Journal of Translational Medicine, ‘Cancer metabolism’. Tumor cells reprogram nutrient acquisition and metabolic pathways to support abnormal growth and metastasis, a dysfunctional process that is a hallmark of cancer. This new section will offer a solid framework to promote and disseminate innovative studies examining metabolic reprogramming in cancer cells, including fatty acid, mitochondrial, glycolytic and other metabolic networks.We are delighted to announce the launch of a new section in the We are interested in research that investigates and characterizes metabolic alterations in tumors at the molecular level by explaining why this reprogramming occurs and identifying links between such phenomena and cell proliferation and metastasis. We will also consider studies regarding the correlation between genomic mutations, gene expression, epigenetic regulation and metabolic pathway alterations. Of particular interest are investigations focused on translational and clinical aspects and those regarding metabolically targeted pharmacological therapies, with emphasis on patient responsiveness.One of the hallmarks of cancer cells is their ability to reprogram cellular metabolism to comply with their biosynthetic, bioenergetic and redox demands, thus providing a selective growth advantage and sustained proliferative capacity. Metabolic reprogramming is largely a consequence of oncogenic mutations and can be defined as a multitude of pathways that are enhanced or suppressed in cancer cells compared to normal tissues. Such alterations are pivotal to acquire and transform nutrients from the tumor microenvironment, which is often nutrient-depleted. Tumorigenesis-associated metabolic alterations affect various stages of the metabolism circuit, such as increasing metabolite influx, modifying nutrient processing and shaping the tumor microenvironment itself. Overall, biosynthetic networks are a crucial feature in cancer metabolism as they enable tumor cells to synthesize the macromolecules required to sustain proliferation and thus, tumor growth.Many of the aberrantly activated oncogenes that are commonly observed in different types of cancer function in the PI3K-AKT-mTOR pathway. Moreover, MYC gain-of-function is associated with many cancers and leads to the upregulation of genes involved in anabolic growth and nucleotide biosynthesis. Alternatively, p53 is an important regulator of metabolism and oxidative stress, in addition to its role as a tumor suppressor. It has been demonstrated that loss of p53 increases glycolytic flux, promoting the anabolism and redox balance. Indeed, redox homeostasis is essential during both tumorigenesis and metastasis, as the increase in metabolic activity augments the levels of reactive oxygen species (ROS). ROS-dependent signaling leads to further activation of pathways that promote cell proliferation and metabolic adaptation. However, high levels of ROS can also cause cellular damage and require a well-orchestrated machinery to offset their negative effects.Aerobic glycolysis is one of the most well-studied examples of a metabolic pathway that is reprogrammed in cancer cells. Glucose is the most abundant nutrient in the bloodstream, and it is catabolized to pyruvate through oxidative glycolysis in normal tissues, with the final aim of producing energy in the form of ATP. Between 1924 and 1927, Otto Warburg described how cancer cellular metabolism shifts from oxidative to glycolytic even in the presence of physiological levels of oxygen. As a result, pyruvate is converted into lactate and secreted into the extracellular microenvironment. Tumors are often characterized by a significant increase in glucose consumption due to genomic mutations (oncogene activation or tumor suppressor loss-of-function) or hypoxic areas. Hypoxia is often associated with poor prognosis in patients and dramatically reshapes the transcriptional landscape, which profoundly impacts metabolic networks. Hypoxia or anoxia are caused by lack of an appropriate vascular system in the tumor core, which also implies reduced nutrient accessibility for cancer cells.de novo, non-essential amino acids and polyamines which are nitrogen-containing molecules. After glucose and glutamine, serine is the third most consumed metabolite by cancer cells. Serine is the main contributor of one-carbon units in malignant cells, which are used in the biosynthesis of thymidine monophosphate and purines. Moreover, part of the serine pool is also converted into glycine. One-carbon molecules are also required for methylation processes, which affect epigenetic regulation and gene expression. Finally, another crucial amino acid is arginine, which is imported into cells and converted into ornithine, a precursor of polyamine synthesis.Therefore, alternative adaptive forms of metabolism are essential to ensure the survival and proliferation of malignant cells in these conditions, including oxidation of fatty acids, autophagy and micropinocytosis. Autophagy relies on the recycling of intracellular biomass and, despite promoting cell survival during periods of starvation, it is incapable of supporting tumor growth. Fatty acid synthesis requires acetyl-CoA, and in vitro studies have demonstrated that glucose is an important source of acetyl-CoA. However, glutamine and acetate can offer alternative carbon sources when cells experience hypoxic conditions. Glutamine is a source of nitrogen and carbon, and is the most abundant circulating amino acid in humans. For example, actively dividing cells synthesize nucleotides Journal of Translational Medicine is to offer a connection between basic and clinical science, supporting scientific discussion and diffusion of innovative concepts through open access and a high standard peer-review process. The “Cancer metabolism” section will ensure the publication of high quality and internationally competitive studies. The Editorial Board is looking forward to receiving your manuscripts.The study of cancer cell metabolism is important to provide new insights into possible therapeutic targets and strategies. Metabolic reprogramming profoundly affects the features of the cells and of the tumor microenvironment, creating hostile conditions for T cell proliferation and survival and negatively affecting the host immune response. The objective of the


The discovery of the genome has been one of the greatest advances in the development of tools for the genetic improvement of cattle in the last decades. The use of information derived from genomics has been demonstrated to be an efficient alternative for the validation of kinship, adjusted mating, and conservation strategies to generate dairy cattle with desirable traits. However, the impact of the genetic improvement strategies in breeding programs has closed bloodlines in several milk breeds, consequently generating inbreeding depression. Thus, to mitigate the negative effects of inbreeding, several tools associated with genomic analysis are currently available that may be useful to reverse undesirable genetic trends. The present review will provide an understanding of the importance of genomic analysis and how it can be used to mitigate the negative effects of inbreeding depression on different traits related to production, reproduction, health, animal welfare, linear type traits, and adaptability in dairy cattle. A historical overview of genomics and future perspectives will also be covered.Genomics comprises a set of current and valuable technologies implemented as selection tools in dairy cattle commercial breeding programs. The intensive progeny testing for production and reproductive traits based on genomic breeding values (GEBVs) has been crucial to increasing dairy cattle productivity. The knowledge of key genes and haplotypes, including their regulation mechanisms, as markers for productivity traits, may improve the strategies on the present and future for dairy cattle selection. Genome-wide association studies (GWAS) such as quantitative trait loci (QTL), single nucleotide polymorphisms (SNPs), or single-step genomic best linear unbiased prediction (ssGBLUP) methods have already been included in global dairy programs for the estimation of marker-assisted selection-derived effects. The increase in genetic progress based on genomic predicting accuracy has also contributed to the understanding of genetic effects in dairy cattle offspring. However, the crossing within inbred-lines critically increased homozygosis with accumulated negative effects of inbreeding like a decline in reproductive performance. Thus, inaccurate-biased estimations based on empirical-conventional models of dairy production systems face an increased risk of providing suboptimal results derived from errors in the selection of candidates of high genetic merit-based just on low-heritability phenotypic traits. This extends the generation intervals and increases costs due to the significant reduction of genetic gains. The remarkable progress of genomic prediction increases the accurate selection of superior candidates. The scope of the present review is to summarize and discuss the advances and challenges of genomic tools for dairy cattle selection for optimizing breeding programs and controlling negative inbreeding depression effects on productivity and consequently, achieving economic-effective advances in food production efficiency. Particular attention is given to the potential genomic selection-derived results to facilitate precision management on modern dairy farms, including an overview of novel genome editing methodologies as perspectives toward the future. The discovery of the genome has been one of the greatest advances in the development of tools used for the genetic improvement of cattle in the last decades. The use of high-performance genomics in the selection of dairy cattle represents a remarkable advance to increase the biological and genetic progress of different features and indices in several animal production systems .Recently, several techniques have been developed for the genomic analysis of dairy cows genetic traits related to production, reproduction, health, animal welfare, linear type traits, and adaptability. Several authors highlight the perspectives of using the genomic approach as a crucial genomic selection method for several dairy breeds and stress the great relevance and applicability of this knowledge in breeding programs to incorporate additional alternative features into the traditional pool of traits . In addiAlthough the use of techniques based on genomic analysis has produced important changes in selection and genetic improvement programs in common dairy cattle breeds , for theThe objective of the present review is to provide an understanding of the importance of genomic analysis and how it can be used to mitigate the negative effects of inbreeding depression on different traits related to production, reproduction, health, animal welfare, linear type traits, and adaptability. From here, we will address several topics covering the use and possible influence of genomic analysis on genotypic traits in dairy cattle as well as its evolution from origins to the present and future perspectives. This document is designed and organized as follows: In the first part, it will briefly describe the chronology and evolution of genomics. In the second part the use of genomics as a predictive tool will be described, as well as the different methodologies used for its application. In the third part, the incidence/impact/effect of genomics as a predictive tool for different parameters associated with production, reproduction, health, animal welfare, evaluation of linear type traits, and adaptability to different production systems and environments will be covered, including causal and highly predictive genetic variants that are key to the prediction of other complex traits.This review will describe how genomic selection based on genomic analysis represents a promising tool that will improve the predictive accuracy and genetic gain of genomic traits. However, challenges lie ahead in the integration of genomic analysis models covering all the genetic traits and expressing an ideal genomic selection method to adequately regulate inbreeding rates. Therefore, the perspectives to increase genetic gains based on the production and reproductive