damaged maternal kidney. Moreover, the culture of kidney cells showed GPF- positive cells by immuno- fluorescence microscopy. The acute renal scar was repaired and the integrity of dam- aged kidney reached to near normal levels in experimental group as shown in DMSA scan. However, no significant improvement was observed in control group.FSC seems to be the main mechanism in repairing of the maternal renal injury during pregnancy as indicated by Y chromosome and GFP-positive cells in the sub-cultured medium. Fetal maternal cell trafficking (FMCT) can be defined as the presence of cells originating from genetically distinct individual without evidence of immunological response. FMCT is considered to be the trafficking of semi-allogenic fetal cells into the maternal circulation that may culminate in a mixtue of both maternal and fetal cells in maternal tissue during and after pregnancy. Several studies have demonstrated the persistence of FMCT in the CD34+ population for more than 30 years after delivery . Male ceHowever, methods to determine the role of fetal progenitor cells in treating damaged organs during the pregnancy are still laborious. The accretion of FSCs in the local damaged organs may be due to the consequence of the disease; or caused by the response to tissue injury. Furthermore, it has been considered that FSCs have the ability to gain tissue specific markers as they migrate to the environment of damaged maternal organ ,4. To daIn the current animal model, we used transgenic male rats expressing green fluorescent protein (GFP) in order to achieve the best sensitivity for better detection of FSCs in the maternal damaged kidney. The aim of the current study was to investigate the multilineage capacity of FSCs in repairing of maternal injured kidney as well as improving its functional capacity in a rat model of FMCT by detecting Y-chromosome and GFP-positive cells in the damaged part of maternal kidney.The local ethics committee approved the experimental protocol. The principles of laboratory animal care  were respected for animal treatment.Eight non-GFP female Sprague-Dawley rats weighting 220-310g were mated with GFP-positive male rats. All rats were maintained in standard single cages on a 12h darkness/12h light cycle with the best access to standard feed and water ad libitum in our laboratory. The rats were examined at 8h interval for detection of vaginal plague. The identification of vaginal plague was considered as gestational day 0 (GD0). Afterwards, female rats were kept in separate cages with consummative diet.®). For right kidney exposure, a lateral incision was made before separating the kidney from the adrenal gland and perirenal fat. Then, the renal damage  was performed on the lateral upper pole of the right kidney. The degree of burning could be precisely controlled by the period of contact with the kidney. In summary, the right upper renal pole was cauterized for a period of 4 seconds by the application of conventional cautery to avoid the incidence of necrosis as an irreversible renal injury. The maintenance of anesthesia was obtained by the injection of 0.75mg/kg Ketamine with 20 minutes intervals. The rats were then kept in specific cages under intensive care for termina-ting the gestational period. The same procedure was also performed in eight non-pregnant rats as control group to compare the role of FSCs in repair of damaged maternal kidney.Renal mass ablation by specially designed diathermy or electrocautery probe has been employed by several studies –11. At G® . In summary, deparaffinized and rehydrated samples underwent antigen retrieval by the application of Tris/EDTA buffer (pH=9.0) and a vegetable steamer. Endogenous peroxidase was blocked with horseradish peroxidase (HRP). In the next step, slides were blocked in 10% normal serum with 1% bovine serum albumin (BSA) in TBS for 2h, after being washed in Tris buffered saline (TBS) plus 0.025% Triton X-100. Ap-propriate dilution of antibodies was applied for overnight incubation of the slides.After 3 months of delivery, the rats underwent a nephrectomy surgery by the same surgeon in order to dissect the injured kidney. The kidneys were perfused with phosphate buffered saline to remove the red blood cells and were stained with anti-GFP antibody for identification of the FSCs in the non-GFP maternal damaged kidney. Anti-GFP antibody was purchased from DakoThe fresh frozen samples were also analyzed by immunofluorescent microscopy to verify the cell trafficking of GFP-positive cells in the kidney of non-GFP rat. Other organs, including liver and lung were also analyzed by immunofluorescent microscopy in order to detect homing of GFP-positive cells in undamaged tissues.® DNA kit  after nephrectomy of the damaged kidneys. The ubiquitous β-globin gene was amplified by PCR for the purpose of controlling the quality of extracted DNA and the lack of PCR inhibitors  were transferred to a tube containing 20mL of warm collagenase-trypsin solution. The tissue was incubated in an orbital shaker with gentle agitation within an incubator at 37°C for 1h. Subsequently, 20mL of basal medium contai-ning 0.05mg/mL DNase was added and the supernatant was gathered at 20-min intervals with gentle trituration. The collected supernatant was diluted with an equivalent volume of complete culture medium, dispensed into aliquots in 50mL tubes, and centrifuged at 100g for 15 min. Then, each pellet was resuspend in 45mL of complete medium and seeded 15 into one 75cm® SP-1 computer was applied to produce images representing slices through the kidneys in different planes in four rats of each control and experimental groups. The images obtained by SPECT are functional in nature rather than being purely anatomical.After induction of renal damage and three months after delivery in experimental group and three months after renal damage in control group, Technetium-99m-DMSA solution