The loss of parenchymal brain volume per normative age comparison is a distinctive feature of brain atrophy. While the condition is the most prevalent to elderly, it has also been observed in pediatric ages. Various causes such as trauma, infection, and malnutrition have been reported to trigger the loss of brain tissues volume. Despite this literature based knowledge of risk factors, the magnitude of brain atrophy in pediatric age group is scantly addressed in most developing countries including Tanzania. The current study aims to understand the magnitude of brain atrophy in children residing in Northern Zone, Tanzania.A cross-sectional hospital survey was performed in which 455 children who were presented with various brain pathologies from the year 2013 to 2019 and whose brains examined by Computerized tomography (CT)-Scanners were recruited in the study. The brain statuses were examined using three linear radiological methods including the measure of sulcal-width, Evans index, and lateral ventricular body width.Results showed a significant number of atrophied brains among children in Northern Tanzania and that the condition was observed to have a 1:1 male to female ratio. The prevalence of pediatric brain atrophy was found to be 16.04%.The cortical subtype of brain atrophy presented as the most prevalent type of brain volume loss. The findings of this study suggest existence of considerable trends of brain atrophy in children which need special attention and mitigation plans. Brain atrophy being unusual reduction in brain volume than expected in the normal development and involution of brain at a particular age, is an important clinical condition in the current and future perspectives of neuroscience and related disciplines in studying neuro-cognitive functions and anatomical structures of the human brain. When fully developed, the atrophied brain manifests with reduction in parenchymal volume with the corresponding enlargement of sulcal spaces, basal cisterns and ventricles which are occupied by increased cerebrospinal fluid volume as a result of ex-vacuo response . Though Subjects and image acquisition: we studied 455 young patients who were presented in radiology departments of health facilities in the Northern Tanzania and performed brain CT scan examinations between the years 2013 to 2019. All patients underwent CT scan brain using primary axial cuts with slice thickness of 5mm and increment of 2mm. All the images were taken along the standard radiological baseline.CT scan image analysis: brain CT scan images were examined using the three known radiological linear methods to determine the presence or absence of brain atrophy. The similar measurements were used to differentiate the subtypes of brain atrophy including global atrophy, central atrophy, cortical atrophy, focal atrophy and hemi-atrophy. While the Evans index (EI) and lateral ventricle body width (VW) measure the central type of brain atrophy [ atrophy , the cor atrophy . The atr atrophy  and a lo atrophy . Global  atrophy .Statistical analysis: data were analyzed by R-statistical package where mean and p-values for age and gender association, and brain ventricular size and sulcal width were obtained. The percentage of subjects with brain atrophy or normal brain was calculated.Demography: the demographic characteristics showed that the predominant population involved children above 10 years of age, while the under five constituted a relatively lower population  and lateral ventricular width (VW) had P-value <0.05 suggesting significant association with age while the Evans Index (EI) had P-value of 0.431 indicating less significant age association  showed various brain areas of infarcts in some children who were abused, all those children in the study developed brain atrophy [et al. (2012) showed the plasticity of pediatric skull contributes to specific patterns of injuries of young children brains [Vulnerability levels seem to be closely associated with age such that the under-five years were more affected by brain atrophy compared with children between 4 and 10 years of age. The children above 10 years were least affected by this condition. This could be due to more risk factors such as low immunity to infection, incidence of trauma  and birt atrophy , Similarn brains  and thern brains . Life inn brains . TherefoBrain atrophy is paradoxically becoming common in childhood, pressing for the needful adoption and use of quantitative neuro-imaging techniques to attain correct diagnosis and classification of this condition. There is unprecedented significant number of children with childhood brain atrophy such that the current prevalence of pediatric brain atrophy in Northern Tanzania is 16.04% bearing a1:1 male to female ratio. Cerebral Cortical Atrophy is the most prevalent type of brain atrophy in childhood while Brain Hemi-Atrophy is the rarest form of brain atrophy sub-type in the studied population. The under-five children are more vulnerable in developing brain atrophy; hence putting in place programs for early neuro-imaging investigations may revolutionize the current clinical practice in developing countries by virtual of early depiction, grading and mitigation of disease progression into severe forms. The preferential selection of imaging modalities with non-ionizing radiation such as trans-cranial sonography and Magnetic Resonance Imaging stands to be the best consideration in young pediatric age groups.The brain atrophy is a condition found most commonly in elderly and may however be found in children especially related to malnutrition, head injury and central nervous system infection;Most causes of brain atrophy are currently irreversible.Pediatric brain atrophy is not uncommon. In the Northern Tanzania a prevalence of 16.04% has been depicted;Vulnerability to brain atrophy has been shown to be higher in children under-five years of age;Among the subtypes of brain atrophy, the cerebral cortical pattern of atrophy is most prevalent in the studied population.The authors declare no competing of interests.


The ability of exosomes to transport different molecular cargoes and their ability to influence various physiological factors is already well known. An exciting area of research explores the functions of exosomes in healthy and pathological pregnancies. Placenta-derived exosomes were identified in the maternal circulation during pregnancy and their contribution in the crosstalk between mother and fetus are now starting to become defined. In this review, we will try to summarize actual knowledge about this topic and to answer the question of how important exosomes are for a healthy pregnancy. Intensive maternal-fetal information exchange is required to establish and to maintain a healthy pregnancy. The placenta is the most important organ in this respect, performing vital functions for the fetus to support its growth and survival and to maintain the pregnancy . The plaExtracellular vesicles (EV) are heterogeneous populations of cell-derived membrane vesicles released by eukaryotic and prokaryotic cells to the extracellular space. The classification of EVs is based on the origin and size of the vesicle  5]. The. The5]. 2, and nutrients takes place between the maternal and the embryonic circulation. The embryonic disc undergoes further differentiation processes that give rise to the gastrula, a trilaminar disc comprised of the three germ layers ectoderm, mesoderm, and endoderm. During the next weeks, early progenitors for all kinds of tissues are formed. The growing embryo detaches from the uterus wall staying connected through the umbilical cord. The cord contains blood vessels of the embryonic circulation and connects it to the placenta where material transfer takes place without direct mixing of the embryonic and maternal blood.After fertilization, the zygote starts to move through the fallopian tube where the first mitoses occur . The firThe crosstalk between fetus and mother can occur as a simple diffusion of molecules through tissue layers or in a better protected manner through extracellular vesicles, especially via exosomes  9,10]. ,10. 9,10Fetal-derived exosomes travel to the maternal side to potentially transmit signals to the uterus and cervix. It was even suggested that inflammatory signals delivered by exosomes could contribute to the onset of parturition . ExosomeThrough the effects on the maternal organism, the released exosomes also support growth and survival of the fetus, therefore one would expect an increased production of exosomes during pregnancy. This hypothesis was confirmed, as the concentration of exosomes in maternal peripheral blood was ~20-fold greater than that observed in non-pregnant women . The conIn general, research techniques to study the placenta-derived exosomes use cell cultures of trophoblasts (primary and cell lines), chorionic villi explants, placental perfusion, plasma and urine from pregnant women. Main areas of interest about exosomes in pregnancy include: demonstration of the exchange of EVs between fetus and mother; the role of EVs in implantation; the establishment of immune tolerance; and the regulation of angiogenesis and endothelial cell migration.The human placenta releases a wide range of molecules and EVs which support the maternal physiology to adapt to fetal requirements during pregnancy ,19,20. POne class of small RNAs transported via extracellular vesicles, the microRNAs (miRNAs), attracted special attention. MiRNAs are post-transcriptional regulators of gene expression by inhibiting the translation of their target mRNAs or inducing mRNA degradation. It was estimated that the expression of 60% of the human genes are regulated by miRNAs . The loaCharacteristic miRNAs, which are highly expressed in human placentas, were identified in the serum of pregnant women. For instance, the concentration of placental miRNA-141 increases in maternal plasma with gestational age . SeveralInterestingly, there is evidence that exosomes can help the fetus to avoid viral infections. Non-placental cells incubated with placental exosomes became more resistant to viral infection and this effect was mediated by the delivery of specific miRNAs . RecentlA metadata analysis of the function of extracellular vesicles in the human reproduction system suggested that they contribute to oocyte and sperm maturation, fertilization, prevention of polyspermy, and embryo implantation . During Exosomes derived from human endometrial epithelial cells were shown to be taken up by trophoblasts to enhance their adhesive potential partially by enhanced focal adhesion kinase signaling . In addiEvaluating exosomes in the uterine luminal fluid of sheep