efficiency, health, animal welfare, and environmental adaptability of dairy cattle will increase the sustainability/sustainability of the production yields in the immediate future as a means for increasing profit in animal production systems.The discovery of genomics and related applications in the dairy cattle industry has led to a significant improvement in the biological prediction of different genotypic traits. Major dairy-producing countries, like United States, Canada, Great Britain, Ireland, New Zealand, Australia, France, the Netherlands, Germany, and the Scandinavian countries have implemented genomic evaluations in their breeding programs, leading to significant changes in the global dairy industry .Bos taurus) were carried out in Hereford beef cattle [Bos indicus the genome was sequenced as well in Gyr, Girolando and Guzerat dairy breeds [A brief history of genomics is depicted in f cattle . Later, f cattle . Around f cattle ,15. In By breeds . The firy breeds .Later, the release of complete genome sequences granted the use of high-performance genetic markers , thus improving trait predictions. This interesting technology requires high throughput genotyping platforms (DNA arrays or SNP chips) . In addiThere is growing concern about pedigree-based genetic evaluation analyses, as being less efficient and accurate than genomic methods. However, linking pedigree data to information obtained through genomics appears to be advisable, since it may significantly improve the accuracy of predicting traits and indices in dairy breeds . HoweverToday and in the immediate future, genomic evaluation as a functional and everyday tool must also consider the effect of new mutations that can generate a genetic variance for quantitative traits . In thisFinally, to decipher and associate the functionality of new dairy cattle traits, other ”omic” technologies have been integrated into genomics such as epigenomics, transcriptomics, proteomics, metabolomics, metagenomics, and meta-transcriptomics . TherefoThe incidence of the use of genomic selection in recent years has resulted in a significant increase in production yields for most dairy-related traits in Holstein cattle in countries such as the USA . HoweverAnalyses of cattle selection pressure to increase milk production yields suggest a positive association with a higher incidence of mastitis . Whole-gFertility rates in dairy cattle play a critical role in breeding programs . FertiliGenomic analyses of most reproductive traits in females show that they tend to be less heritable than in males . HoweverIn the last decade, genomics has received great attention from the perspective of the study of traits related to animal health and welfare. Hence, several traits such as resistance against infectious and non-infectious diseases as well as adaptability to the environment have been introduced as new selection indices in dairy cattle . Recent Several companies managing commercial herds of the American Holstein breed have recently developed and evaluated genomic predictions methods for the estimation of predicted transmission capabilities (heritability) for welfare traits, respiratory disease, diarrhea, and adaptability ,65. GiveFinally, the epigenetic effect of assisted reproductive technologies (ARTs), such as in vitro embryo production (IVP), has not yet been quantified or evaluated in depth. Much more research is needed to better evaluate and understand genomics and its correlation with IVP potentially affecting bovine health and possible effects during adulthood . FurtherThe environment, directly and indirectly, plays an important role in animal genomics . DetermiBos taurus, genomic introgression was found for the progenitors [Concerning the above and the current climate change scenario, the bovine species has permanently required substantial modifications in terms of adaptability; however, not all breeds have achieved these levels of adaptation . In the genitors . This adgenitors . Consequgenitors . This hagenitors . Considegenitors .One of the major environmental-related advances in genomics over the last decade has been the discovery of traits linked with methane emissions, hypoxia, high altitude adaptation, and heat stress. Several trait association studies primarily emphasize the reduction of negative impacts on the environment . With thBos grunniens) and domestic bovine (Bos taurus) show existing genetic differences in protein domains related to sensory perception, energy metabolism, hypoxic stress, and nutritional metabolism [Comparative analyses regarding the high-altitude adaptation of the genome of domestic yak  can be affected by several factors such as the method of prediction used, the proportion of the categories (sires and cows), or the genetic relationship between traits . SimilarAmong the potential uses of genomic analysis in dairy cattle breeding, direct selection of heritable measures of gene expression is available and specifically called expression-assisted selection and genomic genetic selection of QTL and eQTL (expression quantitative trait loci) . In thisAn improved and more accurate methodology for genomic prediction has recently been proposed for new individuals with a genotype, with and without phenotypes . This meGenomic evaluation is currently present in the leader dairy-producing countries worldwide. The discovery of the genomic analysis in the last years has notably improved production features (genetic merit) in most of the dairy cattle breeds . HoweverAlthough genomic analysis and derived applications have a very important role in the selection of economically relevant traits in dairy cows, the available reports referring to new traits have not yet been clarified in an extensive and detailed manner for specialized meat cattle breeds . In thisThe success of genetic evaluation applying genomic analysis information derived from breeding and selection programs have shown to be an efficient methodology to obtain higher genetic gain rates while reducing selection times and controlling the inbreeding coefficients in dairy cattle populations . LikewisRecently, many applications of genomics have emerged in the animal science field. Several countries leading in genotyping, including the United States through its Department of Agriculture (USDA), have already begun to conduct genomic evaluations of production and health traits in Holstein, Jersey, and Brown Swiss breeds . FurtherIn recent years, several studies dealing with the genotyping of continuous homozygous segments  have been generated, providing a new way to assess the degree of genomic inbreeding . In addiSeveral studies have used genomic selection in high-yielding Holstein and Jersey cows to determine the degree of association of the components of additive variation and dominance for fertility and milk production traits ,128,129,The genomics of nutritional and metabolic traits related to the environment opens possibilities to develop novel concepts for production functionality in dairy cattle populations. Understanding the mechanisms through which genes are expressed and interact with the environment is of remarkable importance because it allows correcting undesirable features and including other traits of zootechnical interest.Genomic analysis has recently proven to be an efficient methodology to estimate dairy cattle traits not traditionally considered important. Consequently, the interest in production traits in the dairy industry has led several researchers to study specific related genes. For example, several genes have been described that are differentially expressed during lactation and participate in biological processes such as the development of the mammary gland, protein and lipid metabolism, signal transduction or cell growth, and differentiation . Other sRegarding metabolic activity, genotyping work has been done in Holstein dairy cattle based on the number of births  highlighting the existence of high genetic correlations between non-protein nitrogen (MUN) and several production  and reprIn countries at the forefront of genomic evaluation, such as Australia, new traits such as feed efficiency and heat tolerance have been included in association with health traits . Thus, tFinally, some authors have established genomic reliabilities for different traits evaluated in dairy cattle breeds between 60 and 75%, and in new traits, they could be under 50% ,142,143.Genome editing comprises a set of genetic engineering tools that allow the direct manipulation of a sequence in the genome of an organism by removing, inserting, or replacing nucleotides for scientific or commercial purposes. This technology employs nucleases (enzymes coined as “molecular scissors”), which hydrolyze or catalyze the double-stranded DNA in a very precise way at specific sites in the genome . Genome The Cas9 protein is derived from the CRISPR Type II bacterial immune systems (CRISPR/Cas9) that can be easily programmed to target new sites by altering the RNA guide sequence . RecentlAnother cutting-edge technology in the field of genome editing besides CRISPR involves the transcription-activating nuclease (TALEN). This system can be artificially engineered to bind to a chosen DNA sequence, allowing it to be cut at desired locations for use in genomic editing or in situ genome editing, a capability shared with other nucleases like zinc finger nucleases (ZFN) that make use of the target proteins that bind to the DNA at specific locations .Homologous recombination has been used to edit genomes in different animal species . One exaRegulatory perspectives may slow the widespread implementation of genome editing in animals. Several international organizations consider that genome editing, used to replicate a “natural” mutation, should not be of regulatory concern because it would be equivalent to natural zootechnical management of existing alleles from the natural gene pool. On the contrary, the US FDA considers that there is a need for regulatory oversight of intentional genomics in animals because the editing of one target allele or nucleotide may indirectly or accidentally result in editing another with potentially detrimental effects. For example, there could be modifications affecting food safety due to changes in protein expression, interruption of protein functions, protein over-expression , or the creation of new expression products . TherefoThe application of genomic analysis in dairy cattle selection provides very interesting predictions for the genetic improvement in the short/medium term while controlling the inbreeding coefficient, reducing the generational interval, and improving the selection of progenitors, thus generating more productive genetic lines. The estimation of the different traits in dairy cattle is more efficient through the use of data derived from genomic analysis than through data derived from the pedigree. However, the association of both methodologies significantly improves the estimates in terms of prediction and accuracy. Genomic trait estimation accuracy must be supported by multi-generation databases to make predictions more consistent. This will be most noteworthy with the incorporation of new traits into the evaluations, considerably improving selection programs based on such estimates at both the individual and population levels. Selection rates based on genomic analysis provide a better understanding of the biology of the animals, new sources of data, novel genome editing methodologies, and pertinence for changing economic and environmental conditions. This becomes highly relevant as estimations derived from dairy cattle genomic data regarding adaptability traits in different environments or settings are more needed than ever. Genomic methodologies are a perfect fit during these challenging times of global climate change.Dairy cattle genomic-based selection will be used in the very near future for genetic selection in the beef cattle industry and other animal species. The frequency of rare alleles, recessive genes, and new mutations could generate genetic variability of desirable economic traits. Thus, the information derived from genomic analysis represents an important alternative for the production of databases of great interest in animal sciences and novel genome editing applications, accelerating the process of genetic evaluations in dairy cattle.