was injected into the tail vein of rats to determine the differential renal function of the damaged kidney in four rats of each control and experimental groups. The mean renal function was obtained in both groups. As DMSA solution needs approximately 4-6 hours to travel around the blood stream to reach the kidneys, anesthesia was maintained during this period. The progress of the DMSA solution through the kidneys was traced with a single-head rotating camera. For SPECT, an ElscintAll rats survived the whole period of our study. The presence of FMCT in renal papilla and tubular epithelial cells was confirmed by immu-nohistochemistry staining and immunofluores-cence analysis of the damaged kidneys. These results indicated that fetal cells were present in the maternal peripheral circulation and they also contributed to the repairing process of maternal kidney by migrating to the damaged parts for further differentiation. As depicted in The presence of Y-chromosome in the damaged maternal kidney was obviously proved by the gradient PCR of female rat purified DNA. Regarding the fact that the exact temperature for PCR of this specific primer was unknown, gradient PCR method was applied for a purified DNA sample at different temperatures. Results of the gradient PCR showed that the most appropriate temperature for this specimen was 54°C. Gradient PCR at 49°C was applied in order to reassure the specific and non-specific binding of the primer.The sub-cultured cells from the damaged part of the non-GFP kidney were viewed with immunofluorescent microscopy. The GFP-positive renal cells were observed in company with undamaged maternal renal cells B.Renal imaging was performed by the application of Technetium-99m-DMSA solution the high percentage of which in the renal cortex results in the high gamma flow. Each of the static scintiphotos was obtained by the application of a high-resolution collimator 1 hour after administration of Technetium-99m-DMSA solution. The result of DMSA scan after induction of renal damage in experimental group revealed decreased renal function in the right kidney in which the damage was induced (41%±0.9% versus 59%±0.8%). However, similar uptake in both damaged (right) and normal kidneys (left) with uniform distribution pattern of renal activity was obtained 3 months after delivery (49%±0.7% versus 51%±0.4%). The results of DMSA scan in control group revealed no signi-ficant improvement in function of right kidney (mean±SD) after 3 months of renal damage in-duction . DMSA scan of pregnant and non-pregnant rats confirmed the role of FSCs in repairing the damaged part of the kidney . The splThe current experimental study provided an overview about the role of FSCs in healing maternal renal damage during pregnancy where the GFP is set as the marker of fetal-origin cells in the maternal damaged part of kidney. The focus of the current study was on determining the role of FCSs in maternal renal function improvement.It has been shown that the frequency of FMCT can be affected by histocompatibility between the mother and fetus . ApproxiFSCs have the capability to increase locally in the injured organ during and after pregnancy. In several maternal damaged tissues such as kidney, specific FMCT have been detected in both mice and humans. In the study of Khosro-tehrani et al. the liver of a pregnant murine model was injured by carbon tetrachloride and it was demonstrated that the number of FSCs increased in the affected organ . In one It has been demonstrated that the morphological appearances of the FSCs within different mater-nal tissues was similar to differentiated maternal cells in mice during pregnancy .Their results were in accordance with the outcomes of the current study in which we revealed that the morphological appearance of FSCs in the damaged kidney after three months of delivery was indistinguishable from the maternal counterparts. In spite of the fact that no specific tissue marker was applied in the present study which is considered as one of the its limitations, the improvement of renal function, and the successful cell culture with the renal cell protocol can approve the differentiation of these FSCs to renal cells.In the study of Perin et al. stem cells from human amniotic fluids were injected into embryonic kidneys of murine. The results demonstrated the potential of these stem cells to differentiate into renal vessels culminating in early development of the kidney . In spitIn the present study, we sought to determine the prolonged persistence of FSCs in damage part of maternal kidney even after the pregnancy. Therefore, we decided to apply GFP-positive trans-genic male rats to identify the FSCs more easily by their green color. Using this method, we achieved high sensitivity detection of FSCs in the section of the injured kidney. The ability of FSCs in migrating to damaged maternal tubular epithelial cells was demonstrated by immune histochemical analysis with anti-GFP antibody. The domiciliation of GFP-positive FSCs was confirmed by immunofluorescent microscopy according to their special immunofluorescent staining.By PCR amplification of Y-chromosome sequences, we confirmed the persistence of FSCs in the blood circulation of the pregnant rat and the migration of Y-chromosome DNA of the male fetus to the damaged region of the kidney for further regeneration. In the present study, we confirm that FMCT may contribute to organ regeneration and repair. FSCs were only detected in injured region of the kidney and no FSC was distinguished in non-injured organs. These outcomes are in accordance with previous studies in which FSCs migrated to the maternal organs after induction of injury , 24. HowWe developed a surgical model of renal injury during pregnancy in order to essay the fetal cell response in maternal damaged kidney. As the number of FSCs rise with the gestational age, we decided to perform the