demonstrated that vesicle transported proteins could affect the implantation and fertility outcomes. Released by the endometrial epithelium, vesicles were taken up by blastocysts and the endometrium around the uterus . A totalThe fetus is antigenically distinct from the mother and therefore it is necessary to establish a tolerant immune state to prevent the rejection of the developing organism . The rolTrophoblast-derived EVs were further shown to induce the differentiation of T cells into Treg (regulatory T) cells . Treg ceIn cattle, placenta-derived exosomes are abundant in the circulation of pregnant cows and exosome-derived miR-499 attenuated the expression of proinflammatory cytokines by inhibiting NF-κB signaling, thereby attenuating inflammatory responses and forming an immune-tolerant microenvironment in the uterus. Inhibition of miR-499 lead to inflammatory deregulation and increased risk of pregnancy failure .A role in preventing the embryo from being attacked by the maternal immune system was described for glycosphingolipids (GSLs) in exosomes . GSL expInflammation is part of an active immune response. Considering the necessity for inducing tolerance for the embryo, is might seem counterintuitive that the implantation requires a slightly inflammatory status. However, this condition does not affect the fetus directly, but facilitates tissue remodeling and embryo implantation . PlacentInitial stages of pregnancy are characterized by spiral artery remodeling and physiological adaptations in the cardiovascular system to provide sufficient supply of nutrients and oxygen to the growing fetus . CoordinExosomes derived from porcine trophectoderm cells induced the proliferation of the maternal endothelial cells and promoted angiogenic processes due to the presence of specific miRNAs. Increased miR-150 levels in the EVs stimulated the proliferation and migration of endothelial cells, thereby exhibiting a pro-angiogenic effect. A reduced expression of porcine miR-150 in the umbilical cord blood-derived exosomes of pigs showed intrauterine growth restriction . MiRNAs Placental exosomes are reported to be involved in the pathology of pregnancy as well ,67. The One of the most serious and common complications of pregnancy is preeclampsia (PE). It occurs especially in the third trimester of pregnancy. PE and associated hypertensive disorders are responsible for nearly 40% of premature births ,73. In tBoth, exosomal miRNAs and proteins, were successfully employed as predictive biomarkers . Small RProteomic studies identified about 400 proteins in syncytiotrophoblast EV samples with 25 proteins  unique to PE compared to healthy pregnant controls . SyncytiAccording to the World Health Organization, an estimated 15 million infants are born too early each year . ComplicIn pregnant mice, the concentration of exosomes was significantly lower in preterm birth induced by inflammation compared to animals with normal delivery . PathwayA range of miRNAs isolated from maternal plasma were described as predictive biomarkers for pre-term birth, however, whether these miRNAs were transported by EVs was not evaluated in these studies ,95,96,97Another important complication of pregnancy, which can harm both the fetus and the mother, is gestational diabetes mellitus (GDM). During normal pregnancy, at the time of the second and third trimester, limited insulin resistance gradually increases as a normal phenomenon to ensure sufficient nutrient supply for the fetus. A pregnant mother with pathologic insulin resistance—acquired or chronic—is not able to compensate for the increased circulating glucose concentrations because of β-cell dysfunction. As a result, maternal hyperglycemia occurs, leading to increased risk of disease for both the mother and the fetus . GDM is A proteomic study identified 78 proteins with significantly altered levels in GDM-derived exosomes compared to normal ones. Pathway analysis revealed that many of these proteins are involved in energy production and inflammation . AnalysiThe contribution of exosomes in fetal-maternal communication during pregnancy has been firmly established . It is e


From ontogenesis to homeostasis, the phenotypes of complex organisms are shaped by the bidirectional interactions between the host organisms and their associated microbiota. Current technology can reveal many such interactions by combining multi-omic data from both hosts and microbes. However, exploring the full extent of these interactions requires careful consideration of study design for the efficient generation and optimal integration of data derived from (meta)genomics, (meta)transcriptomics, (meta)proteomics, and (meta)metabolomics. In this perspective, we introduce the holo-omic approach that incorporates multi-omic data from both host and microbiota domains to untangle the interplay between the two. We revisit the recent literature on biomolecular host-microbe interactions and discuss the implementation and current limitations of the holo-omic approach. We anticipate that the application of this approach can contribute to opening new research avenues and discoveries in biomedicine, biotechnology, agricultural and aquacultural sciences, nature conservation, as well as basic ecological and evolutionary research.  Microbiome; Evolutionary Biology These microorganisms associated with plants and animals are now acknowledged as relevant—even essential—assets to many basic biological processes, including nutrient acquisition , immune genomic properties  can be expanded to the holo-omic level by the incorporation of data from multiple omic levels from both host and microbiota domains transcriptomes, (meta)proteomes, and (meta)metabolomes. The same technologies also enable epigenomic and exposomic profiling, which can further contribute to disentangling the biochemical associations between host-microbiota-environment interactions and their effect on host phenotypes . Howeveenotypes . The anaenotypes . This apenotypes  and metaenotypes , but metIn this context, the technology to generate large amounts of data to be used in a holo-omic context is already available, but the analytical tools to reveal and identify host-microbiota interactions are still limited. As a consequence, only a handful of research groups worldwide have been able to effectively implement the holo-omic approach. To contribute to the development of this new field, in this perspective we first revisit the available evidence for the biological importance of host-microbiota interactions. Second, we present how the holo-omic toolbox can be used to study host-microbiota interactions at varying levels of complexity to guide researchers through applying the holo-omic approach. Third, we showcase the potential provided by the holo-omic approach to host-microbiota interactions in both basic and applied biological sciences and finally we identify the limiting factors that currently prevent the widespread implementation of the holo-omic approach and discuss possible solutions to overcome them.The holo-omic approach to host-microbiota interactions relies on three major assumptions: (1) host-associated microorganisms interact not only with each other but also with their host ; (2) theLactobacillus rhamnosus increases the uptake of fatty acids in zebrafish by down-regulating the transcription of host genes related to cholesterol and triglycerides metabolism   . The orime (IBS)  and it sme (IBS) .All these examples highlight the relevance of acknowledging and understanding the biomolecular interactions occurring between different omic levels of hosts and microorganisms. In the following section we will describe how holo-omics can be implemented by addressing different methodological, experimental, and analytical approaches.amplicon sequencing, targeted RNA-sequencing, and western blotting) to characterize the microbial domain. However, untargeted approaches , which non-selectively provide a snapshot of nucleotides, proteins, and metabolites present in a sample, are progressively complementing or replacing targeted approaches. For instance, coupled untargeted host/microbe data from shotgun sequencing offers advantages over targeted approaches, such as the construction of metagenome assembled genomes (MAGs) from metagenomic data , might provide the required resolution when trying to uncover the interaction between well-defined binary interactions, e.g., the effect of microbiota-produced butyrate on host transcriptomics in epithelial cells  have linked specific loci in the host genome to the presence of pathogenic or beneficial microbes (MGWASs) linking specific genes in the metagenome to phenotypic traits of interest in the host Are host-microbiota interactions relevant in the system under study?(2)Is it meaningful to implement a holo-omic approach?Researchers must assess whether host-microbiota interactions are relevant for understanding the system under study. The impact of microorganisms associated with complex hosts is now regarded as almost universal , but theIt is necessary to evaluate whether the implementation of a holo-omic approach is reasonable given the properties of the biological system and its environment. Holo-omics relies on the premise that genomic and metagenomic differences across individuals, treatments, populations, or species affect biological processes and phenotypic outcomes. Thus, the existence of genomic or metagenomic variation in the system is essential. It is also necessary to bear in mind that the capacity to recover genomic and metagenomic signatures is largely affected by environmental variables . The bac(3)Which omic levels are relevant and how to maximize the amount of useful data derived from them?Assessing the economic and technical feasibility of the study is also paramount. This includes acknowledging the genome size of the host, as the genome of some species can be magnitudes larger than others, e.g., amphibians  versus bLastly, researchers should identify which omic levels are the most relevant, both by considering the biological and technical features of the experimental system and their relevance to the research questions. The omic levels selected for analysis will largely determine the number of samples to include , where aThe methodological, experimental, and analytical approaches mentioned above are challenged by the high costs of data generation and the complexity of downstream analyses. This requires that researchers consider at least three fundamental questions about the system under study before taking on a holo-omic study .Box 2ThrThe holo-omic approach outlined above can be implemented in many basic and applied biological research fields to address