The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a new coronavirus that was recently discovered in 2019. While the world is working hard to overcome and control the coronavirus disease 2019 (COVID-19) pandemic, it is also crucial to be prepared for the great impacts of this outbreak on the development of antimicrobial resistance (AMR). It is predicted that inappropriate and too much use of antibiotics, biocides, and disinfectants during this pandemic may raise disastrous effects on antibiotic stewardship programs and AMR control all around the world. Furthermore, the use of certain antibiotics alone or in combination with antiviral agents or other medications for the treatment of secondary bacterial infections among COVID-19 patients may be regarded as a major factor that negatively affects host immune response by disrupting mitochondrial function and activity. Herein, we suggest that the current management strategies to control AMR and prioritize antibiotic stewardship schemes should be extremely highlighted in relation to the COVID-19 outbreak. The rising concerns about excessive use of antimicrobials and biocides and taking too much hygiene also need to be addressed during this pandemic due to their impacts on AMR, public health, and the environment. Globally, coronavirus disease 2019 (COVID-19) has created a health catastrophe that has had an immense impact on social care systems, economy, and financial resources. The importance of infection control and prevention measures such as social distancing, hand hygiene, and self-isolation has now been advised at a communal level . The COVThe World Health Organization (WHO) is worried that the trend will later be fueled by the wrong consumption of antibiotics during the COVID-19 pandemic. Indeed, when physicians are not aware or do not have access to the laboratory diagnostic findings related to the etiology of the infected patients, they tend to drive higher administration of different antimicrobials . Recent The uncontrolled use of biocide-based products could lead to the unusual release of such antimicrobials into environmental resources and cause the selective survival of bacterial strains carrying resistance genes during the COVID-19 catastrophe . FurtherStaphylococcus aureus (MRSA) in the hospital environment . Based on their approach, empirical antifungal therapy does not seem to be beneficial, and about 21% of patients who received empirical antifungals such as voriconazole, isavuconazole, and caspofungin had adverse effects without providing a survival benefit. Therefore, incorporation of strategic algorithms for accurate diagnosis and effective treatment of invasive fungal disease is vital in critically ill patients with COVID-19 and subsequently may result in a definitive diagnosis and successful antifungal therapy throughout the period of severe respiratory distress.Antimicrobial resistance has been emphasized as the most considerable threat to global health and economy in recent years, but it can hide behind COVID-19 for a while. It is predicted that the current emergence of the COVID-19 pandemic can seriously facilitate the growth rate of AMR worldwide; thus, the continuing and preventive measures against AMR should not be neglected during this global disaster . Unfortuironment . Until nironment . In anotironment , 11% of ironment , putativIstituto Superiore di Sanità in Italy, 84% of the expired SARS coronavirus 2 (SARS-CoV-2)-positive patients used antibiotic therapy, whereas 54% received antiviral therapy and 31% received steroid therapy during hospitalization  and microecological niches where various bacterial species are present is critical. Generally, it is supposed that high concentrations of such microbicidal agents could completely kill most bacterial species especially those that provide beneficial services to the ecosystem and to other living organisms while inhibiting the emergence of drug-resistant microorganisms . On the n health .qac genes in Staphylococcus, Enterococcus, and Escherichia coli (It should be noted that understanding the transferability of disinfectant resistance genes can considerably help to limit the outspread of such mobile genetic elements among microorganisms . Albeit hia coli . Taken tSince there is currently no specific and effective therapeutic agent, medication, or vaccines available to treat COVID-19 patients, the human immune system is considered as the solely active weapon against the virus. Unfortunately, off-target sites of majority of the approved antibiotics include mitochondria, which are evolutionary linked to prokaryote cells . MitochoBefore the emergence of the SARS-CoV-2 pandemic, the world was already demanding immediate, coordinated, and ambitious actions to avert the potentially disastrous AMR crisis and its related economic and health consequences. Now that we are in the midst of a global crisis, which is disrupting normal life for everyone, causing hundreds of thousands of people sick or hospitalized, and killing some of the most high-risk individuals, a better understanding of the pathophysiology of COVID-19 and advocating of potential effective therapeutics for the treatment of secondary bacterial co-infections seem to be critical. Also, prospective monitoring of the co-infections in patients with COVID-19 could help apprehend whether such co-infections affect disease progression and may interfere with antiviral treatment. Furthermore, current management practices and strategies to control AMR and prioritize antibiotic stewardship schemes should be extremely highlighted in relation to the COVID-19 outbreak. The concerns about consumption of “too many antimicrobials and biocides” and taking “too much hygiene” also need to be addressed during this pandemic due to their impacts on AMR, public health, and the environment.SR and AY contributed significantly to the literature review and wrote the draft of the manuscript. AY worked on concept and design of the study and interpreted the collected information. BH, HA, and MZ provided clinical advice and guidance for the improvement of the manuscript. AY critically revised the final version of the manuscript. All authors approved the final version of the manuscript and the authorship list.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.


Biodiesels and biolubricants are synthetic esters produced mainly via a transesterification of other esters from bio-based resources, such as plant-based oils or animal fats. Microwave heating has been used to enhance transesterification reaction by converting an electrical energy into a radiation, becoming part of the internal energy acquired by reactant molecules. This method leads to major energy savings and reduces the reaction time by at least 60% compared to a conventional heating via conduction and convection. However, the application of microwave heating technology alone still suffers from non-homogeneous electromagnetic field distribution, thermally unstable rising temperatures, and insufficient depth of microwave penetration, which reduces the mass transfer efficiency. The strategy of integrating multiple technologies for biodiesel and biolubricant production has gained a great deal of interest in applied chemistry. This review presents an advanced transesterification process that combines microwave heating with other technologies, namely an acoustic cavitation, a vacuum, ionic solvent, and a supercritical/subcritical approach to solve the limitations of the stand-alone microwave-assisted transesterification. The combined technologies allow for the improvement in the overall product yield and energy efficiency. This review provides insights into the broader prospects of microwave heating in the production of bio-based products. Microwaves have been used in various fields, such as food, chemical, medical, and telecommunication industries. Previous studies have also demonstrated the potential of non-thermal aspects of using microwaves in biological, hydrothermal depolymerization, or carbonization processes ,2,3,4. FMicrowave heating has shown potential applications in biodiesel production through a transesterification reaction , syngas The application of microwave heating requires an understanding of the dielectric properties of the microwave reaction materials . The molThe microwave heating requires significantly less energy because of localized heating rather than the use of conventional heating through a conduction and convection. The conventional heating depends on the temperature gradient between the internal reactor wall and the reaction mixture, and the conductivity of the reactor wall. The heating of the reaction mixture is established only when the conductive heat transfer through the wall has occurred, after which the heat is transferred to the reaction media by convection. Factors affecting conduction heating are the properties of the material, the temperature gradient, and the cross-section of the reactor and its thickness. The convection heating starts when the fluid near the wall surface is hot. The molecules then gain energy in the form of internal energy as well as from heat of reaction. When the moving fluid is energized (heated), it expands and the density decreases, which causes it to rise and the process continues until the thermal equilibrium is reached. The ability of the microwave to reduce processing time, save energy and improve production yield is attributed to the intense heating at molecular level . The majSignificant developments in biodiesel and biolubricant synthesis involving the raw materials, catalysts, an optimization of operating conditions, have been underway over the decades. The main objective is to optimize the yield of the product at the lowest overall production cost. The process of transesterification in biodiesel synthesis involves the chemical fragmentation of triglycerides into smaller, straight-chain alkyl ester molecules. Glycerol and fatty acid ester molecules are produced by the reaction of one mole of triglycerides with three molar equivalents of methanol. This reversible transesterification reaction is also known as an alcoholysis or methanolysis. Biolubricant, especially polyol ester, on the other hand, is produced by transesterification of fatty acids or fatty acid esters with polyhydric alcohol.Biolubricant is a safer candidate to mitigate environmental issues contributed by the industrial lubricants in the event of accidental spillage or irresponsible disposal in water or soil . It is aAlkaline catalysts tend to form metal soap during transesterification because of free fatty acids reacting with the metal component of the catalyst. Hence, saponification reactions can affect product separation and conversion . RegardlThis paper reviews developments in the use of combined microwave techniques in biodiesel production, as well as reviews the present state of technology and provides insights into potential technologies. The literature analysis suggests that several review papers have been published regarding microwave-assisted heating combined with acoustic cavitation, vacuum pump, and supercritical/ subcritical approach to improve chemical reactions, but a comprehensive review of the applications of these combined microwave technologies for biolubricant and biodiesel production is still lacking. This review provides guidance