renal damage at GD 11 which is in the middle of gestational period. In the current study, we showed that FSCs improved renal function and increased the proliferative response in the damaged kidney compared to the normal one.This study has also other limitations. Although the trend of improvement in functional parameters can ensure a persistent outcome in tissue repair, renal specific markers were not applied. However, renal function improvement, and detection of Y chromosome and GFP-positive cells in the sub-cultured medium indicated the ability of FSCs in migration from the peripheral circulation to injured maternal kidney and differentiate into renal cells. Considering the fact that serum creatinine and glomerular filtration rate are easier and cheaper modalities in bilateral renal injuries, their application was not feasible in this model in which unilateral renal damage was created. So, we focused on imaging techniques rather than these laboratory tests.While medical research teams are now trying to estimate the advantages and disadvantages of FMCT in human societies, this study demonstrated a perfect result of this phenomenon in functional improvement of the maternal damaged kidney in rat model. Therefore, we concluded that fetal GFP-positive cells have the potential to persist after delivery and domiciliate in injured kidneys with dynamic respond. It can be also realized that FSCs in maternal tissues have the ability to act as a reservoir of stem cells and pregnancy is a protection against susceptibility to several diseases. However, further studies are required to estimate the role of FSCs in repairing the maternal damaged organs by introduction of FSCs to maternal organ after harvesting and expanding the cells in vitro. Additionally, further investigations are required to estimate the efficiency of FMCT in human pregnancies.In this study, we investigated the role of FSC migration and homing in their final destination in different target organs and tissue regeneration. We sought to characterize FSCs with multilineage potential that migrate to the maternal organs. The results of the current study revealed that FSCs play a crucial role in repairing maternal damaged kidney and improve its impaired function without prior in vitro manipulation. However, more studies are required to conclusively demonstrate the role of FMCT in human maternal damaged organs.


Multi-Owned Housing: Law, Power and Practice explores the issues of private governance and management with respect to multi-owned housing. The foundations of the book were laid in 2005 with a conference paper in which the editors analysed multi-owned developments in England and New Zealand. The aim of this book is to provide a thorough conceptual basis on which the governance and management of housing with multiple owners can be examined across different jurisdictions in various countries.The choice of countries was partly based on practical considerations. The editors opted for countries with a common law regime and English language tradition. These rather arbitrary criteria may explain a certain imbalance as regards the book’s contents: five of the twelve country chapters deal with New Zealand and Australia. The other seven refer to England & Wales, Scotland, the United States, Israel, Singapore, Hong Kong and China. Besides its international perspective, interdisciplinarity is a key aspect of this book. As the backgrounds of the contributors suggest, the subject is relevant to scholars from a range of scientific disciplines.The introductory chapter presents a short and pragmatic conceptual framework for studying multi-owned housing. Its key concepts are rights and obligations and its focus is on the way these are allocated to the stakeholders during the life span of a complex. This conceptual framework is not applied consistently throughout the book, however, which makes it difficult to draw comparisons between the countries studied. Nonetheless, the book is informative and the case studies provide good insight in the issues the various countries encounter with regard to multi-owned housing.The book kicks off with England & Wales (Blandy). Following the conceptual framework, Blandy explores a wide range of statutory rules that apply to multi-owned residential projects and their owners’ experiences. Irrespective of the legal framework, individual owners find themselves in a far weaker position than the developers involved. Although this power imbalance has been reduced slightly by legislative reforms, further improvements seem necessary. Besides statutory changes, Blandy advocates initiatives that should lead to better-informed purchasers. A similar conclusion is drawn for Scotland (by Robertson). Although major reforms have recently changed the property system, the interests of developers and managers are still better safeguarded than those of the occupants. Robertson also goes into the negative effects that statutory requirements have on housing conditions. The Scottish legal arrangements are inadequate for proper governance, and this directly contributes to the very poor quality and largely unaccountable management services offered to owners of multi-owned complexes. The situation in the US (McKenzie) seems to be just the opposite. The relations between homeowner associations (CID’s) and the occupants are viewed as a private matter. Based on case studies in three US states, McKenzie concludes that these private relationships require more public oversight and a stronger regulatory framework.The chapter on Israel (Alterman) zooms in on the quality of multi-owned housing. She elaborates on the effectiveness of ‘simple’ (in Israel) versus ‘enhanced’ (in the US state of Florida) condominium laws to ensure long-term financing for long-term maintenance. The overall conclusion is not clear-cut: neither of the two types is adequate. Although enhanced laws provide a far better