relevant scientific questions concerning host-microbiota interactions. In the following, we showcase and discuss the application of the holo-omic approach in diverse fields of biological sciences. For some fields, we include boxes containing case examples to better illustrate its potential implementation.Box 3(1)Are host-microbiota interactions relevant in the system under study?Chickens are an important source of high-quality protein for a large proportion of the human population. The gut microbiota of broilers (chicken bred for meat production) is highly variable since they are slaughtered before reaching an age in which the microbial community dynamics stabilize . Althoug(2)Is it meaningful to implement a holo-omic approach?The controlled environmental conditions in intensive poultry production systems, which use the same feeding strategy and environment for all individuals, indicate that the likely reason behind variation in the chicken performance and their gut microbiota when administered pre- and probiotics might be explained by microbial founding effects and microgenomic variation of broilers ((3)Which omic levels are relevant and how to maximize the amount of useful data derived from them?Pre- and probiotics interact with native gut microorganisms as well as with the host. The gut microbiota of broilers is relatively simple because of the closed environment where the broilers are reared. The genetic diversity of conventionally bred broilers is low, yet even small interindividual differences can be crucial and might have wide implications on the response to pre- and probiotics. These system properties allow the successful application of the holo-omic framework for obtaining relevant microbe-microbe and microbe-host interactions, which can help researchers optimize feed additives design, production, and administration, thereby preventing production inefficiency driven by gut dysbiosis.If associations are detected between the (meta)genome and host phenotypic traits, the study of transcriptome, metatranscriptome, and metabolome can unveil the nature of microbe-microbe and host-microbe interactions and how they affect the host. Detecting molecular pathways that are activated or deactivated in the presence of pre- and probiotics can enable researchers to identify production-related phenotypic changes in the host.The holo-omic approach could prove a meaningful tool in developing animal and plant production as microorganisms are increasingly considered essential assets to improve efficiency and sustainability . Among oThe holo-omic approach could also contribute to developing and optimizing biotechnological solutions. For instance, it could be used to better understand host-microbiota systems capable of enzymatically degrading complex polysaccharides  in the sBox 4(1)Are host-microbiota interactions relevant in the system under study?The use of fecal transplants, i.e., transferring fecal material from a healthy donor to a patient with a gastrointestinal disorder, is now becoming a promising treatment for multiple gastrointestinal disorders . Althougepigenotype of the human host ((2)Is it meaningful to implement a holo-omic approach?The success of fecal transplants relies on the capability of beneficial microbes from the donor fecal sample being able to colonize and establish themselves in the gut environment of the recipient. One can hypothesize that the probability of success relates to differences among patient gut “environments” that depends not only on the existing microorganism community but also on the genotype or Clostridioides difficile ((3)Which omic levels are relevant and how to maximize the amount of useful data derived from them?The information gained from a holo-omic approach will ultimately lead to more efficient treatments by, for example, optimizing the biological match between a fecal donor and recipient. For example, a holo-omics analysis in a controlled cohort can reveal concrete genotypes of a host that are associated with the gut microbiota composition. Then, once we have accumulated knowledge about specific candidate genes directly associated with composition and function of gut microbiota, we can screen these genes to optimize the genetic match between donor and recipient, thereby improving the odds that the recipient is likely to adopt the healthy microbes from the donor and thereby counteract the negative effect from microbes such as ifficile .(3)WhichA holo-omic approach to identify the factors underlying the differential success of fecal transplants could include the patients' genomic and epigenomic features coupled with transcriptomic, metagenomic, metatranscriptomic, and metabolomic variation before and after a fecal transplant. Associating these features with the success of the treatment, and with each other, would shed light on the functional changes introduced by the transplant, which would enable identifying the factors leading to a success or failure of the treatment.Incorporating the holo-omics approach to biomedical research offers an exciting new avenue toward better treatments of many modern human diseases. Most people in industrialized societies exhibit depauperate gut microbiotas , which ihologenome theory of evolution  feeds almost exclusively on conifer needles rich in resin and phenol and low in nutrients ((1)Are host-microbiota interactions relevant in the system under study?In winter, the Western capercaillie (utrients . It has utrients . Failureutrients .(1)Are h(2)Is it meaningful to implement a holo-omic approach?The highly specialized diet with many hard-to-digest components of the capercaillie suggests that the digestion of these compounds might be facilitated by the microbiota. An initial screening using shallow shotgun sequencing will indicate microbial differences between wild and captive capercaillies to identify taxa and functions related to the degradation of resin and phenol that might be missing in captive individuals.(3)Which omic levels are relevant and how to maximize the amount of useful data derived from them?If the captive bred individuals originate from the same population as they are meant to be released in, then the system is relatively simple with two similar genetic backgrounds (wild and captive-bred from the same wild population). This means that the effect of genetics is roughly the same for wild and captive conspecifics, which will allow researchers to study the impact of the environment  on microbiota functionality transcriptome. Screening both the transcriptome and metatranscriptome will allow conservationists to uncover complementary interactions between host and microbiota genes. If the genes of interest are suddenly present and expressed then the dietary change has been enough to provide the captive capercaillies with a “wild microbiota” and released animals can then be monitored and their fitness compared with control animals with a captive microbiota. If captive individuals fail to acquire the needed functionalities through the gradual change to a more natural diet other vectors of enrichment should be tested .Holo-omics could also be relevant for developing optimal active conservation actions, such as captivity breeding and animal translocations . As captAlthough holo-omics represent a valuable tool for many fields, its implementation is still hampered by economic, technical and biological limitations. A main economic hurdle is the high cost of shotgun sequencing. Targeted sequencing or DNA microarrays approaches can be cost-effective alternatives for characterizing (meta)genomes in some cases, although shallow shotgun sequencing can in some instances recover higher taxonomic resolution at the same cost, while also providing direct inference about functionality . TargeteThe holo-omic approach faces essential challenges, such as those linked to the quantity and complexity of the data to be analyzed. The interactions between different microbes, each synthesizing and metabolizing a variety of molecules, and the interactions between microbes and host cells is extremely complex, with the nature of these interactions being far from uniform and linear. This demands an integrative approach that can account for the different data types under the same inference framework. Generative/mechanistic models exist for many of the individual omics data, such as transcriptome, proteome, and metagenome, but integrating these models under a single inference framework is challenging, given the different data types  and the vastly different biological processes that underlie them. Thus, developing mechanistic models for such data are an active area of research. In addition, in most current studies, the holo-omics data contain a lot of missing values, e.g., the transcriptomics and microbiome data may not come from the same individuals, and the generated data fall under the small N (sample size), large P (features) paradigm. That is, the data contain a limited number of independent observations of a large number of features. In the case of holo-omics data, features can include millions of genomic variants, mRNA quantification for thousands of genes, abundance estimates of hundreds to thousands of taxa in the microbiome, and tens to hundreds of phenotypes such as health parameters and growth rates. Unfortunately, the large number of features (P) are not accompanied by a corresponding increase in sample sizes (N), owing to the high cost of generating such comprehensive data for a large number of individuals. Identifying the important determining features in such datasets can be very challenging given the limited number of independent observations. Statistical advances in the last decade including development of deep learning methods are helping address the challenges posed by the high dimensionality and complex correlation structure of the data. Development of such methods is an area of active research where several advances have been made in integrating host-microbiome data .Although still challenged by many limitations, the feasibility to conduct holo-omic research will only increase in the near future, aided by the continuous publication and improvement of macro- and microorganism genomes, the decrease of costs for DNA/RNA sequencing and mass spectrometry, the increase of computational capacities, and the uninterrupted development of analytical tools to analyze the huge amounts of data generated. These trends will allow a broader range of research groups to conduct holo-omic studies and as the need for detailed information on host-microbiota interactions increases in both applied and basic sciences there is no doubt that the holo-omic approach will gain popularity in the future.