on how the combination of microwave and other techniques can be incorporated into the sustainable production of biodiesel and biolubricants.Currently, microwave irradiation is more common in the production of biodiesel than in the productions of biolubricants. An improved process for the transesterification of biodiesel fuels and fuel additives involving the use of microwave energy had been patented . It was Although a great deal of research has been done on the microwave-assisted synthesis of biodiesel and biolubricants, some issues related to microwaves, such as arching , overheaThe lack of uniformity in a temperature distribution due to the limitation of microwave penetration depth had also been addressed . Other rThree other limitations of microwave technology, including scalability, limited application, and health hazard have been addressed by other study . The wavN-alkylations, and Dimroth rearrangements [Microwave heating is a convenient process for drying polar compounds. The presence of water draws the -OH bond to align and continuously change the magnetic field. This motion leads to molecular friction, leading to heat generation. Heating non-polar molecules by microwave is not effective and therefore requires some modification, such as the addition of polar substances to the reaction mixture. Previous studies have investigated the effects of adding both passive heating element and ionic liquid to heat non-polar substances  ,66. Sintngements . Carbon-ngements . Furtherngements . The limThe transesterifications of biodiesel and biolubricant involve multiphase reaction mixtures, such as the mixture of oil-alcohol for biodiesel synthesis and ester-alcohol for biolubricant synthesis. A chemical process intensification is crucial to improve an emulsification of two immiscible liquids in the reaction media. Moreover, a formation of sufficiently small droplet size of reaction mixture would result in the increase of contact surface area between reactants and thus, intensify the transesterification process. Recently, a combination of two simultaneous reactor technology involving microwave has been explored to enhance the mass and heat transfers and at the same time could reduce the energy consumption during the transesterification reaction.An acoustic cavitation has been widely used to produce biodiesel and accelerate the transesterification reactions with either homogeneous or heterogeneous catalysts ,70,71. TThe combination of microwave with either ultrasonic cavitation or hydrodynamic cavitation see  would reA combination of microwave and ultrasound technologies in sequential or simultaneous mode has been studied for biodiesel production. Gole and Gogate  developeArdebili et al.  conducteFurthermore, Martinez-Guerra and Gude  evaluateWhen using a combined microwave-ultrasound technology, operating power and time of the process play significant roles in enhanced transesterification. Martinez-Guerra and Gude  suggesteEven though combining microwave and acoustic cavitation technologies appears promising, the use of this method is still new to biodiesel. As for biolubricant production, research studies on the use of combined microwave-ultrasound cavitation reactor are still limited. However, the use of a stand-alone acoustic cavitation for the transesterification of bio-based resources had a better exergy efficiency in the biodiesel production, whereby an energy saving around the hydrodynamic cavitation reactor was better than the use of the ultrasound biodiesel reactor . PatiencAlthough the development study of a combined technology of microwave and hydrodynamic cavitation is still limited, the use stand-alone hydrodynamic cavitation-assisted reactor had resulted in 98.1% conversion of methyl ester from waste cooking oil and methanol in 15 min by introducing 21 of 1 mm diameter holes in the reactor orifice. Furthermore, 96.5% conversion of methyl ester from rubber seed oil was also reported by using the reactor in 20 min reaction time with the reactor capacity of 50 L . AlthougJatropha curcas L. (JCL) seed by increasing the porosity of the sample, thus weakening its structure [Transesterification of bio-based resources via a supercritical or subcritical approach is catalytic-free, which simplifies the process, it does not require pretreatment or purification steps and is a wastewater-free process ,85 see . Howevertructure . Furthertructure  using metructure . By usintructure ,88.2 at a milder condition [2 application allowed the reaction to take place at 280 °C, 20 MPa with a yield of 97.8% in 90 min. In addition to the longer reaction time, the ratio of oil to methanol used was as large as 1:60. In addition, Aboelazayem et al. [Further improvement in the process condition by lowering the operating temperature was later reported by introduction of two steps of subcritical acetic acid and oil for fatty acids and triacetin production, followed by using a supercritical methanol for the conversion of FAME in a milder environment. After reaction times in 5 min at 300 °C, 20 MPa and 15 min at 270 °C, 17 MPa, the yield was 96%. This has remarkably shortened the total reaction time . On the ondition  had beenm et al.  bridged A research study was conducted on a combined microwave-subcritical assisted transesterification of palm oil with dimethyl carbonate (DMC) as a solvent . In 2.5 Some researchers have suggested that polar substances reduce their polarity or dielectric constant at increasing temperatures ,92,93,94In the biodiesel production, vacuum pumps are mainly used in the separation of biodiesel and the by-product, methanol recovery and glycerin purifying. The utilization of vacuum pump can shorten the reaction time as it drives the reaction in the forward direction by removing the volatile by-product under continuous vacuum see . In vacuThe use of vacuum in a conventional heating was reported to increase yields of transesterification and esterification . A vacuuWhen combined with a microwave, Khalfan and Dwivedi  reportedAziz et al.  evaluateIn the microwave-assisted heating method, heat transfer via dipole and ionic polarizations are attractive attributes that contribute to an efficient heating. Microwave heating has benefitted from the polarization of alcohol (methanol and ethanol) and oil or fatty acids in conventional biodiesel and biolubricant production methods. With an addition of sufficient free ions as a heating carrier/passage in a sample, higher microwave absorbance characteristic would be achieved and ionic polarization would be intensified. Researchers are recently explored in-depth for the use of microwave and ionic solvent or ionic liquid (IL) as a catalyst to enhance the thermal effect of microwave heating in biodiesel production see .Biodiesel production using ionic solvent under microwave heating was summarized in The comparison of a conventional and microwave heating for the biodiesel production with imidazolium-base IL had been investigated . It tookThe efficiency of combined microwave technologies in the intensification production of biodiesel and biolubricants was recognized by comparing their performance with the conventional and stand-alone microwave heating. The combined technology approach involve undoubtedly new techniques that have a promising potential for applications in the processing of fine chemicals on an industrial scale. Currently, the potential of microwave technology to produce biodiesel and biolubricants are mostly conducted at a laboratory scale using relatively small quantities of raw materials. Nevertheless, previous findings have served as a solid basis for the expansion of the microwave-based transesterification technology to a larger scale. Several studies suggested developing continuous flow and higher capacity system for microwave and other technology enhanced biodiesel production for a more energy-efficient and chemical saving system ,78. ThisThe use of a combined microwave-acoustic cavitation reactor has high potential to shorten the reaction time significantly, with high yield of products, excellent energy savings, and minimal utilization of hazardous chemicals ,112. TheAccording to the most recent studies, the method of transesterification with microwave hybrid technology for biodiesel and biolubricant production is currently at the laboratory validation level ). The production facilities are still working in batch process and not in an entire logistics material chain. The application of microwave hybrid technology is under research and development, similar with various conversion routes such as gasification, hydrodeoxygenation, and pyrolysis. Compared with other biofuel production, such as pyrolysis, the technological maturity of pyrolysis suggests that there is a viable opportunity to further develop this technology as technological developments move towards commercial applications . The chaThis review article provides a summary of research findings in the field of microwave-assisted transesterification processes combined with other technologies. This review has demonstrated the advantages of combining microwave heating and other technologies, such as acoustic cavitation, ionic approach, supercritical/subcritical, and vacuum. As compared to the stand-alone microwave assisted reactor technology, the innovative combined technologies with microwave adapted for biodiesel and biolubricant production has great advantages, such as faster heating, lower energy consumption, shorter processing time, higher production yield, and product quality improvement. In a combined microwave-acoustic cavitation technology, acoustic cavitation could provide a mechanical mixing effect needed by microwaves to improve mass and heat transfer, while microwaves generate an excellent thermal energy. Hence, the assistance of microwave-acoustic cavitation during the synthesis of biodiesel or biolubricant clearly takes advantage of the high-energy impact of microwave and acoustic cavitation to address certain shortcomings of conventional processes. On the other hand, combining a microwave with a vacuum pump drives the reaction in the forward direction by removing the volatile by-product under continuous vacuum; therefore, the reaction time can be shortened. The subcritical/supercritical condition with the microwave approach may simplify the separation and purification steps by omitting the catalyst. Adversely, this strategy is correlated with higher solvent demand and energy usage. Microwave-assisted transesterification of triglycerides with ionic liquid combination may somehow benefitted the reusability of the solvent provided. The combination technology could benefit the industry in terms of production time and cost. However, most previous studies are still in laboratory scale and there are still limited scientific reports on the upscaling application of this method to the industrial level, as the technology is relatively new. Detailed research studies on process design, reaction kinetics, thermodynamics, and biodiesel/biolubricant analytical protocols for the larger production are needed for the future up-scaling process for the industrial production of biodiesel and biolubricant. With strict regulations and an increase in the use of renewable energy, microwave-assisted biodiesel and biolubricant synthesis with combined technologies may become more common in the future.