legal framework in stable economic situations, they are less flexible when facing an economic crisis. The fees the owners have to pay can overburden them. In her main recommendation, Alterman states the obvious: do not build tower condominiums! If policymakers nevertheless want to go that direction, an enhanced law should regulate their maintenance and a public body should monitor the financial means and range of the maintenance fund.The following chapters analyse residential multi-owned housing in Asia. Wang elaborates on the recently instituted but inadequate statutory framework for the management of new multi-owned housing in China. The power relationships between the stakeholders are hugely unbalanced. Again, owners lack power vis-à-vis developers and property management firms. In Hong Kong (Yip) a similar unbalanced legal framework fails to properly empower the occupants. In Singapore (Christudason) the problem lies in the power imbalance of minority residential owners versus majority commercial owners. The current statutory framework offers the minority owners inadequate legal protection in, for instance, decisions to sell mixed-use buildings.The final five chapters explore a variety of issues in multi-owned housing in Australia and New Zealand. The chapters on Australia deal with residential satisfaction in Sydney (Bounds), long-term management (Sherry) and multi-titled tourism accommodations in Queensland . The common denominator of these contributions is that they—again—highlight the unbalanced power situation during the life course of multi-owned projects. The two chapters on New Zealand elaborate on new forms of private governance for multi-owned housing (Dupuis & Dixon) and describe models for private governance of sustainable design features (Dixon & Van Roon).Despite differences in context and jurisdiction, the practical experiences in the countries studied are remarkably similar. The regulatory systems for multiple ownership give developers far more power than owners and occupants. Some serious problems need to be straightened out to allow positive changes in the future. The problems are not confined to the current statutory mechanisms but include the improvement of consumer protection, the development of ways to deal with mutual disagreements and the provision of unambiguous and comprehensible information for prospective purchasers (and other parties involved). An essential topic for the near future is the quality of multi-owned complexes. Although quality aspects are only touched upon in this book, the concluding chapter mentions maintenance as a vital component of any research agenda. The topic is explicitly related to the role of local authorities and the roles and obligations of individual owners. This approach should be broadened. An important task for the near future is to safeguard the physical and energetic quality of the multi-owned housing sector. The sector has grown rapidly over the past few decades and is expected to keep growing. The research focus must not only be on public law and the role of (public) authorities. It should also include the instruments that could be provided by private parties and the roles they could play. Besides physical quality, the future prospects of this sector also deserve attention. In the Netherlands, for instance, a considerable portion of the multi-owned housing sector consists of small dwellings in unattractive neighbourhoods. It is important to prevent these parts of the housing market from spiralling downward. Learning from experiences in other countries and sharing findings and recommendations from national research projects could bring us all further. This book provides a sound and valuable foundation on which the research work of scholars from different disciplines can be based.


We present a case of a 2-year-old boy with chronic suppurative osteomyelitis of the left jaw. A computed tomography (CT) scan demonstrated a periosteal reaction on the left side of the mandible with cortical destruction of the left mandibular head. The diagnosis could be confirmed histologically. During biopsy of the mandibular lesion, a purulent exudate was withdrawn. A two-year-old boy presented with a painless thickening of the left posterolateral portion of the mandible. The swelling had developed progressively over two months. In general, the child was in a good condition. On physical examination, the swelling of the left jaw felt fixed, hard and painless. Intra-oral inspection was unremarkable. Neither tooth decay nor regional lymphadenopathy was present. Laboratory findings showed anemia due to iron deficiency. A computed tomography (CT) scan demonstrated sclerotic thickening of the left mandibular ramus with periosteal new bone formation in a parallel pattern Fig.  and B, aOur case illustrates an example of the rare suppurative form of a primary chronic osteomyelitis of the left jaw in a two-year-old boy. Osteomyelitis present for at least one month is termed chronic; the mandibular swelling was already present for two months in our case . The mosCT of the mandible demonstrated some typical radiologic features of osteomyelitis described in the literature: a sclerotic thickening of the bone with parallel periosteal reaction Fig.  and B anThe laboratory findings in patients with chronic osteomyelitis of the jaw are nonspecific and are mostly limited to an increase of ESR and/or CRP . In our To obtain a definitive diagnosis and to exclude malignancy, we performed a biopsy of the mandibular lesion. The withdrawal of pus during biopsy already suggested an infectious cause. Histologic examination revealed bone trabeculae, fibrous intertrabecular stroma and foci of inflammatory cells (neutrophil granulocytes). Biopsy specimens were conclusive for oral bacteria that were sensitive for amoxicillin/clavulanic acid. The treatment was effective with regression of the mandibular lesion.