Vascular calcification is an abnormal cell-mediated process in which bone-specific hydroxyapatite crystals are actively deposited on the blood vessel wall and is a significant pathological basis for the increased incidence and mortality of adverse cardiovascular events. Macrophages play an important regulatory role in the occurrence, development, and regression of vascular calcification. After the tissue microenvironment changes, macrophages subsequently change their polarity and phenotype or secrete functional substances as an adaptive response. As research on macrophages continue to move into this field, we gain a new understanding of the mechanism of the formation and regression of vascular calcification, which might offer valuable new intervention targets for the prevention and inhibition of vascular calcification. This review summarizes a wealth of research in this field and explores the roles of macrophages in the development process of vascular calcification. Vascular calcification is a prevalent complication of various diseases that increases the incidence of acute coronary cardiovascular events and seriously impacts the quality of life of patients due to increased vascular stiffness and decreased compliance . CurrentVascular calcification was previously considered to be the passive deposition of calcium and phosphorus in the vascular wall after a long term metabolic imbalance and to not be regulated by other factors. At the end of the 20th century, Demer proposed the concept of high ossification of vascular calcification. Currently, it is increasingly recognized that vascular calcification is an active and highly regulatable biological process that shares certain similarities with bone formation . VasculaThe pathogenesis of vascular calcification is complex and manifold, including high calcium and phosphorus levels, a chondrogenic phenotype differentiated from vascular wall cells, decreased levels of natural calcification inhibitors , inflammatory stimulation, apoptosis, and release of pro-calcified particles . The cruVascular calcification shares many similarities with skeletal mineralization. Osteoblasts and osteoclasts maintain a dynamic balance during physiological bone remodeling. Osteoblasts deposit bone matrix proteins that can subsequently become mineralized and coordinate with bone resorption by activated osteoclasts. Once the dynamic balance between osteogenesis and osteolysis is broken, vascular calcification or pathological bone remodeling (such as osteoporosis) can occur .in vitro without altering the integrity of elastin, which is associated with almost all types of vascular calcification. These results showed that osteoclast-like cells can effectively exert their de-mineralization ability outside the bone tissue  and bone marrow derived macrophages (BMDM). YSDM arise from macrophage precursor cells of the yolk sac that migrate through blood vessels to organs and tissues during embryonic development, whereas BMDM are formed by differentiation of monocytes into individual tissues and organs in the peripheral blood after birth . MacrophRemarkable plasticity and functional heterogeneity are important features of macrophages. Macrophage plasticity refers to the transformation of the function of macrophages in a specific direction and the shift of macrophages into different subsets or phenotypes congruent with the alteration of the microenvironment  and “the alternative activated macrophage” (M2) . Bacterivia CD163 in human bleeding plaques. Mhem can play a role in clearing iron and red blood cells through their phagocytosis and can be characterized by increased expression of IL-10 and apolipoprotein E  in vitro, and deletion of ENPP1 can cause aortic calcification in mice , which not only promote differentiation of vascular wall cells into chondrocytes and osteoblasts but also stimulate the production of oxygen free radicals and the expression of bone morphogenetic protein . These pvia the Notch signaling pathway , TNAP, osteoprotegerin (OPN), and bone morphogenetic protein 2 (BMP-2), can also promote the osteogenic process. A macrophage conditioned medium that leads to the overexpression of Wnt10b, BMP-6, and SPHK1  can stimulate migration and differentiation of human mesenchymal stem cells (MSCs) into osteoblast lines . BMP-2 c pathway . Tenipos pathway . Poly [A pathway . OsteogeMVs secreted by macrophages are the nucleation sites of calcium phosphate crystals. In the early stage of vascular calcification, marked accumulation of macrophages can be readily observed. Macrophages release large amounts of calcifying MVs containing the phosphatidylserine-annexin V-S100A9 complex . Sophie Vascular calcification is a dynamic process that represents a balance between osteogenesis and osteolysis. Once the balance is broken, pathological calcification easily occurs .Promotion of the differentiation of macrophages into osteoclast-like cells is conducive to the regression of vascular calcification. Conversely, inhibition of osteoclast differentiation or dysfunction of osteoclastic bone resorption activity may be related to the emergence and persistence of vascular calcification. Giulia et al. found that although the expression of the osteoclast marker CA2 was enhanced after macrophages were treated with IL-4, the expression and activity of CTSK  was decreased . The mecin vitro cellular experiments, and IL-6 may play a crucial role in aberrant differentiation of PC  can reduce the number of SMC-derived macrophages and MSCs . The queon of PC . In a hufication . The glufication . In summVascular calcification is a complex process that involves multiple factors. Disorder of the calcium and phosphorus metabolism balance, the imbalance between mineralization inducers and inhibitors, and the imbalance between osteoclast‐mediated bone resorption and osteoblast‐induced bone formation all have significant impacts on the formation and regression of vascular calcification. Macrophages are closely related to vascular calcification. A research boom has occurred based on the role of macrophages in the process of vascular calcification. The mechanisms of how macrophages can intervene in the progression and regression of vascular calcification have become increasingly clear. However, the phenotypic plasticity and functional heterogeneity of macrophages also lead to their pleiotropic effects. The question of how to precisely regulate the differentiation of macrophages into a phenotype in a manner that is conducive to calcification regression may become an urgent problem to be solved. Therefore, it is significant in theory and in practice to further identify and characterize macrophage subsets using single-cell technology. Understandably, additional questions currently exist relative to these potential roles of macrophages in pathological vascular biology: Can we achieve the treatment purpose of vascular calcification by targeting and regulating the M1 shift into M2 or promoting macrophages around calcified areas to differentiate into functional osteoclast-like cells while limiting undesired macrophage functions? Which other mechanisms might participate in the inhibition of bone resorption capacity of macrophage-derived osteoclasts surrounding calcium deposits? What mechanisms initiate and mediate the release/enrichment of macrophage-derived MVs? By which mechanisms do macrophages and their active substances interact and communicate with vascular wall cells? The current research has not yet supplied a clear-cut answers to these questions. Due to the existence of the bone-vascular axis and calcification paradox, it is necessary to further explore whether the reversal of vascular calcification by targeting macrophages could lead to bone loss in bone tissue. Many anti-calcification drugs have been restricted from clinical application due to various potential side effects. Active development of drugs such as teniposide, which can effectively inhibit vascular calcification in a tissue-dependent manner but had no effect on bone structure, is expected to be a boon for the treatment of vascular calcification. In conclusion, further exploration of the potential molecular mechanism of how macrophages regulate the progression and regression of vascular calcification is expected to reveal a new entry point for the prevention and treatment of vascular calcification, with far-reaching implications.ZW conceived and designed review; YL, ZS, LZ and LJ wrote and revised manuscript; YL, ZS, LZ, JY, CS, LJ, LL and ZW approved final version of manuscript.This work was supported by the foundations as follows: the National Natural Science Foundation of China ; the related Foundation of Jiangsu Province ; the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions; Postgraduate Research & Practice Innovation Program of Jiangsu Province (SJKY19_2585); Zhenjiang Cardiovascular Clinical Research Center Project (SS2018008).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.


This study aimed to investigate the influence of oral health on nutritional status, self-perception of oral health and health related quality of life of institutionalized elders. p<0.05). A cross-sectional study was conducted with 193 institutionalized elders living in the metropolitan region of João Pessoa (Brazil). The independent variables included were: 1) caries experience (DMFT index and its components); 2) use and need of dental prostheses; and 3) type of edentulism. The dependent variables included were related to nutritional status, self-perception of oral health and health-related quality of life. Data were submitted to a descriptive and comparative analysis, through correlation, association and difference tests, considering a significance level of 5% (p>0.05). Individuals who did not need prosthesis had higher scores for nutritional status. Self-perception of oral health and health-related quality of life did not vary significantly according to the studied variables. No statistical significant correlations or associations between the oral health status and nutritional status and quality of life were found (The oral health status has a limited impact on the nutritional status, and does not impact the self-perception of oral health and quality of life of the institutionalized elders. Key words:Nursing homes, oral health, quality of life. A significant demographic transformation has been observed in the last decades, characterized by the increase in the proportion of the elders worldwide . In thisInstitutionalized elders have greater difficulty to access dental care compared to non-institutionalized, which predisposes the formers to a greater risk of deterioration of their oral health. The high cost of dental treatments, lack of knowledge related to oral care, mobility difficulties and the fear of dental interventions could be factors associated with lower access to dental care. Besides that, the lack of interest of dentists and dental practitioners in providing dental services may further aggravate the oral status of institutionalized elders .Previous studies have shown that elderly people who live in nursing homes have an even worse state of oral health ,5. FactoThe evaluation of self-perception in oral health provides information on how individuals perceive their own oral health status with regards the functional, social and psychological dimensions, being those significantly associated with clinical findings and treatment needs . HoweverThere is evidence of an association between the presence of oral diseases and an increased risk of malnutrition . The higConsidering the growing population living in long-term institutions and the importance of oral health status in general health and quality of life of institutionalized elders, the aim of this study was to verify the impact of oral health on nutritional status, self-perception in oral health and health-related quality of life of institutionalized elders. In this study, the hypothesis was that the oral health influenced the nutritional status, the self-perception of oral health and the quality of life of the institutionalized elders.