Deep Eutectic Solvents/Complex Salts-Based Electrolyte for Next Generation Rechargeable Batteries focuses on the effects of electrolytes on the electrochemistry/chemistry of rechargeable batteries and cells.Recent years have seen an expansion of renewable energy technologies driven by global demands for energy alongside social and environmental concerns. One of the most significant solutions, rechargeable batteries have promising features which include high capacity, energy density, rate capability, long lifetime, and cost-effectiveness. As the key component in energy storage devices, the electrolyte has had a major impact on the chemistry/electrochemistry of rechargeable batteries/cells for a number of reasons. These include its potential window, which limits the redox potential of an electrochemical reaction. Its electrochemical activity and conductivity also influence the electrochemical reaction and consequently the battery performance. The composition, as well as the stability, of rechargeable batteries, shapes the electrolyte-electrode interface. Furthermore, its corrosivity cannot be neglected. For these reasons, researchers are highly motivated toward breakthroughs in battery performance, exploring the fundamental properties of electrolytes based on novel formulation/synthesis. Hence, this special issue of Pan et al.). Other submissions discuss the compositing strategy for a polyethylene oxide-based solid state electrolyte for flexible device and lithium metal batteries . Another study considers the emerging ionic conductor for an Mg ion solid state electrolyte , where another describes how electrolyte modification can be used to depress zinc dendrite in zinc-based flow batteries . The electrolyte concentration, as well as cation/proton types of diffusion kinetics in a vanadium flow battery  are also interest. Another contribution examines the molecular design of organic electroactive molecules via functional group modification for redox flow batteries . Furthermore, the electrolyte-electrode interface is also emphasized in this topic, where its material characteristics and stability strongly influence the kinetics and cycling stability of batteries/cells for example in an in situ Scanning Electrochemical Microscopy study, which revealed that the SEI formation on carbon depends on voltage range , or, as described elsewhere, regulating the nature of ionic liquid and the interface compatibility . Another study designed a novel 3D MoSe2 template for constructing stable SEI for reversible stripping/plating of metallic Mg , where others discuss regulating the interface between electrolyte and Zn for preventing zinc dendrites , or investigating the interfacial charge transfer in aqueous zinc electrolytes for zinc ion batteries .In this Research Topic, representative types of electrolytes are discussed in relation to next-generation rechargeable batteries, including ionic liquid, solid state electrolyte (polymeric and inorganic), multivalent electrolyte (aqueous and non-aqueous) for multivalent ion batteries, aqueous, and a novel organic electroactive molecule-based electrolyte for flow batteries. Recent developments build upon the correlation between design and synthesis of the electrolyte, its properties, and the related electrochemical properties and performance of batteries and cells. From the perspective of electrolyte design, contributions include exploration of formulating ionic liquid toward designed ion separation for tuning the performance of supercapacitors (We would like to thank all the authors for their valuable contributions and the reviewers for their thoughtful insights and suggestions. It is essential to further examine and share the fundamentals of the chemistry and electrochemistry of electrolytes, which serves as the basis for the successful creation of next-generation rechargeable energy storage devices.Although the papers included here provide significant insights, there are still important developments and points for the future development of this area of research. Safety is a long-term pursuit in rechargeable batteries, and further insights into safe electrolyte design with promising electrochemical properties are essential. The pursuit of high performance and long lifetime in rechargeable batteries will also need to go further than constructing a database of electrolytes, and future explorations on the relation between electrolytes and battery chemistry are essential. It would also be promising to uncover the correlation between electrolyte and charge storage mechanism for any novel electrodes designs, which will synergistically guide the development of new generation battery systems. Moreover, the complexity of the electrolyte-electrode interface creates room for exploring battery chemistry, which will drive cutting-edge characterization and analysis tools and in parallel, boost the development of new battery systems, especially those for utilizing metal anode based or multivalent ion batteries.We hope that this special issue will inspire future research in the development of novel electrolytes and drive the development of new generation rechargeable energy storage devices. These endeavors will pave the way for realizing a green and sustainable society.DY, GC, CJ, and HZ co-edited this special issue. All authors contributed to the article and approved the final 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.


Traditionally, gross cardiac anatomy has been described mainly based on the findings in the dissection suite. Analyses of heart specimens have contributed immensely towards building a fundamental knowledge of cardiac anatomy. However, there are limitations in analyzing the autopsied heart removed from the thorax. Three-dimensional imaging allows visualization of the blood-filled heart in vivo in attitudinally appropriate fashion. This is of paramount importance for not only demonstration of cardiac anatomy for educational purposes, but also for the detailed anatomical evaluation in patients with acquired and congenital heart disease. In this review, we discuss the advantages of three-dimensional imaging, specifically focusing on virtual dissection, a volume rendering-based reconstruction technique using computed tomographic data. We highlight examples of three-dimensional imaging in both education and guiding patient management. The modern understanding of cardiac anatomy is incomplete without referencing the works of Professor Robert H. Anderson. He continues to be a pillar to which our knowledge of cardiac anatomy in both normal as well as malformed hearts is supported. During his long illustrious career, he has enriched the field of cardiac development and anatomy, the latter of which has largely depended on analysis of heart specimens. The autopsy-based analyses, however, have inherent disadvantages of loss of in vivo blood-filled shape and attitudinally appropriate relationship to surrounding structures. Besides, delineation of cardiac anatomy mandates dissection, which results in deformation and permanent loss of its original state.Modern computed tomography scanners acquire high-resolution datasets. Simultaneous improvement in computational capabilities now permits high-quality three-dimensional reconstruction of computed tomographic datasets. The three-dimensional reconstructions, particularly when achieved by volume rendering, permit visualization of cardiac anatomy equivalent to the dissection of heart specimens with the additional benefit of viewing in attitudinally appropriate orientation. Such dissection performed virtually has an added advantage of evaluating cardiac anatomy in vivo. Furthermore, the feasibility of reconstructing in any plane relevant to the cardiologist, radiologist or surgeon makes virtual dissection an ideal tool for education and clinical decision making. The ease and accuracy of cardiac anatomy using virtual dissection has prompted Professor Anderson to consider it as the gold standard for analysis of cardiac anatomy in living patients ,2,3. We Recent advances in visualization have permitted much needed three-dimensional reconstruction of datasets obtained from computed tomography and magnetic resonance imaging. Although commonly performed using commercial software packages, three-dimensional reconstruction can also be achieved using free open-source software ,5,6,7,8.Commonly, contrast enhancement is used for detailed cardiac evaluation to visualize the chambers, vessels, and/or coronary arteries. When the contrast agent is used, it is the enhanced chamber and/or the lumen of these vessels that are usually the focus of reconstruction . We callIn this material, we mainly focused on the computed tomographic images as the spatial resolution of clinical datasets is commonly superior to that obtained from cardiac magnetic resonance imaging. However, three- or four-dimensional analysis is also feasible for datasets obtained from magnetic resonance imaging, which have advantages in its non-invasiveness, temporal resolution, tissue characterization, perfusion analysis, and flow analysis ,20.Virtual dissection is nothing but a modification of the volume-rendering technique wherein the focus is shifted to the walls of the cardiac chambers and vessels. While endocast images can be extremely useful, especially when visualizing the great vessels and their relationships to the other intrathoracic structures or relating to angiographic imaging, they become less informative when aiming to understand the detailed intracardiac anatomy and how this relates to the proximal great vessels. It is often the walls, septums, and valves of the chambers and/or vessels which are targeted for transcatheter or surgical intervention , informaDepending on the interest of visualization, reconstruction using the volume-rendering method requires a combination of various concepts of imaging .The main advantage of virtual dissection from computed tomographic datasets is that it can provide a high-resolution image of the blood-filled heart without removing it from the thorax . Unlike It is well understood that “form begets function”. Therefore, it is not surprising that the foundation of medical education often begins with instruction in anatomy followed by (patho)physiology. While medical students are taught cardiac anatomy in their first years of medical education, supplemented by cadaveric dissection and inspection, the cardiology and cardiothoracic surgery trainees are often starved of such experiences. This relates to several factors, including the scarcity of preserved heart archives, competing educational requirements in a relatively limited time, and demanding clinical schedules ,29,30. SWhile three-dimensional imaging lacks the tactile advantage of the printed model of the heart, an excellent demonstration of cardiac anatomy in attitudinally appropriate orientation makes up for this deficiency to a large extent. Three-dimensional imaging has the potential to serve as the mainstay of imparting knowledge of cardiac anatomy in the absence of heart specimens . Even atFor educational and diagnostic purposes related to the heart, it is not always sufficient to understand only the cardiac anatomy. Often, it is more important to clarify relationships with the adjacent structures in the thoracic cavity ,37,38. VFor more specific purposes, volume-rendered images, involving both endocast and virtual dissection images, can be also used to educate fluoroscopic anatomy to cardiac interventionists and electrophysiologists as their procedures are commonly performed under fluoroscopic guidance and with angiography . VirtualSimilar applications of three-dimensional reconstructions can be applied towards congenital heart disease. This includes not only in the evaluation of unrepaired congenital heart disease which may involve complex abnormal three-dimensional relationships of chambers and vessels, deficiencies in septation, or abnormalities of atrioventricular or semilunar valves but also in the evaluation of their repaired and palliated forms. A combination of both endocast and virtual dissection images is often beneficial to provide a comprehensive three-dimensional understanding. There is increasing application towards the adult congenital population, given the complexity of evaluating many of their repaired and palliated lesions as well as the common scenario of poor acoustic windows limiting complete echocardiographic evaluation .Relationship of the great arteries to an interventricular communication in the setting of double outlet right ventricle for surgical planning ;Unicuspid aortic valve for surgical repair planning ;Obstruction in surgically constructed pulmonary venous baffles for surgical planning  16];;16];Right ventricular outflow tract obstruction in repaired tetralogy of Fallot to guide transcatheter pulmonary valve replacement .In addition to cases demonstrated in Three-dimensional imaging is currently used to create files for converting the image into three-dimensional printing  and virtThere is no doubt three-dimensional printed models and the increasing use of augmented and virtual reality can be extremely useful for education, procedural simulation and planning ,43,44,45Furthermore, for the analysis of cadaveric hearts, three-dimensional imaging plays an important role to clarify three-dimensional arrangement of the myocardial mesh and conduction system using diffusion tensor magnetic resonance imaging and microcomputed tomography ,53.When creating three-dimensional images based on the volume-rendering method, it should be always kept in mind that the quality of the final image depends on the quality of the raw data. In this regard, the collaboration of radiologists and radiological technologists is an important prerequisite to optimize image acquisition with appropriate radiation dose and contrast material . While rThe volume-rendered image might not precisely reflect the real anatomy. For example, the visualized anatomy can be enlarged or contracted from the real thing, depending on the software and window level/width applied. Thin or moving structures, such as the floor of the oval fossa (primary septum), membranous septum, and valvar leaflets are sometimes difficult to reconstruct, and can easily be mistaken for a defect. Some software can virtually create structures that are not present, such as vessels, as if it were a part of the real anatomy. Smoothing is commonly applied to some extent, which may distort the original information in the raw data. Therefore, measurements should not be performed on the volume-rendered image but should be measured using two-dimensional multiplanar reconstruction images. This can then be overlaid on the three-dimensional image for an improved understanding of these measurements related to the three-dimensional structure . Thus, tThe obtained three-dimensional data from each patient should be fully utilized to create images to show comprehensible clinical cardiac anatomy critical to the clinical diagnosis and treatment. In this regard, virtual dissection, a three-dimensional volume-rendering imaging technique, is emerging as the gold standard for understanding detailed cardiac anatomy in the living patient. This technique is useful for both medical education as well as a detailed clinical evaluation of the complex three-dimensional heart, whether demonstrating normal anatomy for educational purposes or evaluating the patient with acquired or congenital heart disease. Any reconstruction depends both on the source imaging data as well as the anatomical understanding of the reconstructor. It is a requirement for radiologists, anatomists, cardiologists, interventionalists and surgeons to keep abreast of the benefits and limitations of these techniques and to fully utilize the three-dimensional anatomical data obtained to optimize patient management and outcomes.