The interaction could lead to enhanced immunogenicity of PCV as a result of a phenomenon called carrier priming, whereby DTP is given some weeks before the first dose of PCV. This phenomenon could be implemented in the immunisation schedule in developing countries and among vulnerable populations to enhance the immunogenicity of PCV, reduce the number of doses required, and produce a more cost-effective immunisation program in developing countries.Pneumococcal conjugate vaccine (PCV) has the potential to interact with other vaccines containing diphtheria toxin-like antigens (such as those found in the DTP vaccine) upon sequential administration. This is attributed to the similarity of the diphtheria toxoid antigen to the carrier protein used to make PCV,  to diphtheria toxin  Streptococcus pneumoniae is a leading cause of pneumonia and death in children worldwide [orldwide . It is eorldwide . Vaccinaorldwide . Howeverorldwide . The cosorldwide . The besorldwide , which iorldwide . Therefoorldwide .The use of the 3 + 0 schedule is supported by randomised controlled clinical trials for prevention of pneumonia and invasive pneumococcal disease (IPD) in developing countries , and hasThe 13-valent PCV utilises carrier protein cross-reacting material 197 (CRM197) of diphtheria toxin, which is antigenically similar to the diphtheria toxin in DTP. It is believed that priming via the carrier molecule enhances the response to conjugate vaccines by increasing the number of carrier-specific T lymphocytes; these can provide the necessary support for the expansion and differentiation of polysaccharide specific B lymphocytes . Several


Metacommunity theory has provided many insights into the general problem of local versus regional control of species diversity and relative abundance. The metacommunity framework has been extended from competitive interactions to whole food webs that can be described as spatial networks of interaction networks. Trophic metacommunity theory greatly contributed to resolving the community complexity-stability debate by predicting its dependence on the regional spatial context. The meta-ecosystem framework has since been suggested as a useful simplification of complex ecosystems to apply this spatial context to spatial flows of both individuals and matter. Reviewing the recent literature on metacommunity and meta-ecosystem theories suggests the importance of unifying theories of interaction strength into a meta-ecosystem framework that captures how the strength of spatial, species, and ecosystem fluxes are distributed across location and trophic levels. Such integration predicts important feedback between local and regional processes that drive the assembly of species, the stability of community, and the emergence of ecosystem functions, from limited spatial fluxes of individuals and (in)organic matter. These predictions are often mediated by the maintenance of environmental or endogenous fluctuations from local to regional scales that create important challenges and opportunities for the validation of metacommunity and meta-ecosystem theories and their application to conservation. Identifying and inferring the relative importance of local and regional processes for predicting the number and relative abundance of species, as well as productivity and stability of local communities, have become defining goals of community ecology. Extending the concept of metapopulations to metacommunities2, defined as a set of discrete communities partially connected through movement of individuals, has greatly contributed to the study of local versus regional control. It provided a spatially discrete framework to the broader problem of scale3. Metacommunity theory also allowed studying regional processes as the collective property of local communities rather than as an external control5. In that respect, it contributed to understanding reciprocal rather than strictly hierarchical effects linking local and regional patterns and processes6.Natural ecosystems display patterns of variation in the diversity and abundance of species over a broad range of scales. One pragmatic approach to understanding these patterns contrasts local mechanisms regulating the assembly and growth of species to regional processes of immigration and speciation7. However, metacommunity theory has contributed not only a rich set of models but also experiments predicting regional patterns of intra-guild diversity, of food-web structure, and of their relationship to community stability and productivity8. It is also through the metacommunity framework that theories of local and regional control have included important progress in food-web theory, phylogenetics, and spatial dynamics, including spatially explicit, individual-based, and stochastic10 models. Finally, the metacommunity framework has inspired a similar spatially discrete approach to the study of cycling of matter and energy at local and regional scales. The relative importance of local cycling of matter and of regional subsidies has led to the development of meta-ecosystem theories, integrating spatial flows of individuals and of (in)organic matter over regional scales and integrating species interactions and the recycling of matter within local ecosystems.Theories of local versus regional control of spatially structured communities have been developed independently from the metacommunity frameworkBy reviewing recent progress, I will emphasize the importance of temporal fluctuations, both environmental and endogenous, and whole-ecosystem dynamics. I will also argue that such integration would be most successful within the broader theoretical framework of meta-ecosystems, where both individuals and (in)organic matter contribute to linking local and regional processes. Recent studies suggest that addressing these challenges is a key step toward resolving the reciprocal control of local and regional dynamics in natural ecosystems.11. In spatially implicit models assuming a homogeneous  connectivity network, intermediate movement reveals the reciprocal effects of local and regional processes: both regional and local diversity and relative species abundance