-Ethical aspectsThis study was in accordance with the ethical standards proposed by the national health council, as well as the Declaration of Helsinki (1964) and its subsequent modifications. The institutional ethical committee approved this study under protocol CAAE: 66122917.6.0000.5188. All participants signed a written and informed consent for information and research purposes.-Subjects, design and location of the studyA cross-sectional study was conducted with 193 institutionalized elders living in the metropolitan region of João Pessoa . This municipality is located in the Northeast region of Brazil and has 880,323 inhabitants, with a Human Development Index (HDI) of 0.763 and a Gross Domestic Product (GDP) per capita of R$23,169. A total of seven long-term institutions were identified in the metropolitan region of João Pessoa, all of them philanthropic. Most of the costs of these institutions are covered through the retirement income of the residents themselves.Elders who were able to assimilate the proposed methodological instruments and who agreed to participate were included in the study. The participants answered a questionnaire and were then submitted to an oral examination by a group of previously calibrated researchers (Kappa ≥ 0.85).-Instrument and variablesThe independent variables analyzed in this study were: 1) dental caries experience ; 2) use and need of dentures; and 3) type of edentulism . Nutritional status, self-perception in oral health and health-related quality of life (HRQL) were considered as dependent variables in this study. Previously validated methodological instruments were used to collect the data.-Nutritional status assessmentThe Mini Nutritional Assessment - Short Form (MNA-SF) was used o evaluate the nutritional status of the participants. This instrument allows the identification of elderly individuals who are malnourished or at risk of malnutrition. The higher the score , the better the nutritional status of the elders . Partici-Evaluation of self-perception in oral healthThe self-perception in oral health of the elders was evaluated using the Geriatric Oral Health Assessment Index (GOHAI). This validated questionnaire is composed of 12 items that analyze self-perception in oral health in the following domains: physical function, psychosocial function and pain or discomfort. The participants answered the questions as follows: never (3 points), sometimes (2 points) and always (1 point); and poor perception (<34 points).-Assessment of health related quality of life (HRQL)The SF-12 instrument was used to evaluate the health related quality of life related of institutionalized elders. The instrument consisted of 12 questions involving the following domains: physical, emotional, mental, pain, general health condition, vitality and social relations . Partici-Statistical analysisData was tabulated and analyzed using program for this purpose . A descriptive analysis of the data was conducted to calculate the absolute and relative frequencies.The median value of the Health Related Quality of Life (HRQL – median = 62) was considered as a reference for the dichotomization of this variable. For the other dependent variables included , the reference values were obtained according to the score established by the questionnaires related to nutritional status (MNA threshold = 8 points) and self-perception in oral health (GOHAI threshold = 34 points). Statistical tests for correlation (Spearman), association (Chi-square or Fisher) and difference  were used to verify the relationship between the independent and dependent variables of this study, considering 5% level of significance.The population of this study was majoritarian female  and above 80 years . No statistically significant correlation  was obsep<0.05). Self-perception in oral health and HRQL did not vary significantly as a function of the studied variables   . These fDiscussion The findings of this study show that, overall, oral health status did not interfere with the risk of malnutrition, poor self-perception in oral health, and poor health-related quality of life of institutionalized elders. Based on that, the null hypothesis was accepted.Possibly, other health problems experienced by institutionalized elders with regards to psychological, physical and social domains would interfere more significantly with the dependent variables under study. Based on that, oral health would have a secondary role on the nutritional status and on HRQL. In addition, the poor relationship between oral status and self-perception on oral health suggests that institutionalized elders do not perceive appropriately their poor oral status, or do not give enough importance to it .Although the National Oral Health Survey provides data referring to the elderly population in Brazil with an age range of 65-74 years, there is no data of population-based study on oral health status of Brazilian institutionalized elders . PreviouIn this study, the oral health condition did not influence the self-perception in oral health of the elderly participants. Even though oral health problems were prevalent among institutionalized older adults, their perception of oral health was positive . PossiblThe poor oral health has been considered a risk factor for malnutrition, mainly in hospitals and nursing care homes . A previThe findings of this study revealed that 114 institutionalized elders were classified as total edentulous, a cause for alarming and concern. The absence of occluding pairs and lower masticatory function was therefore expected to perform a significant impact on nutritional status . HoweverThe institutions vary in relation to the minimum number of people destined to the elderly care, being rare the inclusion of the dental staff within the professional team. Although oral health is an important issue within the general health, caregivers have multiple concerns with regards to elderly care. Since most part of elders does not have natural teeth and elders have good perception of their own mouth, caregivers do not give enough importance oral health . None ofFor a long time, dentistry was based on healing and mutilating practices, which did not value the preservation of teeth. It is important to consider that a significant portion of the elders investigated attribute little, if any, importance to the oral health. In part, these elderly people face health problems that go beyond the difficulty of chewing or shame when smiling. Theses findings are of relevance for better comprehension of oral health during elderly life, mainly for institutionalized people. The restriction to perform daily living activities and limited family-life may be much more involved in the poor quality of life of these elderly individuals. Besides that, low social interactions and high medication intake experienced in these institutions makes the aesthetic perception of the smile underestimated. Qualitative studies should be designed to better understand the psychological aspects involved in the quality of life of these individuals.Although the sample is composed of 40% of the universe of participants, this study is representative for the elderly living in nursing care homes that have little or no cognitive impairment. These results can be applied to other institutions located in the northeast or institutionalized elders in other countries with similar socioeconomic and cultural conditions.Oral health status did not affect the risk of malnutrition, self-perception in oral health, and the quality of life of the institutionalized elders. This is of relevance because oral health status at the end of life does not impair health-related outcomes. Interventions on oral health among those individuals may be balanced with their potential benefits.


The design and synthesis of functional nanomaterials have been extensively explored over the last decade, primarily due to their exceptional physico-chemical properties. Various nanomaterials have been used for the development of nanocomposite membranes due to the vast potential in harnessing their compositional and structural advantages. The desired physical and chemical properties endowed by nanomaterials at the molecular level can potentially address the limitations and bottlenecks of conventionally used polymeric and ceramic membranes. The development of nanocomposite membranes involves multidisciplinary strategies that include the innovative design and applications of nanocomposite membranes. The structural and separation properties of membranes can be feasibly tailored and fine-tuned through the introduction of various natural and engineered nanomaterials. When introduced on a surface or within a matrix of membranes, nanomaterials characterized by different dimensional and structural properties render synergetic effects that are unmet by conventionally fabricated membranes.The Special Issue of “Advances in Nanocomposite Membranes” aims to bring together membrane scientists and engineers to look into multiple aspects associated with the development of state-of-the-art nanocomposite membranes. This Special Issue covers important topics associated to the design and fabrication of nanocomposite membranes, their functionalization and modifications approaches, and the applications of these advanced membranes in niche separation processes, including gas separation, wastewater treatment, and desalination. In total, nine contributions, comprising seven research articles and two reviews, have been successfully compiled in this Special Issue. This collection of research articles and reviews focuses on the synthesis, design, optimization, and applications of advanced nanocomposite membranes, and it also looks into the recent advancements and future trends of this realm. 2) and carbon-based nanomaterials such as graphene oxide (GO) have been extensively used for the development of nanocomposite membranes due to their desired surface hydrophilicity, as well as their textural and structural properties. Based on three types of TiO2-incorporated polyvinylidene fluoride (PVDF) membranes, Teow et al. [2 exhibited promising anti-fouling and UV-defouling properties, primarily ascribed to its smooth surface enriched with abundant reactive surface groups. By coupling sol–gel and modified phase inversion techniques, Peixoto et al. [2) and TiO2. In the former nanocomposite membrane, the covalently bound TiO2–SiO2 was bound to CA through the interaction between SiO2 and CA, while in the latter, the TiO2 was directly bound to CA. The binding site significantly affected the permeability of the resultant nanocomposites, where the introduction of single TiO2 rendered an improvement of hydraulic permeability by more than two folds when compared to the neat CA membrane. On the other hand, the presence of SiO2 resulted in a neglected change in membrane permeability. Metal oxide-based nanomaterials such as titania  has become a new tool for treating various wastewaters and for desalination application. The performance of FO processes greatly depends on the characteristics of the membranes. Suzaimi et al.  providedThe findings and critical discussion from these nine contributions have evidenced the roles and importance of nanocomposite membranes as valuable tools to heighten the performances of various membrane-based separation processes. The flexibility in coupling a wide range of nanomaterials and conventionally used membrane materials offers resultant nanocomposites with high versatility. The authors of these contributions have delved into the important aspects of this topic and addressed many important fundamental questions. The findings have led to a better understanding of material design and the possibilities to fill the research gaps. It is envisioned that this Special Issue could illustrate some beneficial guidelines and future trends in the development of more exciting and high-performance nanocomposite membranes.