Immunotherapy with checkpoint blockers (ICBs), aimed at unleashing the immune response toward tumor cells, has shown a great improvement in overall patient survival compared to standard therapy, but only in a subset of patients. While a number of recent studies have significantly improved our understanding of mechanisms playing an important role in the tumor microenvironment (TME), we still have an incomplete view of how the TME works as a whole. This hampers our ability to effectively predict the large heterogeneity of patients' response to ICBs. Systems approaches could overcome this limitation by adopting a holistic perspective to analyze the complexity of tumors. In this Mini Review, we focus on how an integrative view of the increasingly available multi-omics experimental data and computational approaches enables the definition of new systems-based predictive biomarkers. In particular, we will focus on three facets of the TME toward the definition of new systems biomarkers. First, we will review how different types of immune cells influence the efficacy of ICBs, not only in terms of their quantification, but also considering their localization and functional state. Second, we will focus on how different cells in the TME interact, analyzing how inter- and intra-cellular networks play an important role in shaping the immune response and are responsible for resistance to immunotherapy. Finally, we will describe the potential of looking at these networks as dynamic systems and how mathematical models can be used to study the rewiring of the complex interactions taking place in the TME. Tumor cells are able to activate several mechanisms to evade the immune response by disguising themselves as “self” cells. Binding to inhibitory checkpoint molecules  they can block antitumor activities of the immune system. Immunotherapy with immune checkpoint blockers (ICBs) uses antibodies to target immune checkpoints, such as PD1, PD-L1, and CTLA-4, unleashing the immune response. In clinical trials, ICB therapy has been shown to achieve durable therapeutic response and to increase patient survival in different cancer types, although still a small number of ICBs are FDA-approved . Even ifThe investigation of mechanisms supporting immune resistance has provided a great opportunity for biomarker discovery of patient response to ICBs . Two bioDespite being promising, these biomarkers also present some limitations. For instance, IHC enables measuring PD-L1 expressed on tumor cells, however the expression of this biomarker fluctuates over time and varies between different tumor sites. This variability undermines the ability to evaluate PD-1/PD-L1 therapies effectiveness based on IHC, as reviewed in Topalian et al.  and CamiAbove-mentioned examples shed light upon the conceptual problem of looking only at individual components of the TME. While the characterization of different parts playing a role in the interaction between tumor and immune system has been essential to elucidate the most important actionable mechanisms, further research is required to define biomarkers harnessing a more coordinated joint action of these mechanisms. Predictive biomarkers for immunotherapy with ICBs have been extensively reviewed previously , 23, 25.It is well-known that different types of immune cells can play a different role in the response to ICBs . For exaApart from quantification of immune cells, their spatial localization also plays a pivotal role in the response to immunotherapy . For insAnother important factor that affects patients' response to ICBs is the functional state of the different immune cells . DysfuncAnalysis on the immune infiltrate quantification, functionality, and localization can help both to explain the diversity of the tumor immune milieu and develop informative biomarkers for ICBs , 52, 53.The functional state of cells in the TME is defined by a complex system of communication between molecules within the cells (intra-cellular networks) and among different cells (inter-cellular networks). Looking at intra- and inter-cellular networks can provide a more holistic perspective of the TME and inform a new class of biomarkers for immunotherapy and its potential combination with other targeted therapies .Intra-cellular signaling pathways play a part in shaping the interaction with the immune system , 66; Fig. AbnormaIn addition, cancer cells receive signals from other cells in the TME through ligand-receptor interactions. These inter-cellular communications lead to changes in the phenotype of the regulated cells thus playing an important role in both progression and prognosis of cancer , 74. An in vitro and ex vivo investigation of intra-cellular communication, and to study their effect on the response to ICBs  from what is called pseudoprogression, i.e., temporary progression followed by a response to the treatment (patient should be kept on ICB). The latter behavior has been described using a model of immune activation incorporating the dynamics of antigen presentation . Based oresponse , 112, thresponse , 114 forCurrent limitations in identifying predictive biomarkers for ICB therapy are partially due to overlooking the complexity of the TME. Following the advancements in technologies to measure multi-omics data, measurements of bulk populations, individual cells, and spatial information have paved the way to a more comprehensive view of the TME. Recent efforts are focused on searching for signatures of response to ICBs that consider quantification, localization, and functionality of different immune cells in the TME, showing improved predictive power with respect to simpler biomarkers . HoweverThere is compelling evidence that the interplay of the immune system, tumors, organs, and external environment, harmonizes antitumor immune responses . TherefoÓL-S and FE wrote and edited the article. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The impact caused by the Coronavirus Disease 2019 (COVID-19) pandemic has been strong enough to force all aspects of medical services to innovate. Although initially call into doubt, virtual medical services emerge to be one of the most promising options for orthopedic services globally. Considering the widespread implementation of isolation, quarantine, and lockdown, the incidence of motor vehicle accidents and traumatic injuries is markedly reduced which leads to a decrease in emergency surgeries . Also, most elective procedures, including orthopedic treatments , are deferred indefinitely, resulting in a drastic reduction in orthopedic service  transmission from patient to surgeon and the team through inhalation of the infected virus in aerosols or surgical smokes . A carefP = 0.29)  .Patient-reported satisfaction and health outcomes were found to be not significantly different between audiovisual-guided sessions and traditional doctor-patient appointments, and a high proportion of patients preferred virtual meeting as their next consultation . ConsideThe classification of surgical procedures based on the degree of emergencies is highly important, with certain conditions  requiring direct examination and immediate action by the surgeons and nurses. However, some elective cases can be adequately managed through consultation and rehabilitation at a distance . During In Indonesia, most orthopedic services are still provided through conventional face-to-face meetings, with many elective procedures being postponed indefinitely during such pandemic. A lot of people are still reluctant or afraid to visit hospital to seek treatment, while a portion of the population in the remote areas cannot reach healthcare facilities or depend on more traditional treatments, such as bone setters, soft tissue massage and manipulation, and traditional medicines . This isDuring the COVID-19 lockdown, many individuals became accustomed to sedentary behavior since they spent more time at home or even worked from home. They rarely exercise or participate in sports activities partly because of the inability to go outside. Physical inactivity is a serious concern other than an ever-increasing pandemic, where a surge of non-communicable diseases can emerge shortly . ConsideBoth types of telemedicine delivery, real-time (synchronous) and store-and-forward (asynchronous), bring significant benefits to patients undergoing rehabilitation . This teAs with any new system, the practice of virtual examination and assessment in orthopedic and rehabilitation is not without problems. Technological constraints, as well as individual skepticism and reluctance, both from providers and patients, are considered as the major obstacles in delivering teleorthopedic and telerehabilitation . Many inCOVID-19 is changing the delivery of orthopedic and rehabilitation care. In the future, there will be more virtual care. Designing standardized virtual examinations and measurements is the next move to enable doctors and physiotherapists to provide the same quality of medical services. Guidelines and recommendations for reinstalling orthopedic activity had been developed, including classification of procedures based on the emergency and urgency levels, stratification of the patient's risk of infection for the eligibility of surgery, recommendations for PPE use in the operating area, and strategy for resuming elective orthopedic surgery in four phases . At presML and RP confirm being the only contributors of this work and have approved it for publication. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The rubella virus (RV) was the first virus shown to be teratogenic in humans. The wealth of data on the clinical symptoms associated with congenital rubella syndrome is in stark contrast to an incomplete understanding of the forces leading to the teratogenic alterations in humans. This applies not only to RV, but also to congenital viral infections in general and includes (1) the mode of vertical transmission, even at early gestation, (2) the possible involvement of inflammation as a consequence of an activated innate immune response, and (3) the underlying molecular and cellular alterations. With the progress made in the development of pluripotent stem cell-based models including organoids and embryoids, it is now possible to assess congenital virus infections on a mechanistic level. Moreover, antiviral treatment options can be validated, and newly emerging viruses with a potential impact on human embryonal development, such as that recently reflected by the Zika virus (ZIKV), can be characterized. Here, we discuss human cytomegalovirus (HCMV) and ZIKV in comparison to RV as viruses with well-known congenital pathologies and highlight their analysis on current models for the early phase of human development. This includes the implications of their genetic variability and, as such, virus strain-specific properties for their use as archetype models for congenital virus infections. In this review, we will discuss the use of induced pluripotent stem cells (iPSC) and derived organoid systems for the study of congenital virus infections with a focus on their prominent aetiologies, HCMV, ZIKV, and RV. Their assessment on these models will provide valuable information on how human development is impaired by virus infections; it will also add new insights into the normal progression of human development through the analysis of developmental pathways in the context of virus-induced alterations. These are exciting perspectives for both developmental biology and congenital virology. Historically, malformations associated with congenital RV infection, as first described by Sir Norman Gregg in 1941, have formed our current concept of teratogenicity [-/- female mice with wild-type males [-/+ fetal cells in the placenta were facing IFNAR-/- cells in