then result from the balance between local sorting and immigration from the regional metacommunity. This balance can, for example, predict the contributions of both local species richness  and regional species turnover (beta diversity) to community stability, through local compensatory effects, and to the maintenance of regional asynchrony in species composition12.One of the main predictions of metacommunity theory is the regional distribution of species diversity based on both local competition and regional movement of species. In competitive metacommunity models, sorting, or environmental filtering, is the local process that operates through competitive exclusion, and sorting can lead to regional patterns of diversity through spatial heterogeneity in the environment. Movement among local communities is the regional process that redistributes, and eventually homogenizes, local communities, working against local sorting. Sorting is purely local in the absence of movement, whereas strong movement leads to regional sorting that favors the best competitor in the average habitat3 based on the relative importance of species movement and habitat heterogeneity as driving processes . The proposed mechanisms implicitly assumed demographic stochasticity, which stimulated the integration of a metacommunity framework to the neutral-versus-niche debate. Demographic stochasticity was treated explicitly as drift, affecting community dynamics along with competition, dispersal, and speciation14. Recent studies have provided a coherent framework reconciling competition and metacommunity theories15, whereas others have refined statistical methods for the inference of neutral- and niche-based mechanisms of species diversity16. Another important integration has recently happened with theories of biodiversity-ecosystem functions, tying into a long-standing empirical debate about the role of species richness for productivity and stability17. Within that debate, the competitive metacommunity framework can reconcile apparent contradictions in observed biodiversity-ecosystem function relationships. Dispersal limitation can, for example, either prevent or saturate local sorting processes and affect the strength of species complementarity underlying the stability and productivity of local communities8. Metacommunity dynamics can also resolve the combined and sometimes opposite effects of species diversity on multiple ecosystem functions18.The balance between local and regional processes in heterogeneous metacommunities was used to propose four mechanisms controlling biodiversity in competitive metacommunitiesa priori-defined. Also, when the local scale approaches the individual level, local and regional (meta)communities become arbitrarily defined along a continuum, based on assumptions about the relevant scales of processes at both ends of the continuum: competitive exclusion in local communities versus speciation and immigration over the metacommunity19. This is illustrated by the application of the metacommunity framework to neutral theory. In early neutral models, the metacommunity was independent from the local community, ignoring feedbacks from local to regional levels21. Later studies integrated both scales through spatially explicit models where metacommunity processes emerge from the collective behavior of connected local communities22. More recently, this scale continuum has been used to improve the testability of neutral predictions using patterns of spatial autocorrelation23.Spatially explicit metacommunities have blurred the distinction between local and regional scales because spatiotemporal heterogeneity in the distribution of species can emerge at multiple scales rather than being25. However, the emphasis on sorting through competitive interactions has limited the understanding of other traits and interactions, and of their complexity, as drivers of species diversity and function. The study of trophic interactions allowed researchers to extend this understanding beyond the effects of competitive exclusion to networks of species interactions that are more compatible with the complexity of natural systems.The competitive metacommunity framework emphasizes spatial heterogeneity among communities and as the regional mechanism of coexistence. It has also integrated the role of temporal environmental variability as a mechanism of coexistence26. One major implication of trophic metacommunities is the generalization of species traits involved in the local and regional control of community structure and dynamics. This generalization has recently led to a deeper contribution of metacommunities to the community complexity-stability relationship, involving the integration of evolutionary and non-equilibrium dynamics to the framework.The extension of competitive metacommunity theories to trophic metacommunities leads to a ‘spatial network of interaction networks’27, trait-mediated and density-dependent movement28, or covariance between trophic position and movement31. It has recently led researchers to revisit the hump-shaped relationship between dispersal and diversity in competitive metacommunities. When multiple trophic levels with heterogeneous dispersal rates are considered, this expected balance between immigration and sorting can instead give rise to monotonic increase in diversity with dispersal30  and intra-specific  synchrony. Using a similar approach, Pedersenet al.31 considered heterogeneous dispersal across trophic levels and showed that tri-trophic food webs are stabilized by the formation of patterns in the presence of both weak and strong dispersal across trophic levels organic matter is key to understanding food-web stability50 and essential to extending local-regional theories to a broad range of ecosystem functions. This understanding is a long-standing goal of ecosystem ecology and more recently has been addressed through meta-ecosystem theory.Both competitive and trophic frameworks are readily applicable to evolutionary ecology where both sorting and selection interact with dispersal to drive the assembly of species traits in relation to their environment52 and has been further