The carotico-carotid foramen occurs due to the ossification of the carotico-carotid ligament or dural fold stretching between the anterior and middle clinoid processes. The foramen is closely related to the cavernous sinus, pituitary gland, sphenoidal air sinus, and internal carotid artery irrigating the major part of the cerebrum (part of the brain). Due to the close association of the carotico-clinoid foramen with the aforementioned intracranial structures that may be affected by the formation of an anomalous foramen creating various neurological complications. Therefore, the study was carried out. The aim of the study is to consolidate all the data relating to the carotico-clinoid foramen to make it available to neurosurgeons as a ready reference.For this, the literature was surveyed using various databases, and the terms related to the carotico-clinoid foramen and associated clinical significance have been elucidated. The literature survey brought out that this foramen is congenital in origin and it is of three types, of which the third type is the most dangerous, as it may cause severe hemorrhage, creating a plethora of complications. The other two types may compress the internal carotid artery, causing ischemic changes in the brain.The information provided by this study will be of utmost use to neurosurgeons to carry out surgical interventions in the vicinity of the carotico-clinoid foramen. The lesser wing of the sphenoid is a very crucial bone, as it is frequently used as a landmark in neurosurgery around the middle cranial fossa. The projecting medial ends of the lesser wing are known as the anterior clinoid process (ACP); they are connected to the middle clinoid process by the carotico-clinoid ligament or dural fold. Complete ossification of the ligament or dural fold culminates into the carotico-clinoid foramen (CCF) . CCF is The ICA is an important source of irrigation to the major part of the cerebrum of the brain. The ICA that passes through the cavernous sinus follows its path along the medial to anterior clinoid process. Thus, the ICA is intimately related to the anterior clinoid process in addition to structures mentioned in the preceding paragraph. Therefore, neurosurgery involving the anterior clinoid process and thereby CCF is likely to damage the ICA and surrounding intracranial structures. Injury to ICA may disrupt regular arterial supply to the brain and injury to intracranial structures in close proximity of the CCF may create a plethora of neurological complications to the extent of fatality in many cases. The presence of complete or partial CCF may further compress or impinge the ICA causing degeneration or bleeding. Moreover, the cavernous sinus, sphenoid sinus, and pituitary gland situated in close proximity of the CCF may be affected, making radiological interpretation subjective and neurosurgical access and intervention difficult. This may enhance the chances of iatrogenic injuries.Therefore, considering the immense serious clinical complications of ICA due to compression or impingement by bony spurs of a variant morphology of CCF, the study has been carried out to consolidate the anatomical variations associated with the CCF to facilitate neurosurgeons for ready reference.Classification and incidence of carotico-clinoid foramenThe carotico-clinoid ligament ossifies partially or fully to form a complete or incomplete foramen. Depending on the degree of ossification of the carotico-clinoid ligament/dural fold, Keyes classified these foramina into three categories: Type-I: Complete foramen Figure , Type-IICCF has been extensively studied by various investigators. Ozdogmus et al. studied 50 autopsy bodies (100 sides) and observed a complete foramen on 27 sides and incomplete on 18 sides . Three mA study revealed the complete carotico-clinoid canal in 4.1% and incomplete in 11.6% in 73 Korean skulls . Freire Etiology of foraminaThe ossification of ligaments is normally an age-related physiological process. The carotico-clinoid foramen and the interclinoid bony bridge were demonstrated in fetal and infant skulls, suggesting CCFs as the congenital anomalies -21 develClinical implications of the carotico-clinoid foramenThe ICA is located medially to the anterior and middle clinoid process. Type-I and II may compress or stretch the ICA, causing irregular ischemic changes producing headaches and other neurological inconveniences resulting from inadequate blood supply to the brain ,11,13. AType-III CCF is more dangerous, as spurs formed due to incomplete ossification may impinge on ICA during the pulsating of the ICA, causing severe bleeding.Various types of CCF may create a misinterpretation of radiographs if this anomaly is unknown. Thus, detailed and thorough awareness of various forms of CCF is essential to neurosurgeons to avoid injury to various structures during neurosurgical interventions and to radiologists to improve their interpretation of radiographs. The anterior clinoid process is usually removed in order to approach the cavernous sinus and related structures in neurosurgery. The presence of the CCF and interclinoid osseous bridge creates obstacles while approaching the anterior clinoid process in regional neurosurgery and increases the risks, especially if an aneurysm of the internal carotid artery is present. Further, a high incidence of these structures has been reported in subjects with hormonal disturbance, the developmentally disabled, criminals, and epileptics. Therefore, detailed anatomical knowledge of the carotico-clinoid foramen and interclinoid osseous bridge is of paramount importance to obtain successful results while operating on this region for various neurological conditions.


This study is concerned with the development of a new unidirectional cellular (UniPore) copper structure with multiple concentric pipe layers. The investigated UniPore structures were grouped into three main types, each having a different number of pipes  and different pore arrangements. The specimens were fabricated by explosive compaction to achieve tightly compacted structures with a quasi-constant cross-section along the length of the specimens. The bonding between copper pipes was observed by a metallographic investigation, which showed that the pipes and bars were compressed tightly without voids. However, they were not welded together. The mechanical properties were determined by quasi-static compressive testing, where the typical behaviour for cellular materials was noted. The study showed that porosity significantly influences the mechanical properties, even more so than the arrangement of the pipes. Cellular metals are some of the most suitable materials for modern lightweight engineering structures due to their mechanical and thermal properties, which result in diverse functionalities. Ashby et al.  describeThe idea of unidirectional cellular metals was initiated by the development of Gasar structures based on the metal–gas eutectic reaction by Shapovalov . These sThe UniPore structures were proposed by Hokamoto  to avoidUtsunomiya et al.  proposedThis study is concerned with the development of a unidirectional cellular metal structure (UniPore) with multiple pipe layers and longitudinal cells and characterisation of its mechanical properties. The study takes advantage of the UniPore structure’s fabrication method and initial concentric assembly of wires (bars and pipes) as fabricated by Utsunomiya, which are described above. The leaching step is minimised by substituting the aluminium wires with polymer bars. The UniPore fabrication method enables fabrication of longer specimens in the order of several metres with minimal changes in the pipe wall thickness due to radial compaction. The newly developed specimens, using an upgraded UniPore structure fabrication method, were thoroughly analysed by metallographic analysis, while their mechanical properties and deformation mechanism were characterised with compressive experimental tests.Preparation of the specimen in the form of an assembly of metal pipes or bars, where the voids are filled with low-melting-point material ;Placement into the explosive container, where the specimen is surrounded by the primary explosive;Explosive compaction, with the main explosive detonated by the booster;Removal of the low-melting-point material by thermal treatment.The fabrication procedure for unidirectional cellular metal structures with longitudinal cells (UniPore) consists of the following steps :PreparatThe specimens were assembled with concentric layers of copper pipes, where the distance between the pipes was controlled by layers of copper and polymer bars . PolymerCopper (JIS-C1220) pipes with different diameters and wall thicknesses were used in this study to fabricate specimens with different cross-section arrangements, varying wall thicknesses, and varying levels of porosity of the final UniPore structure. Layers of copper and polymer bars were placed between the pipes. The solid copper (JIS-C1220) bars measured 2 mm in diameter, while the polymer filler (acryl or glue sticks) bars measured 2 mm, 8 mm, or 14 mm in diameter. The length of the specimens was 100 mm. Three types of UniPore structures were fabricated, with differing numbers of pipes: type 1 contained 5 concentric copper pipes, type 2 contained 4 concentric copper pipes, and type 3 contained 3 concentric copper pipes. Two different assemblies of copper and polymer bars were prepared for each specimen type. The numbers of layers, bars, and pipes and the actual dimensions of the prepared assemblies are listed in Each specimen assembly was positioned centrally in the explosive container made of polymethyl methacrylate , which measured 165 mm in height and 83 mm in diameter c, surrouThe UniPore specimens were then cut into smaller specimens suitable for metallography and mechanical testing. The cross-sections with different topologies of obtained specimens with respect to their initial assemblies  are showThe cross-sections of the fabricated specimens were also visually analysed. It was observed that the cross-sections did not vary through the length of the specimens, assuring a constant cross-section along the length of the specimens, with a constant wall thickness and cell size and shape. The porosity of the specimens was evaluated before  and after  explosive compaction of the specimens. The theoretical porosity was calculated as the ratio between the cross-sectional area of polymer bars and the cross-sectional area of copper pipes, copper bars, and polymer bars. In contrast, the actual porosity was determined by digital image correlation of the specimens’ cross-sections . The numThe porosity of fabricated specimens varied between 32% and 49%. The lowest porosity was observed for type 1 specimens, while the highest porosity was measured for type 3 specimens. For comparison, the porosity values of previous copper  and alumThe metallographic analysis of the UniPore structures with multiple pipe layers was performed using the Nikon UM-2 light microscope device . The standard metallographic methods  were applied to prepare the samples. The transversal  and longitudinal (only type 1 specimens) cross-sections were microscopically investigated with a magnification factor of up to 50.The deformation behaviour and mechanical properties of the fabricated UniPore specimens were evaluated by uniaxial quasi-static compressive testing using the servo-hydraulic Shimadzu AutoGraph IS-10T testing machine . The UniPore specimens were cut perpendicular to the cell orientation  into cylindrical compressive specimens measuring 10 mm in thickness. Due to the expected repeatability (constant cross-section) of the results, only 2 specimens were prepared from each fabricated UniPore specimen. The specimens  were