maternal tissue. In comparison to their homozygous (IFNAR-/-) counterparts, the placental damage caused by ZIKV infection was reduced. Moreover, these heterozygote (IFNAR-/+) pups were partially protected from high viral burden, especially in the brain [The rubella virus (RV) was not only the first human teratogen identified, and as such the first human pathogenic virus that has been classified as a teratogen, it is still one of the most efficient teratogenic viruses. A teratogen is defined as a physical, chemical, or infectious agent associated with physical or functional birth defects, including growth retardation and pregnancy loss, that result from abnormal embryonal or fetal development  by the initial contact of the trophoblast cells of the blastocyst with the maternal uterine epithelium, the endometrium, which converts into the decidua. In utero transplacental transmission of pathogens can occur through passage from maternal endothelial microvasculature to endovascular extravillous CTBs or through transfer from infected immune cells to placental syncytiotrophoblasts . DiffereIn contrast to HCMV and ZIKV, the mode of transplacental passage of RV is rather ill defined and not well studied. Transplacental passage of RV is linked to viremia, as the appearance of the rash in the infected mother can be linked to congenital rubella A. InfectAs outlined above, RV can be teratogenic from early pregnancy onwards. Experimental proof is provided for in utero transmission of ZIKV before formation of the placenta is initiated and even before implantation of the blastocyst . AlthougRV infection during pregnancy can lead to miscarriage and a wide range of manifestations referred to as congenital rubella syndrome (CRS). As summarized elsewhere , the mosFor the majority of the aforementioned teratogenic viruses, an essential determinant of the clinical symptoms of the newborn is the gestational age at which infection occurs, e.g., ,36,38. HSo far, an efficient vaccine is only available for RV, which was implemented in an effective vaccination program and led to a dramatic decrease in the incidence of CRS. However, there are still developing countries in Africa and Asia lacking routine rubella vaccination . Thus, fViral pathogenesis and disease-associated characteristics, such as those occurring during congenital infections, can differ among clinical isolates of human-pathogenic viruses. Such strain-specific differences occur in vitro as well as in vivo through viral adaptation to its host cell or human host, respectively. In the case of HCMV, these adaptations can occur at a fast rate. A recent review summarizes how this could hamper research on HCMV. Low-passaged HCMV strains are usually cell-associated, which restricts their use in experimental characterizations. During cell culture adaptation, HCMV strains acquire mutations and a reduced level of cell association and, thus, produce higher titers . Moreovewww.who-rubella.org/files/wer8832.pdf), [The two ZIKV lineages, Asian and African, display differences in their associated pathology and, especially infections with the Asian lineage, may result in severe malformations and developmental defects . However32.pdf), . In many32.pdf), ,60,61) a32.pdf), . This epThe application of experimental animal models for congenital infections in humans reveals essential disease mechanisms, but comes with several restrictions due to human-specific aspects. In the case of HCMV, the newborn mouse and guinea pig model were employed in assessment of virus-induced brain pathology  and vaccComparable to other viruses that are transmitted by arthropod vectors, IFNAR−/− mice lacking the receptor for type I interferons (IFN-α and -β) are a suitable experimental model for ZIKV . Any typAlthough transplacental passage of RV was demonstrated for some animal species, including rhesus monkeys ,76, nonewww.hdbr.org). Moreover, for technical and ethical reasons, the use of embryos and the samples provided by these tissue banks is fairly limited. Nevertheless, they could be used to verify findings on pluripotent stem cells . ESCs and iPSCs are especially a cell culture platform with unprecedented possibilities to address human development. Thus, a highly promising approach based on pluripotent stem cells is, not only moving into the focus of developmental biology, but also of congenital virology. While the mouse model in general has provided valuable information for human development and, more specifically as outlined above, on congenital ZIKV syndrome (CZS) and the associated pathology, it faces important restrictions. The interspecies differences between human and animal development and the awareness of the three Rs (3Rs) principle for animal research  ask for alternative research approaches. On the one hand, humans and mice share a hemochorial placenta, 99% of their genes, and most of their protein-coding genes ,78. On tAs highlighted in Pluripotent stem cell-based models mimic early human development through the application of various defined protocols to initiate differentiation. This can be done in two or in three dimensions, as exemplified by the formation of stem cell aggregates, the so-called embryoid bodies (EBs). In vitro differentiation can be spontaneous (undirected), as in the case of EB formation, or directed along a specific developmental route. Directed differentiation can also be initiated in adherent cell cultures or EBs through the addition of defined growth factors and pharmacological compounds ,88. Line-/- mouse model confirmed experimentally in the vertical transmission mouse model that pre-implantation infection of the dam can be transmitted to pre-implantation blastocysts [In the context of the implications that pluripotent stem cell-based models may have for our current concept of congenital virology, a justified question may arise on their relevance for addressing research questions on congenital infections. Human embryonic stem cell lines originate from the inner cell mass of human blastocysts, which were derived through cultivation of embryos donated by participants of in vitro fertilization programs . The appstocysts . This emstocysts .The implication of the two pluripotent stem cell lines, ESCs and iPSCs, as a suitable model system for congenital virology is two-fold. First, the outer-lining of the blastocyst, the trophoblast, can be modelled through trophoblast differentiation of pluripotent stem cells. Second, the epiblast, which gives rise to the three embryonic germ layers, can be recapitulated through two- and three-dimensional differentiation approaches and reflect early gastrulation-like developmental stages. Already in 2002, the first protocol on differentiation of hESCs into trophoblasts was published . While tSeveral studies were conducted on the impact of virus infections on mouse as well as on human pluripotent stem cells. Here, we want to focus on the findings related to the impact of HCMV, ZIKV, and RV on human development. Early reports on embryonal carcinoma cells suggested that HCMV permissiveness could change during the transition of the undifferentiated (pluripotent) to the differentiated state . The susIn contrast to infectivity by HCMV, iPSCs are susceptible and permissive to ZIKV and RV ,106. AmoThe unique ability of RV to replicate in iPSCs without cytopathogenic alterations enabled passaging of RV-infected iPSCs and their differentiation into embryonic germ layer cells . HoweverHuman ESCs and iPSCs share almost all characteristics of pluripotent cells. However, gene expression profiles of different iPS cell lines are very similar, but not identical to human ESCs . Some isPluripotent stem cell-based in vitro models grow increasingly complex. Moreover, the current technological advancements in the research on early human embryogenesis offer great possibilities for the understanding of diseases and for finding new therapies. Organoid cultures and engineered tissue cultures might reach a complexity that is comparable to the one found in tissues and organs of the adult human body. As a technical advance in the field, three-dimensional bioprintings of human alveolar cells were infected with the seasonal influenza virus to mimic the clustered infection pattern that is also observed in the lung of human beings . AccordiCurrently, rodents are broadly applied for the modelling of infectious diseases including congenital viral infections. One can conclude that species-specific differences most probably represent a major road block for the development of drugs and vaccines besides toxicological testing. However, species differences could also broaden our view on the evolution of humans and the co-evolution of congenital virus infections. Through the application of the iPSC technology to various species, including members of the great apes, we could address species-specific differences in congenital virus infections. Similar studies have already been conducted to address questions on human evolution. The comparison of endodermal differentiation that was induced in human and chimpanzee iPSCs revealed conserved, but also divergent evolutionary circuits . In turnPluripotent stem cells enable the differentiation of mature and functional cells, tissues, and organoids mimicking the (1) differentiation and maturation status of cells, (2) the specific cell composition of a certain tissue, (3) physical impulses necessary for maturation, and (4) the three-dimensional structure of certain tissues. However, there are technical limitations for reconstructing embryonal and developing tissues and organs for a more precise analysis of congenital virus infections. Accordingly, there is a need for pluripotent stem cell-based models for advanced three-dimensional in vitro models that closely resemble the in vivo situation. The importance of such three-dimensional scaffolds is exemplified by brain organoids that, in contrast to neuronal cell culture, reflect at least some parts of the architecture in early embryonal brain tissue. The recent review by Chen et al. does not only summarize the advancements in brain organoid technology over the past five years, but also their application in the study of human diseases, including ZIKV microcephaly . This reWith this review, we have addressed the possibilities offered by novel stem cell-based cell culture models for the study of the clinical presentation following congenital virus infections. We also want to stress the potential use of viruses such as HCMV, ZIKV, and RV, with their well-defined clinical symptoms during congenital virus infections, for the identification of novel gene functions and regulatory networks during human embryonal development. The combination of congenital virology with the continuous advancements of stem cell-based technology will complement developmental biology and extend our current way of getting information on the function of genes that are active in human development. Our knowledge of human development is, so far, to a large extent fueled by our knowledge from naturally occurring mutations. Data gained on congenital virus infections in stem cell-based models will also help us to better understand deleterious pregnancy outcomes. Moreover, the validation of iPSC-based cell culture models for the study of human development through infection with teratogenic viruses with well-known aetiologies will provide a framework for pathogens with unknown teratogenic potential. We will be able to meet the constraints posed by emerging viruses and to confirm the safety of viral vaccines for their application during pregnancies. We have never been so close to the experimental assessment of the open questions from teratogenic alterations during human pregnancies. Despite the remaining challenges, this is an exciting outlook.