developed in recent years.When both spatial and interaction networks are partially closed to flows of matter, metacommunities are better described as meta-ecosystems . Partial53. However, the meta-ecosystem concept has emerged as a more specific integration of trophic metacommunities and landscape ecosystem ecology52. It is partly based on the extension of cross-habitat subsidies developed in landscape ecology56 to reciprocal subsidies58. As such, meta-ecosystem theory integrates the cycling of matter across scales and can directly predict local versus regional control of community structure and ecosystem functions through the interplay between local recycling and regional fluxes of matter and their implications for the emergence of both local and regional community stability and productivity. For instance, explicit ecosystem dynamics have profound impacts on colonization-extinction dynamics in metapopulation and metacommunity models. As mentioned above, metacommunity theory predicts the local sorting of species in suitable habitats and their transient occupancy of unsuitable habitats through migration. In meta-ecosystem models, spatial fluxes and cycling of matter can become the driver of habitat suitability and result in dynamic habitat properties that are affected by both species interactions and ecosystem processes60. The dynamics of habitats become coupled to that of species over both local and regional scales and can lead to the emergence of facilitative interactions between competitors59. These facilitative interactions result from local cycling and regional fluxes and predict cascades of extinctions following habitat destruction60.Spatial ecosystem dynamics is not a new concept and has been studied as part of ecosystem ecology through flows of matter across macro-habitats and through large-scale quantitative ecosystem modelset al.61 showed in experiments and in models that increasing the frequency of such perturbations leads to a transient increase in subsidies but eventually to the collapse of recipient populations that are unable to recover fast enough. Feedbacks between trophic interactions and recycling also force us to reconsider the steady-state nature of ecosystem functions. Recent studies of non-equilibrium meta-ecosystems point to the importance of recycling and spatial flows of matter for the maintenance of endogenous spatiotemporal heterogeneity that predict both strong local fluctuations and regional stability. Meta-ecosystem models where only nutrients are flowing between local ecosystems lead to the destabilization of local consumer-resource dynamics62. Spatial fluxes of matter drive this destabilization, but recycling can facilitate the transition from equilibrium to cyclic dynamics. In simple two-patch meta-ecosystems, phase-locked cycles emerge from the positive feedback between recycling and passive movement of nutrients: more movement into a local ecosystem is correlated with increasing growth of the primary producer, thus destabilizing primary production. This prediction can be generalized to larger (many patch) meta-ecosystems63 where the top-down control of biomass storage into inorganic form can stabilize a meta-ecosystem in the face of nutrient enrichment. For more complex finite and irregular topologies, the eigenvalues of the connectivity matrix, rather than more common topological metrics, predict the minimum movement of nutrients or of individuals that can destabilize the meta-ecosystem64. Applying the meta-ecosystem framework to non-equilibrium dynamics has further revealed the relevance of endogenous spatiotemporal patterns of matter for the local versus regional control of ecosystem functions such as nutrient limitation: differences in movement rates between nutrients and other compartments lead to the emergence of nutrient co-limitation in growth over regional scales despite the lack of local mechanisms of nutrient co-limitation65. These recent studies illustrate how novel metrics of landscape heterogeneity, combined with a knowledge of spatial and species interaction strength, can lead to predictions that are tailored to specific empirical study systems. Finally, non-equilibrium meta-ecosystem theories are also relevant to conservation endeavors by predicting how the emergence of multiple scales of spatiotemporal patterns of abundance can inform decisions on the optimal size and spacing of local protected areas within regional reserve networks67.Recent studies have emphasized the importance of temporal variability in reciprocal subsidies of matter and the importance of the resulting non-equilibrium dynamics of meta-ecosystems. In simpler cases, this variability can be caused by external perturbations resulting in the release of matter associated with mortality. Harvey68. This challenge could constitute an important hurdle to reaching regional conservation goals.Metacommunity theory has broadened its range of predictions by integrating trophic interactions, spatially explicit movement, evolutionary processes, and phylogenetic history. By giving rise to meta-ecosystem theories through the integration of recycling and movement of matter, it has further emphasized the interactions between local and regional scales for the regulation of community structure and ecosystem functions. Overall, recent metacommunity and meta-ecosystem theories have benefited from the simplified representation of ecosystems as ‘networks of networks’. This approach has stressed the importance of differential movement rates across ecosystem compartments for predicting regional ecosystem stability. It more generally calls for the integration, both conceptually and methodologically, of spatial and trophic interaction strength. One fundamental challenge that materialized from these efforts is the potential for non-equilibrium dynamics and emerging spatiotemporal patterns that can drastically affect both local and regional community structure and ecosystem functions. Indeed, metacommunity and meta-ecosystem theories predict when local fluctuations, far from averaging out over regional scales, propagate and cause variability in these properties across spatial and temporal scales. This is a fundamental challenge for the development of meta-ecosystem theories and for testing their predictions using experimental metacommunity approaches that are based on steady states and that ignore the recycling and spatial fluxes of matter