subjected to quasi-static loading conditions with a cross-head rate of 5 mm/min. A thin polytetrafluoroethylene (PTFE—Teflon) layer was positioned between the specimens and the loading and support plates to reduce the friction. The displacement and force during compression were measured by the testing device, while the sample deformation behaviour was captured by a digital camera. The measured force–displacement relationship was used to determine the mechanical properties of the new UniPore structures , while observation of the video recordings allowed for evaluation of the deformation mechanisms during the loading process. The force plateau was defined as the average force in the plateau region of the force–displacement diagram. The energy absorption was determined by integrating the force along with the displacement. The energy absorption efficiency was evaluated as the ratio between the actual and ideally absorbed energy.The connectivity between the individual components (copper pipes and bars) of the UniPore structure depends on the colliding conditions during the fabrication process based on explosive compaction. The connectivity is related to the velocity and inclination angles of the copper pipes and bars required for explosive welding with tight bonding . A metalThe investigated segments of the longitudinal cross-sections are shown in Figure 6. A wavy interface, which is typical for strong interfacial bonding due to intense fluidisation of the metals, was not observed. From the top left image in The transversal cross-section segments of the specimens are shown in The UniPore structures can also be used as structural load-carrying components. Thus, it is vital to understand and characterise their deformation and collapse mechanisms and their responses when subjected to mechanical loading. The uniaxial quasi-static compressive tests in the transversal direction of the fabricated specimens were performed for this purpose.Type 1 specimens represent the group with the lowest average porosity. Their deformation behaviour is shown in Uniform deformation throughout the loading process was observed for both specimens. Some intermetallic bonds between bars and pipe surfaces failed in the initial loading phase of specimen #1.1. It is interesting to notice that the bonding failure appears on the outer surface in all three cases. The surfaces of two inner pipes come into contact almost simultaneously at a displacement of 5 mm. Additionally, the outer two copper pipes begin to buckle, which is even more pronounced with larger displacements. In contrast, specimen #1.2 (with a slightly lower porosity) experienced separation of the bonded surfaces throughout the loading process. Contact between pipe surfaces appeared at a displacement of 10 mm due to the lower porosity of the sample. However, local buckling of the inner pipe was observed at a displacement of 5 mm.The compressive force–displacement relationship of the type 1 specimens is shown in The type 2 specimens have fewer pipes and a bigger hole in the centre of each specimen than the type 1 specimens. The average porosity of type 2 specimens is higher than of type 1 specimens. Their deformation mechanism can be observed in The compressive force–displacement relationship of the type 2 specimens is shown in The type 3 specimens have the least number of pipes and the biggest hole in the center of the specimen of all investigated specimens. Consequently, they have the highest average porosity. Their deformation sequence during the compressive loading is shown in A single contact between the surfaces of two inner pipes was observed at a displacement of 7 mm. A very similar deformation behaviour  of the inner pipe at displacement lengths of 2 mm and 5 mm can be observed in the case of specimen #3.2. The local bending (and later buckling) results in the debonding between the pipe and the bar. The dense and regular arrangement of bars again prevents contact between pipes until a displacement of 7 mm.The compressive force–displacement relationship of the type 3 specimens is represented in The observed force plateau and calculated energy absorption values of the analysed specimens are listed in The porosity significantly influenced the mechanical properties, even more so than the arrangement of the pipes. The specimens with the lowest porosity values  gave the highest force plateau values, which were comparable to the force plateau value of the original UniPore structure. Consequently, their energy absorption capacity values were also the highest.The study describes the development of a new unidirectional cellular copper structure with concentric multiple pipe layers. The specimens were fabricated by explosive compaction to achieve bonding between single pipe components. Different arrangements of inner pipes and layers  were analysed experimentally. The bonding between copper pipes was observed by a metallographic investigation, which showed that the pipes and bars were tightly compressed without voids. However, they were not welded together. The mechanical response was analysed by quasi-static compressive tests, where typical behaviour for cellular materials was noted. The specimens with the highest numbers of layers and lowest porosity values (type 1) showed the shortest force plateaus, while the specimens with the lowest numbers of layers and highest porosity values (type 3) exhibited the most prolonged constant force plateaus. The study has shown that porosity significantly influences the mechanical properties, even more so than the arrangement of pipes. The specimens with the lowest porosity values  provided the highest force plateau values, which were comparable to the force plateau of the original UniPore structure. Consequently, their energy absorption capacity values were also the highest. The observation of the deformation mechanism provided an insight into the debonding process, bending and buckling of pipe walls, and contact between constituents, which is directly connected to the initiation of the densification.


Cellular communication can be mediated by the exchange of biological information, mainly in the form of proteins and RNAs. This can occur when extracellular vesicles, such as exosomes, secreted by a donor cell are internalized by an acceptor cell. Exosomes bear specific repertoires of proteins and RNAs, indicating the existence of mechanisms that control the sorting of molecules into them. Knowledge about loadings and processes and mechanisms of cargo sorting of exosomes is essential to shed light on the physiological and pathological functions of these vesicles as well as on clinical applications involving their use and/or analysis. In this review, we will discuss the molecular mechanisms associated with exosome secretion and their specific cargo sorting, with special attention to the sorting of RNAs and proteins, and thus the outcome and the emerging therapeutic opportunities of the communication between the exosome-producer and recipient cells. Fig. 1) Almost all cells are capable for secreting various types of extracellular vesicles (EVs). In accordance with the origin, functions, and biogenesis, a series of EVs were classified into the following three types: exosomes, microvesicles (MVs)/shedding particles, and apoptosis bodies The isolation of exosome is critical to understanding their mechanisms. Isolation strategies typically used include centrifugation, ultrafiltration, size-exclusion chromatography (SEC), polymer-based precipitation, affinity capture on antibody-coupled magnetic beads, and microfluidics-based technologies One of the most vexing problems in exosome biology is how to accurately measure and assess exosome purity Fig. 1). The classic process of exosome generating involves double invagination of the plasma membrane and the formation of intracellular MVBs containing ILVs The best studied pathway of exosome formation by far is the classic pathway . The mechanisms and pathways associated with exosome uptake Upon formed, exosomes pinch off from the plasma membrane and prime them for secretion, including autocrine, paracrine and endocrine Recently, it has reported the membrane proteins transfer during direct cell-cell contact The eukaryotic and prokaryotic cells both release exosome, which are cup-shaped particles enclosed by a phospholipid bilayer Fig. 1). Rab5 and Rab7 play an essential role in delivering cargo to the early endosomes The process of exosome release may also be subjected to further modulation by Rabs, SNAP and SNARE proteins Fig. 1) Once release, exosomes fuse with the plasma membranes of recipient cells to deliver their contents Upon secreted, exosomes deliver their contents to adjacent or distant cells and play a role of regulating of gene expression and modificating of phenotypes and biology The exosomal protein content reflects their origin and varies depending on the types of host cell The plentiful RNA content could result in the greatest source of signaling diversity. Many researches have demonstrated that mRNAs, miRNAs, and other noncoding RNAs contain in exosome Exosomes carry certain lipids, which play an essential role in maintaining the biological activity Fig. 2) Exosomes are enriched with a unique content of proteins, RNAs, and lipids that can vary from the parent cell. The intraluminal content of the exosomal membrane depends on the host cells, and on their cytoplasmatic composition in particular Fig. 2) The ESCRT complex has been well acknowledged as a main cargo sorting mechanism. The ubiquitination and ubiquitin-like modifiers represent one of regulatory systems in protein location, stability and function Table 1). In summer, deciphering the sophisticated ESCRT-dependent pathway is necessary to unveil specific functions in the selective import of proteins into exosomes.However, not all the sorting of proteins depends on ubiquitination. The recent studies show that ubiquitinated proteins can evade deubiquitination. Ubiquitination drives the endocytosis and sorting of MHC II to the luminal vesicles of MVBs for lysosomal degradation PTMs can direct exosomes loading and specific PTMs can control the selective mechanisms of protein cargo sorting and promote some proteins to enrich in exosomes On the other side, growing evidences indicate that the existence of ESCRT-independent mechanisms also play an essential role, including lipid raft, tetraspanin and ceramide-mediated mechanisms. The ESCRT-dependent and independent pathways were not necessarily contradictory, but rather showed the presence of heterogeneous populations of exosome. It has been found that the pathway for intraendosomal transport of proteolipid protein (PLP) may be independent on ESCRT The exosomal RNAs are not random and the RNAs sorting process is highly selective Table 2) 1) The sumoylated heterogeneous nuclear ribonucleoproteins (hnRNPs)-dependent mechanism 2) The 3'-end of the miRNA sequence-dependent pathway 3) The miRNA induced silencing complex (miRISC)-related pathway 4) Membrane proteins related pathway. Membrane proteins like Vps4A and nSMase2 are worked in the miRNA sorting into exosomes. A ceramide-dependent secretory machinery could release some exosomal miRNAs and this process is independent on the ESCRT system 5) Other RNA-binding proteins related pathway. The studies proved that specific proteins may control miRNA sorting through recognizing and binding to specific RNAs sequences 6) Other potential pathway. Another ESCRT-independent mechanism may depend on raft-based microdomains for the lateral segregation of cargoes As RNA and DNA sorting is still a relatively novel field, it is possible that the current literature only represents studies that have positive findings. Additionally, RNA and DNA sorting is a complicated phenomenon and it is diffcult determine whether the sorting proteins are part of independent sorting mechanisms or whether they involved in a unified larger sorting mechanism.Fig. 3).Exosomes exist in all biological fluids and are secreted by all kinds of cells, making them attractive as minimally invasive liquid biopsies with the potential for sampling to follow disease progression Given their presence in most bodily fluids, exosomes have been wildly known as potential biomarkers for many diseases in vivo and in vitro package therapeutic agents, and membrane modifications to enhance tissue-specific homing in vivo for the specific targeting of disease. Specially, exosomes have a protective effect on craniocerebral diseases. They have ability to accross the blood-brain barrier, and enhance neural and motor function, which opens a wide variety of possibilities for treatment of neurodegenerative diseases As the function of exosome cargo has become more widely recognized, exosomes have been proposed as a potential alternative to cell-based therapies. The multifaceted nature of exosome and their compositions underscores the advantage of exosome therapy