As the fibre reinforced plastic composites gain larger and larger share in industry, the problem of joining them with metal elements becomes significant. The current paper is the first part of the literature review, which gathers and evaluates knowledge about methods suitable for mechanical joining of composite and metal elements. This paper concerns bolted joining, because this method of mechanical joining is widely used for joining composite materials. The paper describes failure modes of bolted joints in composite materials, the influence of the bolt clamping torque, the clearance between the bolt and the hole and aging on the performance of the joint, drilling techniques used in composite materials in order to minimize damages, different fastener types, inspection techniques, and finally, the techniques that have been developed in order to improve the strength of the bolted joints in composites. Since the hole drilled in a composite material in order to perform bolted joining is a weak point of the structure, those techniques: bonded inserts, titanium foil internal inserts, fibre steering, additional reinforcement, and moulded holes, mainly aim to improve the strength of the hole in the composite. The techniques have been discussed in details and compared with each other in the summary section. The amounts of composite laminates used in the Boeing 787 Dreamliner and Airbus A350 exceed 50% of the vehicle weight ,2. Even the stress concentrations caused by the fastener hole,the disruption of the fibre continuity during drilling, which severely reduces the load carrying capacity ,25,26,undesirable damages  inducedMoreover, the choice of an adhesive which works properly with both composite and metal elements and the proper way of surface treatment for adhesive joining is a demanding and tedious task ,22,23. Triveting,cinching,form-locked joints,pin joints,loop joints.In consequence, the bolted joint effectiveness in composites is worse than in metals, which detract from the weight advantage of composites ,29. Aparmodes of failure of bolted joints focusing on the mechanisms characteristic for composite materials,the influence of the bolt clamping torque, the clearance between the bolt and the hole and aging on the performance of the joint,drilling techniques used in composite materials in order to minimize damages,different fastener types,inspection methods,techniques that have been developed in order to improve the strength of bolted joints in composites.The number and complexity of mechanical methods of joining composite and metal materials makes the choice of the proper method tedious and difficult. Thus, the current paper along with an accompanying paper  providesbonded inserts,titanium foil internal inserts,fibre steering,additional reinforcement,moulded holesmainly aim to improve the strength of the hole in the composite. As this last part is particularly interesting from the point of view of a designer looking for new solutions to enhance the performance of composite structures, those techniques have been discussed in details and compared with each other in the summary section.Since a hole drilled in a composite material in order to perform bolted joining is a weak point of the structure, those techniques:transverse matrix cracks,axial splits (fibre/matrix shearing),delaminations between adjacent layers around the highly stressed regions of the hole edgecontribute to that stress concentration relief in composites .Although the failure modes of bolted joints are similar in metal and composite materials, the phenomena occurring during the failure are different. The joints in metals are known to outperform the joints in composites ,29. Accobearing,net-tension,shear-out.The basic failure modes in bolted fibre reinforced materials are ,17,24,37the normal stresses at the net-tension plane,the shear stresses at the shear-out plane,the radial bearing stresses at the bearing planeas illustrated in In addition, out-of-plane loading can cause partial or complete pull-out of the bolt from the composite structure . TypicalThe bearing failure is also the most complicated failure mode in the case of composite laminates. Fibre micro-buckling, matrix cracks, delamination, and shear caused by the compressive load imposed by the bolt result in bearing failure of the laminate ,40. Figujoint configuration,bolt torque,laminate and bolt materials,laminate lay-up,temperature.The joint strength depends on different parameters, such as :joint co8]S and [±45/08]S glass-epoxy laminates made of UD tapes is about 20% higher than that of the bolted joints in the [902/08]S laminate S2 and two clearance values 1.55% and 3.05% of the nominal hole diameter were investigated. The bearing strength at 4% hole deformation was shown to decrease down by 19% for the thinner laminate and by 6% for the thicker laminate as the clearance was increased to 3.05%. However, the ultimate bearing strength remained almost constant and independent of the clearance between the bolt and the hole. The application of lateral clamping load was found to increase the bearing values of the laminates by approximately 20% in comparison to laminate loaded without any clamping force . Th. Th93]. Another way to decrease the drilling-induced delamination may be the use of ultrasonic oscillation assisted drilling. Guputa et al. presented a research aiming to evaluate the effect of such drilling on the delamination in quasi-isotropic carbon fibre reinforced composite material with bismaleimide matrix . The ultAnother problem which arises during drilling of composite materials is the heat generated by the drilling. The thermo-mechanical conditions during the drilling process are severe due to very poor thermal conductivity of the composite materials compared to metals . It can carbon fibre/polyimide,carbon fibre/PEEK,carbon fibre/carbon composites.Many materials can be used to manufacture fasteners. However, the most obvious ones, such as aluminum alloys and steel cannot be used to join carbon fibre reinforced composites without protecting coating or shielding, because of galvanic corrosion arising due to the difference in the potential conductivity of the composite and the aforementioned metal alloys ,121. A sStarikov and Schon conducted two researches in which strength of composite joints with composite bolts (PEEK/carbon) and several types of titanium bolts was evaluated in static  and fatiAnother question is the shape of the bolt head. Two main types of fasteners are used in aerospace structures: with protruding head and with countersunk head. The latter type is usually used in the case of outer surface joining in the areas when the aerodynamic smoothness is of great importance . The couAlthough a circular cross-section of a fastener is an obvious choice from the manufacturing point of view, there are a few theoretical works which indicate the possibility that the non-circular cross-section would be a better solution in terms of the joint strength. Wang et al.  and Persultrasonic C-scans—according to ASTM E2580: Standard practice for ultrasonic testing of flat panel composites and sandwich core materials used in aerospace applications,shearography—according to ASTM E2581: Standard practice for shearography of polymer matrix composites and sandwich core materials in aerospace applications,thermography—according to ASTM E2582: Standard practice for infrared flash thermography of composite panels and repair patches used in aerospace applications,acoustic emission—according to ASTM E2661: Standard practice for acoustic emission examination of plate-like and flat panel composite structures used in aerospace applications,radiography—according to ASTM E2662: Standard practice for radiographic examination of flat panel composites and sandwich core materials used in aerospace applicationsare dedicated for flat panels and are difficult to perform in the case of bolted joints due to the inaccessibility of the joint stigated . The resstigated ,141. Thestigated . Shimamustigated . The eledetected . The advdetected . On the Thostenson and Chou and Friedrich et al. employed unique capability of carbon nanotube networks as in situ sensors for sensing local composite damage and bolt loosening in mechanically fastened glass/epoxy composite joints ,134. DueDue to many problems arising in composite bolted joining, which deteriorate the performance of the joints, some ways to improve their efficiency have been developed. Those ways are described in the following sections.σrr, is normalized using the bearing stress σb, defined as the ratio of maximum bearing force to the cross-section area of the bolt hole S glass/polyester woven-roving composites in tensile test [S exhibits larger failure strength, smaller initial stiffness and larger failure strain for moulded holes than for drilled holes  inducedbonding metal inserts around the hole,substitution of some composite plies around the holes by titanium foil,fibre steering technique which consists in adding roving yarns shaped along principal stress lines or load paths around the holes in composite,adding reinforcement through the thickness or in plane of the composite around the holesmanufacturing moulded holes in composite, which assure the fibre continuity around the holes and prevents drilling-induced damages.In the present work, the techniques developed in order to improve the performance of the bolted joints in composite materials are also described. Those techniques include:Since all of those techniques have advantages and disadvantages, they were summarised in The analysis of The present article is the first part of the literature review concerning the mechanical methods suitable to join composite and metal elements. It focuses on the various aspects of bolted joining and methods developed to improve the performance of bolted joints in composite materials. The presented advantages and disadvantages of those solutions along with the results of the experiments conducted in order to evaluate their usefulness and feasibility should constitute a guide suitable for researchers and engineers looking for novel solutions to enhance the performance of composite-metal and composite-composite bolted joints.