The accepted review manuscripts have described the enzymes and proteins which contributed to IDILI and the various assays and current trends in developing comprehensive models. They represent the important research finding and will contribute to the development of toxicology.With the development of the society, the xenobiotics have brought high potential risk to human and animal. The study focused on the adverse health effect of xenobiotics which plays an important role in addressing public health challenge. The special issue has focused on the toxicity of xenobiotics. We call for manuscripts describing recent findings and future perspectives in the toxic field of xenobiotics on the human and animal health. Until the deadline of the special issue, 20 manuscripts have been received worldwide. After the review process, 7 manuscripts have been accepted by this special issue. The accepted research manuscripts have focused on the insecticides  and Bulldock 125®SC  and reported cases of intoxication responsible for approximately 2.2 million deaths. The number of incidents of exposure to pyrethroid insecticides has increased. Organochlorinated compounds (OCPs) are one of the important pesticides and have been used globally for many years. OCPs are not efficiently eliminated from the human body and may accrue in tissues because of enterohepatic reabsorption and affinity to adipose tissue. There has been limited work on interventions to facilitate elimination of OCPs. Evidence confirmed that enhanced mobilization and excretion via induced perspiration reduced the body burden of hexachlorobenzene (HCB) and polychlorinated biphenyl congeners. Removal of OCPs has also been facilitated by specific interventions which interrupts the enterohepatic circulation. To confirm that, sweat analysis may be useful in detecting some accrued OCPs, not found in regular serum testing. This method supplements the detection method of the OCPs. But perspiration is hard to collect. It maybe restricts its extensive use. Pyrethroid insecticides are another one of the important pesticides. Pyrethroid insecticides represent more than 30% of the worldwide insecticide market. Furia 180 SC is an isomer of cypermethrin insecticide. Bulldock 125 SC is an isomer of cyfluthrin. However, there is little information concerning the toxic effects of zeta-cypermethrin. But the research on the toxic effect of the Furia 180 SC (Zeta-cypermethrin) and Bulldock 125 SC (β-cyfluthrin) is still necessary in the future.Sclerosing agents as zinc gluconate-based chemical sterilants (Infertile®) are used for chemical castration, but there is not enough evidence of its safety profiles for the receivers. Infertile at 60 mM presented risk for animal health. It suggests that the optimal dose must be less than 6 mM. The future studies must be continued in order to clarify the safety of Infertile on cells and tissues, in order to better understand the pathophysiological mechanisms of this drug.The toxic component of plant often attracts the attention from food safety. Usnic acid is a compound of natural origin resulting from lichen secondary metabolism. It is considered one of the most important biologically active metabolites with important pharmacological properties: antitumor, antibiotic, antiviral, antioxidant, tuberculostatic, anti-inflammatory, and molluscicide. But limited studies showed toxicological potential of usnic acid. The teratogenic effect of usnic acid is in the period of organogenesis. It reveals the importance of evaluating the toxic effects of natural substances for elucidating the care in their indication as drug, particularly during pregnancy. Swainsonine is the primary toxin in locoweeds which is toxic plants. Its toxicity often occurs during pregnancy in livestock, and it may have different effects on embryonic development depending on the conceptus phase and maternal conditions during acute intoxication. Limited data showed the potential reproductive and developmental toxicity caused by swainsonine. The swainsonine caused reproductive and developmental toxicity in both parents and offspring mice. It suggests that severe reproductive toxicity and developmental toxicity are associated with swainsonine poisoning.Ethyl maltol and iron complexes are products of ethyl maltol and the iron found in the cooking pots used to prepare the Chinese dish, hot-pot. Limited data showed their safety. The ethyl maltol and iron complexes were toxic, and the targeted endocrine organs were the liver and kidneys. It suggests that alternative strategies for preparing the hot-pot, including the use of non-Fe-based cookware, need to be developed and encouraged to avoid the formation of the potentially toxic complexes. This experiment focused on the toxic of ethyl maltol and iron complexes, which is related to the Chinese dish, hot-pot. It is significant and it will attract more attention on these complexes.Idiosyncratic drug-induced liver injury (IDILI) is a significant source of drug recall and acute liver failure (ALF) in the United States. While current drug development processes emphasize general toxicity and drug metabolizing enzyme- (DME-) mediated toxicity, it has been challenging to develop comprehensive models for assessing complete idiosyncratic potential. The enzymes and proteins contributed to IDILI, including ones that affect phase I and phase II metabolism, antioxidant enzymes, drug transporters, inflammation, and human leukocyte antigen (HLA) which are important. It is imperative to develop various assays to detect individual reactions focusing on each of the mechanisms. The current trends in developing comprehensive models focused on examining the mechanisms. This review about IDILI is comprehensive and deeply showed the literature of potential mechanisms and predictive assays in idiosyncratic drug-induced liver injury. This review is significant and presents high value.