The within-herd relationship among the individual age categories as well as the influence of the herd size on nDDDch per animal measured as kept sows were analyzed by simple linear regression. The evaluation of the treatment days showed that 50% of the nDDDch were used in piglets, 44% for weaners, and 3% each for fattening pigs and sows. Compared to the other age categories, the examination of the number of nDDDch per animal showed a significantly higher number for sows, whereas for fattening pigs the number was significantly lower (P < 0.01). The farm-based analysis using linear regression showed a relationship between antimicrobial usage in sows and piglets . Similarly, a significant relationship between larger herd size and increased antimicrobial usage was observed . The present study provides an insight into the antimicrobial treatment dynamics of farrow-to-finish farms. In particular, the age categories piglets and sows—with their higher number of treatment days in total or per animal—are of interest regarding the potential reduction in antimicrobial usage. Likewise, larger farms with higher management requirements were found to be of particular importance for the reduction of antimicrobial usage. Monitoring programs should therefore evaluate different age categories separately to identify problems for individual farms.In the Swiss pig sector, the usage of antimicrobials has been recorded, evaluated and systematically reduced on a voluntary basis since 2015. This monitoring has been carried out using various methods thereby enabling continuous national scrutiny as well as international comparisons. To gain a better understanding of the dynamics of the antimicrobial usage on Swiss farms, consumption data of farrow-to-finish farms were analyzed for (i) the within-herd relationships among different age categories and (ii) the influence of the herd size. The data were collected on 71 farms for the year 2017, encompassing the amount of active ingredients and number of defined daily doses Switzerland (nDDDch) in total, and stratified for the different age categories of piglets, weaners, fattening pigs, and sows. The differences in nDDDch per animal among the age categories were determined by a Wilcoxon test and subsequent  Awareness of the spread of antimicrobial-resistant genes is an important issue at the public health interface between human and veterinary medicine . The genIn addition to treatment guidelines, the first important step is setting up a powerful AMU monitoring system, which allows for correlations with resistance data  and is lAccording to this description, nationally defined doses (DDDch/DCDch) were developed for the pig sector in Switzerland and compared with the values of the EMA in the field , 23. TheIn addition to comparing different types of farms, the relationship among AMU and different age categories within farrow-to-finish farms has also been examined and described with other monitoring systems . This stThe objectives of this study were (1) to investigate the association among AMU and animal age category on farrow-to-finish farms and (2) to investigate the association between AMU and herd size on farrow-to-finish farms. The hypotheses of this study were that (1) AMU would differ among age categories on farrow-to-finish farms, and (2) AMU would differ among herd sizes on farrow-to-finish farms. AMU was measured using DDDch, and herd size was measured in the number of sows kept.In collaboration with the Swiss Pig Health Service (SSHS), AMU data were collected from 71 Swiss farrow-to-finish pig farms in 2017. The study farms joined a nationwide voluntary program known as Suissano/Safety+, which evaluates the AMU on their farms to improve transparency along the pig production process. Overall, 598 out of 6,406 pig farms throughout Switzerland participated in the Suissano/Safety+ program in 2017, which represents a proportion of 9% . Out of To quantify AMU, the amount of prescribed antimicrobial ingredient from the study farms during 2017 was divided by the defined daily doses Switzerland (DDDch) of each corresponding antimicrobial classes multiplied by the standard weights of the different age categories as defined by the European Surveillance of Veterinary Antimicrobial Consumption (ESVAC)  .The required information describing the doses in mg/kg was taken from the product leaflets which are available in the Swiss Veterinary Medicines Drug Compendium . The detThe number of DDDch (nDDDch) and the amount of prescribed antimicrobial ingredients were calculated in total, as were the different age categories and the different used antimicrobial classes.For the evaluation of the AMU among different herd sizes and to compare the consumption by different categories, the number of kept (sows) or produced pigs  in the year 2017 were considered according to the following formula:This calculation was performed separately for each prescribed antimicrobial ingredient used on the farm, and the resulting numbers were added up for each age category. Finally, the results of the different age categories were summarized as the total nDDDch per pig in 2017. The herd size was defined as the number of sows kept.https://cran.r-project.org). Differences among the tested groups having a P ≤ 0.05 were considered statistically significant. The data for calculated AMU on the farm level were tested for normal distribution and the differences among the individual age categories were investigated using the Kruskal-Wallis test followed by a post-hoc pairwise analysis (Bonferroni correction). The relationship between the herd sizes and the nDDDch/animal/year on farms was examined by a simple linear regression. Both were set as continuous variables with herd size as the predictor and nDDDch/animal/year as the response variable. Similarly, simple linear regression was used to compare the nDDDch/animal/year for the different age categories. The nDDDch/animal/year of sows or the respective younger age category was determined as the predictor variable and was examined with the nDDDch/animal/year of another age category as the response variable. The nDDDch/animal/year of the six different models of predictor-response variables  was therefore analyzed. Homoscedasticity tests and plots were examined to check whether assumptions of normality and homoscedasticity were fulfilled and scatterplots were prepared to visualize the results.The preparation of all operating farm data and the calculation of the number of defined doses were carried out using Microsoft Excel 2016 . The statistical analysis and preparation of graphs to visualize the results were performed with R   . The anaP = 0.02).As shown in This study found major differences in AMU among the age categories of piglets, weaners, fatteners, and sows, which varied depending on whether active ingredients or the number of defined daily doses are used as method of measurement. Due to the different weights of the age categories, 9.8 kg (10%) of all active ingredients in piglets represent 50% of the number of defined daily doses, whereas 36.1 kg (36%) in sows represents only 3% of the total number of these doses. For the weaners, the percentage for both the active ingredients and the number of defined daily doses was 44%. As previous studies have shown, the proportion of AMU measured by defined daily doses in weaners is comparatively high , 34. A fThis could also partly explain that in weaners, in contrast to the other age categories, it is not penicillins, often used as an injection solutions, but sulfonamides and tetracyclines that were the most frequently observed antimicrobial classes within the amount of active ingredients. Both sulfonamides and tetracyclines are potentially part of the premixes, which are used as group treatments in weaners in Switzerland . This stThis study shows that for HPCIAs, the calculated number of defined daily doses was 21% of the overall consumption. Similar results with similar measuring methods have already been shown in earlier studies, which could also demonstrate the potential for reduction over time , 39. SimThese differences in the output of different measurement methods were mentioned in theoretical reviews , 40 and The average herd size of the farms in this study was 79 kept sows, which is comparable to earlier studies describing the usage of antimicrobials on Swiss pig farms . Since pMycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae and free of the porcine reproductive and respiratory syndrome (PRRS) virus, there are some indications missing which have been linked to increased antibiomicrobial usage in growing pigs in other countries (This study identified significant differences in the number of defined daily doses per animal among the different age categories. Fatteners were treated less frequently, which has already been shown in a previous study  and coulountries , 47. Theountries .Escherichia coli has been observed previously (In contrast to the lower AMU in fatteners, the AMU in sows measured as defined daily doses per animal was significantly higher compared to all other age categories. This finding was observed also by other authors , 38, alteviously , 52; inceviously  and Sjöleviously , this waSimilarly, a significant relationship between larger herd size measured as kept sows and increased AMU was observed. This underlines the findings of previous studies , 28 and Our study demonstrates that AMU monitoring programs should evaluate different age categories separately to identify specific points of concern and suggest possible solutions for individual farms. The categories of piglets and sows are especially important on Swiss pig farms due to their high and related AMU either in absolute terms or per animal. Likewise, a link between larger farms and increased usage was observed and should be considered through improved on-farm health management.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher.Ethical review and approval was not required for the animal study because since this was only data that had no influence on the actual treatment of the animals, an animal welfare permit was not required. No veterinary manipulations or similar interventions were carried out on any animals by the research team. Written informed consent was obtained from the owners for the participation of their animals in this study.TE drafted the manuscript. TE, DK, and XS designed and directed the study. CM supported the pharmacological aspects of the used measurement methods. TE prepared the data processing and statistical analysis. All authors have read and approved the manuscript.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.